il 23  (PeproTech)

 
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    Name:
    Recombinant Human IL 23
    Description:
    IL 23 is a proinflammatory heterodimeric protein composed of two subunits a unique p19 subunit and a p40 subunit which is shared with IL 12 IL 23 is secreted by activated dendritic cells and macrophages and signals though a receptor comprised of IL 23R complexed with IL 12Rβ2 IL 23 has been shown to enhance proliferation of memory T cells It also stimulates the production of IFN γ in NK cells induces IL 17 production and drives Th17 mediated responses Recombinant IL 23 is a 53 5 kDa heterodimeric protein consisting of two subunits p19 170 amino acids and p40 306 amino acids
    Catalog Number:
    200-23-100UG
    Price:
    1,800.00
    Category:
    Recombinant Proteins
    Source:
    (BTI‐Tn‐5B1‐4) Hi‐5 Insect cells
    Reactivity:
    Mouse
    Purity:
    95.0
    Quantity:
    100UG
    Buy from Supplier


    Structured Review

    PeproTech il 23
    <t>IL-23</t> plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    IL 23 is a proinflammatory heterodimeric protein composed of two subunits a unique p19 subunit and a p40 subunit which is shared with IL 12 IL 23 is secreted by activated dendritic cells and macrophages and signals though a receptor comprised of IL 23R complexed with IL 12Rβ2 IL 23 has been shown to enhance proliferation of memory T cells It also stimulates the production of IFN γ in NK cells induces IL 17 production and drives Th17 mediated responses Recombinant IL 23 is a 53 5 kDa heterodimeric protein consisting of two subunits p19 170 amino acids and p40 306 amino acids
    https://www.bioz.com/result/il 23/product/PeproTech
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    il 23 - by Bioz Stars, 2021-05
    95/100 stars

    Images

    1) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    2) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    3) Product Images from "Salt generates antiinflammatory Th17 cells but amplifies pathogenicity in proinflammatory cytokine microenvironments"

    Article Title: Salt generates antiinflammatory Th17 cells but amplifies pathogenicity in proinflammatory cytokine microenvironments

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI137786

    TGF-β governs the reciprocal enhancement of pro- versus antiinflammatory Th17 cell functions by NaCl in vitro and in vivo in an EAE mouse model. ( A ) Naive murine CD4 + T cells from 2D2 mice were cultured in vitro in neutral (Th0) or various Th17-polarizing conditions (Th17), including TGF-β in low- or high-NaCl conditions, and analyzed by FACS on day 3.5 for IL-17A and FoxP3 expression. Representative dot plots show intracellular IL-17A and FoxP3 expression after restimulation of the cells with PMA and ionomycin. ( B ) Intracellular IL-17A and FoxP3 expression was quantified as in A . ( C ) Clinical EAE scores (0–5) were determined after adoptive transfer of T cells (1.5 × 10 6 ) that were polarized in vitro for 3.5 days with TGF-β and IL-6 (both 25 ng/mL) in high- or low-NaCl conditions. ( D and E ) Naive murine CD4 + T cells were cultured as in A in the absence of exogenous TGF-β. Representative dot plots and summary plots are shown. ( F ) Clinical EAE scores after adoptive transfer of T cells (2 × 10 6 ) that were polarized in vitro for 3.5 days with IL-1β, IL-6, and IL-23 in high- versus low-NaCl conditions. * P
    Figure Legend Snippet: TGF-β governs the reciprocal enhancement of pro- versus antiinflammatory Th17 cell functions by NaCl in vitro and in vivo in an EAE mouse model. ( A ) Naive murine CD4 + T cells from 2D2 mice were cultured in vitro in neutral (Th0) or various Th17-polarizing conditions (Th17), including TGF-β in low- or high-NaCl conditions, and analyzed by FACS on day 3.5 for IL-17A and FoxP3 expression. Representative dot plots show intracellular IL-17A and FoxP3 expression after restimulation of the cells with PMA and ionomycin. ( B ) Intracellular IL-17A and FoxP3 expression was quantified as in A . ( C ) Clinical EAE scores (0–5) were determined after adoptive transfer of T cells (1.5 × 10 6 ) that were polarized in vitro for 3.5 days with TGF-β and IL-6 (both 25 ng/mL) in high- or low-NaCl conditions. ( D and E ) Naive murine CD4 + T cells were cultured as in A in the absence of exogenous TGF-β. Representative dot plots and summary plots are shown. ( F ) Clinical EAE scores after adoptive transfer of T cells (2 × 10 6 ) that were polarized in vitro for 3.5 days with IL-1β, IL-6, and IL-23 in high- versus low-NaCl conditions. * P

    Techniques Used: In Vitro, In Vivo, Mouse Assay, Cell Culture, FACS, Expressing, Adoptive Transfer Assay

    4) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    5) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    6) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    7) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    8) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    9) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    10) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    11) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    12) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    13) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    14) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    15) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    16) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    17) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    18) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    19) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    20) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    21) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    22) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    23) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    24) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    25) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    26) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    27) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    28) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    29) Product Images from "The NOTCH-HES-1 axis is involved in promoting Th22 cell differentiation"

    Article Title: The NOTCH-HES-1 axis is involved in promoting Th22 cell differentiation

    Journal: Cellular & Molecular Biology Letters

    doi: 10.1186/s11658-021-00249-w

    Naïve CD4 + T cells could be differentiated into Th22 cells with multiple cytokines in vitro. Naïve CD4 + T cells were collected from mice and cultured for 6 days in conditions designed to induce Th22 differentiation (anti-CD3/CD28 Abs, anti-IFN-γ Ab, anti-IL-4 Ab, IL-6, TNF-α, and IL-23). a Representative plots of naïve CD4 + T cells stimulated for 6 days under optimal Th22 conditions. CD4 + T cells and Th (CD4 + IFN-γ − ) were gated by flow cytometry to analyze the Th22 cells. b Percentage quantitation of Th22 cells. c The alterations of Nicd , Hes-1 , Ahr , and Il-22 mRNAs were evaluated by RT-PCR. d Western blotting of the expression of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 in total protein lysates from different treatment cells. e–i Representative densitometric quantification of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 expression in T cells, β-ACTIN was used as an endogenous control for protein expression. The results show a typical experiment; each bar represents the mean ± S.E.M. of at least three independent experiments. ** p
    Figure Legend Snippet: Naïve CD4 + T cells could be differentiated into Th22 cells with multiple cytokines in vitro. Naïve CD4 + T cells were collected from mice and cultured for 6 days in conditions designed to induce Th22 differentiation (anti-CD3/CD28 Abs, anti-IFN-γ Ab, anti-IL-4 Ab, IL-6, TNF-α, and IL-23). a Representative plots of naïve CD4 + T cells stimulated for 6 days under optimal Th22 conditions. CD4 + T cells and Th (CD4 + IFN-γ − ) were gated by flow cytometry to analyze the Th22 cells. b Percentage quantitation of Th22 cells. c The alterations of Nicd , Hes-1 , Ahr , and Il-22 mRNAs were evaluated by RT-PCR. d Western blotting of the expression of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 in total protein lysates from different treatment cells. e–i Representative densitometric quantification of p-STAT3, STAT3, NICD, HES-1, AHR, and IL-22 expression in T cells, β-ACTIN was used as an endogenous control for protein expression. The results show a typical experiment; each bar represents the mean ± S.E.M. of at least three independent experiments. ** p

    Techniques Used: In Vitro, Mouse Assay, Cell Culture, Flow Cytometry, Quantitation Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

    30) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    31) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

    32) Product Images from "IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders"

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders

    Journal: Theranostics

    doi: 10.7150/thno.41378

    IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays a critical role in aging-mediated bone loss. ( A ) Micro-CT images of the trabecular bone in the distal femoral metaphysis of 12-month-old male WT mice and age-sex matched IL-12p35 -/- and IL-12p40 -/- mice. n = 4 per group. Scale bar, 1mm (upper and middle panel), 100 µm (lower panel). ( B ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), bone volume/tissue volume (BV/TV), trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4 per group. ( C ) The H E staining of femur sections from 12-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4 per group. Scale bar, 50 µm. ( D ) Trabecular percentage (%) was quantified via H E images from the groups described in C . n = 4 per group. ( E , F ) The immunohistochemical analysis ( E ) and quantification ( F ) of ALP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ( G , H ) The immunohistochemical analysis ( G ) and quantification ( H ) of TRAP expression in the distal femoral metaphysis. n = 4 per group. Scale bar, 50 µm. ALP, alkaline phosphatase; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining, Immunohistochemistry, Expressing

    IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 deficiency improves calvarial defect repair and enhances bone mass. ( A ) Microcomputed tomography (micro-CT) three-dimensional reconstruction of bone repair in mouse calvarial defects. Red dotted lines indicate the periphery of the defect site. n = 4-5 per group. Scale bar, 500 µm (upper panel), 1 mm (lower panel). ( B ) Defect area coverage (%) was calculated via micro-CT images from WT, IL-12p35 -/- , and IL-12p40 -/- mice. ( C-E ) Micro-CT analysis including ( C ) bone volume (BV), ( D ) bone volume/tissue volume (BV/TV), and ( E ) healing score for the extent of bony bridging and bone union. ( F ) The H E staining of calvarial bone defects. n = 4-5 per group. Scale bar, 100 µm. ( G ) Representative images showed trabecular architecture by micro-CT three-dimensional reconstruction in distal femurs of 2-month-old mice. n = 4-5 per group. Scale bar, 1mm (upper and middle panel), 500 µm (lower panel). ( H ) Micro-CT measurements for the indicated parameters in distal femurs. Bone mineral density (BMD), BV/TV, trabecular numbers (Tb.N), and trabecular thickness (Tb.Th) were determined by micro-CT analysis. n = 4-5 per group. ( I ) The H E staining of femur sections from 2-month-old WT, IL-12p35 -/- , and IL-12p40 -/- mice were shown. n = 4-5 per group. Scale bar, 50 µm. ( J ) Trabecular percentage (%) was quantified via H E images from the groups described in I . WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Micro-CT, Mouse Assay, Staining

    IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 affect bone formation and resorption in vivo . ( A ) The expression levels of osteoblast-related genes ( ALP and OCN ) in cranium from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 8 per group. ( B ) Serum levels of OCN. n = 8 per group. ( C ) ALP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 20 µm. ( D ) Quantification of ALP activity was shown via images from the groups described in C . ( E ) The expression levels of osteoclast-specific genes ( TRAP and RANKL ) in calvarial bone from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 6 per group. ( F ) Serum levels of CTX-1. n = 6 per group. ( G ) TRAP staining of femur sections from WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4 - 5 per group. Scale bar, 20 µm. ( H ) Quantification of TRAP activity was shown via images from the groups described in G . ALP, alkaline phosphatase; CTX-1, collagen type I cross-linked C-telopeptide; OCN, osteocalcin; RANKL, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: In Vivo, Expressing, Mouse Assay, Staining, Activity Assay

    IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-12 and IL-23 regulate BMMSC osteogenesis and osteoclast differentiation. ( A , B ) BMMSCs mixed with β-TCP plus IL-12 or IL-12+IL-23 were subcutaneously implanted into immunocompromised mice for 8 weeks. ( A ) New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B ) Quantitative analysis of new bone volume. n = 4-5 per group. ( C-E ) Bone histomorphometric measurements among each group, including ( C ) osteoblast number (N.Ob), ( D ) osteoblast number per bone surface (N.Ob/BS), and ( E ) osteocyte number (N.Ot). ( F ) Compared to untreated CD4 + T cell culture supernatants, IL-12- or/and IL-23-stimulated supernatants showed reduced capacities to form mineralized nodules of BMMSCs, as assessed by Alizarin Red staining. ( G ) Quantitative analysis of the calcium mineralization in F . ( H ) The blockade of IFN-γ or/and IL-17 in IL-12- and IL-23-stimulated supernatants rescued reduced capacities to form mineralized nodules in BMMSCs. ( I ) Quantitative analysis of the calcium mineralization in H . ( J ) RAW264.7 cell proliferation analysis treated with RANKL (75 ng/mL) and different cytokines. All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. ( K ) Effects of different cytokines (IL-12, IL-23, IFN-γ, and IL-17) on the RANKL-induced osteoclast differentiation of RAW264.7 cells. ( L, M ) The number of TRAP + osteoclasts per field ( L ) and the average size of osteoclasts ( M ). All: IL-12 + IL-23 + IFN-γ + IL-17. *: compared with control group; #: compared with IL-12 + IL-23 group; x: compared with All group. B, bone; CT, connective tissue; Con, control; CM, control medium; IFN-γ, interferon γ; OM, osteogenic medium; RANKL, receptor activator of NF-κB ligand. Results are shown as mean ± S.D. * p

    Techniques Used: Mouse Assay, Staining, Cell Culture

    IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p
    Figure Legend Snippet: IL-23 plays an essential role in inflammation-mediated inhibition of bone regeneration. ( A ) BMMSCs mixed with β-TCP were implanted into the dorsal surface of WT and nude mice for 8 weeks. A substantial amount of bone was formed in nude mice, as detected by H E staining. n = 4-5 per group. Scale bar, 50 µm. ( B , C ) IL-12p40 expression levels in implants after 7 days of implantation. Representative images ( B ) and quantification ( C ) of immunohistochemical staining of IL-12p40. n = 4-5 per group. Scale bar, 50 µm. ( D ) BMMSCs mixed with β-TCP plus IL-12 and IL-23 or no cytokine were implanted into the dorsal surface of WT and IL-12p40 -/- mice. New bone formation was detected with H E staining. n = 4-5 per group. Scale bar, 50 µm. ( E-G ) Bone histomorphometric measurements among each group, including ( E ) osteoblast number (N.Ob), ( F ) osteoblast number per bone surface (N.Ob/BS), and ( G ) osteocyte number (N.Ot). ( H ) Representative images and quantification of ectopic bone formation in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. ( I-K ) Bone histomorphometric measurements, including ( I ) N.Ob, ( J ) N.Ob/BS, and ( K ) N.Ot. ( L ) Osteocalcin (OCN) expression was increased in IL-12p40 -/- mice. Representative images and quantification of immunohistochemical staining of OCN were shown in WT, IL-12p35 -/- , and IL-12p40 -/- mice. n = 4-5 per group. Scale bar, 50 µm. B, bone; CT, connective tissue; WT, wild-type. Results are shown as mean ± S.D. * p

    Techniques Used: Inhibition, Mouse Assay, Staining, Expressing, Immunohistochemistry

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    Isolation:

    Article Title: Inherited salt-losing tubulopathies are associated with immunodeficiency due to impaired IL-17 responses
    Article Snippet: Electrolyte concentrations in unadjusted XVIVO15 media were measured by automated analyser (Cobas 8000 modular analyser series, Roche). .. Th17 and Tc17 polarisation analysis PBMCs were isolated from SLT patients (n = 27), HCs (n = 21) and disease controls (n = 19) and fresh cells stimulated with anti-human CD3 (10 µg/ml plate bound), anti-human CD28 (1 µg/ml), IL-1β (12.5 ng/ml; Peprotech, cat#200-01B), IL-6 (25 ng/ml; Peprotech, cat#200-06), IL-21 (25 ng/ml; Peprotech, cat#200-21), IL-23 (25 ng/ml; Peprotech, cat#200-23), and TGFβ (5 ng/ml; Peprotech, cat#100-36E) in XVIVO15 media for 7 days. .. On day 7, cells were restimulated for 4 h with PMA (50 ng/ml) and ionomycin (1 µg/ml; Sigma, cat#I9657) in the presence of brefeldin A (5 µg/ml).

    Recombinant:

    Article Title: IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders
    Article Snippet: CD4+ T cells were isolated by magnetic sorting from splenocytes with a mouse CD4+ T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and stimulated with 5 μg/mL anti-CD3 antibody and 2 μg/mL anti-CD28 antibody (eBioscience, San Diego, CA, USA). .. The cultures were supplemented with recombinant mouse IL-12 (10 ng/mL) and IL-23 (25 ng/mL). .. BMMSCs were stimulated with the supernatants and rat IgG (0.3 µg/mL)/IFN-γ neutralizing antibody (0.3 µg/mL)/IL-17 neutralizing antibody (0.3 µg/mL) in osteogenic differentiation.

    Article Title: IL23 Promotes Antimicrobial Pathways in Human Macrophages, Which Are Reduced With the IBD-Protective IL23R R381Q Variant
    Article Snippet: Monocyte differentiation was conducted in 10 ng/mL macrophage colony-stimulating factor (Shenandoah Biotechnology Systems, Warwick, PA) for 7 days to generate MDMs. .. MDM StimulationMDMs were treated with 10 ng/mL recombinant human IL23 (200-23) or IL12p70 (200-12) (Peprotech, Rocky Hill, NJ) or 100 μg/mL muramyl dipeptide (Bachem, Torrance, CA) for 48 hours. .. In some cases, cells were given neutralizing anti-IL23p19 (4 μg; HNU2319) or anti-IL12p35 (4 μg; BT21) antibody (eBioscience, San Diego, CA) for 1 hour before treatment.

    other:

    Article Title: Effect of interleukin (IL)-35 on IL-17 expression and production by human CD4+ T cells
    Article Snippet: One is induction by a cytokine cocktail including TGF-β , IL-6, IL-1β , anti-IFN-γ Ab, anti-IL-2 Ab and anti-IL-4 Ab; the second involves addition of rIL-23.

    Blocking Assay:

    Article Title: IL‐6‐specific autoantibodies among APECED and thymoma patients
    Article Snippet: Briefly, the serum samples were pre‐incubated with 40 μg/ml of recombinant human (rh) IL‐6, (PeproTech EC Ltd, London, UK) or rhIFN‐γ (Miltenyi Biotec, Bergisch Gladbach, Germany). .. To test for IL‐23‐ blocking, the thymoma serum samples were pre‐incubated with 40 μg/ml of rhIL‐23 (PeproTech EC Ltd, London, UK), rhIL‐12 (PeproTech EC Ltd, London, UK), or rhIL‐6 (PeproTech EC Ltd, London, UK). .. The samples were rotated for 2 h at room temperature and centrifuged for 15 min at 16,000g and supernatants were transferred into new Eppendorf tubes before performing LIPS assay as above.

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  • 97
    PeproTech recombinant human il 4
    Induction of <t>IL-4</t> responsiveness following tgD-CD154 binding to bovine B cells. Supernatants from (a) mock-transfected, (b) pSLIAtgD-transfected, or (c) pSLIAtgD-CD154-transfected Cos-7 cells were incubated with bovine B cells. Bound protein was detected with a gD-specific mAb cocktail, followed by FITC-conjugated goat anti-mouse immunoglobulin. (d) Triplicate cultures of bovine B cells (4 × 10 4 cells/well) were incubated with medium or with pSLIAtgD- or pSLIAtgD-CD154 transfected Cos-7 cell supernatants. Cultures were incubated for 72 hr with either 100 μl/well of 10 ng/ml recombinant human IL-4 (+IL-4) or medium (–IL-4). Cell proliferation was detected by adding methyl-[ 3 H]thymidine during the last 18 hr of culture. Data are shown as average c.p.m. + SEM for triplicate cultures.
    Recombinant Human Il 4, supplied by PeproTech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human il 4/product/PeproTech
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    86
    PeproTech hril 23
    <t>hrIL-23</t> regulates growth of hepatoma cells Cells were treated with hrIL-23 (0 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml) for 48 h. ( A ) CCK8 proliferation assay was performed. Data were expressed as fold change in proliferation rate in hrIL-23-treated cells versus untreated ones (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. ( B ) Cells were seeded at 500 cells/cm 2 and incubated for 5–7 days to form colonies. A representative experiment was shown. The experiment was performed two times yielding similar results. ( C ) Flow cytometric analysis of DNA content in hepatoma cells in the presence of hrIL-23. The percentages of cells in each cycle phases were shown. ( D , E ) Cells were cultured in serum-free medium containing different concentration of hrIL-23 for 48 h. Then cells were stained with Annexin V and PI to detect the percentage of apoptotic cells by FCM (D) and Bcl-2 level was detected by realtime PCR ( E ). Data were expressed as fold change in Bcl-2 expression in hrIL-23-treated versus untreated cells (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. * P
    Hril 23, supplied by PeproTech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hril 23/product/PeproTech
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    hril 23 - by Bioz Stars, 2021-05
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    86
    PeproTech il 3
    <t>IL‐3</t> stabilizes ICOS‐L on mast cells via ERK signalling. Bone marrow‐derived mast cells (BMMCs) were cultured as indicated, and ICOS‐L expression was assessed on CD117 + BMMCs by flow cytometry. (A‐C) BMMCs were generated from total bone marrow cells of C57BL/6 mice in medium containing supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3. Differentiation was monitored over 8 weeks ( n = 12 independent biological replicates). FACS plots in (A) show representative results after 1–2 weeks or after 7–8 weeks of BMMC generation. Generation of CD117 + BMMCs is summarized in (B). ICOS‐L expression among CD117 + BMMCs is summarized in (C). (D) BMMCs were cultured in the presence of supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3 with or without a blocking anti‐IL‐3 antibody (10 µg/ml) for 24 h. CD117 + BMMCs and ICOS‐L + cells among CD117+ BMMCs are summarized in the diagrams ( n = 6 independent biological replicates). (E) BMMCs were cultured in medium alone or medium containing either 50 ng/ml recombinant IL‐3, 50 ng/ml recombinant IL‐33 or 50 ng/ml recombinant SCF for 24 h. Frequencies of ICOS‐L + cells among CD117 + BMMCs are summarized ( n = 3 independent biological replicates). (F‐I) BMMCs were starved from IL‐3 for 60‐min prior stimulation with 50 ng/ml IL‐3 for the indicated time periods. Phosphorylation of MAP kinases ERK1/2, JNK1/2 and p38 was analysed by Western blotting. Representative Western blot bands are shown in (F). Phosphorylation of MAP kinases ERK1/2 (G), p38 (H) and JNK1/2 (I) was quantified ( n = 4 biologically independent experiments). (J) BMMCs were cultured in the presence of recombinant IL‐3 and DMSO or with JNK inhibitor (SP600125, n = 3 biological replicates), p38 inhibitor (SB203580, n = 6 biological replicates), or ERK inhibitor (UO126, n = 6 biological replicates) for 24 h. Subsequently, frequencies of ICOS‐L + cells among CD117 + BMMCs were analysed and summarized in the diagram. (K) BMMCs were starved from IL‐3 for 30 min and incubated in the presence of DMSO or UO126 for further 30 min. Subsequently, cells were cultured in medium alone or in the presence of 50 ng/ml IL‐3 for 24 h. Phosphorylation of MAP kinases ERK1/2 was analysed by Western blotting. Representative Western blot bands are shown, and quantification of ERK phosphorylation is summarized in the diagram below ( n = 3 independent biological replicates). Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p
    Il 3, supplied by PeproTech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/il 3/product/PeproTech
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    il 3 - by Bioz Stars, 2021-05
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    95
    PeproTech il 13
    IL-4Rα-expression by epithelial cells (ECs) and smooth muscle cells (SMCs) is required in IL-4Rα flox/− mice for maximal airway hyperresponsiveness (AHR) induced by exogenous <t>IL-13.</t> IL-4Rα flox/− mice were treated i.t. with 3 µg of IL-13 daily for 7 days. A. Invasive measurement of AHR. B. Absolute eosinophil count of BAL fluid. C. Quantitation of goblet cell metaplasia (GCM). D. Distribution of residual GCM in club cell 10 kDa protein-Cre + (CC10-Cre + ) mice and CC10-Cre + α-smooth muscle actin-Cre + (SMP8-Cre + ) mice. E. Absolute eosinophil count of BAL fluid in CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high or low residual GCM. F. Percentage of eosinophils in BAL fluid in CC10-Cre + mice and CC10-Cre + SMP8-Cre + that had high or low residual GCM. G, AHR when CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high residual GCM were excluded. The results represent 12 pooled experiments with panels A-D showing 17, 14, 26, 22 and 47 mice for groups: Cre − + IL-13, SMP8-Cre + + IL-13, CC10-Cre + + IL-13, Both Cre + + IL-13, all genotypes + saline, respectively. Panel G represents 14 and 12 mice for groups: CC10-Cre + + IL-13 and both Cre + + IL-13, respectively. NS, not significant; *, p
    Il 13, supplied by PeproTech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Induction of IL-4 responsiveness following tgD-CD154 binding to bovine B cells. Supernatants from (a) mock-transfected, (b) pSLIAtgD-transfected, or (c) pSLIAtgD-CD154-transfected Cos-7 cells were incubated with bovine B cells. Bound protein was detected with a gD-specific mAb cocktail, followed by FITC-conjugated goat anti-mouse immunoglobulin. (d) Triplicate cultures of bovine B cells (4 × 10 4 cells/well) were incubated with medium or with pSLIAtgD- or pSLIAtgD-CD154 transfected Cos-7 cell supernatants. Cultures were incubated for 72 hr with either 100 μl/well of 10 ng/ml recombinant human IL-4 (+IL-4) or medium (–IL-4). Cell proliferation was detected by adding methyl-[ 3 H]thymidine during the last 18 hr of culture. Data are shown as average c.p.m. + SEM for triplicate cultures.

    Journal: Immunology

    Article Title: Modulation of immune responses to bovine herpesvirus-1 in cattle by immunization with a DNA vaccine encoding glycoprotein D as a fusion protein with bovine CD154

    doi: 10.1111/j.1365-2567.2004.01877.x

    Figure Lengend Snippet: Induction of IL-4 responsiveness following tgD-CD154 binding to bovine B cells. Supernatants from (a) mock-transfected, (b) pSLIAtgD-transfected, or (c) pSLIAtgD-CD154-transfected Cos-7 cells were incubated with bovine B cells. Bound protein was detected with a gD-specific mAb cocktail, followed by FITC-conjugated goat anti-mouse immunoglobulin. (d) Triplicate cultures of bovine B cells (4 × 10 4 cells/well) were incubated with medium or with pSLIAtgD- or pSLIAtgD-CD154 transfected Cos-7 cell supernatants. Cultures were incubated for 72 hr with either 100 μl/well of 10 ng/ml recombinant human IL-4 (+IL-4) or medium (–IL-4). Cell proliferation was detected by adding methyl-[ 3 H]thymidine during the last 18 hr of culture. Data are shown as average c.p.m. + SEM for triplicate cultures.

    Article Snippet: Briefly, bovine PBMCs were grown for 2–3 weeks in the presence of recombinant human IL-4 (Peprotech EC, London, UK) and murine cells expressing membrane-bound CD154.

    Techniques: Binding Assay, Transfection, Incubation, Recombinant

    hrIL-23 regulates growth of hepatoma cells Cells were treated with hrIL-23 (0 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml) for 48 h. ( A ) CCK8 proliferation assay was performed. Data were expressed as fold change in proliferation rate in hrIL-23-treated cells versus untreated ones (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. ( B ) Cells were seeded at 500 cells/cm 2 and incubated for 5–7 days to form colonies. A representative experiment was shown. The experiment was performed two times yielding similar results. ( C ) Flow cytometric analysis of DNA content in hepatoma cells in the presence of hrIL-23. The percentages of cells in each cycle phases were shown. ( D , E ) Cells were cultured in serum-free medium containing different concentration of hrIL-23 for 48 h. Then cells were stained with Annexin V and PI to detect the percentage of apoptotic cells by FCM (D) and Bcl-2 level was detected by realtime PCR ( E ). Data were expressed as fold change in Bcl-2 expression in hrIL-23-treated versus untreated cells (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. * P

    Journal: Oncotarget

    Article Title: IL-23 enhances the malignant properties of hepatoma cells by attenuation of HNF4α

    doi: 10.18632/oncotarget.24875

    Figure Lengend Snippet: hrIL-23 regulates growth of hepatoma cells Cells were treated with hrIL-23 (0 ng/ml, 5 ng/ml, 10 ng/ml, 20 ng/ml, 40 ng/ml) for 48 h. ( A ) CCK8 proliferation assay was performed. Data were expressed as fold change in proliferation rate in hrIL-23-treated cells versus untreated ones (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. ( B ) Cells were seeded at 500 cells/cm 2 and incubated for 5–7 days to form colonies. A representative experiment was shown. The experiment was performed two times yielding similar results. ( C ) Flow cytometric analysis of DNA content in hepatoma cells in the presence of hrIL-23. The percentages of cells in each cycle phases were shown. ( D , E ) Cells were cultured in serum-free medium containing different concentration of hrIL-23 for 48 h. Then cells were stained with Annexin V and PI to detect the percentage of apoptotic cells by FCM (D) and Bcl-2 level was detected by realtime PCR ( E ). Data were expressed as fold change in Bcl-2 expression in hrIL-23-treated versus untreated cells (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. * P

    Article Snippet: A low serum (2% FBS) culture medium was added to the plates to inhibit cell proliferation. hrIL-23 was supplemented at different concentrations.

    Techniques: Proliferation Assay, Incubation, Flow Cytometry, Cell Culture, Concentration Assay, Staining, Polymerase Chain Reaction, Expressing

    hrIL-23 induces motility and invasivity of hepatoma cells ( A ) Wound healing assays. Cells were plated in 12-well plates and a scratch was made 24 h later. And then cells were cultured in 2% FBS medium containing hrIL-23 for 24 h and 48 h to check wound closure with an inverted microscope. ( B ) Cell motility assays. 1 × 10 5 HepG2 or Huh-7 cells pretreated with increasing doses of hrIL-23 (0–40 ng/ml) for 24 h were seeded in the upper chamber in serum free medium and 2.5% FBS medium was added to the lower chamber. 24 h later, migrated cells were fixed with 4% paraformaldehyde and then stained with 0.1% Crystal Violet. ( C ) Realtime PCR analysis for MMP9 expression in hrIL-23 treated hepatoma cells. Data were expressed as fold changes in MMP9 expression in hrIL-23-treated versus untreated cells (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. * P

    Journal: Oncotarget

    Article Title: IL-23 enhances the malignant properties of hepatoma cells by attenuation of HNF4α

    doi: 10.18632/oncotarget.24875

    Figure Lengend Snippet: hrIL-23 induces motility and invasivity of hepatoma cells ( A ) Wound healing assays. Cells were plated in 12-well plates and a scratch was made 24 h later. And then cells were cultured in 2% FBS medium containing hrIL-23 for 24 h and 48 h to check wound closure with an inverted microscope. ( B ) Cell motility assays. 1 × 10 5 HepG2 or Huh-7 cells pretreated with increasing doses of hrIL-23 (0–40 ng/ml) for 24 h were seeded in the upper chamber in serum free medium and 2.5% FBS medium was added to the lower chamber. 24 h later, migrated cells were fixed with 4% paraformaldehyde and then stained with 0.1% Crystal Violet. ( C ) Realtime PCR analysis for MMP9 expression in hrIL-23 treated hepatoma cells. Data were expressed as fold changes in MMP9 expression in hrIL-23-treated versus untreated cells (arbitrary value = 1). Each value represents the mean ± SD for triplicate samples. * P

    Article Snippet: A low serum (2% FBS) culture medium was added to the plates to inhibit cell proliferation. hrIL-23 was supplemented at different concentrations.

    Techniques: Cell Culture, Inverted Microscopy, Staining, Polymerase Chain Reaction, Expressing

    hrIL-23 promotes the “stemness” gene CD133 expression in hepatoma cells Cells were treated with hrIL-23 at different concentrations as indicated for 24 h or 48 h. CD133 expression was analyzed by realtime PCR ( A ) and flow cytometer ( B ). Bar graphs ( C ) are the percentage and MFI of CD133 expression. Each column and bar represents the mean ± SD of three independent experiments. * P

    Journal: Oncotarget

    Article Title: IL-23 enhances the malignant properties of hepatoma cells by attenuation of HNF4α

    doi: 10.18632/oncotarget.24875

    Figure Lengend Snippet: hrIL-23 promotes the “stemness” gene CD133 expression in hepatoma cells Cells were treated with hrIL-23 at different concentrations as indicated for 24 h or 48 h. CD133 expression was analyzed by realtime PCR ( A ) and flow cytometer ( B ). Bar graphs ( C ) are the percentage and MFI of CD133 expression. Each column and bar represents the mean ± SD of three independent experiments. * P

    Article Snippet: A low serum (2% FBS) culture medium was added to the plates to inhibit cell proliferation. hrIL-23 was supplemented at different concentrations.

    Techniques: Expressing, Polymerase Chain Reaction, Flow Cytometry, Cytometry

    IL‐3 stabilizes ICOS‐L on mast cells via ERK signalling. Bone marrow‐derived mast cells (BMMCs) were cultured as indicated, and ICOS‐L expression was assessed on CD117 + BMMCs by flow cytometry. (A‐C) BMMCs were generated from total bone marrow cells of C57BL/6 mice in medium containing supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3. Differentiation was monitored over 8 weeks ( n = 12 independent biological replicates). FACS plots in (A) show representative results after 1–2 weeks or after 7–8 weeks of BMMC generation. Generation of CD117 + BMMCs is summarized in (B). ICOS‐L expression among CD117 + BMMCs is summarized in (C). (D) BMMCs were cultured in the presence of supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3 with or without a blocking anti‐IL‐3 antibody (10 µg/ml) for 24 h. CD117 + BMMCs and ICOS‐L + cells among CD117+ BMMCs are summarized in the diagrams ( n = 6 independent biological replicates). (E) BMMCs were cultured in medium alone or medium containing either 50 ng/ml recombinant IL‐3, 50 ng/ml recombinant IL‐33 or 50 ng/ml recombinant SCF for 24 h. Frequencies of ICOS‐L + cells among CD117 + BMMCs are summarized ( n = 3 independent biological replicates). (F‐I) BMMCs were starved from IL‐3 for 60‐min prior stimulation with 50 ng/ml IL‐3 for the indicated time periods. Phosphorylation of MAP kinases ERK1/2, JNK1/2 and p38 was analysed by Western blotting. Representative Western blot bands are shown in (F). Phosphorylation of MAP kinases ERK1/2 (G), p38 (H) and JNK1/2 (I) was quantified ( n = 4 biologically independent experiments). (J) BMMCs were cultured in the presence of recombinant IL‐3 and DMSO or with JNK inhibitor (SP600125, n = 3 biological replicates), p38 inhibitor (SB203580, n = 6 biological replicates), or ERK inhibitor (UO126, n = 6 biological replicates) for 24 h. Subsequently, frequencies of ICOS‐L + cells among CD117 + BMMCs were analysed and summarized in the diagram. (K) BMMCs were starved from IL‐3 for 30 min and incubated in the presence of DMSO or UO126 for further 30 min. Subsequently, cells were cultured in medium alone or in the presence of 50 ng/ml IL‐3 for 24 h. Phosphorylation of MAP kinases ERK1/2 was analysed by Western blotting. Representative Western blot bands are shown, and quantification of ERK phosphorylation is summarized in the diagram below ( n = 3 independent biological replicates). Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Journal: Immunology

    Article Title: IL‐3 is essential for ICOS‐L stabilization on mast cells, and sustains the IL‐33‐induced RORγt+ Treg generation via enhanced IL‐6 induction

    doi: 10.1111/imm.13305

    Figure Lengend Snippet: IL‐3 stabilizes ICOS‐L on mast cells via ERK signalling. Bone marrow‐derived mast cells (BMMCs) were cultured as indicated, and ICOS‐L expression was assessed on CD117 + BMMCs by flow cytometry. (A‐C) BMMCs were generated from total bone marrow cells of C57BL/6 mice in medium containing supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3. Differentiation was monitored over 8 weeks ( n = 12 independent biological replicates). FACS plots in (A) show representative results after 1–2 weeks or after 7–8 weeks of BMMC generation. Generation of CD117 + BMMCs is summarized in (B). ICOS‐L expression among CD117 + BMMCs is summarized in (C). (D) BMMCs were cultured in the presence of supernatant of a X63Ag‐653 BPV‐rmIL‐3 stably transfected cell line equivalent to 20 ng/ml IL‐3 with or without a blocking anti‐IL‐3 antibody (10 µg/ml) for 24 h. CD117 + BMMCs and ICOS‐L + cells among CD117+ BMMCs are summarized in the diagrams ( n = 6 independent biological replicates). (E) BMMCs were cultured in medium alone or medium containing either 50 ng/ml recombinant IL‐3, 50 ng/ml recombinant IL‐33 or 50 ng/ml recombinant SCF for 24 h. Frequencies of ICOS‐L + cells among CD117 + BMMCs are summarized ( n = 3 independent biological replicates). (F‐I) BMMCs were starved from IL‐3 for 60‐min prior stimulation with 50 ng/ml IL‐3 for the indicated time periods. Phosphorylation of MAP kinases ERK1/2, JNK1/2 and p38 was analysed by Western blotting. Representative Western blot bands are shown in (F). Phosphorylation of MAP kinases ERK1/2 (G), p38 (H) and JNK1/2 (I) was quantified ( n = 4 biologically independent experiments). (J) BMMCs were cultured in the presence of recombinant IL‐3 and DMSO or with JNK inhibitor (SP600125, n = 3 biological replicates), p38 inhibitor (SB203580, n = 6 biological replicates), or ERK inhibitor (UO126, n = 6 biological replicates) for 24 h. Subsequently, frequencies of ICOS‐L + cells among CD117 + BMMCs were analysed and summarized in the diagram. (K) BMMCs were starved from IL‐3 for 30 min and incubated in the presence of DMSO or UO126 for further 30 min. Subsequently, cells were cultured in medium alone or in the presence of 50 ng/ml IL‐3 for 24 h. Phosphorylation of MAP kinases ERK1/2 was analysed by Western blotting. Representative Western blot bands are shown, and quantification of ERK phosphorylation is summarized in the diagram below ( n = 3 independent biological replicates). Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Article Snippet: Coculture For cocultures, 100 000 BMMCs were seeded with 25 000 MHC‐II− CD4+ CD25+ Treg in the presence of 1.5 µg/ml soluble anti‐mouse CD3ε (clone 2C11, Institute for Immunology) in the presence of IL‐33 (50 ng/ml, Peprotech), with or without IL‐3 (50 ng/ml, Peprotech), with or without SCF (50 ng/ml, Peprotech) in a 96‐well plate for 3 days.

    Techniques: Derivative Assay, Cell Culture, Expressing, Flow Cytometry, Generated, Mouse Assay, Stable Transfection, Transfection, FACS, Blocking Assay, Recombinant, Western Blot, Incubation

    IL‐3 blocks the IL‐33‐induced IL‐2 production by BMMC, which selectively enhances the RORγt − Helios + T reg maintenance. (A, B) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or 50 ng/ml recombinant IL‐2 was added. Transcription factor expressions of FoxP3 among CD4 + , and RORγt and Helios among FoxP3 + CD4 + Tregs were analysed by flow cytometry in 2 experiments with BMMCs from 6 independent donors. (A) Representative FACS plots of RORγt + and Helios + cells among FoxP3 + CD4 + T regs . (B) Diagrams summarize RORγt + Helios − , RORγt + Helios + , RORγt − Helios + and RORγt − Helios − cells among FoxP3 + CD4 + T regs . (C) BMMCs were cultured in medium alone or with 50 ng/ml recombinant IL‐33 for 24 h. IL‐2 levels in the supernatants were analysed by ELISA. If indicated, BMMCs were costimulated with 50 ng/ml recombinant IL‐3. All data points were normalized to IL‐33‐induced levels of IL‐2. Data were summarized from 4 independent experiments with BMMC of 19 independent donors. (D) Splenic CD25 + CD4 + T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or 50 ng/ml recombinant SCF was added. Transcription factor expressions of RORγt and Helios among FoxP3 + CD4 + Tregs were analysed by flow cytometry in 2 experiments with BMMCs from 3 independent donors. Ratio of RORγt − Helios + Tregs among Th cells related to conditions with IL‐33 alone is shown. Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Journal: Immunology

    Article Title: IL‐3 is essential for ICOS‐L stabilization on mast cells, and sustains the IL‐33‐induced RORγt+ Treg generation via enhanced IL‐6 induction

    doi: 10.1111/imm.13305

    Figure Lengend Snippet: IL‐3 blocks the IL‐33‐induced IL‐2 production by BMMC, which selectively enhances the RORγt − Helios + T reg maintenance. (A, B) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or 50 ng/ml recombinant IL‐2 was added. Transcription factor expressions of FoxP3 among CD4 + , and RORγt and Helios among FoxP3 + CD4 + Tregs were analysed by flow cytometry in 2 experiments with BMMCs from 6 independent donors. (A) Representative FACS plots of RORγt + and Helios + cells among FoxP3 + CD4 + T regs . (B) Diagrams summarize RORγt + Helios − , RORγt + Helios + , RORγt − Helios + and RORγt − Helios − cells among FoxP3 + CD4 + T regs . (C) BMMCs were cultured in medium alone or with 50 ng/ml recombinant IL‐33 for 24 h. IL‐2 levels in the supernatants were analysed by ELISA. If indicated, BMMCs were costimulated with 50 ng/ml recombinant IL‐3. All data points were normalized to IL‐33‐induced levels of IL‐2. Data were summarized from 4 independent experiments with BMMC of 19 independent donors. (D) Splenic CD25 + CD4 + T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or 50 ng/ml recombinant SCF was added. Transcription factor expressions of RORγt and Helios among FoxP3 + CD4 + Tregs were analysed by flow cytometry in 2 experiments with BMMCs from 3 independent donors. Ratio of RORγt − Helios + Tregs among Th cells related to conditions with IL‐33 alone is shown. Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Article Snippet: Coculture For cocultures, 100 000 BMMCs were seeded with 25 000 MHC‐II− CD4+ CD25+ Treg in the presence of 1.5 µg/ml soluble anti‐mouse CD3ε (clone 2C11, Institute for Immunology) in the presence of IL‐33 (50 ng/ml, Peprotech), with or without IL‐3 (50 ng/ml, Peprotech), with or without SCF (50 ng/ml, Peprotech) in a 96‐well plate for 3 days.

    Techniques: Recombinant, Flow Cytometry, FACS, Cell Culture, Enzyme-linked Immunosorbent Assay

    IL‐3 enhances the IL‐33‐induced RORγt + T reg development independent from ICOS‐L. (A‐C) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone, or in the presence of 50 ng/ml recombinant IL‐3 ± 50 ng/ml recombinant IL‐33 for 3 days. Transcription factor expression of FoxP3 among CD4 + , and RORγt among FoxP3 + CD4 + T regs was analysed by flow cytometry. Representative FACS plots and histograms are shown in the right panels. Diagrams on the right summarize data from separate experiments representing BMMCs from n > 10 biologically independent donors. (A) FoxP3 + cells among CD4+ Th cells. (B) RORγt + FoxP3 + expression among CD4 + Th cells. (C) RORγt + FoxP3 + expression among CD4 + Th cells in the presence of blocking anti‐ICOS‐L antibody. (D, E) BMMCs were cultured in the presence of recombinant 50 ng/ml IL‐3 ( n = 12 different donors) and 50 ng/ml IL‐33 for 2 ( n = 2 different donors), 6 ( n = 2 different donors) or 24 h ( n = 12 different donors). Frequencies of ICOS‐L + cells were analysed among CD117 + BMMCs by flow cytometry. (D) Representative FACS plots. (E) Data from all cultures of BMMCs are summarized. (F) BMMCs were cultured in medium ( n = 6), medium containing 50 ng/ml recombinant IL‐3 alone ( n = 6) or with additional 50 ng/ml recombinant IL‐33 ( n = 3). Transcript levels of Icosl and Gapdh were quantified by qRT‐PCR after 24 h. Relative expressions of Icosl related to Gapdh are summarized from the indicated numbers of independent biological replicates. Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Journal: Immunology

    Article Title: IL‐3 is essential for ICOS‐L stabilization on mast cells, and sustains the IL‐33‐induced RORγt+ Treg generation via enhanced IL‐6 induction

    doi: 10.1111/imm.13305

    Figure Lengend Snippet: IL‐3 enhances the IL‐33‐induced RORγt + T reg development independent from ICOS‐L. (A‐C) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone, or in the presence of 50 ng/ml recombinant IL‐3 ± 50 ng/ml recombinant IL‐33 for 3 days. Transcription factor expression of FoxP3 among CD4 + , and RORγt among FoxP3 + CD4 + T regs was analysed by flow cytometry. Representative FACS plots and histograms are shown in the right panels. Diagrams on the right summarize data from separate experiments representing BMMCs from n > 10 biologically independent donors. (A) FoxP3 + cells among CD4+ Th cells. (B) RORγt + FoxP3 + expression among CD4 + Th cells. (C) RORγt + FoxP3 + expression among CD4 + Th cells in the presence of blocking anti‐ICOS‐L antibody. (D, E) BMMCs were cultured in the presence of recombinant 50 ng/ml IL‐3 ( n = 12 different donors) and 50 ng/ml IL‐33 for 2 ( n = 2 different donors), 6 ( n = 2 different donors) or 24 h ( n = 12 different donors). Frequencies of ICOS‐L + cells were analysed among CD117 + BMMCs by flow cytometry. (D) Representative FACS plots. (E) Data from all cultures of BMMCs are summarized. (F) BMMCs were cultured in medium ( n = 6), medium containing 50 ng/ml recombinant IL‐3 alone ( n = 6) or with additional 50 ng/ml recombinant IL‐33 ( n = 3). Transcript levels of Icosl and Gapdh were quantified by qRT‐PCR after 24 h. Relative expressions of Icosl related to Gapdh are summarized from the indicated numbers of independent biological replicates. Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Article Snippet: Coculture For cocultures, 100 000 BMMCs were seeded with 25 000 MHC‐II− CD4+ CD25+ Treg in the presence of 1.5 µg/ml soluble anti‐mouse CD3ε (clone 2C11, Institute for Immunology) in the presence of IL‐33 (50 ng/ml, Peprotech), with or without IL‐3 (50 ng/ml, Peprotech), with or without SCF (50 ng/ml, Peprotech) in a 96‐well plate for 3 days.

    Techniques: Recombinant, Expressing, Flow Cytometry, FACS, Blocking Assay, Cell Culture, Quantitative RT-PCR

    IL‐3‐enhanced IL‐33‐induced RORγt + T reg generation at the expense of RORγt − Helios + T regs is dependent on IKK2 and on the p38‐MK2/3 module. (A‐F) BMMCs were cultured in medium alone or with 50 ng/ml recombinant IL‐33 for 24 h. IL‐6 levels in the supernatants were analysed by ELISA. If indicated, BMMCs were costimulated with 50 ng/ml recombinant IL‐3. All data points were normalized to IL‐33‐induced levels of IL‐6. Data are pooled from independent BMMC cultures. (A) Preincubation of the BMMC with IKKi7 ( n = 4). (B) Preincubation of the BMMC with UO126 ( n = 17). (C) Preincubation of the BMMC with SP600125 ( n = 15). (D) Preincubation of the BMMC with SB203580 ( n = 10). (E) MK2/3 dKO BMMCs were used ( n = 4). (F) Preincubation of the BMMC with PF3644022 ( n = 18). (G‐J) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or blocking anti‐IL‐6 antibody was added. Transcription factor expression of FoxP3 among CD4 + , and RORγt and Helios among FoxP3 + CD4 + T regs was analysed by flow cytometry in 2 independent experiments with BMMCs from 8 independent donors. (G) Representative FACS plots show FoxP3 + cells among CD4 + Th cells. (H) Diagram summarizes FoxP3 + cells among CD4 + Th cells. (I) Representative FACS plots of RORγt + and Helios + cells among FoxP3 + CD4 + T regs . (J) Diagrams summarize RORγt + Helios − , RORγt + Helios + , RORγt − Helios + and RORγt − Helios − cells among FoxP3 + CD4 + T regs . Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Journal: Immunology

    Article Title: IL‐3 is essential for ICOS‐L stabilization on mast cells, and sustains the IL‐33‐induced RORγt+ Treg generation via enhanced IL‐6 induction

    doi: 10.1111/imm.13305

    Figure Lengend Snippet: IL‐3‐enhanced IL‐33‐induced RORγt + T reg generation at the expense of RORγt − Helios + T regs is dependent on IKK2 and on the p38‐MK2/3 module. (A‐F) BMMCs were cultured in medium alone or with 50 ng/ml recombinant IL‐33 for 24 h. IL‐6 levels in the supernatants were analysed by ELISA. If indicated, BMMCs were costimulated with 50 ng/ml recombinant IL‐3. All data points were normalized to IL‐33‐induced levels of IL‐6. Data are pooled from independent BMMC cultures. (A) Preincubation of the BMMC with IKKi7 ( n = 4). (B) Preincubation of the BMMC with UO126 ( n = 17). (C) Preincubation of the BMMC with SP600125 ( n = 15). (D) Preincubation of the BMMC with SB203580 ( n = 10). (E) MK2/3 dKO BMMCs were used ( n = 4). (F) Preincubation of the BMMC with PF3644022 ( n = 18). (G‐J) CD25 + CD4 + splenic T regs were cocultured with BMMCs in medium ± 50 ng/ml recombinant IL‐33 alone for 3 days. If indicated, 50 ng/ml recombinant IL‐3 or blocking anti‐IL‐6 antibody was added. Transcription factor expression of FoxP3 among CD4 + , and RORγt and Helios among FoxP3 + CD4 + T regs was analysed by flow cytometry in 2 independent experiments with BMMCs from 8 independent donors. (G) Representative FACS plots show FoxP3 + cells among CD4 + Th cells. (H) Diagram summarizes FoxP3 + cells among CD4 + Th cells. (I) Representative FACS plots of RORγt + and Helios + cells among FoxP3 + CD4 + T regs . (J) Diagrams summarize RORγt + Helios − , RORγt + Helios + , RORγt − Helios + and RORγt − Helios − cells among FoxP3 + CD4 + T regs . Diagrams show average values of all experiments ± SEM. Statistics were done with Student's t‐ test: n.s . not significant; * p

    Article Snippet: Coculture For cocultures, 100 000 BMMCs were seeded with 25 000 MHC‐II− CD4+ CD25+ Treg in the presence of 1.5 µg/ml soluble anti‐mouse CD3ε (clone 2C11, Institute for Immunology) in the presence of IL‐33 (50 ng/ml, Peprotech), with or without IL‐3 (50 ng/ml, Peprotech), with or without SCF (50 ng/ml, Peprotech) in a 96‐well plate for 3 days.

    Techniques: Cell Culture, Recombinant, Enzyme-linked Immunosorbent Assay, Blocking Assay, Expressing, Flow Cytometry, FACS

    IL-4Rα-expression by epithelial cells (ECs) and smooth muscle cells (SMCs) is required in IL-4Rα flox/− mice for maximal airway hyperresponsiveness (AHR) induced by exogenous IL-13. IL-4Rα flox/− mice were treated i.t. with 3 µg of IL-13 daily for 7 days. A. Invasive measurement of AHR. B. Absolute eosinophil count of BAL fluid. C. Quantitation of goblet cell metaplasia (GCM). D. Distribution of residual GCM in club cell 10 kDa protein-Cre + (CC10-Cre + ) mice and CC10-Cre + α-smooth muscle actin-Cre + (SMP8-Cre + ) mice. E. Absolute eosinophil count of BAL fluid in CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high or low residual GCM. F. Percentage of eosinophils in BAL fluid in CC10-Cre + mice and CC10-Cre + SMP8-Cre + that had high or low residual GCM. G, AHR when CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high residual GCM were excluded. The results represent 12 pooled experiments with panels A-D showing 17, 14, 26, 22 and 47 mice for groups: Cre − + IL-13, SMP8-Cre + + IL-13, CC10-Cre + + IL-13, Both Cre + + IL-13, all genotypes + saline, respectively. Panel G represents 14 and 12 mice for groups: CC10-Cre + + IL-13 and both Cre + + IL-13, respectively. NS, not significant; *, p

    Journal: Mucosal immunology

    Article Title: IL-4Rα expression by airway epithelium and smooth muscle accounts for nearly all airway hyperresponsiveness in murine allergic airway disease.

    doi: 10.1038/s41385-019-0232-7

    Figure Lengend Snippet: IL-4Rα-expression by epithelial cells (ECs) and smooth muscle cells (SMCs) is required in IL-4Rα flox/− mice for maximal airway hyperresponsiveness (AHR) induced by exogenous IL-13. IL-4Rα flox/− mice were treated i.t. with 3 µg of IL-13 daily for 7 days. A. Invasive measurement of AHR. B. Absolute eosinophil count of BAL fluid. C. Quantitation of goblet cell metaplasia (GCM). D. Distribution of residual GCM in club cell 10 kDa protein-Cre + (CC10-Cre + ) mice and CC10-Cre + α-smooth muscle actin-Cre + (SMP8-Cre + ) mice. E. Absolute eosinophil count of BAL fluid in CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high or low residual GCM. F. Percentage of eosinophils in BAL fluid in CC10-Cre + mice and CC10-Cre + SMP8-Cre + that had high or low residual GCM. G, AHR when CC10-Cre + and CC10-Cre + SMP8-Cre + mice that had high residual GCM were excluded. The results represent 12 pooled experiments with panels A-D showing 17, 14, 26, 22 and 47 mice for groups: Cre − + IL-13, SMP8-Cre + + IL-13, CC10-Cre + + IL-13, Both Cre + + IL-13, all genotypes + saline, respectively. Panel G represents 14 and 12 mice for groups: CC10-Cre + + IL-13 and both Cre + + IL-13, respectively. NS, not significant; *, p

    Article Snippet: For IL-13-based experiments, 2 or 3 µg IL-13 were administered i.t. in 40 µl daily for 7 treatments ( ).

    Techniques: Expressing, Mouse Assay, Quantitation Assay

    A single functional IL-4Rα allele is sufficient for induction of maximal GCM and IL-4 and IL-13 responses, but not AHR. A–C: mice were treated i.t. with 3 µg IL-13 daily for 7 days. A. Invasive measurement of AHR. B. Absolute eosinophil count of BAL fluid. C. Quantitation of GCM. D-G. Mice were treated i.t. with 16 µg HDM extract (produced in our lab) every other day over 14 days. D. Invasive measurement of AHR. E. Absolute eosinophil count of BAL fluid. F. Quantitation of GCM. G. Measurement of cytokine production in vivo for the ~24hrs following the seventh i.t. treatment with HDM or saline. A–C. 8, 17 and 18 mice for the experimental groups: WT + IL-13, IL-4Rα flox/− + IL-13 and both genotypes + saline, respectively. D–F. 17, 24 and 33 mice for the experimental groups: WT + HDM, IL-4Rα flox/− + HDM and both genotypes + saline, respectively. G. 5, 8, 7 and 8 mice for the groups: WT + saline, IL-4Rα +/− + saline, WT + HDM and IL-4Rα +/− + HDM, respectively. Because of breeding constraints, WT mice and IL-4Rα +/− mice were not littermates to IL-4Rα flox/− or to themselves. NS, not significant; *, p

    Journal: Mucosal immunology

    Article Title: IL-4Rα expression by airway epithelium and smooth muscle accounts for nearly all airway hyperresponsiveness in murine allergic airway disease.

    doi: 10.1038/s41385-019-0232-7

    Figure Lengend Snippet: A single functional IL-4Rα allele is sufficient for induction of maximal GCM and IL-4 and IL-13 responses, but not AHR. A–C: mice were treated i.t. with 3 µg IL-13 daily for 7 days. A. Invasive measurement of AHR. B. Absolute eosinophil count of BAL fluid. C. Quantitation of GCM. D-G. Mice were treated i.t. with 16 µg HDM extract (produced in our lab) every other day over 14 days. D. Invasive measurement of AHR. E. Absolute eosinophil count of BAL fluid. F. Quantitation of GCM. G. Measurement of cytokine production in vivo for the ~24hrs following the seventh i.t. treatment with HDM or saline. A–C. 8, 17 and 18 mice for the experimental groups: WT + IL-13, IL-4Rα flox/− + IL-13 and both genotypes + saline, respectively. D–F. 17, 24 and 33 mice for the experimental groups: WT + HDM, IL-4Rα flox/− + HDM and both genotypes + saline, respectively. G. 5, 8, 7 and 8 mice for the groups: WT + saline, IL-4Rα +/− + saline, WT + HDM and IL-4Rα +/− + HDM, respectively. Because of breeding constraints, WT mice and IL-4Rα +/− mice were not littermates to IL-4Rα flox/− or to themselves. NS, not significant; *, p

    Article Snippet: For IL-13-based experiments, 2 or 3 µg IL-13 were administered i.t. in 40 µl daily for 7 treatments ( ).

    Techniques: Functional Assay, Mouse Assay, Quantitation Assay, Produced, In Vivo