Journal: International Journal of Nanomedicine

Article Title: Hollow Hydroxyapatite Microspheres Loaded with rhCXCL13 to Recruit BMSC for Osteogenesis and Synergetic Angiogenesis to Promote Bone Regeneration in Bone Defects

doi: 10.2147/IJN.S408905

Figure Lengend Snippet: Analysis of osteogenic differentiation in BMSCs co-cultured with scaffolds. ( A ) Quantitative RT-qPCR analysis of osteogenic gene expression (Runx2, ALPL, OCN) in BMSCs co-cultured with each group. GAPDH was used as housekeeping genes. The results were expressed as mean ± SD. ( B ) Relative protein expression of RUNX2 and OCN in BMSCs co-cultured with various treatment groups. ( C ) Representative images of Alizarin red-stained BMSCs co-cultured with various scaffolds for 21 days in 6-well plates (scale bar = 100μm). ( D ) Quantitative analysis of Alizarin Red Staining in BMSCs co-cultured with various treatment groups. Statistical analysis was performed using ANOVA and LSD tests, with results expressed as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001, n=3).

Article Snippet: Sections were blocked for one hour with the blocking solution at room temperature and then transferred to RUNX2 (ABclonal, China), OCN (ABclonal, China), COLI (ABclonal, China), BMP-2 (ABclonal, China) of primary antibodies overnight at 4°temperature.

Techniques: Cell Culture, Quantitative RT-PCR, Gene Expression, Expressing, Staining

Journal: International Journal of Nanomedicine

Article Title: Hollow Hydroxyapatite Microspheres Loaded with rhCXCL13 to Recruit BMSC for Osteogenesis and Synergetic Angiogenesis to Promote Bone Regeneration in Bone Defects

doi: 10.2147/IJN.S408905

Figure Lengend Snippet: Immunofluorescence Staining Analysis of Osteogenic Differentiation in BMSCs Co-Cultured with Scaffolds. ( A ) Immunofluorescence staining of BMSCs (COLI, BMP-2, RUNX2, and OCN) when co-cultured with scaffolds (( A and B ). ( B ) Quantitative analysis of immunofluorescence staining for COLI, BMP-2, RUNX2, and OCN performed by Image J. ( a1, a2, b1 and b2 ). Statistical analysis was performed using ANOVA and LSD tests, with results expressed as mean ± SD (*p<0.05, **p<0.01, ***p<0.001, n=3).

Article Snippet: Sections were blocked for one hour with the blocking solution at room temperature and then transferred to RUNX2 (ABclonal, China), OCN (ABclonal, China), COLI (ABclonal, China), BMP-2 (ABclonal, China) of primary antibodies overnight at 4°temperature.

Techniques: Immunofluorescence, Staining, Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Hollow Hydroxyapatite Microspheres Loaded with rhCXCL13 to Recruit BMSC for Osteogenesis and Synergetic Angiogenesis to Promote Bone Regeneration in Bone Defects

doi: 10.2147/IJN.S408905

Figure Lengend Snippet: Heat Map, Protein-Protein Interaction (PPI) Network, and KEGG Up-Pathway Enrichment Analyses in Various Treatment Groups. ( A ) Heat Map of differentially expressed genes related to cellular activity, osteogenesis, angiogenesis, and BMSC recruitment in different groups comparisons. ( B1 and B2 ) PPI analysis revealing the function of DEG-encoded proteins in different groups. ( C1-C3 ) KEGG up-pathway enrichment analysis in different groups comparisons. The red symbol in ( C1 ) indicates that the KEGG up-pathway enrichment analysis of differentially expressed genes mainly concentrated in the PI3K-AKT signaling pathway. ( D ) Changes in the relative gene expression levels of RUNX2 and OCN after treatment with PI3K-AKT inhibitor BEZ235 in BMSCs from all groups. ( E ) Representative alizarin red staining image of BMSCs co-cultured with rhCXCL13-HHM/CS scaffold after 21 days of osteogenic differentiation treated with PI3K-AKT inhibitor BEZ235 (scale bar = 100μm, n=3). ( F ) Quantitative analysis of alizarin red staining of BMSCs in different treatment groups (***p<0.001, n=3).

Article Snippet: Sections were blocked for one hour with the blocking solution at room temperature and then transferred to RUNX2 (ABclonal, China), OCN (ABclonal, China), COLI (ABclonal, China), BMP-2 (ABclonal, China) of primary antibodies overnight at 4°temperature.

Techniques: Activity Assay, Gene Expression, Staining, Cell Culture

Journal: BMC Musculoskeletal Disorders

Article Title: Nodular osteochondrogenic activity in soft tissue surrounding osteoma in neurogenic para osteo-arthropathy: morphological and immunohistochemical study

doi: 10.1186/1471-2474-5-46

Figure Lengend Snippet: Immunological study of serial sections : a: Type II Collagen immunolabellingcountre stained with h&e: Slide 69: Type II collagen had the same pattern of ALP activity of nodule showing prehypertrophic chondrocytes in nonmineralized zone. b: Type I collagen Immunolabelling countre stained with h&e: Slide 72; Type I collagen expression encercled the mineralized nodule. c: Type X collagen immunolabelling countre stained with h&e:Slide 73: Type X collagen was expressed by most of cells and distributed in their matrix. d: OCN Immunolabelling countre stained with h&e: Slide 64: OCN was expressed by hypertrophic chondrocytes. e: OCN immunolabelling counter stained with h&e: Activated osteoblasts (OB) and woven bone (W) are strongly labelled. Some capillaries (C) near by these areas express osteocaline too. f: VEGF immunolabelling counter stained with h&e: VEGF was expressed by some hypertrophic chondrocytes (H). Activated and non activated osteoblasts-like cells (OB) lining the cartilage express also VEGF. Inset: Clustered and isolated cells in the matrix, surrounding hypertrophic chondrocytes, which could be destinated to capillary or osteoblast formation, are also labelled by VEGF.

Article Snippet: The anti-human monoclonal mouse antibodies against type II collagen (NeoMarkers Inc., CA, USA), OCN (Interchim, Montlucon, France), anti-human polyclonal goat antibody against type I collagen (Santacruz Biotechnology, CA, USA), anti-rat polyclonal rabbit antibody against type X collagen (Calbiochem-Novabiochem, CA, USA) and anti-human polyclonal rabbit antibody against VEGF (Santacruz Biotechnology, CA, USA) were used at 1 mg/ml.

Techniques: Staining, Activity Assay, Expressing, Isolation

Journal: Bioactive Materials

Article Title: Pretreated exosomes by electrical stimulation accelerate bone regeneration

doi: 10.1016/j.bioactmat.2025.04.019

Figure Lengend Snippet: Histological staining of the distal femur defect site at 6-weeks post-injection. (A) Hematoxylin and eosin (H&E) staining, (B) Masson's trichrome staining, and (C) Immunohistochemical staining for OCN. D) The relative proportion of collagen from the quantitative analysis of Masson staining. E) Quantitative analysis results of IHC Results are shown as mean ± SD. ns: non-significant differences, ∗∗p < 0.01.

Article Snippet: They were then co-incubated with the primary antibody, specifically the OCN primary antibody from Beyotime, China.

Techniques: Staining, Injection, Immunohistochemical staining

Journal: Stem Cell Research & Therapy

Article Title: A NOTCH1 / LSD1 / BMP2 co-regulatory network mediated by miR-137 negatively regulates osteogenesis of human adipose-derived stem cells

doi: 10.1186/s13287-021-02495-3

Figure Lengend Snippet: NOTCH1 inhibitor (tangeretin) reverses the effects of miR-137 knockdown on osteogenesis and downstream genes expression. a , b ALP staining ( a ) and quantification ( b ) of transfected hASCs after a 7-day culture in PM or OM (scale bar = 100 μm). c , d ARS staining ( c ) and quantification ( d ) of transfected hASCs after a 14-day culture in PM or OM. e Relative expression analyses of RUNX2 , ALP , and OCN by qRT-PCR in transfected hASCs on 3 days, 7 days, and 14 days. f Relative expression analyses of NOTCH1 , HES1 , LSD1 , BMP2 , and SMAD4 by qRT-PCR in transfected hASCs on 3 days, 7 days, and 14 days. g , h Western blotting ( g ) and band intensity analyses ( h ) of NOTCH1, HES1, LSD1, BMP2, SMAD4, and p-SMAD1/5 in transfected hASCs on 7 days. Data are shown as mean ± SD of three independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001 versus respective NC group

Article Snippet: The rabbit anti-OCN primary antibodies diluted to 1:100 (Servicebio, Wuhan, Hubei, China; GB11233) were used for immunohistochemical (IHC) staining.

Techniques: Expressing, Staining, Transfection, Quantitative RT-PCR, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: A NOTCH1 / LSD1 / BMP2 co-regulatory network mediated by miR-137 negatively regulates osteogenesis of human adipose-derived stem cells

doi: 10.1186/s13287-021-02495-3

Figure Lengend Snippet: HES1 knockdown promotes osteogenesis of hASCs in vitro. a , b ALP staining ( a ) and quantification ( b ) of transfected hASCs after a 7-day culture in PM or OM (scale bar = 100 μm). c , d ARS staining ( c ) and quantification ( d ) of transfected hASCs after a 14-day culture in PM or OM. e Relative expression analyses of HES1 , RUNX2 , ALP , and OCN by qRT-PCR in transfected hASCs on 3 days, 7 days, and 14 days. Data are shown as mean ± SD of three independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001 versus respective NC group

Article Snippet: The rabbit anti-OCN primary antibodies diluted to 1:100 (Servicebio, Wuhan, Hubei, China; GB11233) were used for immunohistochemical (IHC) staining.

Techniques: In Vitro, Staining, Transfection, Expressing, Quantitative RT-PCR

Journal: Regenerative Biomaterials

Article Title: Combination of graphene oxide and platelet-rich plasma improves tendon–bone healing in a rabbit model of supraspinatus tendon reconstruction

doi: 10.1093/rb/rbab045

Figure Lengend Snippet: Primer sequences used for RT-PCR

Article Snippet: Samples were fixed with 4% paraformaldehyde for 30 min, treated with 0.5% Triton X-100 for 15 min and blocked with 2% BSA (Solarbio) for 40 min. After washing with PBS, the samples were stained against an OCN primary antibody (1:100; GeneTex, USA) or a Col II primary antibody (1:100; Proteintech, China) overnight.

Techniques: Sequencing

Journal: Regenerative Biomaterials

Article Title: Combination of graphene oxide and platelet-rich plasma improves tendon–bone healing in a rabbit model of supraspinatus tendon reconstruction

doi: 10.1093/rb/rbab045

Figure Lengend Snippet: ( A ) The expressions of genes related to osteogenic differentiation analyzed by qRT-PCR. ( B ) Immunofluorescence staining images of OCN expression at 14 d. ( C ) The expressions of genes related to chondrogenic differentiation analyzed by qRT-PCR. ( D ) Immunofluorescence staining images of col II expression at 21 d. Scale bar = 50 μm. * P < 0.05 and ** P < 0.01 vs PRP

Article Snippet: Samples were fixed with 4% paraformaldehyde for 30 min, treated with 0.5% Triton X-100 for 15 min and blocked with 2% BSA (Solarbio) for 40 min. After washing with PBS, the samples were stained against an OCN primary antibody (1:100; GeneTex, USA) or a Col II primary antibody (1:100; Proteintech, China) overnight.

Techniques: Quantitative RT-PCR, Immunofluorescence, Staining, Expressing

Journal: Dentistry Journal

Article Title: In Vivo Evaluation of Regenerative Osteogenic Potential Using a Human Demineralized Dentin Matrix for Dental Application

doi: 10.3390/dj12030076

Figure Lengend Snippet: Representative histological images of (H&E) ( A , D , G , J ), Masson’s trichrome ( B , E , H , K ) and osteocalcin immune staining ( C , F , I , L ) are shown at 4 weeks post-treatment. Coronal sections through the mid-point of the defects were prepared from decalcified specimens. Fibrous connective tissues were seen to be mostly filled in the defect areas in the control group ( A – C ). The group treated with SA hydrogel as the only grafted material showing fibrous and adipose tissue ( D – F ). lacuna spaces were observed in the newly formed lamellar bone in NHH with a wide bone marrow space ( G – I ). DDMH-treated groups showed considerable new bone trabeculae formation ( J – L ). BT: bone trabeculae; FT: fibrous tissue. Arrows represent MT- and OCN-positive reactions. Scale bar: 100 μm.

Article Snippet: Subsequently, the samples were embedded in paraffin; and after staining with hematoxylin and eosin (H&E), Masson’s trichrome (MT) (Cat# HT15, Sigma Aldrich), and immunohistochemical staining with a polyclonal anti-rabbit osteocalcin antibody (OCN; Cat# GB11233, diagnostic biosystems) at a 1:150 dilution, histological examination was conducted using an Olympus BX-51 optical microscope (Olympus Co., Tokyo, Japan).

Techniques: Staining, Control

Journal: Dentistry Journal

Article Title: In Vivo Evaluation of Regenerative Osteogenic Potential Using a Human Demineralized Dentin Matrix for Dental Application

doi: 10.3390/dj12030076

Figure Lengend Snippet: Representative histological images of (H&E) ( A , D , G , J ), Masson’s trichrome ( B , E , H , K ) and osteocalcin immune staining ( C , F , I , L ) are shown at 8 weeks post-treatment. Rather than the newly created bone, the defect locations in the control group were found to be primarily filled with fibrous connective tissues and adipose tissue ( A – C ). Circular new bony islands with the presence of early woven bone were observed in the group treated with SA as the only grafted material ( D – F ). Newly formed lamellar bone was observed in NHH with a wide bone marrow space ( G – I ). In contrast, DDMH-treated groups showed considerable new bone formation at 8 weeks with relatively narrow marrow spaces ( J – L ). BT means bone trabeculae; FT: fibrous tissue. Arrows represent MT- and OCN-positive reactions. Scale bar: 100 μm.

Article Snippet: Subsequently, the samples were embedded in paraffin; and after staining with hematoxylin and eosin (H&E), Masson’s trichrome (MT) (Cat# HT15, Sigma Aldrich), and immunohistochemical staining with a polyclonal anti-rabbit osteocalcin antibody (OCN; Cat# GB11233, diagnostic biosystems) at a 1:150 dilution, histological examination was conducted using an Olympus BX-51 optical microscope (Olympus Co., Tokyo, Japan).

Techniques: Staining, Control