Journal: International Journal of Endocrinology

Article Title: MEK1/2 Inhibitor (GDC0623) Promotes Osteogenic Differentiation of Primary Osteoblasts Inhibited by IL-1 β through the MEK-Erk1/2 and Jak/Stat3 Pathways

doi: 10.1155/2021/5720145

Figure Lengend Snippet: Osteoblastic gene expression profiles in osteoblasts after treatment with GDC0623. (a) Osteoblasts were maintained in an osteogenic differentiation medium containing IL-1 β or IL-1 β plus GDC0623 for 7 days. qPCR analysis of osteogenic gene expression in osteoblasts was performed, relative to internal control GAPDH and normalized to control cells. (b) Immunocytochemical staining of Runx2, OCN, and Col I in osteoblasts after treatment with IL-1 β or GDC0623 plus IL-1 β for 7 days. Data are expressed as mean ± SD for n = 3. ∗ p < 0.05 versus the control group, # p < 0.05 versus the IL-1 β groups.

Article Snippet: Slides were incubated in the blocking solution for 10 min (QuickBlockTM kit, Beyotime), followed by overnight incubation at 4°C in the presence of primary antibodies, ColI (ARG21965, Taiwan, China), and OCN (AF6297, Beyotime, China).

Techniques: Gene Expression, Control, Staining

Journal: International Journal of Nanomedicine

Article Title: Nano-Hydroxyapatite Coating Promotes Porous Calcium Phosphate Ceramic-Induced Osteogenesis Via BMP/Smad Signaling Pathway

doi: 10.2147/IJN.S216182

Figure Lengend Snippet: HE histological analysis of in vivo ectopic bone formation ability for BCP ( A ) and nHA-coated BCP ( B ) scaffolds after implanted in back muscles of rabbits for 90 days. Yellow arrows: inflammatory cells; black arrows: bone formation as evidenced by osteocytes settled in bone lacuna. Immunofluorescent staining of osteocalcin (OCN) for BCP ( C ) and nHA-coated BCP ( D ) groups. Green and blue colors represented osteocalcin and DAPI-stained nuclei, respectively.

Article Snippet: The others were subjected to co-staining with DAPI, rabbit-anti-mouse osteocalcin primary antibody, and goat-anti-rabbit fluorescent secondary antibody (Servicebio, China), and visualized under a fluorescence microscope (Leica, Germany) for immunofluorescence analysis.

Techniques: In Vivo, Muscles, Staining

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

Journal: International Journal of Molecular Sciences

Article Title: Ascorbic Acid Attenuates Senescence of Human Osteoarthritic Osteoblasts

doi: 10.3390/ijms18122517

Figure Lengend Snippet: Effect of ascorbic acid-2-phosphate (AA) on the outgrowth and proliferation of osteoarthritic osteoblasts (OB). ( A ) The number of outgrowing cells from subchondral bone chips from sclerotic (Sc_OB) and non-sclerotic regions (N_OB) was quantified and normalised per gram of bone tissue in standard culture medium and in the presence of AA (+AA; for n = 5–7 donors/experimental group, analysed in triplicates for each donor); ( B ) Expression profile determined by FACS of alkaline phosphatase (ALP) and osteocalcin (OC) in isolated Sc_OB and N_OB, following outgrowth (P0) in the absence (CM) or presence of AA (+AA); ( C ) Proliferation rate, defined by the number of population doublings (PD)/day, of expanded Sc_OB and N_OB ( n = 7 donors, analysed in duplicates for each donor); ( D ) Total number of population doublings before ceasing proliferation by the end of passage 4; ( E ) Expression of the proliferation marker Ki67 at the mRNA level (normalised to the housekeeping gene GAPDH) and ( F ) at the protein level in P2 expanded cells, determined by immunofluorescence. Scale bar = 100 µm. Data in ( A , C – D ) are presented as mean ± SD (standard deviation). Sc_OB = sclerotic osteoblasts; N_OB = non-sclerotic osteoblasts; CM = culture medium; +AA = culture medium supplemented with 0.1 mM ascorbic acid. * indicates statistically significant differences ( p < 0.05).

Article Snippet: Osteocalcin was stained by unconjugated rabbit anti-osteocalcin antibodies (AB1857, Merck, Burlington, MA, USA), followed by Atto 550-conjugated goat anti-rabbit-IgG (43328, Sigma).

Techniques: Expressing, Isolation, Marker, Immunofluorescence, Standard Deviation