Journal: Biomaterials

Article Title: The use of heparin/polycation coacervate sustain release system to compare the bone regenerative potentials of 5 BMPs using a critical sized calvarial bone defect model

doi: 10.1016/j.biomaterials.2022.121708

Figure Lengend Snippet: Effects of 5 different BMPs on the osteogenic differentiation of hMDSCs at day 28 (3D pellet culture). A. MicroCT 3D images showed that the mineralized pellets in 5 BMPs groups were larger than the CTL group. Scale bar=1mm. B. Quantification of mineralized pellet volume. All BMPs groups showed higher mineralized pellet volume compared to the CTL group. The mineralized pellet volume of the BMP2 group was significantly higher than the BMP6 and BMP7 groups. *P<0.05, ** P<0.01, ****, P<0.0001. C. Mineralized pellet density quantification. No statistical difference between BMP groups and the CTL group was found. D. Von Kossa staining. Brown-black color indicates mineralization. All pellets have regions peeled off due to high mineralization in the white region of pellets. E. Immunohistochemistry of osteocalcin for osteogenic pellets. Brown color indicates osteocalcin expression. The mineralized parts of the pellets demonstrate crystal purple color intermingled with osteocalcin brown staining. Scale bars=100μm.

Article Snippet: Osteocalcin immunohistochemistry using a mouse anti-human osteocalcin primary antibody (MAB1419, 1:100, R&D Systems) was also performed, as previously described [ 27 ].

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Journal of pharmacological sciences

Article Title: Farrerol suppresses osteoclast differentiation and postmenopausal osteoporosis by inhibiting the nuclear factor kappa B signaling pathway.

doi: 10.1016/j.jphs.2023.12.011

Figure Lengend Snippet: Fig. 6. Histological and immunohistochemical analyses of distal femur sections from mice. (A) Representative images of H&E, Masson’s trichrome, and TRAP staining. The trabecular bone was sparser in OVX mice, but the density improved in response to treatment with 25 and 50 mg/kg farrerol. Images were scanned and presented at low magnification ( × 2 magnification, scale bar, 500 μm) and high magnification ( × 20 magnification, scale bar, 50 μm). (B) Quantification of TRAP staining. Farrerol treatment (25 and 50 mg/kg) effectively prevented in vivo osteoclast formation. (C) Serum CTX-1 levels. TRAP-positive multinucleated osteoclasts in the trabecular bone region immediately below the whole growth plate were evaluated. Results of TRAP staining and ELISA were obtained from six independent experiments. (D) Representative images of the immunohistochemical staining of p-P65 and p-P38 ( × 40 magnification, scale bar, 25 μm). Quantitative analysis of OCN expression levels were obtained from 10 fields from six independent experiments. **P < 0.01. n.s., not significant.

Article Snippet: The slices were evaluated using hematoxylin and eosin (H&E), Masson’s trichrome, and TRAP staining, as reported previously.11 For IHC analyses, the slices were treated overnight with primary antibodies against OCN (1:500; 23418-1-AP, Proteintech), p-P65 (1:50; ab31473, Abcam) or p-P38 (1:100; #4511, Cell Signaling Technology) at 4 ◦C.

Techniques: Immunohistochemical staining, Staining, In Vivo, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Journal of pharmacological sciences

Article Title: Farrerol suppresses osteoclast differentiation and postmenopausal osteoporosis by inhibiting the nuclear factor kappa B signaling pathway.

doi: 10.1016/j.jphs.2023.12.011

Figure Lengend Snippet: Fig. 7. Farrerol did not affect osteoblastic bone formation in vitro or in vivo. (A) Representative ALP staining images (scale bar, 500 μm). (B) Representative ARS staining images (scale bar, 500 μm). (C) Quantitative analysis of ALP-positive areas. (D) Quantitative analysis of ARS-positive areas. (E) Representative images of the immunohistochemical staining of OCN. Images were scanned and presented at low magnification ( × 2 magnification, scale bar, 500 μm) and high magnification ( × 20 magnification, scale bar, 50 μm). (F) Quantitative analysis of OCN expression levels. (G) Serum P1NP level. (H) Representative images of calcein-alizarin red S double labeling (scale bar, 20 μm). Compared to that in the OVX group, farrerol did not affect the MS/BS (I) or MAR (J). The results of ALP and ARS staining were obtained from nine randomly selected visual fields from three independent experiments. In the in vivo study, data were obtained from 18 fields from six independent experiments. *P < 0.05. n.s., not significant. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The slices were evaluated using hematoxylin and eosin (H&E), Masson’s trichrome, and TRAP staining, as reported previously.11 For IHC analyses, the slices were treated overnight with primary antibodies against OCN (1:500; 23418-1-AP, Proteintech), p-P65 (1:50; ab31473, Abcam) or p-P38 (1:100; #4511, Cell Signaling Technology) at 4 ◦C.

Techniques: In Vitro, In Vivo, Staining, Immunohistochemical staining, Expressing, Labeling

Journal: Life (Basel, Switzerland)

Article Title: Inhibition of Pathological Increased Matrix Metalloproteinase (MMP) Activity for Improvement of Bone Regeneration in Diabetes.

doi: 10.3390/life12020134

Figure Lengend Snippet: Figure 5. Immunohistochemistry for osteocalcin. An increase in osteocalcin level was detected in healing bones of db−/db−mice after inhibition of MMP activity seven days postoperatively, albeit it was not significant. Values are depicted as ±SD. Scale bar: 50 µm.

Article Snippet: Briefly, primary antibody against osteocalcin (rabbit, polyclonal, SantaCruz Biotechnology, sc-30045, 1:50) was applied.

Techniques: Immunohistochemistry, Inhibition, Activity Assay

Journal: Scientific Reports

Article Title: Viable cryopreserved human bone graft exhibit superior osteogenic properties in mandibular lateral augmentation

doi: 10.1038/s41598-023-28170-6

Figure Lengend Snippet: Association between OCN ( A ), COL1 ( B ), OPN ( C ), TRAP ( D ) and graft type at each time point. A-HBG acellular-human bone graft, VC-HBG viable cryopreserved-human bone graft, OCN osteocalcin, COL1 collagen type 1, OPN osteopontin, TRAP tartrate-resistant acid phosphatase.

Article Snippet: Immunohistochemical reactions were performed using the following primary antibodies to assess bone formation and remodeling: anti-OCN (Osteocalcin, clone: ab10911, Millipore, Massachusetts, USA, 1:200 dilution), anti-OPN (Osteopontin, clone: Akm2A1, Santa Cruz Biotechnology, Inc, 1:100 dilution), anti-COL 1 (Collagen type 1, clone: ab90395, Abcam, Massachusetts, USA, 1:100 dilution) and anti-TRAP (tartrate resistant acid phosphatase, clone: EPR15556, Abcam, Massachusetts, USA, 1:200 dilution).

Techniques:

Journal: Tissue Engineering. Part C, Methods

Article Title: Development of Modular, Dual-Perfused Osteochondral Constructs for Cartilage Repair

doi: 10.1089/ten.tec.2018.0356

Figure Lengend Snippet: Osteocalcin expression in osteogenic microbeads. (A) IHC revealed osteocalcin (brown) within osteogenic microbeads cultured under both osteogenic and control conditions (scale bar = 100 μm). (B) Osteocalcin was detected through ELISA in conditioned medium of control (agarose only) microbeads cultured under osteogenic conditions (*p ≤ 0.05 compared with other groups). ELISA, enzyme-linked immunosorbent assay; IHC, immunohistochemistry. Color images are available online.

Article Snippet: Next, sections were blocked and subsequently incubated overnight at 4°C in a mouse anti-rat osteocalcin primary antibody (R&D Systems) in tris-buffered saline (TBS).

Techniques: Expressing, Cell Culture, Control, Enzyme-linked Immunosorbent Assay, Immunohistochemistry

Journal: Tissue Engineering. Part C, Methods

Article Title: Development of Modular, Dual-Perfused Osteochondral Constructs for Cartilage Repair

doi: 10.1089/ten.tec.2018.0356

Figure Lengend Snippet: Osteogenic differentiation in constructs as indicated by osteocalcin IHC (brown staining). Dotted lines mark the physical interface between phases (scale bar = 100 μm). Color images are available online.

Article Snippet: Next, sections were blocked and subsequently incubated overnight at 4°C in a mouse anti-rat osteocalcin primary antibody (R&D Systems) in tris-buffered saline (TBS).

Techniques: Construct, Staining