primary antibodies against osteocalcin (ocn)  (Affinity Biosciences)

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Irisin Promotes Osteogenesis by Modulating Oxidative Stress and Mitophagy through SIRT3 Signaling under Diabetic Conditions

doi: 10.1155/2022/3319056

Figure Lengend Snippet: Irisin improved the osteogenic differentiation of ASCs exposed to AGEs in vitro. (a–d) Osteogenic differentiation was assessed by ALP staining on day 7 (a) and semiquantitative analysis of ALP staining (b) and ARS on day 21 (c). Calcium deposition was assessed by measuring the optical density at 570 nm (d). (e) qPCR analysis of the expression of the osteogenesis-specific genes Runx2 , Col1 , Ocn , and Opn ( β -actin was used as the internal control). All results are representative of three independent experiments, and values are presented as mean ± SEM. Scale bar = 400 μ m. ALP: alkaline phosphatase; ARS: Alizarin red staining. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: Further, specimens were also permeabilized in 0.3% Triton X-100 and then blocked in 5% normal goat serum, incubated overnight with primary antibodies against osteocalcin (OCN) (1 : 200) and Collagen I (Col1) (1 : 200) (Bioss, Beijing, China) at 4°C, and stained with HRP-conjugated secondary antibodies (ZSJQ-BIO, Beijing, China).

Techniques: In Vitro, Staining, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Irisin Promotes Osteogenesis by Modulating Oxidative Stress and Mitophagy through SIRT3 Signaling under Diabetic Conditions

doi: 10.1155/2022/3319056

Figure Lengend Snippet: Irisin alleviated AGE-induced suppression of cell viability and osteogenic differentiation by increasing SIRT3 expression in vitro. (a, b) The gene and protein expression levels of SIRT3 were assessed by qPCR (a) and WB analysis (b) with different concentrations of irisin treatment. (c) The gene and protein expression levels of SIRT3 in ASCs treated with or without irisin under AGE-induced conditions. (d) Cells were transfected with lentiviruses that either overexpress or silence SIRT3, and cell viability was evaluated by CCK-8 assay. (e, f) Osteogenic differentiation was determined by ALP staining on day 7 and semiquantitative analysis of ALP staining (e) and ARS on day 21. Calcium deposition was assessed by measuring the optical density at 570 nm (f). (g) qPCR analysis of the expression of the osteogenic-specific genes Runx2 , Col1 , Ocn , and Opn ( β -actin was used as the internal control). All results are representative of three independent experiments, and values are presented as mean ± SEM. Scale bar = 400 μ m. Lv-Control: control lentivirus; Lv-SIRT3: lentivirus-mediated SIRT3 overexpression; Lv-shSIRT3: lentivirus-mediated SIRT3 knockdown. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: Further, specimens were also permeabilized in 0.3% Triton X-100 and then blocked in 5% normal goat serum, incubated overnight with primary antibodies against osteocalcin (OCN) (1 : 200) and Collagen I (Col1) (1 : 200) (Bioss, Beijing, China) at 4°C, and stained with HRP-conjugated secondary antibodies (ZSJQ-BIO, Beijing, China).

Techniques: Expressing, In Vitro, Transfection, CCK-8 Assay, Staining, Over Expression

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Irisin Promotes Osteogenesis by Modulating Oxidative Stress and Mitophagy through SIRT3 Signaling under Diabetic Conditions

doi: 10.1155/2022/3319056

Figure Lengend Snippet: Transplantation of gels encapsulating ASCs and irisin promoted bone healing in CSDs in diabetic rats. (a) Timeline for the operation process in SD rats. (b) Representative micro-CT reconstruction images of rat CSDs (scale bar = 1 mm). (c) The microstructural parameters of the ROI, including BV/TV and BMD ( n = 6). (d) Representative images of calvarial defect sections stained with HE (scale bar = 1 mm in upper images; scale bar = 200 μ m in lower images). The lower panels show magnifications of the insets in each group. (e) Representative images of calvarial defect sections stained with Masson's trichrome staining (scale bar = 1 mm) (the blue color indicates the regenerated bone, collagen fibers, or osteoid, while the red color indicates the mature bone). (f) Representative images of calvarial defect sections with immunohistochemical staining of OCN. The dark brown color represents the positive staining for OCN protein, with black arrows indicating OCN-positive staining. (g) Representative images of calvarial defect sections with immunohistochemical staining of Col1. The dark brown color represents the positive staining for Col1 protein. Scale bar = 1 mm in upper images, scale bar = 50 μ m in lower images in (f, g), and the lower panels show magnifications of the insets in each group. BV/TV: bone volume/total volume; BMD: bone mineral density. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001.

Article Snippet: Further, specimens were also permeabilized in 0.3% Triton X-100 and then blocked in 5% normal goat serum, incubated overnight with primary antibodies against osteocalcin (OCN) (1 : 200) and Collagen I (Col1) (1 : 200) (Bioss, Beijing, China) at 4°C, and stained with HRP-conjugated secondary antibodies (ZSJQ-BIO, Beijing, China).

Techniques: Transplantation Assay, Micro-CT, Staining, Immunohistochemical staining

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Knockdown of enhancer of zeste homolog 2 inhibits steo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that the expression of enhancer of zeste homolog 2 (EZH2) was inhibited in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed the knockdown of EZH2 in hSCAPs; C: Knockdown of EZH2 decreased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis demonstrated that knockdown of EZH2 inhibited mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that knockdown of EZH2 downregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs. GAPDH and ACTB was used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Staining

Journal: World Journal of Stem Cells

Article Title: EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells

doi: 10.4252/wjsc.v17.i4.103482

Figure Lengend Snippet: Overexpression of enhancer of zeste homolog 2 enhances osteo/dentinogenic differentiation potential of human apical papillary stem cells. A: Quantitative polymerase chain reaction showed that enhancer of zeste homolog 2 (EZH2) was overexpressed in human apical papillary stem cells (hSCAPs); B: Western blot analysis confirmed overexpression of EZH2 in hSCAPs; C: Overexpression of EZH2 increased alkaline phosphatase activity in hSCAPs; D and E: Alizarin red staining and quantitative calcium analysis results demonstrated that overexpression of EZH2 enhanced mineralization in hSCAPs; F-H: Quantitative polymerase chain reaction showed that overexpression of EZH2 upregulated mRNA expression levels of bone sialoprotein (F), dentin sialophosphoprotein (G), and osteocalcin (H) in hSCAPs; I: Hematoxylin-eosin staining and quantitative measurement showed that overexpression of EZH2 promoted bone/dentin-like tissue formation. Scale bar = 100 μm (B: Bone/dentin-like tissues; HA: Hydroxyapatite tricalcium carrier; CT: Connective tissue); J: Immunohistochemical staining and quantitative analysis of dentin sialophosphoprotein and bone sialoprotein. GAPDH and ACTB were used as the internal controls. Data are presented as the mean ± SD ( n = 3). Statistical analysis was performed using Student’s t -test. a P ≤ 0.05, b P ≤ 0.01, c P ≤ 0.001. EZH2: Enhancer of zeste homolog 2; BSP: Bone sialoprotein; DSPP: Dentin sialophosphoprotein; OCN: Osteocalcin.

Article Snippet: Immunohistochemical analysis followed established protocols[ ] using primary antibodies against osteocalcin (OCN) (Bioss bs-4917R) and DSPP (Bioss bs10316R).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Staining, Expressing, Immunohistochemical staining

Journal: Biomaterials Translational

Article Title: Ångstrom-scale silver particle-infused hydrogels eliminate orthopedic implant infections and support fracture healing

doi: 10.12336/biomatertransl.2025.01.007

Figure Lengend Snippet: Abbreviations: AgÅPs: Ångstrom-scale silver particles; OCN: Osteocalcin; TRAP: Tartrate resistant acid phosphatase.

Article Snippet: Immunohistochemical staining was performed using primary antibodies against osteocalcin (OCN) (Cat no.: GB11233, Servicebio, China) with a dilution of 1:300, tumor necrosis factor-alpha (TNF-α) (Cat no.: GB11188, Servicebio, China) with a dilution of 1:800, and interleukin-1 beta (IL-1β) (Cat no.: GB11113, Servicebio, China) with a dilution of 1:400.

Techniques:

Journal: Biomaterials Translational

Article Title: Ångstrom-scale silver particle-infused hydrogels eliminate orthopedic implant infections and support fracture healing

doi: 10.12336/biomatertransl.2025.01.007

Figure Lengend Snippet: Note: Statistical significance determined at * p < 0.05 and ** p < 0.01. Abbreviations: AgÅPs: Ångstrom-scale silver particles; E. coli: Escherichia coli ; Gel: Hydrogel; IM: Induced medium ; OCN: Osteocalcin; TRAP: Tartrate resistant acid phosphatase.

Article Snippet: Immunohistochemical staining was performed using primary antibodies against osteocalcin (OCN) (Cat no.: GB11233, Servicebio, China) with a dilution of 1:300, tumor necrosis factor-alpha (TNF-α) (Cat no.: GB11188, Servicebio, China) with a dilution of 1:800, and interleukin-1 beta (IL-1β) (Cat no.: GB11113, Servicebio, China) with a dilution of 1:400.

Techniques: