rabbit ocn pab  (Bioss)

Journal: Journal of Nanobiotechnology

Article Title: Drynaria roosii -derived exosome-like nanovesicles promote alveolar socket healing via activation of ITPR3-mediated calcium flux

doi: 10.1186/s12951-026-04530-y

Figure Lengend Snippet: In vivo imaging and in vitro osteogenic activity of DRDENs. ( A ) Whole body imaging of the mice at 2, 6, 12, 24, and 48 h after administration of DiR-labeled DRDENs. ( B ) Distribution of DiR-labeled DRDENs in cranio-maxillary bone, mandible, tongue, brain, and tibia at 48 h. ( C - D ) Alkaline phosphatase activity at day 7 and alizarin red S staining at day 21-28 after treatment with different concentrations of DRDENs in MC3T3-E1 cells and hDFSCs, and the corresponding quantitative analysis. ( E - F ) ALP, RUNX2, and OCN mRNA expression levels in DRDENs-incubated MC3T3-E1 cells and hDFSCs were determined by qRT-PCR

Article Snippet: Subsequently, the membranes were incubated with primary antibodies against ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) (ABclonal, China), and β-Actin (Abmart, China) at 4 °C overnight.

Techniques: In Vivo Imaging, In Vitro, Activity Assay, Imaging, Labeling, Staining, Expressing, Incubation, Quantitative RT-PCR

Journal: Journal of Nanobiotechnology

Article Title: Drynaria roosii -derived exosome-like nanovesicles promote alveolar socket healing via activation of ITPR3-mediated calcium flux

doi: 10.1186/s12951-026-04530-y

Figure Lengend Snippet: In vitro cytotoxicity and osteogenic activity of naringenin chalcone. ( A - B ) Cell viability of naringenin chalcone-treated MC3T3-E1 cells and hDFSCs in 24, 48, and 72 h. ( C ) Hemolysis experiment of naringenin chalcone. ( D - E ) Alkaline phosphatase activity and at day 7 and alizarin red S staining at day 21-28 after treatment with different concentrations of naringenin chalcone in MC3T3-E1 cells and hDFSCs, and the corresponding quantitative analysis. ( F - G ) ALP, RUNX2, and OCN mRNA expression levels in naringenin chalcone-treated MC3T3-E1 cells and hDFSCs, analyzed by qRT-PCR

Article Snippet: Subsequently, the membranes were incubated with primary antibodies against ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) (ABclonal, China), and β-Actin (Abmart, China) at 4 °C overnight.

Techniques: In Vitro, Activity Assay, Staining, Expressing, Quantitative RT-PCR

Journal: Journal of Nanobiotechnology

Article Title: Drynaria roosii -derived exosome-like nanovesicles promote alveolar socket healing via activation of ITPR3-mediated calcium flux

doi: 10.1186/s12951-026-04530-y

Figure Lengend Snippet: Histological staining and ELISA assay showing alveolar bone healing in tooth extraction mice with DRDENs and naringenin chalcone treated. ( A ) H&E staining in the M1 alveolar socket (Scale bar: 100 μm). ( B ) Masson’s trichrome staining in the M1 alveolar socket (Scale bar: 100 μm). ( C - D ) Immunohistochemistry staining of RUNX2 and OCN in the M1 alveolar socket (Scale bar: 100 μm). ( E ) Quantitative analysis of inflammatory cell infiltration area in H&E staining and collagen area of new bone in Masson’s trichrome staining. ( F ) Quantitative analysis of positive stained area of RUNX2 and OCN. ( G ) PINP and TRACP-5b levels in mice serum measured by ELISA

Article Snippet: Subsequently, the membranes were incubated with primary antibodies against ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) (ABclonal, China), and β-Actin (Abmart, China) at 4 °C overnight.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Extraction, Immunohistochemistry

Journal: Journal of Nanobiotechnology

Article Title: Drynaria roosii -derived exosome-like nanovesicles promote alveolar socket healing via activation of ITPR3-mediated calcium flux

doi: 10.1186/s12951-026-04530-y

Figure Lengend Snippet: Activation of ITPR3 by naringenin chalcone released by DRDENs ( A ) Volcano plots of differentially expressed genes between the negative control group vs. DRDENs group and the negative control group vs. naringenin chalcone group, analyzed by transcriptome sequencing of the tooth extraction sockets at 14d post procedure. ( B ) Venn diagram showing overlapping genes between the negative control group vs. the DRDENs group and the negative control group vs. the naringenin chalcone group. ( C ) Gene Ontology (GO) enrichment analysis for genes in the purple module of the Venn plot. ( D ) 3D visualization analysis of molecular docking between ITPR3 and naringenin chalcone. ( E ) Interaction between ITPR3 and naringenin chalcone measured by SPR. ( F ) Sequencing validation of L2523Y and E2526R mutated hDFSCs constructed by the CRISPR-Cas9 technique. ( G ) ALP, RUNX2, and OCN mRNA expression levels in the WT hDFSCs group and the mutated hDFSCs group. ( H ) Western blot analysis in the WT hDFSCs group and the mutated hDFSCs group with and without naringenin chalcone treatment. ( I - J ) Calcium flux in the WT hDFSCs group and the mutated hDFSCs group treated with different concentrations of naringenin chalcone

Article Snippet: Subsequently, the membranes were incubated with primary antibodies against ALP, runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) (ABclonal, China), and β-Actin (Abmart, China) at 4 °C overnight.

Techniques: Activation Assay, Negative Control, Sequencing, Extraction, Biomarker Discovery, Construct, CRISPR, Expressing, Western Blot

Journal: Fundamental Research

Article Title: Harmine ameliorates steroid-induced osteonecrosis of the femoral head by modulating metabolism and ferroptosis via HIF1-α

doi: 10.1016/j.fmre.2023.08.015

Figure Lengend Snippet: Harmine restored BMSC activity under MP inhibition. (a) Structural formula of Hm and CCK-8 analysis with various concentrations of Hm in BMSCs. (b) Immunofluorescence images of intracellular ROS with 100 µM MP and 10 µM Hm (Hm-low) or 50 µM Hm (Hm-high) treatment. (Scale bar: 200 µm) (c) The mean fluorescence intensity of ROS was detected by flow cytometry. (d) PI staining was analyzed by flow cytometry. (e) Runx2 was detected with immunofluorescence staining. (Runx2 stained with Alexa Fluor® 647. Scale bar: 25 µm.) (f) Immunofluorescence staining of OCN. (OCN stained with Alexa Fluor® 488 and F-actin stained with rhodamine. Scale bar: 50 µm.) (g) PI positive cells in flow cytometry analysis. (h) MFI of Runx2. (i) MFI of OCN. (All bar graphs are the mean ± SD; * p < 0.05 and ** p < 0.01 compared with the 100 µM MP treatment group; ns: no significant difference; # p < 0.05 and ## p < 0.01 compared with the control group; n = 3 per group).

Article Snippet: After treatment with osteogenic induction medium for 5 days, BMSCs were incubated with primary antibodies against osteocalcin (OCN) (1:200, ABclonal, China: A6205) or runt-related transcription factor 2 (Runx2) (1:200, Abcam: #ab192256) and then stained with immunofluorescence secondary antibodies.

Techniques: Activity Assay, Inhibition, CCK-8 Assay, Immunofluorescence, Fluorescence, Flow Cytometry, Staining, Control

Journal: Fundamental Research

Article Title: Harmine ameliorates steroid-induced osteonecrosis of the femoral head by modulating metabolism and ferroptosis via HIF1-α

doi: 10.1016/j.fmre.2023.08.015

Figure Lengend Snippet: Harmine recovered the osteogenic differentiation potential of BMSCs through the HIF1-α pathway. (a) Runx2 expression was detected with immunofluorescence staining. (Runx2 stained with Alexa Fluor® 647. Scale bar: 25 µm.) (b) OCN expression was detected with immunofluorescence staining. (OCN stained with Alexa Fluor® 488 and F-actin stained with rhodamine. Scale bar: 50 µm.) (c) ALP staining. (ALP activity was detected with an ALP kit. Scale bar: 200 µm) (d) ARS staining. (ARS was dissolved in 10% cetylpyridinium chloride, and the solution was analyzed with a spectrophotometer at 562 nm. Scale bar: 200 µm) (All bar graphs are the mean ± SD; * p < 0.05 and ** p < 0.01 compared with the MP group; ns: no significant difference; # p < 0.05 and ## p < 0.01; n = 3 per group).

Article Snippet: After treatment with osteogenic induction medium for 5 days, BMSCs were incubated with primary antibodies against osteocalcin (OCN) (1:200, ABclonal, China: A6205) or runt-related transcription factor 2 (Runx2) (1:200, Abcam: #ab192256) and then stained with immunofluorescence secondary antibodies.

Techniques: Expressing, Immunofluorescence, Staining, Activity Assay, Spectrophotometry

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA expression is upregulated in CAVD. A H&E staining showing the normal and calcific human aortic valve tissues and immunohistochemical staining showing P21 and MSRA expression in the normal and calcific human aortic valve tissues ( n = 6 for each group), scale bar = 500 μm. B Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in calcified VICs and controls ( n = 6 for each group). C Immunofluorescence staining and quantification of RUNX2 (red) and MSRA (green) in calcified VICs and controls ( n = 6 for each group), scale bar = 20 μm. D Flow cytometry analysis and the mean fluorescence intensity of RUNX2 and MSRA in calcified VICs and controls ( n = 4 for each group). Data are presented as mean ± SEM and compared by Student’s t -test. MSRA, methionine sulfoxide reductase A; CAVD, calcific aortic valve disease; H&E, hematoxylin-and eosin; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Expressing, Staining, Immunohistochemical staining, Western Blot, Immunofluorescence, Flow Cytometry, Fluorescence

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates osteogenic differentiation of human VICs in vitro. A Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in VICs under osteogenic medium or control medium following MSRA silencing ( n = 6 for each group). B Immunofluorescence staining and quantification of RUNX2 (red) under osteogenic medium or control medium after MSRA silencing ( n = 6 for each group), scale bar = 20 μm. C Flow cytometry analysis and the mean fluorescence intensity of RUNX2 in VICs under osteogenic medium or control medium after MSRA silencing ( n = 4 for each group). D Western blot analysis and quantification of OCN, RUNX2, OPN, and MSRA protein expression in VICs under osteogenic medium or control medium following MSRA overexpression ( n = 6 for each group). E Immunofluorescence staining and quantification of RUNX2 (red) under osteogenic medium or control medium after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. F Flow cytometry analysis and the mean fluorescence intensity of RUNX2 in VICs under osteogenic medium or control medium after MSRA overexpression ( n = 4 for each group). Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Staining, Flow Cytometry, Fluorescence, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates VIC senescence in vitro. A Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group). B Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA silencing ( n = 6 for each group), scale bar = 20 μm. C Western blot analysis and quantification of P21, OCN, RUNX2, OPN, and MSRA protein expression in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group). D Immunofluorescence staining and quantification of P21 (magenta) and RUNX2 (red) in VICs under H 2 O 2 stimulus after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; P21, cyclin-dependent kinase inhibitor 1 A; H 2 O 2 , hydrogen peroxide; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; VICs, valvular interstitial cells; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: In Vitro, Western Blot, Expressing, Immunofluorescence, Staining, Over Expression

Journal: GeroScience

Article Title: Age-associated methionine sulfoxide reductase A protects against valvular interstitial cell senescence and valvular calcification

doi: 10.1007/s11357-025-01675-w

Figure Lengend Snippet: MSRA alleviates osteogenic differentiation of VICs by inhibiting the TLR2/NF-κB pathway. A Western blot analysis and quantification of OCN, RUNX2, and OPN protein expression when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 (0.1 μg/mL) after MSRA overexpression ( n = 6 for each group). B Immunofluorescence staining and quantification of RUNX2 (red) when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 20 μm. C Flow cytometry analysis and the mean fluorescence intensity of RUNX2 when human VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 4 for each group). D Alkaline phosphatase staining when VICs were cultured in osteogenic medium with or without TLR2 agonist Pam3CSK4 after MSRA overexpression ( n = 6 for each group), scale bar = 5 mm and 500 μm. Data are presented as means ± SEM and compared by Student’s t -test or one-way analysis of variance followed by Bonferroni post hoc test. MSRA, methionine sulfoxide reductase A; VICs, valvular interstitial cells; OM, osteogenic medium; OCN, osteocalcin; RUNX2, runt-related transcription factor; OPN, osteopontin; DAPI, 4′,6-diamidino-2-phenylindole. NS, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against osteocalcin (OCN) (A20800, Abclonal, Wuhan, China, dilution 1:1000), runt-related transcription factor 2 (RUNX2) (A11753, Abclonal, Wuhan, China, dilution 1:1000), osteopontin (OPN) (ab214050, Abcam, Cambridge, USA, dilution 1:1000), cyclin-dependent kinase inhibitor 1 A (P21) (10,355–1-AP, Proteintech, Wuhan, China, dilution 1:1000), MSRA (14,547–1-AP, Proteintech, Wuhan, China, dilution 1:1000), TLR2 (66,645–1-Ig, Proteintech, Wuhan, China, dilution 1:1000), phosphorylated NF-κB p65 (3033, CST, Danvers, USA, dilution 1:1000), NF-κB p65 (8242, CST, Danvers, USA, dilution 1:1000), and GAPDH (A19056, Abclonal, Wuhan, China) were used.

Techniques: Western Blot, Expressing, Cell Culture, Over Expression, Immunofluorescence, Staining, Flow Cytometry, Fluorescence