hydroxyapatite tricalcium phosphate hatcp based scaffold  (Zimmer Biomet)

Journal: Journal of Advanced Research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: P2rx7 promoted the bone formation of MSCs. a The CFU-F in MSCs derived from the control and the P2rx7 -/- MSCs. b Mineralized nodules formed by MSCs of the control and the P2rx7 -/- mice were analyzed by alizarin red staining. c, d The expressions of osteogenesis marker genes in the control and the P2rx7 -/- MSCs, as assessed by western blotting ( c ) and qPCR ( d ). e, f Transplants consisting of MSCs from control and P2rx7 -/- mice mixed with HA/TCP were analyzed by H&E staining ( e ), and immunofluorescent staining of CD31 and ACAN ( f ), n = 5 per group. g Mineralized nodules formed by MSCs under osteogenic induction when exposed to 0, 40 nM or 100 nM BzATP were showed by alizarin red staining. h The expressions of osteogenic markers Runx2, ALP, and OCN in MSCs exposed to 0, 40 nM or 100 nM BzATP, as assessed by western blotting. i, j Transplants consisting of MSCs treated with 0, 40, 100 nM BzATP from control mice mixed with HA/TCP were analyzed by H&E staining ( i ), and immunofluorescent staining of CD31 and ACAN ( j ), n = 5 per group. Scale bar: e, f, i, j: 50 μm. e, i yellow, new bone; black, blood vessel; blue, new cartilage. Data was presented as mean ± SD. The data (a-d, g) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test (a, b, e, f) and one-way ANOVA (d, g, i, j). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Then MSCs were mixed with 40 mg hydroxyapatite tricalcium phosphate (HA/TCP, framing, Inc.) ceramic powder as scaffold (Zimmer Inc., Warsaw, IN, USA).

Techniques: Derivative Assay, Control, Staining, Marker, Western Blot, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: The purinergic receptor P2rx7 mediated ATP sensing is required to prevent bone aging by directing mitochondrial fitness of MSCs

doi: 10.1016/j.jare.2025.06.026

Figure Lengend Snippet: P2rx7 regulated mitochondrial fusion through Mfn1. a The mitochondria (top) labeled by Mitotracker Red and mitochondrial morphology (bottom) based on Mitochondrial Analyzer after grey processing, analyzed by Airyscan confocal images in the control and P2rx7 -/- MSCs. b The expression of genes associated with mitochondrial dynamics in the control and the P2rx7 -/- MSCs as assessed by western blotting. c Representative micrographs visualizing P2rx7 (arrows) on the mitochondria. d Reprehensive Airyscan confocal images of mitochondrial morphology (Mitotracker Red, top) and mitochondrial morphology (bottom) based on Mitochondrial Analyzer after grey processing in the MSCs treated with 0, 40 nM and 100 nM BzATP. e The expression of genes associated with mitochondrial dynamics in the MSCs treated with 0, 40 nM and 100 nM BzATP as assessed by western blotting. f Western blotting analysis of mitochondrial (left) and cytosolic (right) fractions of the MSCs treated with 40 nM and 100 nM BzATP. g IHC staining of Mfn1 and Opa1 proteins in the control and the P2rx7 -/- MSCs that were subcutaneously implanted into immunocompromised mice with HA/TCP (n = 5 per group). Scale bar: a, d:10 μm (top), 5 μm(bottom); g: 100 μm; h: 500 μm. Data was presented as mean ± SD. The data (a, b, d-f) are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs con by two-tailed Student’s t test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Then MSCs were mixed with 40 mg hydroxyapatite tricalcium phosphate (HA/TCP, framing, Inc.) ceramic powder as scaffold (Zimmer Inc., Warsaw, IN, USA).

Techniques: Labeling, Control, Expressing, Western Blot, Immunohistochemistry, Two Tailed Test