Yoda1对人施耐德膜来源间充质细胞成骨分化影响的实验研究OA

Objective:To investigate the effect of the PIEZO1-specific agonist Yoda1 on osteogenic differentiation of mesenchymal cells derived from human Schneiderian membrane(hSM).Methods:hSM-derived cells were isolated and cultured by enzymatic digestion.Cell surface markers were identified by flow cytometry,and the expression of keratin 14(KRT14)and cathepsin K(CTSK)was detected by immunofluorescence staining.PIEZO1 protein expression was evaluated by Western blotting,and a calciumion fluorescent probe was used to detect activation of the PIEZO1 channels by Yoda1.The effects of different concentrations of Yoda1 on the proliferation and migration abilities of hSM-derived mesenchymal cells were evaluated using CCK-8 and wound healing assays,respectively.Following osteogenic induction,the expression levels of alkaline phosphatase(ALP)gene ALPL and Runt-related transcription factor 2(RUNX2)were analyzed by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blotting.ALP staining and alizarin red S staining were performed to evaluate the osteogenic differentiation ability of the cells.Results:hSM-derived cells expressed mesenchymal cell surface markers and co-expressed KRT14 and CTSK proteins.PIEZO1 protein was detected in these cells,and Yoda1 effectively activated the PIEZO1 channels.Low concentrations of Yoda1 had no significant effect on cell proliferation,whereas a high concentration(10.0 µmol/L)significantly inhibited proliferation(P<0.05).No significant effect on cell migration was observed.During osteogenic induction,low-concentration Yoda1(0.1 µmol/L)upregulated ALPL expression and enhanced ALP activity,while no significant change was observed in RUNX2 expression.Western blotting results were consistent with mRNA expression trends.ALP staining and alizarin red S staining indicated that low-concentration Yoda1 exerted a modest promoting effect on osteogenic differentiation.Conclusion:Yoda1 exhibits a limited promoting effect on osteogenic differentiation of hSM-derived mesenchymal cells,and low concentrations may promote its early osteogenic differentiation.