FIGURE 5.

Piezo1 knockdown hampers the proliferation, migration and osteogenic differentiation of BMSCs. (A) Fluorescence intensity of Ca²⁺ after Piezo1 knockdown or treatment with 1 μmol/L Yoda1 as measured by the Fluo‐3 AM probe under a fluorescence microscope. (B) Effects of 1 μmol/L Yoda1 or knockdown of Piezo1 in macrophages on vertical migration of BMSCs under static 2‐h tension, as shown by ×4 micrographs of BMSCs in different groups at the bottom of the migration transwell culture chamber. (C) Cell Counting Kit 8 (CCK8) was performed to explore BMSCs proliferation following treatment with 1 μmol/L Yoda1 or knockdown of Piezo1. (D) The protein expression of Runx2, Osx and Opn in BMSCs were detected by western blot. GAPDH was used for normalization. (E) Col1, Osx, Runx2, Ocn, Opn and Alp in BMSCs were measured by real‐time RT‐PCR. GAPDH was used for normalization. (F) ALP expression in BMSCs was measured by the ALP staining method. The top are gross scanning images (scale bar: 1 mm), and the lower are enlarged images (magnification: ×250, scale bar: 160 μm). (G) After 21 days, the osteogenic differentiation of BMSCs was evaluated by Alizarin Red staining. The top are gross scanning images (scale bar: 1 mm), and the lower are enlarged images (magnification: ×250, scale bar: 160 μm). Data represent the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.