Figure 3.

sEV-miR-324 derived from osteoclasts can be delivered into MSCs and induce osteogenic differentiation

(A) The expression profile of miRNAs in sEVs secreted during osteoclast differentiation. sEVs from BMMs were used as a normalization control. Red color represents higher expression, and blue color represents lower expression relative to the control. (B) ALP activity assay shows that overexpression of miR-324 promoted osteogenic differentiation; n = 3. (C and D) Relative expression levels of (C) miR-324 and (D) pri-miR-324 in MSCs cultured with sEVs from miR-324 overexpression (miR-324-sEVs) or knockdown (anti-miR-324-sEVs) osteoclasts; n = 3. (E) Cell viability evaluation of MSCs cultured with miR-324-sEVs or anti-miR-324-sEVs using a CCK-8 test at 1, 3, 5, and 7 days; n = 5. (F) Relative mRNA expression levels of Col1a1, Alpl, Sp7, and Runx2 in MSCs cultured with miR-324-sEVs or anti-miR-324-sEVs; n = 3. (G) Western blot analysis of COL1A1, RUNX2, ALP, and β-actin in MSCs cultured with miR-324-sEVs or anti-miR-324-sEVs. (H) Representative images of alizarin red staining of MSCs cultured with miR-324-sEVs or anti-miR-324-sEVs. Scale bar represents 100 μm. (I) Quantification analysis of calcium deposit of MSCs in indicated groups; n = 5. Data in the figures represent the averages ± SD. ∗p < 0.05, ∗∗p < 0.01, for differences between the treatment and control groups.