Fig. 4

Systemic SHED-EV infusion rescued the functions of mBMMSCs in recipient OVX mice. a Representative images of mineralized nodules after Alizarin Red staining 4 weeks after osteogenesis. The Alizarin Red⁺ cells. Bars = 10 mm. The graph shows Alizarin Red-positive (Alizarin Red⁺) areas. b The ratio of the expression of osteoblast markers, including Runt-related transcription factor 2 (Runx2) and bone gamma carboxyglutamic acid protein (Bglap), and osteoprotective marker semaphorin-3a (Sema3a) by RT-qPCR 1 week after the induction of osteogenesis. The data are shown as a ratio to the expression in the mBMMSCs derived from sham-operated mouse (Sham = 1). c The expression of osteoblast markers, including RUNX2 and BGLA by Western blot analysis 1 week after the induction of osteogenesis. ACTB actin, beta. d The expression of SEMA3A in the conditioned medium by enzyme-linked sorbent immunoassay (ELISA) 1 week after the induction of osteogenesis. a–d Sham, sham-operated group; OVX, PBS-infused OVX group; MOCK-EV, MOCK-treated SHED-EV-infused OVX group; RNase-EV, RNase-retreated SHED-EV-infused OVX group. n = 7 for all groups. Comparisons between two groups were analyzed by an independent two-tailed Student’s t test. Multiple group comparisons were performed by a one-way analysis of variance followed by Tukey’s post hoc test.*P < 0.05, **P < 0.01, ***P < 0.005, ns no significance. Graph bars represent mean ± SEM