Fig. 1

Multipotential differentiation of MSCs. a Experimental protocol for the in-vivo study. To isolate diseased MSCs (d-MSCs), wild-type C57BL/6 N mice received both Dex and Zol administered intravenously for 2 weeks with tooth extraction. Control MSCs (c-MSCs) were isolated from normal mice, which received no treatment (n = 4 × 2 groups). b (Top left) Osteogenic differentiation of MSCs. After culture under osteogenic differentiation conditions for 4 weeks, osteogenic differentiation was determined by Alizarin Red S staining. Quantification of the Alizarin Red S dye content in differentiated osteoblasts from independent experiments is shown (mean ± SD). Scale bar, 50 μm. (Top middle) Adipogenic differentiation of MSCs. After culture under adipogenic differentiation conditions for 2 weeks, adipocyte differentiation was determined by Oil Red O staining. Zol treatment decreased adipogenic differentiation of MSCs, as indicated by the decreased number of Oil Red O-positive cells (mean ± SD). *P < 0.05. (Top right) Endothelial differentiation of MSCs. After culture under endothelial differentiation conditions for 2 weeks, differentiation was determined by the formation of typical vessel-like-structures. Scale bar, 50 μm. (Bottom left) MSC migration determined by fluorescence microscopy. Larger numbers of c-MSCs went through the transwell insert compared with d-MSCs. (Bottom middle) Rate of proliferation as determined by the EdU assay. The number of EdU-positive cells was significantly decreased in the d-MSCs from the Zol-treated group compared with c-MSCs from the normal group. (Bottom right) MSCs from normal mice generated fewer CFU-Fs compared with MSCs from MRONJ-like mice. CFU-F colony-forming unit-fibroblast, d days, EdU 5-ethynyl-2′-deoxyuridine, i.p. intraperitoneal, i.v. intravenous