Fig. 8
Ultrasound-powered hydrogel facilitates BMSCs osteogenic differentiation by increasing Ca2+ influx and active PI3K/AKT and MEK/ERK pathways. (a) Detections of Vm in different groups. Scale bar represents 50 μm. (b) Quantitative analysis of immunofluorescence intensity of Vm (n = 3). (c) Immunofluorescent staining of intracellular Ca2+ (green) of BMSCs in GO, 0.1KBGO, and 0.1KBGO + US groups. Scale bar represents 100 μm. (d) Quantitative analysis of immunofluorescence intensity of intracellular Ca2+ (n = 3). (e) Immunofluorescent staining of intracellular Ca2+ (green) of BMSCs in GO, 0.1KBGO, and 0.1KBGO + US groups after Ca2+ influx inhibited by GdCl3. Scale bar represents 100 μm. (f) Quantitative analysis of immunofluorescence intensity of intracellular Ca2+ after Ca2+ influx inhibited by GdCl3 (n = 3). (g) Western blot assay of the phosphorylation levels of PI3K, AKT, MEK, and ERK in BMSCs in different groups. (h) Western blot assay of the phosphorylation levels of PI3K, AKT, MEK, and ERK and osteogenesis-related proteins Runx2 and OCN in BMSCs after Ca2+ influx inhibited by GdCl3. (i) Schematic illustration of the molecular mechanism of the ultrasound-powered hydrogel facilitating osteogenesis. ANOVA followed by Tukey’s post hoc test was performed for statistical analysis (*p < 0.05, **p < 0.01, ***p < 0.001)