Fig. 4

Macrophages treated with the combination of Ex-4 and ED-71 promote the osteogenic differentiation of BMSCs. A Identification of BMSCs by flow cytometry. CD73 and CD105 were positive, while CD34 and CD45 were negative. B Pattern of co-culture system. BMSCs were inoculated in the upper chamber and RAW264.7 cells were cultured in the lower chamber. C Pattern of co-culture system. RAW264.7 cells were inoculated in the upper chamber and BMSCs were cultured in the lower chamber. D Crystal violet staining. Using the pattern of co-culture system "BMSCs up", it was found that Ex-4 and ED-71 promote the migration of BMSCs by acting on macrophages. Bar, 200 μm. E ALP staining. Using the pattern of co-culture system "RAW264.7 up", it was illustrated that Ex-4 and ED-71 promoted the osteogenic differentiation of BMSCs by acting on macrophages. Bar, 500 μm. F AR staining. Using the pattern of co-culture system "RAW264.7 up", it was shown that Ex-4 and ED-71 promoted the osteogenic differentiation of BMSCs by acting on macrophages. Bar, 1 mm. G The results of crystal violet staining, ALP staining and AR staining were quantitatively evaluated. *p < 0.05, **p < 0.01, ***p < 0.001. CON control, Ex-4 exendin-4, ED-71 eldecalcitol, Ex-4 + ED-71 exendin-4 + eldecalcitol, BMSCs bone marrow mesenchymal stem cells osteogenesis, ALP alkaline phosphatase, AR alizarin red