Fig. 4
Osteoclastic differentiation effect of Raw264.7 cells conditioned medium. a TRAP staining of BMMs cultured in conditioned medium for 5 days. b The bone resorption assay of BMMs cultured in conditioned medium for 5 days: claret indicates eroded area. c F-actin rings staining of BMMs cultured in conditioned medium for 5 days. d, e Quantitative analysis of the number and area of osteoclasts in TRAP staining. f Quantitative analysis of the eroded area in bone resorption assay. g Quantitative analysis the total number of osteoclasts with more than three nuclei in F-actin rings staining. h–k RT-qPCR results of Acp5, c-Fos, Oscar, and Sema after BMMs cells cultured in conditioned medium for 5 days. (n = 3; *, # and + represent P < 0.05 when compared with Ti/SF, Ti/SF/MOF and Ti/SF/I, respectively; **, ## and ++ represent P < 0.01)