Fig. 6.

Osteoclast differentiation of rBMMs after coculture with different nanomaterials in medium supplemented with the indicated concentrations of M-CSF and RANKL. (a) Determination of the apoptotic effect of different nanomaterials on rBMMs using a flow cytometry assay after staining with an Annexin V-FITC Detection kit. (b) Observation of cells after being fixed with 4% PFA and stained with an acid phosphatase staining kit, followed by a quantitative analysis of the area and number of TRAP-positive multinuclear cells. (c) Relative mRNA levels of RANKL-induced expression of osteoclast-specific genes (TRAP, CTR, Cathepsin K and NFATc1) in rBMMs at 1, 3 and 5 d determined by RT-PCR. *p < 0.01 compared with the GaNPs and GO/Ga groups, **p < 0.05 compared with the other groups.