Figure 5.

Simvastatin-induced osteogenic differentiation in an animal model of ectopic bone formation was reduced by cytoskeletal alteration agents.

Notes: Immunofluorescence staining revealed that the F-actin, staining with phalloidin, was increased by simvastatin treatment and reduced by cotreatment with biochemical agents, scale bar represents 50 µm (A). Inhibiting actin filament organization or decreasing cell rigidity significantly inhibited simvastatin-induced osteogenic gene expression (B). Representative masses of ectopic bone formation were stained with Alizarin Red S, the 100× figures are represented as the boxed area in the 40× line, and the mineralization of masses was quantified using Image-Pro Plus software (C). The results are the mean values of three independent measurements, error bars: SEM. **P<0.01 vs control; ^#P<0.05; ^##P<0.01 vs SIM (1 µM).

Abbreviations: SIM, simvastatin; Bleb, blebbistatin; CD, cytochalasin D; SEM, standard error of the mean; OC, osteocalcin.