FIG. 3.

Alizarin red S staining. Mineralized nodule formation. Calcium nodules (red) were observed using Alizarin red S staining. LIPUS-treated cell culture. Heat-shock-treated cell culture. Treatment 1 followed the experimental time schedule indicated in Figure 1. In treatment 2, the cells received the stimuli at day 0 when the cells were seeded on the dishes. Control cells were cultured in the osteogenic medium without the stimulations. Quantitative analysis of Alizarin red S staining was performed by measuring the absorbance of destained Alizarin red S at 562 nm. The results are expressed as mean ± standard deviation (SD) (n = 3). *, versus A1 and C1. p < 0.05 was recognized to be significant.