Figure 3.

Biomineralization assay for ALC and LS8 cells. (A) Alizarin Red staining was detected at levels significantly higher than background at day 3 post inoculation, and this is characteristic of calcified nodule formation. In ALC cells, stain accumulation increased over a period of 21 days, with no significant change beyond 21-days. Only background level staining was observed in LS8 cells, when compared to NIH3T3 cells (negative control) at day 1 and also over this same time period. Scale bar 100.0 μm. (B) Alizarin Red stain in NIH3T3, LS8, and ALC cell culture were quantified using Osteogenesis quantification kit at OD₄₀₅. Graph shows significant increase in red staining in ALC cells only. Significance indicated by ^* for p < 0.05 for differences in individual cell lines.