Figure 3.

Alizarin red S staining of human osteoblasts (SaOs-2) after incubation with calcium chloride or calcitonin. Cells were incubated with increasing calcium chloride (1, 2.5, 5, and 10 mM) or calcitonin (1, 2.5, 5, and 10 nM) concentrations for three (left panel) and four weeks (right panel), and subsequently stained with alizarin red S. Each condition was assessed in triplicate in three independent experiments (n = 9). ODM− served as a negative control (n = 6), ODM+ (n = 12) as reference, and acetic acid (n = 6) was used as an additional control as calcitonin was dissolved in acetic acid. Representative pictures of transmitted light microscopy at 20x magnification are shown. Scale bar is 50 µm. ODM; osteoblast differentiation medium.