Fig. 3

Mineralization and gene expression of osteoblast markers. a–e Cells were treated with α-MEM, OM, OM + EGF and OM + bFGF for 21 days and stained with alizarin red S (ARS), micrographs show extracellular calcium deposition. Original magnification 10x, scale bar =100 μm. f Calcified areas were quantified. Total calcium content was significantly increased with EGF treatment compared to all groups (p < 0.001). Error bars indicate mean ± SD (n = 3), asterisk indicate statistical significance (p < 0.001). g Total RNA was extracted from induced osteoblast-like cells. mRNA expression of the osteogenic markers, alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN) and osteopontin (OPN), was examined by RT-PCR. The housekeeping gene β-actin was used as a control for the PCR reaction. The results of this study confirm the participation of these genes in regulating the mineralization process of the extracellular matrix. All treatments were performed in triplicate