Comparison of the osteogenic effect of different growth factors with that of CME.

<p>(A) MG-63 cells grown in OIM with or without various concentrations (1, 5 and 20 ng/mL) of bFGF, TGF β-1, EGF alone or their combination and CME (100–800 μg/mL). On day 9, Calcium concentration was compared by performing the calcium assay: F+T: bFGF + TGFβ-1, F+T+E: bFGF + TGFβ-1+ EGF. (B) MG-63 cells were cultured in OIM with or without high concentration of BMP2 (50–500 ng/mL), bFGF (50–500 ng/mL), and TGFβ-1 (50–500 ng/mL) alone or their combinations (bFGF + TGFβ-1 and bFGF + TGFβ-1+ BMP2) and CME (100–800 μg/mL). The cells were treated with the indicated concentrations of the growth factors and CME every 2–3 days and were cultured for 18 days. ECM mineralization was determined by performing Alizarin red S staining and by measuring the absorbance of solubilized Alizarin red S at 570 nm: F+T: bFGF + TGFβ-1, F+T+B: bFGF + TGFβ-1+ BMP2. Scale bars represent 100 μm. (C) Human MSCs were cultured in OIM with or without various concentrations (1 and 10 ng/mL) of bFGF, TGFβ-1, BMP2 and CME (100–400 μg/mL). ALP activity was measured at day 7. Data are presented as the mean ± SD of multiple repeated experiments; **<i>p</i>< 0.01 and ^#<i>p</i>< 0.001 versus OIM, indicates statistical significance.