Figure 7

Comparison of the FB osteogenic differentiation potential at P3 versus P10. (A) Evaluation of osteogenic differentiation potential at P3 with alizarin red s. The osteogenic differentiation increased over the time course. At day 21 the inhibition of the TGF-β signaling with SB431542 (OM + SB) was significantly superior to the inhibition of BMP signaling with dorsomorphin (OM + DM) and even better than the inhibition of TGF-β signaling with the simultaneous use of BMP-2 (OM + BMP + SB). The differentiation with BMP-2 and simultaneous inhibition of the BMP signaling with dorsomorphin (OM + BMP + DM) was significantly better than the osteogenic differentiation with BMP-2 alone (OM + BMP). (B) Evaluation of the osteogenic differentiation potential at P10 with alizarin red s. At day 21 the osteogenic differentiation with BMP-2 with the simultaneous inhibition of the BMP signaling (OM + BMP + DM) was the best and significantly superior to the treatment with dorsomorphin only (OM + DM). Inhibiting the TGF-β signaling with SB431542 and the additional use of BMP-2 (OM + BMP + SB) was significantly superior to the inhibition of TGF-β signaling with SB431542 (OM + SB). Black bars are FB differentiated with osteogenic standard differentiation media (OM). Grey bars are FB differentiated with OM and dorsomorphin (OM + DM). Stacked grey bars were FB differentiated osteogenically and treated with SB431542 (OM + SB). White bars were FB coincubated with OM and BMP-2 with the inhibitor dorsomorphin (OM + BMP + DM). Stacked white bars are FB differentiated with OM, BMP-2 and the inhibitor SB431542 (OM + BMP + SB). Bars represent mean ± SD of three donors. *p < 0.05 as compared to the respective sample of the culture of P10. (C) Visualized comparison of osteogenic differentiation potential of DSC at P3 and P10. Evaluation was performed with alizarin red s on day 21. Shown is one representative illustration of at least six identical results. The image scales represent a length of 200 µm.