Figure 5

Comparison of the DSC osteogenic differentiation potential at P3 versus P10. (A) Evaluation of osteogenic differentiation potential at P3 with alizarin red s. At day 21 the inhibition of TGF-β signaling with SB431542 (OM + SB) was significantly superior to the inhibition of BMP-2 signaling with dorsomorphin (OM + DM). If TGF-β signaling was inhibited with SB431542 and the BMP-2 signaling was induced with BMP-2 (OM + BMP + SB) the osteogenic differentiation was significantly superior to a solely inhibition of TGF-β signaling (OM + SB). (B) Evaluation of osteogenic differentiation potential at P10 with alizarin red s. At day 21 the inhibition of TGF-β signaling in the course of osteogenic differentiation with SB431542 (OM + SB) was significantly superior to the inhibition of BMP-2 signaling with dorsomorphin (OM + DM), and superior (in the contrary to P3) to the inhibition of TGF-β signaling with SB431542 and inducing BMP signaling with BMP-2 (OM + BMP + SB). Additionally, the inhibition of the BMP-2 signaling with dorsomorphin (OM + DM) was superior to the osteogenic differentiation with dorsomorphin and BMP-2 (OM + BMP + DM). Black bars represent DSC differentiated with osteogenic standard differentiation media (OM). Grey bars demonstrate DSC differentiated with OM and dorsomorphin (OM + DM). Stacked grey bars are DSC differentiated osteogenically and treated with SB431542 (OM + SB). White bars are DSC coincubated with OM and BMP-2 with the inhibitor dorsomorphin (OM + BMP + DM). Stacked white bars are DSC differentiated with OM, BMP-2 and the inhibitor SB431542 (OM + BMP + SB). Bars represent mean ± SD of three donors. ***p < 0.001 as compared to the respective sample. (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.