Fig. 5

Effect of Trolox on the OB differentiation potential of hDPSCs. A Representative images of differentiating hDPSCs ± 500 μM Trolox (administered since the induction start) at days 7, 14 and 21 shown as phase-contrast (left half part of the panel) and photographs of ALP staining (right half part of the panel). Scale bar, 80 μm. The graph on the right displays normalized densitometry analysis (Image J software) of ALP staining along with statistical evaluation; bars are means ± SEM of 4 biological replicates under each condition **P < 0.01 versus untreated cells. B q-RT-PCR analysis of the osteogenic markers Dspp and Ocn of DPSCs cultured under osteogenic conditions ± 500 μM Trolox. The values are means ± SEM of normalized transcript levels of four independent biological experiments, *P < 0.05; **P < 0.01; ***P < 0.001 versus untreated cells. C Protein expression levels of Runx2, assayed by western blotting in undifferentiated (CTRL), untreated (-) and 500 μM Trolox-treated cells after 21 days from osteogenic induction. Left panel: a representative immunoblot. Full-length blots are presented in Additional file 1: Fig. S5. Right bar histogram: densitometry analysis normalized to β-actin as means ± SEM of three independent experiment; *P < 0.05 versus CTRL; ***P < 0.001 versus untreated differentiated cells. D Analysis of mineral matrix deposition assayed by Alizarin Red (red staining) in undifferentiated hDPSCs (CTRL), untreated (−) and 500 μM Trolox-treated cells after 21 days from osteogenic induction. The graph on the right shows quantitative analysis of alizarin red staining carried out by a densitometric analysis (Image J software); *P < 0.05 versus CTRL; **P < 0.01 versus untreated differentiated cells