Fig. 4

5′-HA stimulates BMP-induced osteoblast differentiation. a Dose-dependent stimulatory effect of 5′-HA on BMP2-induced osteoblast differentiation of mBMSCs as assessed by quantification of ALP activity after 7 days of induction and (b) quantitative Alizarin red staining for matrix mineralization after 12 days of induction. Cells were induced to osteoblast differentiation using BMP2 (50 ng/ml) in the absence (0) or the presence of different concentrations of 5′-HA. Representative images were shown for Alizarin Red staining. c 5′-HA stimulates the expression of BMP2-related osteogenic markers in a dose-dependent manner as measured by qPCR analysis. Each target gene was normalized to reference genes and represented as fold change over induced control (0) without 5′-HA. Values are mean ± SD of three independent experiments, (*p < 0.05, **p < 0.005 compared to control induced cells without 5′-HA)