The Conditioned Medium of Calcined Tooth Powder Promotes the Osteogenic and Odontogenic Differentiation of Human Dental Pulp Stem Cells via MAPK Signaling Pathways

Figure 2

The effects of CTP-CM on osteo/odontogenic differentiation of hDPSCs. Cells were cultured with CTP-CM for up to 0, 3, and 7 days in the concentration of 200 μg/mL. The protein levels were determined by western blot (a). Quantitative analysis of protein bands was done by ImageJ. The data were normalized by GAPDH (b). Total RNA was isolated and analyzed by real time RT-PCR (c), normalized to GAPDH, and quantified relative to the control. Osteogenic differentiation of hDPSCs was assessed by Alizarin Red staining (d) and (e). Images of Alizarin Red S (ARS) staining in hDPSCs in the control group, 200 μg/mL CTP-CM, MM, and MM + 200 μg/mL CTP-CM were scanned directly and taken by a microscope (d). Scale . The intensity of Alizarin Red staining was determined by optical density (e). Osteogenic differentiation of hDPSCs was assessed by ALP staining (f). Scale . The expressions of DSPP, OPN, and Runx2 in the CTP-CM-treated hDPSC and control groups were detected by immunofluorescence staining (g–i). Scale . and .

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