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Objective·To explore the effect of all-trans retinoic acid (ATRA) of different concentrations on osteogenic differentiation of jaw bone mesenchymal stem cells (jBMSCs) in rats.Methods·jBMSCs from 4-week-old Sprague-Dawley (SD) rats were isolated and cultured with whole bone marrow adherence method. The surface antigens were identified by using flow cytometry. Alkaline phosphatase (ALP) staining/alizarin red staining, oil red O staining and alcian blue staining were used to prove the multilineage differentiation potential of jBMSCs after osteogenic, adipogenic and chondrogenic induction respectively. jBMSCs were induced in osteogenic medium with ATRA of concentration of 0.01, 0.1, 1, 5, 10, 20 μmol/L in vitro, and dimethyl sulfoxide (DMSO) was used as control group. Cell viability of jBMSCs in different groups were determined by CCK8. ALP staining and alizarin red staining were used to investigate the osteogenic ability of jBMSCs in each group and screened the concentrations for subsequent experiments. Quantitative real-time polymerase chain reaction (qPCR) and immunofluorescence staining were used to analyze the expressions of osteogenesis-related genes and proteins in jBMSCs of different concentrations.Results·The flow cytometry analysis showed that more than 98% of P1 jBMSCs were positive for CD29+CD90+CD31-CD45-, which was congruent with the characteristics of bone mesenchymal stem cells. The results of ALP staining/alizarin red staining, oil red O staining and alcian blue staining indicated that the P1 jBMSCs had the multilineage differentiation potential of osteogenesis, adipogenesis and chondrogenesis. The results of ALP staining/alizarin red staining showed that the osteogenic activity and mineralization ability of jBMSCs in 0.01, 0.1 and 1 μmol/L ATRA groups were increased compared with those in the control group, while the osteogenic activity and mineralization ability were decreased when the concentration of ATRA increased, especially higher than 5 μmol/L (all P |
