Biological Effects Of Hydroxyl Safflor Yellow A On Stemness And Osteogenic Differentiation Of Mesenchymal Stem Cells From Human Craniofacial Bone

Female orthodontic patients oftensuffer from accelerated bone resorptiondue to periodontitis and low hormone levels,resulting in alveolar height and width reduction,which lead to orthodontic treatment difficulties.Therefore,the key to ensure the right progress of orthodontic treatment and aesthetic effect is to take effective measures to reduce the absorption of alveolar ridge,and even reverse the absorption of alveolar ridge.The bone loss in jaws is closely related to the proliferation and osteogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs).However,the characteristics of human bone mesenchymal stem cells from alveolar bone(hAB-BMSCs)and their potential regulatory mechanisms are still not entirely clear.Hydroxyl safflor yellow Acould prevent the occurrence of steroid-induced avascular necrosis of the femoral head,promote the differentiation,transformation and proliferation of bone marrow mesenchymal stem cells,and antagonize the hormone-induced adipogenic differentiation and promote osteogenic differentiationof BMSCs.In this study,we investigated the role of HSYA in the regulation of sternness and osteogenic differentiation of hAB-BMSCs on the basis of comparative analysis of the biological characteristics of human alveolar bone BMSCs in vitro.OBJECTIVE:To analyze the biological characteristics of BMSCs in normal young women,and to explore the related mechanism of HSYA in regulating the stemness and osteogenesis differentiation of hAB-BMSCs.METHODS:(1)hAB-BMSCs were isolated and cultured by whole bone marrow adherent culture and the nonadherent cells were removed by changing the culture medium.Cells were passaged when the monoclonal colony formed and attached each other;(2)The effect of HSYA on the proliferation of hAB-BMSCs was analyzed by CCK-8 method;(3)The expression of stemnessassociated genes in hAB-BMSCs was detected by real-time quantitative PCR and Western blot;(4)The hAB-BMSCs were cultured for 8 days by chromatin staining.The clone formation ability was analyzed;(5)Real-time PCR,Western blots,and alizarin redstain were used to detect the multi-differentiation ability;RESULTS:(1)The effect of different concentrations of HSYA on the proliferation of hAB-BMSCs,10-1 mg/ml HSYA could promote the proliferation of hAB-BMSCs.(2)The results of clonal formation showed that the number of clones in the experimental group was significantly more than that in the control group.Western blot showed that the bands of NANOG,SOX2 and OCT4 were gradually deepened in the 10-1mg/ml group compared with the control group,and RT-PCR showed that in the 10-1mg/ml group the mRNA expression levels of NANOG,SOX2 and OCT4 were increased.(3)Theexpression of osteogenic genes:RUNX2,OCN,OPN and OSX in OB + HSYA group were not only increased at the protein level,but also increased at mRNA level and increased calcium nodule formation.CONCLUSION:hAB-BMSCs were used to study the effect of 10-1 mg/ml HSYA on the sternness of hAB-BMSCs.At the same time,the osteogenic differentiation of hAB-BMSCs was induced by RUNX2,OCN,OPN and OSX.