Effect Of GYY4137 On Osteogenic Differentiation Of BMMSC In Postmenopausal Osteoporosis Patients

Objective:The proliferation and osteogenic differentiation were studied about the BMMSC from postmenopausal osteoporosis patients culturing with exogenous H₂S?GYY4137?and its possible mechanism was explicated.Methods:Twenty patients were chosen from who fractured due to postmenopausal osteoporotic,and their bone marrow was taken from the proximal femur during they were performed the operation of total hip replacement for the cell culturing.Cell was cultured using the method of cell-complete-adherence,and was identified using flow cytometry,and then was purified by the method of limited dilution.The proliferation of cell was measured by MTT after the cell being intervened by GYY4137?a donor of H₂S?.Simultaneously,according to the result of MTT,the best concentration of GYY4137 was sifted out for the further experiments.The cells of the third generation from culturing were divided into control group?only h-BMMSC?and experimental group?h-BMMSC plus GYY4137?.The expression level of RNA from the cells in the two groups was collected using the method of lysis when they were cultured to the day of 1,7,14 and 21 respectively.Then,the changes of expression from genes which were related to osteoblasts in the two groups were detected by RT-PCR.Furthermore,the formation of mineralized substance from the both of groups was observed by staining of alizarin and ARS quantitative analysis was performed after the cell was cultured up to 7,14 and 21 days.At last,the mechanism of proliferation and osteogenesis from effect H₂S on h-BMMSC was discussed by a large number of literatures were summarized.The data from the trials was inputted into the software of SPSS in edition 25.0 for statistical analysis.Results:The results of flow cytometry showed that more than 90%of cells were positive by staining of double reagents of CD_90/73,but were negative in CD_34/45.According to the result of value of OD,the concentration of GYY4137 at 100?M was the best for the cell in proliferation,and there was a positive relationship between the proliferation of cell and time of the stimulation from GYY4137.The results of RT-PCR indicated that the expression of bone-related genes in the experimental composition was significantly higher than that in the control group on days 7 and 14?P<0.01?,nevertheless,the expression of genes above was reduced at 21days in the two groups?P<0.01?.After being stained with alizarin red,the amount of mineralized nodules was significantly increased in the experimental group compared with the control group at 7,14,21 days.ARS quantitative analysis results were compared between groups(OD values:F_7th=6.88,F_14th=38.58,F_21th=38.56;P<0.05).Summarized literatures suggested that Wnt/?-catenin signaling pathway may be the main regulatory mechanism of GYY4137 on the osteogenic differentiation of h-BMMSC.Conclusions:The proliferation and osteogenic-differentiation of BMMSC can be promoted by exogenous H₂S?GYY4137?in the patients of postmenopausal osteoporosis.Moreover,the mechanism of this effect might be achieved by regulating Wnt/?-catenin signaling pathway.