Effects of tension force on osteogenic differentiation of mouse bone marrow mesenchymal stem cells and its mechanism
Objective:To explore the effect and mechanism of tension on osteogenic differentiation ability of mouse bone marrow mesenchymal stem cells(BMSCs).Methods:The primary mouse BMSCs were isolated and cultured,and the BMSCs were divided into control group,tension group for 6 h,12 h,and 12 h + rapamycin(Rap).Dynamic tension was applied using a multiple-unit cell stretching device for intervention.Biochemical method was used to detect alkaline phosphatase(ALP)activity in cells.ELISA method was used to detect the con-tent of periostin(POSTN)in the supernatant of cell culture medium.RT-qPCR was used to detect the mRNA ex-pression levels of runt related transcription factor 2(RUNX2),osteocalcin(OCN),osterix and osteopontin(OPN)in cells.Western blot was used to detect the protein expression levels of RUNX2,OCN,Osterix,OPN,POSTN,mammalian target of rapamycin(mTOR)and phospho mTOR(p-mTOR)in cells.Results:Compared with the con-trol group,the activity of ALP and the mRNA expression levels of RUNX2,Osterix,OCN and OPN in the 6 h ten-sion and 12 h tension groups were significantly increased(P<0.05),while the POSTN content in supernatant liq-uid,the expression level of POSTN protein and p-mTOR/mTOR protein ratio were also significantly increased(P<0.05),and the 12 h tension group being more pronounced.Compared with the 12 h tension group,the ALP activi-ty and the expression levels of RUNX2,OCN,Osterix and OPN mRNA and protein in the 12 h tension +Rap group cells were significantly reduced(P<0.05),while the POSTN content in the supernatant of the cell culture medium showed no significant difference(P>0.05).Conclusion:Tension regulates the activation of the mTOR signaling pathway by mediating POSTN expression,thereby promoting osteogenic differentiation in BMSCs.
distraction osteogenesisbone marrow-derived mesenchymal stromal cellsperiostinmammalian tar-get of rapamycinosteogenic differentiation
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