The Experimental Study On The Osteogenic Differentiation Of Bone Marrow Mesenchymal Stem Cells By The Combination Of Adipose-derived Stromal Vascular Fraction Cells(SVF) And Platelet-rich Plasma(PRP)

Objective: In plastic surgery,the effective reconstruction of bone defect is still a serious challenge for clinicians.There is still no effective solution.The rapid development of regenerative medicine has brought new hope for the treatment of clinical bone defects.The key of bone regeneration is to achieve the directional osteogenic differentiation of stem cells.In some prospective clinical trials,cell therapy based on SVF and its combination with PRP are still in the early stage of clinical application transformation from preclinical research to clinical application,which requires long-term follow-up clinical trials.Further research on microenvironment,differentiation mechanism and regulation mechanism of SVF transplantation will provide new theoretical basis for clinical application and transformation of SVF.Base on the studies of the combination of SVF and PRP,study on the deeply mechanism of bone marrow mesenchymal stem cells(BMSCs)still have a great sense in vitro.Methods:1.Under aseptic conditions,the adipose tissue was removed from the groin of SD rats and blood samples were extracted from the left atrium of rats.Extract and culture SVF cells and prepare PRP.2.Transwell chamber was used to establish a co-culture system of SVF and BMSCs to investigate the effect of SVF paracrine on BMSCs proliferation and osteogenic differentiation,as well as the combined application of SVF and PRP.Results:1.CCK-8 results showed that BMScs proliferation rate increased in SVF group,PRP group and SVF+PRP combined group,with SVF+PRP group being the highest.2.On day 2:Compared with BMSCs group,the expression levels of Alp,Runx2 protein in PRP group and SVF+PRP group were significantly increased.There was no significant difference in ALP protein between the SVF group and the BMSCs group on day 2,and the expression of RUNX2 protein was increased.3.On day 7:The results showed that ALP and RUNX2 protein expression was significantly higher in the other three groups than in the BMSCs group.Additionally,the expression of osteoblasts in SVF + PRP group was the most obvious.Conclusion:Both SVF and PRP can promote the proliferation and osteogenic differentiation of BMSCs,and the combination of the two can enhance significantly.In bone regeneration and repair,SVF injection can repair bone defects not only because of its own osteogenic differentiation,but also because it can promote the proliferation and osteogenic differentiation of BMSCs around bone defects.Combined with PRP,SVF injection can achieve stronger repair effect.