In vitro study on the effect of platelet-rich plasma on osteogeniccapability of bone marrow mesenchymal stem cells

AIM: To investigate the effect of platelet-rich plasma(PRP) on the osteogenic capability of in vitro cultured rat bone marrow mesenchymal stem cells(rBMSCs).METHODS: rBMSCs were separated,cultured and identified in vitro.Cellular proliferation was detected with MTT.Cytochemical method was adopted for alkaline phosphate(ALP) activity detection under ostoegenic induction.RealTime-PCR was used to assay mRNA expression of Col-3 and OPN.Scanning electron microscope was used for cell attachment and ultrastructure observation.RESULTS: The subcultured rBMSCs grew well and most rBMSCs presented shuttle shape observed under inverted phase contrast microscope.Identification of cell surface antigen by flow cytometry showed that CD29 and CD90 expressed positively and CD34,CD45 expressed negatively in the 2nd passage of rBMSCs.Adipogenic and osteogenic differentiation ability of the cultured rBMSCs were confirmed after induction.MTT assay showed that at 1,3,5,7 and 9 days,cell proliferation was significantly higher in the PRP group than that of control.ALP assay showed that after 7 d and 14 d,cells were cultured by ostoegenic induction medium,ALP activity was lower than ostoegenic induction medium and PRP group.The ALP activity was higher on 14d than on 7d(P0.05).Gene expressions of Col-3 and OPN in the cells stimulated by 10% PRP was slightly lower than the cells stimulated by ostoegenic induction medium,but higher than the cells stimulated by α-MEM medium only.Col-3 and OPN expression in the cells stimulated by both ostoegenic induction medium and PRP were much higher than other groups(P0.05).The results of SEM showed promoted cell attachment and stretching area under 10% PRP stimulation.CONCLUSION: PRP can promote proliferation,osteogenic differentiation,collagen synthesis of the rBMSCs,as well as their attachment capability to the biomaterials.