The Experimental Study Of Different Methods Of Platelet-rich Fibrin Extracting Solution On Adipose-derived Stem Cell In Vitro

Objective: Platelet-rich fibrin(PRF) were prepared with different centrifugation speed and time to compare the levels of growth factors particularly platelet-derived growth factor-AB and transforming growth factorbeta 1 in the platelet-rich fibrin and the OD value of adipose-derived stem cells(ADSCs) in vitro. The study will provide an experimental basis for optimized preparation of PRF and clinical studies in soft tissue healing.Methods: To primary culture of ADSCs cells from New Zealand white rabbits, adipose tissue in groin with the enzyme digestion method and growth condition of ADSCs was observed; ADSCs at passage 3 were cultured in adipogenic, osteogenetic and neurospheres differentiation medium to induced for day7,day14,day28 and underwent identification by oil red O staining, Von Kossa staining and immunofluorescence in vitro. PRF were respectively produced by three different centrifugal speed(2700 r/min, 3000 r/min, 3300 r/min)and centrifugal time(10 min, 15 min, 20 min) from the vein of New Zealand white rabbits to test the quantity of PDGF-AB and TGF-β1 at day 7,day 14,day 21 with ELISA kit. Different concentration of PDGF-AB and TGF-β 1 was observed in each group. To investigate the cell proliferation ability of ADSCs in platelet-rich fibrin produced by different methods with Cell Counting Kit-8 in vitro.Results:Most ADSCs at passage 3 were spindle under the inverted microscope. The differenliation of adipogenic, osteogenetic and eurospheres were positive. It proved that the cultured cells were ADSCs. The levels of PDGF-AB and TGF-β 1 in the PRF should be made in 2700r/min were higher than 3000r/min and 3300r/min(P<0.05). PDGF-AB and TGF-β 1 levels in the PRF prepared by 15 min were higher than 10 min and 20 min, the statistics showed significant.(P<0.05). CCK-8 method revealed the optical density value of PRF(2700 r/min) were greater than 3000 r/min and 3300 r/min. There were significant differences between two groups(P<0.05).OD value in the PRF prepared by 15 min were greater than 10 min and 20 min. The differences were significant between two groups(P<0.05).Conclusion: ADSCs will be produced by the collagenase digestion method and primary cultured. PRF were prepared by appropriate methods can promote the proliferation of adipose stem cells in vitro. The excellent PRF were expected to be applied to soft tissue wound healing.