| Background: Platelet-rich fibrin(PRF),as a new generation of platelet products,has been used in a variety of fields due to its simple preparation,unique three-dimensional fibrous network structure,the ability to carry a variety of growth promoting factors,and the lack of anticoagulant required by platelet-rich plasma.With the gradual development of PRF,more and more researchers focus on how to obtain platelet-rich fibrin with more stable structure,more convenient use and better biological efficacy,and gradually derived a variety of PRF based on different relative centrifugal force and centrifugal time conditions.There are also differences in the amount of growth factors and biological properties of different PRF products.Objective: To observe and compare the effects of platelet-rich fibrin(PRF)prepared under different centrifugal conditions on growth factor release ability,internal fibrin structure,fibroblast growth and wound healing in diabetic rats.Methods: 1.Comparison of biological properties of platelet-rich fibrins: The whole blood of SD rats was obtained by cardiac blood collection method.leukocyte-PRF(L-PRF),advanced PRF(A-PRF)and advanced PRF+(A-PRF+)were obtained by centrifugation procedures of 708 g 12 min,208 g 14 min and 208 g 8 min,respectively.PRF membrane was prepared and placed in DMEM/F12 medium.The medium was collected at 1 h,6 h,1 d,3 d and 5 d,respectively.The concentrations of platelet-derived growth factor(PDGF-Ab),transforming growth factor β(TGF-β)and vascular endothelial growth factor(VEGF)in the medium were detected by ELISA method.Scanning electron microscopy was used to observe the internal fibrin reticular structure and cellular components of the above PRFs.2.Platelet-rich fibrin effects on cell growth: Skin fibroblasts from the back of SD rats were isolated and cultured.The cells were divided into L-PRF group,A-PRF group,A-PRF+ group and blank group.The cells were cultured in L-PRF,A-PRF,A-PRF+ conditional medium and 10%FBS medium,respectively,CCK 8 cell proliferation experiment test cell proliferation ability,Transwell little room experiment test cell migration ability.3.Observation of the effect of platelet-rich fibrin on wound healing in diabetic rats: Healthy SD rats were selected and injected with streptozotocin(STZ)to make diabetic rat model.The rats were divided into blank control group,L-PRF group,A-PRF group,and A-PRF+group with a skin opening of 1cm in diameter.The fibrin prepared by the above PRF preparation method was placed in the wound,and the wound healing on the 5th,9th and 14 th day was observed ang recorded,and HE staining,picric acid-sirius red staining and CD31 immunohistochemical staining were performed.Results:We found that a-PRF + prepared by centrifugation at 208 g and 8 min had A three-dimensional environment more conducive to the release of growth factors.ELISA results showed that the overall release level of target growth factors in A-PRF and A-PRF+ medium was higher than that in L-PRF medium.In cell experiments,A-PRF+ conditioned medium showed better ability of cell proliferation and cell migration.In the experiment of wound healing,A-PRF+ showed A more prominent role in promoting healing,promoting collagen growth and maturation,and promoting the growth of new blood vessels.Conclusion:A-PRF + prepared by centrifugation at 208 g and 8 min has A more excellent three-dimensional reticular fibrin structure,which can more effectively promote the wound healing of diabetic rats. |
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