Preparation And Concentration Detection Of Some Growth Factors Of Rabbit PRF And A-PRF

Objective: through the comparative analysis of human and rabbit hemorheological part of indicators,screening the method of preparing rabbit platelet-rich fibrin(PRF)and Advanced platelet-rich fibrin(A-PRF)gel.Studying the release rule of partial growth actors of PRF and A-PRF in rabbit standard,in vitro to provide data reference for animal experiment and clinical research.Methods: 5ml Elbow vein blood of 12 healthy volunteers(male and female of the 6)and Middle ear artery blood of 12 healthy Japanese rabbits(male and female of the 6)were collected to analyze Statistical indexes of blood rheology.Choicing 17 kinds of centrifugal methods to prepare rabbit PRF gel and 16 kinds of centrifugal methods to prepare rabbit A-PRF gel,which are observed by light microscopy and scanning electron microscopy(SEM).Selecting the preparation method of rabbit PRF and A-PRF which is the closest to the microscopic structure of human PRF and A-PRF.Prepareing rabbit PRF,A-PRF by the optimum of preparation method in vitro to observe the release of growth factors as IL-1,IL-4,PDGF,TGF,TNF-?,VEGF,at 20 min,1h,4h,1days,3 days,5 days,7 days,14 days,21 days,28 days which can promote the regeneration of soft and hard tissueResults:(1)the rabbit whole blood viscosity in High,medium and low shear rate,the Whole blood High and low shear reduced viscosity,the whole blood high and low shear relative index and Red cell aggregation index were significantly lower than those of the human blood(P < 0.05),The other indexes were not statistically significant(P > 0.05).(2)The PRF prepared by 17 kinds of centrifugal method and A-PRF prepared by 16 kinds of centrifugal method were different in each group.The structure of the clot,the number of living cells and the arrangement of the fiber protein were different in each group.Among them,the rabbit PRF prepared by 3250 rpm/min,10 min is the closest to the microscopic structure of human PRF.The rabbit A-PRF prepared by 2000 rpm/min,24 min is the closest to the microscopic structure of human A-PRF.(3)The Highest release values of all growth factors of PRF and A-PRF stilled culture In vitro was PDGF,but there was no significant difference between the two groups.The growth factor IL-1 released by A-PRF was significantly higher than that in PRF group at 7d,21 d and 28 d,but no significant difference of the IL-1 total protein content was found between the two groups in the whole experimental period.The growth factor IL-4 released by A-PRF group was significantly higher than that in PRF group at 3days and 21 days,and the total protein content of IL-4 was significantly higher than that in PRF group,the difference was statistically significant.The growth factor TGF-?1,TNF-?,VEGF in A-PRF were higher than that in PRF group,the difference was statistically significant,but there was no significant difference in the two groups at each time period.Conclusion: the whole blood viscosity,Whole blood reduced viscosity,Whole blood relative index and red blood cell aggregation index were significantly lower than those of human blood,and there was no significant difference in other indexes.The rabbit PRF prepared by 3250 rpm/min,10 min is the closest to the microscopic structure of human PRF.The rabbit A-PRF prepared by 2000 rpm/min,24 min is the closest to the microscopic structure of human A-PRF.The Highest of total growth factor of PRF and A-PRF stilled culture in vitro was PDGF,but there was no significant difference between the two groups.The growth factor IL-1 released by A-PRF was significantly higher than that in PRF group at 7d,21 d and 28 d.The growth factor IL-4,TGF-?1,TNF-?,VEGF in A-PRF were higher than that in PRF group,the difference was statistically significant.