The Effects Of Autologous Concentrated Growth Factors Fibrin Extract On MC3T3-E1 Cells

Objective: Through the preparation of CGF extract,observing the effects on the proliferation and differentiation of MC3T3-E1 cells,and to provide the experimental basis for the clinical application of CGF.Methods:(1)The blood collected from volunteers was prepared with Medifuge centrifuge for CGF,then preparation of CGF extract.(2)The experimental groups used 5%,10% and 20% CGF conditioned medium,and the control group used α-MEM complete medium to culture MC3T3-E1 cells.At 1,3,5,7d of culture,the proliferation of the cells was measured by CCK-8.(3)The experimental group used CGF conditioned medium and the control group used α-MEM complete medium to culture MC3T3-E1 cells.At 1,3,5,7d of culture,the activity of ALP of the cells were examined;at 3,7d of culture,the expression of Runx2 and Osx were quantified by qPCR.Result:(1)According to the standardized program for preparation CGF,the blood can be successfully divided into three layers,the CGF gel is larger,soft,tough and strong.(2)At 1,3,5,7d of culture,the absorbance(OD)of the experimental group was significantly higher than the control group by CCK-8(P <0.05).The effect of 10% CGF on cell proliferation was significantly higher than that of 5% CGF and 20% CGF group at the same time(P <0.05).(3)The results of ALP showed that the OD values of the experimental group and the control group increased with the extension of the experiment time(P <0.05).There were no significant difference on the OD values between the two groups at the first day(P> 0.05).But the OD values of the experimental group were significantly higher than the control group at 3,5,7 d(P <0.05).The expression levels of Runx2 and Osx in MC3T3-E1 cells both increased with the extension of the experiment time(P <0.05),and the expression levels of Runx2 and Osx of the experimental group were significantly higher than the control group at each time(P <0.05).Conclusion:(1)CGF extract can promote the proliferation of MC3T3-E1 cells,but the effect of 10% CGF extract on cell proliferation was the most obvious.(2)The growth factors of the CGF extract can increase the activities of ALP in MC3T3-E1 cells,also can promote the differentiation and maturation of the osteoblasts.(3)The growth factors of the CGF extract can effectively promote the expression of genes which are associated with osteogenesis in MC3T3-E1 cells.