基于β-catenin通路研究富血小板纤维蛋白复合β-磷酸三钙对小鼠成骨细胞成骨分化的影响  

The effect of platelet-rich fibrin on differentiation into osteoblasts combined withβ-tricalcium phosphate throughβ-catenin pathway

作　　者：付冬梅[1] 周婧 王浪 罗阳 兰红 李素兰 王劲 FU Dong-mei;ZHOU Jing;WANG Lang;LUO Yang;LAN Hong;LI Su-lan;WANG Jin(Department of Stomatology,Yanjiang District People's Hospital,Ziyang,Sichuan 641300)

机构地区：[1]资阳市雁江区人民医院口腔科,四川资阳641300

出　　处：《赣南医学院学报》2022年第1期5-11,共7页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：2020年度资阳市科技计划项目(Zykjjsc20-yyjc-2020-09)。

摘　　要：目的:探究不同浓度富血小板纤维蛋白(Platelet-rich fibrin,PRF)对小鼠成骨细胞增殖与分化的影响,并观察最优浓度PRF复合β-磷酸三钙对小鼠成骨细胞成骨分化的影响。方法:首先实验分为6组,分别用0%、10%、20%、30%、40%、50%浓度的PRF培养MC3T3-E1成骨细胞,通过CCK-8法检测观察细胞增殖情况,ELISA检测细胞碱性磷酸酶(Alkaline phosphatase,ALP)活性,划痕实验检测细胞迁移能力,茜素红染色观察细胞钙结节形成情况。随后实验分为2组,对照组无PRF培养细胞,实验组为最佳浓度PRF培养细胞,两组成骨细胞均复合于β-磷酸三钙,在1 d、3 d和7 d采集细胞,通过ELISA检测骨钙素(OC)和Ⅰ型胶原蛋白(COL-1)的表达,通过WB检测β-catenin、LEF-1、RunX2、Osterix和OPN的蛋白表达。最后,使用β-catenin抑制剂XAV-939处理成骨细胞,通过ELISA检测细胞ALP活性,WB检测β-catenin、LEF-1、RunX2、Osterix和OPN的蛋白表达。结果:10%~50%浓度的PRF均促进了MC3T3-E1成骨细胞的增殖能力和迁移能力,并提高了成骨细胞ALP活性和钙化结节数量,50%PRF是实验中的最佳浓度。此外50%PRF提高复合β-磷酸三钙成骨细胞的OC和COL-1的表达及β-catenin、LEF-1、RunX2、Osterix和OPN的蛋白表达。抑制β-catenin可以降低成骨细胞ALP活性及降低β-catenin、LEF-1、RunX2、Osterix和OPN的蛋白表达。结论:PRF促进成骨细胞增殖分化,浓度越高,效果越明显,50%PRF是本实验中的最佳浓度,其次PRF促进成骨细胞在β-磷酸三钙支架上的骨生成,其与β-catenin通路激活有关。Objective:To investigate the effect of different concentrations of platelet-rich fibrin(PRF)on the prolifera⁃tion and differentiation into osteoblasts,and to observe the effect of optimal concentration of PRF on differentiation of osteoblasts combined withβ-tricalcium phosphate.Methods:MC3T3-E1 osteoblasts were cultured(6 groups)with PRF of 0%,10%,20%,30%,40%and 50%,respectively.Cell proliferation was detected by CCK-8 assay,cell alkaline phosphatase(ALP)activity was detected by ELISA,and cell migration was detected by scratch assay.The formation of calcium nodules was observed by alizarin red staining.The following experiments were divided into two groups:Control group and PRF group.No PRF cells were cultured in the control group.The cells in PRF group were cultured with PRF of the best concentration and both cells were combined withβ-tricalcium phosphate in two groups.The cells were collected at 1 d,3 d,and 7 d,and the expression of OC and Col-1 was detected by ELISA.The protein expressions ofβ-catenin,LEF-1,Runx2,Osterix,and OPN were detected by WB.Finally,MC3T3-E1 were treated with aβ-catenin inhibitor(XAV-939),and ALP activity was detected by ELISA and the protein expressions ofβ-catenin,LEF-1,RunX2,Osterix and OPN were detected by WB. Results: PRF from 10% to 50% concentration significantly promoted the prolifera⁃tion and migration ability of MC3T3-E1 osteoblasts,and increased the ALP activity and the number of calcified nodules,meanwhile 50% PRF was performed best in the test. In addition,50% PRF significantly increased the expression of OCand Col-1 and the protein expressions of β-catenin,LEF-1,Runx2,Osterix,and OPN in osteoblasts combined withβ-tricalcium phosphate. The inhibition of β-catenin decreased the ALP activity and the protein expressions of β-catenin,LEF-1,RunX2,Osterix,and OPN in MC3T3-E1 cells. Conclusion: PRF could promote the proliferation and differenti⁃ation of osteoblasts,and the higher the concentration,the more obvious the effect is,which indicated that 50% PRFperformed

关 键 词：富血小板纤维蛋白 Β-磷酸三钙 骨生成 成骨细胞 骨分化 

分 类 号：R783.3[医药卫生—口腔医学]