Fe_3O_4涂层多巴胺修饰PET人工韧带对MC3T3-E1细胞粘附、增殖和分化影响的实验研究  

Fe_3O_4 and Dopamine Coating Improves the Adhesion, Proliferation and Differentiation of MC3T3-E1 Cells on the Surface of PET

作　　者：俞斌[1] 丁惠锋[1] 杨超华[1] 禹宝庆[1] 黄建明[1] 

机构地区：[1]复旦大学附属浦东医院骨科,上海201399

出　　处：《中国运动医学杂志》2016年第5期438-444,共7页Chinese Journal of Sports Medicine

基　　金：浦东新区卫生和计划生育委员会卫生科技项目(PKJ2015-Y37);浦东新区卫生系统重点学科群建设资助(PWZxq2014-03)

摘　　要：目的:观察MC3T3-E1细胞在Fe_3O_4涂层多巴胺修饰聚对苯二甲酸乙二醇酯(PET)人工韧带材料表面的粘附、增殖及成骨分化情况。方法:按PET材料表面涂层情况分为4组:PET组,即PET表面未处理组;多巴胺(DA)组,即PET表面进行多巴胺涂层组;100 Fe_3O_4组和200 Fe_3O_4组,分别为PET经多巴胺处理后表面涂有100μg/ml和200μg/ml的Fe_3O_4溶液组。在上述4组PET材料上分别培养MC3T3-E1细胞,进行各项检测。扫描电镜(SEM)和X射线能谱分析(EDS)材料涂层情况。细胞在材料上培养3天和7天后行荧光染色,观察细胞粘附情况。培养1、3、7天后,行CCK-8检测MC3T3-E1细胞的增殖。诱导培养基培养7天检测碱性磷酸酶活性;培养21天行茜素红染色,检测细胞矿化情况;培养14天行Real Time PCR检测骨钙蛋白和胶原I基因的表达。结果:SEM和EDS检测验证涂层成功。荧光染色显示涂层组有效增加了细胞的粘附。细胞增殖实验示涂层组对细胞增殖有促进作用,各涂层组与未涂层组差异具有统计学意义(P<0.05)。涂层材料组碱性磷酸酶OD值明显高于未涂层组(P<0.05)。茜素红染色示各组都有较多矿化结节形成,定量分析显示100 Fe_3O_4组和200 Fe_3O_4组与PET组比较差异有统计学意义(P<0.05)。PCR检测显示涂层组骨钙蛋白和I型胶原基因的表达都较未涂层组增加,骨钙蛋白:Fe_3O_4组与PET组比较差异有统计学意义(P<0.01);I型胶原:DA组、Fe_3O_4组与PET组比较差异具有统计学意义(P<0.05)。结论:经多巴胺修饰四氧化三铁涂层处理的PET人工韧带材料,能明显促进MC3T3-E1细胞的粘附、增殖和分化。Objective To observe the adhesion,proliferation and differentiation of MC3T3-E1 cells on the surface of polyethylene terephthalate（PET）coasted by Fe3O4 and dopamine. Methods According to the materials coated on the surface of PET,four experiment groups were established：PET group（without any coating）, DA group（coating with dopamine）,100 Fe3O4group（coating with nano Fe3O4 of 100 μg/ml）and 200Fe3O4group（coating with nano Fe3O4 of 200 μg/ml）. The MC3T3-E1 cells were cultured on the surface of the PET,and their adhesion,proliferation and differentiation were observed. The morphology of PET was observed by scanning electron microscope（SEM）and energy dispersive spectroscopy（EDS）. The adhesion of cells to the surface of PET was observed with fluorescence staining at the 3rd and 7th day of culture. The proliferation of cells on the surface of PET was detected by CCK-8 reagent at the 1st,3rd and 7th day of culture,and the alkaline phosphatase（ALP）activity was checked by the end of 7th day of culture. The cell mineralization at the 21 st day of culture was detected by Alizarin red staining. Real time PCR was used to measure the m RNA expression of osteocalcin and collagen I by the end of 14 th day of culture. Results The success of coating PET was verified by SEM and EDS. As compared to the without coating group,the adhesion of MC3T3-E1 cells increased significantly（P0.05）and the proliferation of MC3T3-E1 cells improved significantly（P0.001）in coating groups. The OD value of ALP activity in coating groups were higher than that in without coating group（P0.05）. More formation of mineralized nodule revealed in all groups,and in 100 Fe3O4 group and 200 Fe3O4 group was significantly more than in without coating group（P0.05）. Comparing to the without coating group,the m RNA expression of osteocalcin in Fe3O4 group increased significantly（P 0.001）,as well as the expression of collagen I in DA group and Fe3O4group（P0.05）. Conclusion The adhesion,proliferation and different

关 键 词：聚对苯二甲酸乙二醇酯 多巴胺 四氧化三铁 MC3T3-E1 

分 类 号：R873[医药卫生—运动医学]