转化生长因子-β1对氧化锆表面小鼠胚胎成骨细胞前体细胞黏附、增殖及分化的影响研究  

Response of osteoblast-like cells cultured on zirconia to transforming growth factor β1

作　　者：张冉[1] 任光辉[1] 李蕾[2] 滕腾[1] 柳忠豪[1] 

机构地区：[1]滨州医学院附属烟台市口腔医院,山东烟台264000 [2]济南市口腔医院,济南250001

出　　处：《中国实用口腔科杂志》2015年第5期309-312,共4页Chinese Journal of Practical Stomatology

基　　金：泰山学者海外特聘专家项目(tshw20120233)

摘　　要：目的探讨转化生长因子-β1（TGF-β1）对小鼠胚胎成骨细胞前体细胞（MC3T3-E1）在氧化锆（Zirconia Zr）片表面黏附、增殖及分化的影响,为氧化锆材料在口腔临床上的应用提供理论基础。方法将MC3T3-E1分为4组进行培养,即TGF-β1组、Zr组、Zr＋TGF-β1组、对照组。分别使用MTT法检测各组0、1、2、3、4、5、6、7 d细胞增殖活性;荧光实时定量PCR检测各组成骨细胞中3、7 d后成骨相关蛋白OPG,Runx2,BGP,BMP2 m RNA的表达;激光共聚焦显微镜下观察3、5、7 h后各组细胞骨架及微观结构。结果 MTT检测结果显示,随细胞培养时间的增长,各组细胞数目均逐渐增加。其中,1~3 d各组细胞数目未见明显差异（P〉0.05）,3~7 d Zr＋TGF-β1组与TGF-β1组细胞增殖能力明显高于其他组（P〈0.05）,而Zr组的细胞数目略高于对照组（P〈0.05）;PCR结果显示,3 d后可见Zr＋TGF-β1组细胞的OPG、BMP-2、Runx2、BGP m RNA表达相对明显,高于其余组（P〈0.05）,同时TGF-β1组细胞的OPG、BMP-2、Runx2 m RNA的表达水平也略高于Zr组及对照组（P〈0.05）;7 d后氧化锆＋TGF组相较其余组升高不显著,各组之间差别无统计学意义。激光共聚焦显微镜下观察到各个实验组细胞的细胞骨架较对照组有明显铺展,且形态多不规则,肌动蛋白纤维交错排列,形态更加伸展。结论 TGFβ-1可能对成骨细胞在氧化锆片表面的贴附、增殖及分化具有一定的促进作用。Objective The aim of this study was to explore response of MC3T3 E1 osteoblast was effected on zirconia,and low concentration TGF-β1 on the MC3T3 E1 cell adhesion in the surface of zirconium oxide,proliferation differentiation,mineralization osteogenesis.Methods Mouse preosteoblastic cell line（MC3T3-E1）were cultured on sandblasted zirconia and discs will be divided into 4 groups：TGF-beta 1 group,zirconium oxide（Zr）group,TGF- beta 1 and zirconium oxide（Zr）group,the control group. At 24 hours after seeding MC3T3-E1 incubated in media containing：1、Cell proliferation was assessed by MTT at 1,2,3,4,5,6 and 7 days. Semi-quantitative reverse transcription-polymerase chain reaction（RT-PCR）method was used to examine the expression of OPG,Runx2,BGP and BMP2.3、LSCM observe cell skeleton and microstructure.Results MTT test result that after loading the TGF β1,cellular proliferation on the zirconia discs was increased significantly after day 3 rd（P〈0.05）;Zr group slightly higher than the control group at day 3rd（P〈0.05）. Osteocalcin,Runx-2,Bmp-2and osteoprotegerin expression,that of cells on zirconia with TGF β1 was much more highly increased at day 3（P〈0.05）. The gene expression level of cells cultured on TGF β1 was higher than that on group with Zr at day 3（P〈0.05）. LSCM observed that promote the spread of F-actin cytoskeleton.Conclusion With the stimulation of TGF β1,zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells.

关 键 词：氧化锆 骨结合 TGF-Β1 成骨细胞 

分 类 号：R78[医药卫生—口腔医学]