牙胚细胞与骨形态发生蛋白4转染的骨髓间充质干细胞相互诱导的体外实验研究  

Study on the Mutual Induction between Tooth Germ Cells and Bone Marrow Mesenchymal Stem Cells Transfected by Bone Morphogenetic Protein4 Gene in Vitro

作　　者：王欢欢[1] 沈玉凤[1] 胡杨[1] 韩祥祯 董红宾[1] 何惠宇[1] 

机构地区：[1]新疆医科大学第一附属医院口腔修复科,新疆乌鲁木齐830054

出　　处：《口腔医学研究》2015年第3期237-241,共5页Journal of Oral Science Research

基　　金：新疆维吾尔自治区科技厅科技支疆项目(编号:201291173);新疆维吾尔自治区科技厅自然科学基金(编号:20142110037);新医大一附院组织工程专项(编号:2012ZZGC01)

摘　　要：目的:探讨牙胚细胞和骨形态发生蛋白4(BMP4)转染的骨髓间充质干细胞(BMSCs)相互间的成牙诱导作用。方法:构建骨形态发生蛋白4慢病毒载体,转染SD大鼠骨髓间充质干细胞,并将骨髓间充质干细胞和BMP4转染的骨髓间充质干细胞分别与SD大鼠牙胚细胞按1∶1比例混合培养,同时将细胞分为5组,BMSCs组、BMP4/BMSCs组、牙胚细胞组、BMSCs/牙胚细胞组(混合组1)、BMP4/BMSCs/牙胚细胞组(混合组2),分别采用实时荧光定量PCR和western-blotting检测五组细胞Ⅰ型胶原蛋白、成釉蛋白、牙本质基质蛋白1、同源异型盒基因1成牙相关基因mRNA水平和蛋白水平相对表达量的变化。结果:与混合组1相比,混合组2Ⅰ型胶原蛋白、成釉蛋白、牙本质基质蛋白1、同源异型盒基因1mRNA水平和蛋白水平表达量增多,差异有统计学意义(P<0.05)。结论:牙胚细胞与BMP4转染的骨髓间充质干细胞共培养,促进了成牙相关基因的表达,可作为组织工程牙的备选种子细胞。Objective： To study the mutual induction between tooth germ cells and bone marrow mesenchymal stem cells with bone morphogenetic protein4 gene in vitro. Methods： Lentiviral vectors carrying BMP4 were con- structed to transfect rat bone marrow mesenchymal stem cells（BMSCs） and BMSCs tansfected by BMP4 were co-- cultured with tooth germ cells （1：1）respectively . The cells were divided into five groups： BMSCs group, BMP4/ BMSCs group, tooth germ cells group, BMSCs tooth germ cells group （the mixed groupl）, BMP4/BMSCs tooth germ cells group （the mixed group2） . The five groups were extracted using real--time quantitative PCR and west- ern blotting to detect the mRNA and protein levels of collagen I, enamel protein, dentin matrix protein 1, and ho- meobox gene 1. Results： In the mixed group2, mRNA and protein levels of collagen I, enamel protein,dentin matrix protein 1 and homeobox gene 1 were higher, and the difference was statistically significant （P〈0.05）. Conclusion： The co--culture of tooth germ cells and BMSCs tansfected by BMP4 can increase the expressions of odontogenic genes, and they are suitable as seed cells of tissue--engineered tooth.

关 键 词：牙胚细胞 骨髓间充质干细胞 骨形态发生蛋白4 实时荧光定量PCR western-blotting 

分 类 号：R780.2[医药卫生—口腔医学]