多种生长因子对胚胎面突外胚间充质细胞向成牙本质样细胞分化的诱导作用  

Effects of transforming growth fctor β1(TGFβ-1) and dentin noncollagenous proteins(DNCP) on human embryonic ectomesenchymal cells in a three dimensional culture system

作　　者：邓蔓菁[1] 刘鲁川[1] 陈小红[1] 安建平[1] AJ Smith 金岩 史俊南 

机构地区：[1]第三军医大学野战外科研究所大坪医院口腔科,重庆400042 [2]英国伯明翰大学口腔生物教研室

出　　处：《牙体牙髓牙周病学杂志》2008年第8期438-442,共5页Chinese Journal of Conservative Dentistry

基　　金：第三军医大学科技创新工程(2004);重庆市自然科学基金资助(2005bb5307)

摘　　要：目的:探讨转化生长因子β1(TGFβ1)、牙本质非胶原蛋白(DNCP)、TGFβ1+DNCP、肝素+胰岛素样生长因子1(IGF-1)在诱导面突外胚间充质细胞向成牙本质样细胞分化中的作用。方法:取妊娠12.5 d SD大鼠胎鼠颌突外胚间充质细胞(第4代),在转化生长因子β1(TGFβ1)(2 ng/mL)、牙本质非胶原蛋白(DNCP)(100μg/mL)、TGFβ1(2 ng/mL)+DNCP(100μg/mL)、肝素(1μg/mL)+IGF-1(100 ng/mL)的诱导下,通过形态学观察、免疫组化、RT-PCR等方法,探索生长因子在外胚间充质细胞向成牙本质样细胞分化中的作用。结果:6 d后,在DNCP、TGFβ1+DNCP作用下,胎鼠面突外胚间充质细胞出现明显的形态改变,细胞由成纤维样变为梭形,部分核极化,有很长的突起,部分细胞呈现平行排列趋势。TGFβ1组部分细胞出现极化改变。DNCP、TGFβ1+DNCP诱导组抗牙本质涎蛋白(DSP)呈阳性反应,RT-PCR显示mRNA水平表达成牙本质细胞特异的牙本质涎磷蛋白(DSPP),另外DMP1亦呈阳性表达。TGFβ1组不表达DSPP、DMP1。肝素+IGF-1组在细胞形态和蛋白、mRNA上均未出现成牙本质细胞特异的表达。结论:胎鼠面突外胚间充质细胞在DNCP和TGFβ1+DNCP的作用下能分化为成牙本质样细胞。表明在诱导中起关键作用的是DNCP。TGFβ1可诱导细胞在形态学上分化成类成牙本质细胞。本结果与该细胞在三维体系中的诱导分化结果相似,表明胚胎面突外胚间充质细胞在体外被诱导分化为成牙本质样细胞不是偶然现象,这为研究牙齿的分化和发育提供了良好的模型和实验依据。AIM： To investigate the possibility and procedure of ectomesenchymal cells in embryonic facial process differentiating to odontoblast - like cells induced by transforming growth factorβ1 （ TGFβ - 1 ）, dentin noncol- lagenous proteins（ DNCP）, TGFβ - 1 ＋ DNCP and heparin ＋ insulin - like growth factorβ1 （ IGFβ - 1 ）. METHODS： The forth passage ectomesenchymal cells of rat facial process were isolated and TGFβ - 1 （ 2 μg/mL）, DNCP （ 100μg/mL）, TGFβ - 1 （2 μg/mL） ＋ DNCP（ 100 μg/mL） and heparin（ 1 μg/mL） ＋ IGFβ - 1 （ 100 μg/mL） were supplemented in culture medium. The morphology was observed and immunohistochemistry assay was used to identify the characteristics of cells. RT - PCR was also used to identify the expression of DSPP and DMP1. RESULTS ： After 6 days culture, some cells in DNCP, TGFβ - 1 ＋ DNCP were tall columnar with long process. Some cells polarized.Cells in DNCP and TGFβ - 1 ＋ DNCP groups expressed DSP in immunohistological assay. RT - PCR showed the cells in these two groups expressed specific dentin sialophosphoprotein （DSPP） and dentin matrix protein 1 （ DMP1 ）. Cells in TGFβ - 1 group polarized in some extent but didn＇t express DSP and DSPP. Heparin ＋ IGF - 1 didn＇t induce cytological and functional changes. CONCLUSION： DNCP and TGFβ - 1 ＋ DNCP could induce the ectomesenchymal cells of rat embryonic facial process to differentiate into odontoblast - like cells. DNCP acts as the key effect in this process. This result is similar to the ectomesenchymal cells induced to odontoblast - like cells in three - dimensional culture system.

关 键 词：面突外胚间充质干细胞 成牙本质样细胞 生长因子 分化 

分 类 号：R780.2[医药卫生—口腔医学]