牙胚组织中成釉蛋白表达免疫组织化学和原位杂交的特点  

Characteristics of immunohistochemistry and in situ hybridization of ameloblastin in human developing tooth germ

作　　者：田宇[1] 鲁红[1] 倪龙兴[1] 肖明振[1] 余擎[1] 

机构地区：[1]解放军第四军医大学口腔医学院牙体牙髓病科,陕西省西安市710032

出　　处：《中国临床康复》2006年第41期59-61,I0004,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助(30300386);全军"十五"计划基金重点课题(012089)~~

摘　　要：目的:观察人牙胚组织中成釉蛋白的表达特点。方法:实验于2002-02/04在解放军第四军医大学口腔内科实验室完成。取孕5个月流产男性胎儿(产妇知情同意),截取上、下颌骨,常规方法制备人牙胚组织石蜡切片。用兔抗人AMBN多克隆抗体,对人牙胚组织石蜡切片和正常狗牙周组织石蜡切片进行免疫组织化学染色。标记人成釉蛋白cDNA探针,对人牙胚组织石蜡切片进行原位杂交。结果:①免疫组织化学染色发现成釉蛋白由内釉上皮细胞在分化为成熟成釉细胞前开始表达,分泌期成釉细胞高度表达并持续整个釉质形成期,新形成的牙釉质染色阳性;成牙本质细胞在分泌牙本质基质之前开始表达成釉蛋白,在牙本质形成过程中成牙本质细胞持续表达成釉蛋白,呈弱阳性,前期牙本质中呈弱阳性染色;上皮根鞘细胞染色阳性。狗牙周组织染色发现牙骨质表层细胞呈强阳性染色,新形成的牙骨质中可见阳性染色颗粒,呈网状分布。②对人牙胚组织石蜡切片的原位杂交显示,成釉蛋白在成釉细胞、成牙本质细胞、上皮根鞘细胞中有表达,表达部位与免疫组织化学结果一致。结论:内釉上皮首先表达成釉蛋白,之后前成牙本质细胞达成釉蛋白。成釉细胞、成牙本质细胞、上皮根鞘细胞表达成釉蛋白。AIM： To observe the expression of ameloblastin in human tooth germ. METHODS： The experiment was carried out in the Laboratory of Stomatology Medicine, Fourth Military Medical University of Chinese PLA between February and April in 2002. Male fetuses of 5 months pregnancy were adopted to fetch maxillary bone and mandible. Paraffin sections of human developing tooth germ and dog periodontal tissue were used for immunohistochemical staining using a rabbit anti-human AMBN polyclonal antibody, and paraffin sections of human tooth germ was conducted in situ hybridization using a ameloblastin labeled cDNA probe. RESULTS： （l）Immunohistochemistry result showed that ameloblastin was expressed before the polarization of pre-ameloblasts. The secretory ameloblasts were intensively stained during the whole period of enamel formation. The newly-formed dental enamel matrix was positively stained; The odontoblasts expressed ameloblastin before they secreted dentinal matrix during the whole period of dentinification. The secretory odontoblasts and the pre-dentin were both faintly stained; Epithelial root sheath was positively stained. In dog periodontal tissue, some sells covering the cementum and the newly-formed cementum matrix were stained strongly, and distributed in mesh. Gin situ hybridization of human tooth germ reyealed the same distribution of ameloblastin expressing in ameloblasts, odontoblasts and epithelial root sheath as immanohistochemistry. CONCLUSION： Ameloblastin is expressed in internal enamel epithelial primarily, and then in odontoblasts. Ameloblasts, odontoblasts and epithelial root sheath cells all can express ameloblastin.

关 键 词：牙釉质蛋白质类 人 免疫组织化学 DNA探针 原位杂交 

分 类 号：R318[医药卫生—生物医学工程]