离体牙骨组织界面骨形成过程的细胞学分析  

Cytological analysis of bone formation in the bone interface of isolated teeth

作　　者：郑汉峰 

机构地区：[1]广东省东莞市东城医院,广东省东莞523007

出　　处：《中国口腔种植学杂志》2016年第2期56-59,共4页Chinese Journal of Oral Implantology

基　　金：广东省东莞市科技计划医疗卫生类科研项目。项目名称:离体牙-骨组织的界面识别研究。项目编号:2014105101069

摘　　要：目的:探讨离体牙乳头体外培养下的成牙本质细胞分化及牙骨组织界面的骨形成过程。方法:无菌采集同窝6只出生5d内的小白鼠牙乳头,分为A,B两组。A组剪碎,使用1%胰酶消化,培养于细胞培养皿中,37℃,5%CO2培养箱中培养,常规换液培养8d,用于成牙本质细胞分化成骨细胞,制作细胞爬片进行细胞免疫组化分析OPN蛋白表达;B组直接采用1%胰酶消化,培养于专用不锈钢金属滤网上,培养8天进行4%多聚甲醛固定,石蜡包埋切片,HE染色进行免疫组化特定蛋白分析,通过培养初期及培养第8天细胞学和蛋白学对比分析离体牙乳头骨形成过程。结果:A和B组两组小鼠经细胞分离培养、定向诱导成骨分化及仔鼠乳牙消化培养后免疫组化鉴定,细胞OPN蛋白均为阳性表达,且乳牙牙髓干细胞可以体外分离诱导分化,且可传代至20代生长良好。结论:离体牙骨组织体外培养过程中在诱导培养基培养条件下可以发生成骨分化,且乳牙骨组织消化培养后可生长传代经,经体外诱导分化为成骨细胞,因此离体牙骨组织界面骨形成过程为牙骨髓本质干细胞分化导致。Objective： To investigate the differentiation from bone and teeth bone interface odontoblasts-dental papilla in vitro under formation. Methods： Collecting sterile dental papilla littermate mice born 5d within 6, divided into A, B groups. A group cut into pieces, using a 1% trypsin digestion, cells cul-tured in petri dishes, 37℃, 5%CO_2 incubator, conventional culture medium was changed 8d, for odon-toblast differentiation into bone cells, making the cells crawl sheet for cell immunohistochemistry OPN protein expression; group B directly using a 1% trypsin digestion, cultured on special stainless steel metal mesh, cultured for 8 days 4% paraformaldehyde-fixed, paraffin-embedded sections, HE staining immunohistochemistry analysis of specific proteins by culturing the initial cytology and culture on day 8 and comparative proteomics analysis of dental papilla in vitro bone formation. Results：A group and B group mice were isolated and cultured cells, induced to osteogenic differentiation and deciduous off-spring identified by immunohistochemistry, the cells were positive expression of OPN culture digestion and the dental pulp stem cells can be induced to differentiate in vitro, and may be passaged to 20 gen-erations grow well. Conclusion： From extracted teeth in vitro bone tissue in the process of induction medium culture conditions can occur osteogenic differentiation and bone tissue digestion deciduous culture may be grown by passage through induced differentiation into osteoblasts, Therefore, the bone formation at the bone interface of the isolated teeth is the result of bone marrow stem cell differentiation.

关 键 词：离体牙骨 骨组织界面 细胞学 

分 类 号：R781[医药卫生—口腔医学]