儿童乳牙牙髓干细胞顺硬度基质表面延展和向牙本质分化的潜能  

Extension and dentin differentiation potential of dental pulp stem cells from human deciduous teeth on the stiff matrix surface

作　　者：李章一[1] 刘凤婷 黄建永[3] 苏小营[4] 王志兴[1] 郑全 李艳霞[5] 刘晓智[5] Li Zhangyi;Liu Fengting;Huang Jianyong;Su Xiaoying;Wang Zhixing;Zheng Quan;Li Yanxia;Liu Xiaozhi(Department of Stomatology,the Fifth Central Hospital of Tianjin,Tianjin 300450,China;Tianjin Medical University,Tianjin 300070,China;Department of Mechanics and Engineering Science,Peking University,Beijing 100871,China;Department of Pediatric Stomatology,Tianjin Stomatological Hospital,Tianjin 300041,China;Central Laboratory,the Fifth Central Hospital of Tianjin,Tianjin 300450,China)

机构地区：[1]天津市第五中心医院口腔科,天津市300450 [2]天津医科大学,天津市300070 [3]北京大学力学与工程科学系,北京市100871 [4]天津市口腔医院儿童口腔科,天津市300041 [5]天津市第五中心医院中心实验室,天津市300450

出　　处：《中国组织工程研究》2020年第13期2005-2010,共6页Chinese Journal of Tissue Engineering Research

基　　金：天津市卫生和计划生育委员会科技基金项目(2014KY23)，项目负责人：苏小营;天津滨海新区卫生和计划生育委员会科技基金项目(2015BWKY003)，项目负责人：李章一~~

摘　　要：背景:牙髓干细胞在适宜诱导条件下能够向牙本质分化,是牙齿组织工程的重要种子细胞。然而,以往所使用的诱导剂多为化学制剂,不利于体内应用。近来有报道称间充质干细胞能够顺材质硬度分化,这种由物理特性诱导的细胞分化研究报道较少。目的:观察人源性乳牙牙髓干细胞在硬介质表面的延展特点及向牙本质分化潜能,为牙组织工程提供参考。方法:原代分离培养和鉴定儿童自然脱落的乳牙牙髓干细胞;使用低熔点琼脂糖配制弹性模量为(9.12±0.94),(27.18±3.55),(59.37±4.05)和(86.45±5.33)k Pa4个梯度的固体凝胶基质,二维克隆形成实验和划痕实验检测第4代乳牙牙髓干细胞在上述硬基质表面的延展能力,Western blot方法检测牙本质基质蛋白1、牙本质磷蛋白、牙本质涎蛋白的表达。结果与结论:当人乳牙牙髓干细胞接种于极低和低等硬度凝胶介质表面时,乳牙牙髓干细胞几乎均以边缘整齐的细胞克隆存在,很少见细胞平铺和延展现象;但是当将其接种于中等和高等硬度凝胶介质表面时,乳牙牙髓干细胞克隆边缘则表现出明显的平铺和延展,表现为细胞胞体变大,细胞边缘外伸明显。相似的现象也经细胞划痕实验所验证。人源性乳牙牙髓干细胞在中等和高等弹性模量介质表面培养时,表达较高水平的牙本质基质蛋白1、牙本质磷蛋白、牙本质涎蛋白。结果表明,人源性乳牙牙髓干细胞随着培养基质硬度的增加其延展性及成牙本质分化能力逐渐增强,为未来牙组织工程提供方法借鉴。BACKGROUND: Dental pulp stem cells can differentiate into dentin under appropriate induction conditions, which are important seed cells in dental tissue engineering. However, the commonly used inducers are chemical agents, which are not available for in vivo application. Mesenchymal stem cells can differentiate with the material hardness, and the physical property-induced cell differentiation is little reported. OBJECTIVE: To observe the extension characteristics and dentin differentiation potential of dental pulp stem cells from human deciduous teeth on the stiff matrix surface. METHODS: Dental pulp stem cells from naturally shed deciduous teeth were isolated, cultured and identified. Four solid gel matrixes with elasticity modulus of(9.12±0.94),(27.18±3.55),(59.37±4.05) and(86.45±5.33) kPa were made using low melting point agarose. The extension ability of passage 4 dental pulp stem cells on the surface of the above solid matrixes was detected by two-dimensional clone formation and cell scratch tests. The protein expression levels of dentin matrix protein-1, dentin phosphoprotein and dentin sialoprotein were detected by western blot assay. RESULTS AND CONCLUSION: Dental pulp stem cells from human deciduous teeth seeded on the gel matrix with extremely low and low hardness almost existed as cell clones with neat edges, and cell spreading and extension were rare. When seeded on the gel matrix with moderate and high hardness, the cloned edge of deciduous dental pulp stem cells spread and extended obviously. The cell body became large and the cell edge extended significantly. The cell scratch test revealed the similar phenomenon. When seeded on the gel matrix with moderate and high hardness, dental pulp stem cells from human deciduous teeth exhibited high expression levels of of dentin matrix protein-1, dentin phosphoprotein and dentin sialoprotein. In summary, with the increase of matrix hardness, the abilities of extension and differentiation into dentin of dental pulp stem cells from human deciduous teeth ar

关 键 词：牙髓干细胞 牙本质 细胞迁移 细胞分化 牙本质基质蛋白1 牙本质磷蛋白 牙本质涎蛋白 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]