处理牙本质基质诱导牙囊细胞向成牙本质细胞分化的关键因子筛选及功能研究

Tooth regeneration of tissue engineering method is an ideal dental restoration strategy. To explore the key factors which could promote odontogenic stem cells differentiated into odontolblasts is the key problem in tooth regeneration. In previous study, at the presence of low speed treated dentin matrix, dental follicle cells can differentiate into odontoblasts, and subsequently form dentin tissue; at the presence of high speed treated dentin matrix, dental follicle cells can only differentiate into osteo/cemento-blasts, and subsequently form bone/cementum. The differences of mechanism of two materials induction abilities were unclear. Proposed based on the previous research, this project adopts the high flux differential proteomics and mass spectrometry to further comparative analysis two kinds of materials, to select the key factors which could induce dental follicle cells to differentiate into odontoblasts, to further build a co-culture system including key factors, materials and dental follicle cells and the corresponding cell and animal models, to research the effect and mechanism of key factors to promote cells to differentiate into odontoblasts. Expect to establish a simple and stable induction system. This project also provides certainly experimental basis for the directional differentiation of seeding cells and a new choice for tooth regeneration.

种子细胞复合支架材料行牙齿再生是一种理想的牙齿修复新策略。其中探索可以促进种子细胞向成牙本质细胞定向分化的关键因子及其作用机理是目前牙再生领域亟待解决的关键问题。课题组前期发现低速机械力处理的牙本质基质材料所提供的诱导微环境可以诱导牙囊细胞向成牙本质细胞分化进而形成新生牙本质，而采用高速车针磨除处理的牙本质基质材料只能诱导牙囊细胞向成骨/成牙骨质细胞分化进而形成新生牙骨质结构，两种材料诱导能力差异的作用机理尚不清楚。本研究拟在前期研究基础上，采用高通量差异蛋白质组学及质谱技术对比分析两种牙本质基质材料，以期筛选出可诱导牙囊细胞向成牙本质细胞分化的关键因子，并进一步构建这些关键因子/材料/牙囊细胞共培养体系以及相应细胞、动物模型，研究其促进细胞向成牙本质细胞分化的作用及机理。期望利用关键因子建立简单稳定的诱导体系，为牙齿再生中种子细胞定向分化奠定一定的实验基础，为牙再生组织构建提供新的选择。

种子细胞复合支架材料行牙齿再生是一种理想的牙齿修复新策略。其中探索可以促进种子细胞向成牙本质细胞定向分化的关键因子及其作用机理是目前牙再生领域亟待解决的关键问题。课题组前期发现低速机械力处理的牙本质基质材料所提供的诱导微环境可以诱导牙囊细胞向成牙本质细胞分化进而形成新生牙本质，而采用高速车针磨除处理的牙本质基质材料只能诱导牙囊细胞向成骨/成牙骨质细胞分化进而形成新生牙骨质结构，两种材料诱导能力差异的作用机理尚不清楚。本研究分别制备了低速处理牙本质基质和高速处理牙本质基质。通过动物体内实验验证了两种材料的不同诱导能力，并使用同位素标记相对和绝对定量技术对比分析两种牙本质基质材料蛋白。在两种材料中共鉴定出了487个蛋白，其中382个蛋白具有定量信息，78个蛋白在低速处理牙本质基质中高表达，69个蛋白筛在高速处理牙本质基质中高表达。通过文献检索和筛查，进一步筛选出了一些可能的关键因子，并进一步构建了这些关键因子/材料/牙囊细胞共培养体系以及相应细胞、动物模型，研究其促进细胞向成牙本质细胞分化的作用及机理。期望利用关键因子建立简单稳定的诱导体系，为牙齿再生中种子细胞定向分化奠定一定的实验基础，为牙再生组织构建提供新的选择。