釉质基质蛋白对乳牙牙髓干细胞体外增殖、分化能力影响的研究
发布时间：2019-01-30 10:34
【摘要】：背景：牙齿发育包括一系列至关重要的有序的相互作用，发生在口腔上皮与颅神经嵴间充质细胞之间。研究已经证实原发性上皮带在牙齿发育中的诱导作用，并且这些相互作用，包括生长因子、同源基因和转录因子的高度协调表达，逐步诱导牙原基在牙弓内形成一个具有特定大小、形状和位置的复杂矿化结构。有研究在小鼠体内标记一种基因来追踪神经嵴细胞的迁移与分化，结果证明在蕾状期，颅神经嵴源外胚间充质对于牙乳头和其周围的牙囊的形成有重要作用。研究证明，成牙本质细胞，牙本质基质以及大部分的牙髓组织都来源于颅神经嵴。 牙齿损伤的机制包括诱导细胞凋亡，活化的免疫反应和牙齿组织生理的变化，在多数情况下，凋亡细胞迅速清除。并未诱发炎症反应。而牙齿组织重建的机制类似于牙齿发育的过程。髓腔的结构重建发生于牙齿修复性牙本质沉积。成牙本质层的细胞凋亡明显多于牙髓的其它部位。由于细胞凋亡而消失的成牙本质细胞层有可能产生死亡信号，导致同时临近的祖细胞消失。损伤发生后，，干细胞从邻近的储存处转移至创伤修复区，此处出现大量的干细胞。成牙本质细胞和邻近的祖细胞由于细胞凋亡而消失，这引起牙髓干细胞迁移至损伤区域，在此处它们分化成成牙本质样细胞，从而保证牙髓的重建。 目的：研究釉质基质蛋白（Enamel Matrix Proteins，EMPs）对人脱落乳牙牙髓干细胞（stem cells from human exfoliated dediduous teeth，SHED）体外增殖分化能力的影响。方法：利用酶消化法联合组织块法获得脱落乳牙牙髓干细胞，并进行形态学观察。三氯乙酸法制备EMPs，用不同浓度的EMPs对SHED进行诱导，利用四唑盐比色法（MTT）检测并分析诱导后的SHED增殖活性的变化，检测经诱导后的培养液中碱性磷酸酶（ALP）。RT-PCR检测牙本质涎磷蛋白（dentin sialophosphoprotein,DSPP）及牙本质基质蛋白1(dentin matrix protein1, DMP-1）的mRNA表达。结果：人脱落乳牙牙髓干细胞呈集落生长，并且在体外具有一定的自我增殖能力。EMPs对乳牙牙髓干细胞的增殖无明显影响，而能够显著提高ALP的活性，并呈现一定的剂量依赖性。经EMPs诱导后，细胞相对高表达DSPP、DMP-1mRNA。结论：EMPs对于SHED向成牙本质样分化具有积极作用。
[Abstract]:Background: tooth development includes a series of important ordered interactions between oral epithelium and cranial neural crest mesenchymal cells. Studies have demonstrated the induction of primary upper belt in tooth development, and these interactions, including the highly coordinated expression of growth factors, homologous genes and transcription factors, A complex mineralized structure with a specific size, shape and position was gradually induced to form in the dental arch. A gene was labeled in mice to track the migration and differentiation of neural crest cells. The results showed that the extracellular mesenchymal of cranial neural crest was important for the formation of dental papilla and its surrounding dental sac in the bud stage. It has been shown that odontoblast, dentin matrix and most of dental pulp come from cranial nerve crest. The mechanisms of tooth injury include inducing apoptosis, activated immune response and physiological changes of tooth tissue. In most cases, apoptotic cells are eliminated rapidly. No inflammatory response was induced. The mechanism of tooth tissue reconstruction is similar to the process of tooth development. The structural reconstruction of the pulp cavity occurs when the dentin is restored. The apoptosis of dentin layer was much more than that of other parts of dental pulp. The layer of odontoblast which disappeared due to apoptosis may produce death signal, resulting in the disappearance of adjacent progenitor cells at the same time. After the injury, stem cells are transferred from nearby stores to the wound repair area, where a large number of stem cells are present. The odontoblast and adjacent progenitor cells disappear due to apoptosis, which causes dental pulp stem cells to migrate to the damaged region, where they differentiate into dentin-like cells, thus ensuring dental pulp reconstruction. Aim: to study the effect of enamel matrix protein (Enamel Matrix Proteins,EMPs) on the proliferation and differentiation of human deciduous dental pulp stem cells (stem cells from human exfoliated dediduous teeth,SHED) in vitro. Methods: pulp stem cells of deciduous teeth were obtained by enzyme digestion combined with tissue block method. EMPs, was prepared by trichloroacetic acid and SHED was induced with different concentrations of EMPs. The proliferation activity of SHED was detected and analyzed by (MTT) with tetrazolium salt colorimetry. The expression of mRNA in dentin sialophosphorus protein (dentin sialophosphoprotein,DSPP) and dentin matrix protein 1 (dentin matrix protein1, DMP-1) was detected by alkaline phosphatase (ALP). RT-PCR). Results: human deciduous dental pulp stem cells grew in colony and had the ability of self-proliferation in vitro. EMPs had no obvious effect on the proliferation of deciduous dental pulp stem cells, but could significantly increase the activity of ALP. And showed a certain dose dependence. After induction by EMPs, the expression of DSPP,DMP-1mRNA. in the cells was relatively high. Conclusion: EMPs plays a positive role in the odontoid differentiation of SHED.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R780.2
本文编号：2418041
[Abstract]:Background: tooth development includes a series of important ordered interactions between oral epithelium and cranial neural crest mesenchymal cells. Studies have demonstrated the induction of primary upper belt in tooth development, and these interactions, including the highly coordinated expression of growth factors, homologous genes and transcription factors, A complex mineralized structure with a specific size, shape and position was gradually induced to form in the dental arch. A gene was labeled in mice to track the migration and differentiation of neural crest cells. The results showed that the extracellular mesenchymal of cranial neural crest was important for the formation of dental papilla and its surrounding dental sac in the bud stage. It has been shown that odontoblast, dentin matrix and most of dental pulp come from cranial nerve crest. The mechanisms of tooth injury include inducing apoptosis, activated immune response and physiological changes of tooth tissue. In most cases, apoptotic cells are eliminated rapidly. No inflammatory response was induced. The mechanism of tooth tissue reconstruction is similar to the process of tooth development. The structural reconstruction of the pulp cavity occurs when the dentin is restored. The apoptosis of dentin layer was much more than that of other parts of dental pulp. The layer of odontoblast which disappeared due to apoptosis may produce death signal, resulting in the disappearance of adjacent progenitor cells at the same time. After the injury, stem cells are transferred from nearby stores to the wound repair area, where a large number of stem cells are present. The odontoblast and adjacent progenitor cells disappear due to apoptosis, which causes dental pulp stem cells to migrate to the damaged region, where they differentiate into dentin-like cells, thus ensuring dental pulp reconstruction. Aim: to study the effect of enamel matrix protein (Enamel Matrix Proteins,EMPs) on the proliferation and differentiation of human deciduous dental pulp stem cells (stem cells from human exfoliated dediduous teeth,SHED) in vitro. Methods: pulp stem cells of deciduous teeth were obtained by enzyme digestion combined with tissue block method. EMPs, was prepared by trichloroacetic acid and SHED was induced with different concentrations of EMPs. The proliferation activity of SHED was detected and analyzed by (MTT) with tetrazolium salt colorimetry. The expression of mRNA in dentin sialophosphorus protein (dentin sialophosphoprotein,DSPP) and dentin matrix protein 1 (dentin matrix protein1, DMP-1) was detected by alkaline phosphatase (ALP). RT-PCR). Results: human deciduous dental pulp stem cells grew in colony and had the ability of self-proliferation in vitro. EMPs had no obvious effect on the proliferation of deciduous dental pulp stem cells, but could significantly increase the activity of ALP. And showed a certain dose dependence. After induction by EMPs, the expression of DSPP,DMP-1mRNA. in the cells was relatively high. Conclusion: EMPs plays a positive role in the odontoid differentiation of SHED.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R780.2
【参考文献】
相关期刊论文 前1条
1 邹慧儒;张兰成;秦宗长;杨学斌;Steven Brookes;;釉基质蛋白组成及诱导细胞分化研究进展[J];中国组织工程研究;2012年16期
本文编号：2418041
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