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EMPs作用下人牙龈上皮细胞及牙周膜细胞基因表达差异研究

Analysis of Differential Expressed Gene Profiles in hGECs and hPDLCs Challenged by EMPs

【作者】 杜岩

【导师】 吴织芬; 金岩;

【作者基本信息】 第四军医大学 , 口腔临床医学, 2006, 博士

【摘要】 牙周组织再生一直是牙周病治疗领域追求的目标,理想的牙周组织再生需达到以下要求:1)重建功能性上皮封闭;2)新的Sharpey氏纤维插入重建功能性牙周膜;3)暴露根面上形成新的无细胞性牙骨质;4)牙槽骨高度恢复到距釉牙骨质界2mm以内。但由于对牙周组织再生机理认识还不完全清楚,导致目前牙周组织再生还不能完全达到上述要求。 近年发现釉基质蛋白(Enamel Matrix Proteins,EMPs)也可以促进牙周组织再生,它可诱导出无细胞性牙骨质和生理性排列的牙周韧带,且很少伴随有长上皮结合。牙周组织再生的细胞学基础就在于如何促使牙周膜细胞优先占据缺损区、抑制牙龈上皮细胞的过度增殖。EMPs促进牙周组织再生可能遵循这一细胞学机制。但它作为一种细胞外基质蛋白,如何发挥这种作用还不清楚。对此进行深入研究,不仅有助于我们深入了解牙周组织再生的潜在机制,也能够指导临床应用,而且有希望开发出新的效果更好的牙周缺损治疗技术。 本课题以两种与牙周组织再生密切相关的细胞(牙周膜细胞和牙龈上皮细胞)为研究对象,从基因水平初步分析EMPs对它们发挥不同生物学作用的机理。通过本课题的研究,不仅可能发现介导EMPs对细胞增殖发生影响的关键信号转导通路和关键基因、为进一步研究EMPs如何促进牙周组织再生打下基础;而且可能发现参与牙周组织再生的因子。本课题分为两部分。第一部分 EMPs对人牙龈上皮细胞抑制作用的基因表达差异分析 本部分主要研究了EMPs对人牙龈上皮细胞(Human Gingival Epithelial

【Abstract】 The regeneration of periodontal attachment apparatus destroyed or lost due to periodontitis is a major objective in periodontal therapy. These destroyed tissues should be repaired and regenerated into physiologic status, which is termed as "true regeneration of periodontal tissues". Recently, a derivative of enamel matrix proteins (EMPs) extracted from porcine tooth buds has been introduced as a device to achieve this goal predictably. They can induce the "true" regeneration of acellular cementum, alveolar bone, and periodontal ligament where the collagen fibers arranged physiologically, which are quite different with the histological results of other methods. However, the underlying biological mechanisms are not so clear.In this study, we try to explore the genetic mechanism of EMPs in promoting periodontal regeneration by genechip methods. Two kinds of primary cultured cells, human periodontal ligament cells (hPDLCs) and gingival epithelial cells (hGECs), were used because their important role in periodontal tissues regeneration.Our study would not only map the gene differential expression profiles of hPDLCs or hGECs challenged by EMPs, but also find some key genes and/or signaling pathways, and even some new promoting factors in periodontal regeneration.We divided our study into 2 parts for the convenience of expression. Both of them shared the common investigation stratege including as follows: chosing a proper content of EMPs and test timepoint for gene chip, exploring the gene differential expression profile of hPDLCs or hGECs challenged by EMPs, and

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