Characterisation of Epithelial-Mesenchymal Transition (EMT) in Oral Keratinocytes

口腔内有大量不同的微生物群落。复杂的细菌生物膜在牙齿硬组织（牙齿）上发育，特别是在龈缝的根表面，在那里它们与龈沟上皮密切相关。这种上皮具有与其他身体部位相同的特性，在保护宿主组织免受细菌侵袭方面起着关键作用。事实上，这种上皮可以协调局部炎症反应并释放防御分子，帮助维持其完整性。口腔上皮中发生的一个潜在的新过程是上皮-间充质转化（EMT），它导致上皮细胞向间充质表型转化。虽然EMT在组织形成和修复中是必不可少的，但它在疾病期间的失调可能导致长期和慢性细胞因子释放的发展，使细菌组织入侵永久化，并导致慢性炎症损伤。假设：关键的口腔细菌通过EMT破坏上皮屏障，导致溃疡和无法解决的慢性炎症病变。该项目将涉及使用不朽和原代人类细胞开发EMT模型系统（2D和3D）。随后将使用分子和细胞分析来评估EMT调节，包括基因和蛋白质表达分析、细胞形态和组织学特征、上皮屏障和涉及相关相互作用免疫系统细胞（如中性粒细胞）的细菌入侵研究。EMT细胞内信号通路（如NF-kB， SMAD）也将在使用药物调节剂的模型系统中进行剖析。主要参考文献：1；Milward。张建军，张建军，张建军，张建军。口腔上皮细胞NF-kB基因在牙周病致病菌中的表达与表达。临床试验免疫学杂志2007年5月；[j] .中国科学院学报（自然科学版）；Prime SS, Nixon SV, Crane IJ。人口腔鳞状细胞癌在细胞培养中的行为。J病理学杂志1990；[j] .中国科学院学报（自然科学版）；张建军，张建军，张建军，等。大鼠角化细胞的结构和结构特征研究。[J] .生物医学工程学报。2012年6月26日。Abdulkareem;Milward;库珀;谢尔顿;Landini。牙周病原体在体外促进口腔鳞癌细胞的EMT，被接受发表在2017年细胞粘附与迁移杂志上。

INTRODUCTION: The oral cavity is heavily colonised by a diverse microflora. Complex bacterial biofilms develop on the dental hard tissues (teeth) and in particular on the root surface in the gingival crevice where they become intimately associated with the crevicular epithelium. This epithelium, exhibits properties in common with other bodily sites and plays a key role in protecting the underlying host tissues from bacterial invasion. Indeed this epithelium can orchestrate the local inflammatory response and releases defence molecules which help maintain its integrity. A potential novel process that occurs in the oral epithelia is that of epithelial-mesenchymal transition (EMT) which results in the conversion of epithelial cells to a mesenchymal phenotype. While EMT is essential in tissue formation and repair its dysregulation during disease can result in the development of prolonged and chronic cytokine release perpetuating bacterial tissue invasion and resulting in a chronic inflammatory lesion.HYPOTHESIS: Key oral bacteria disrupt the epithelial barrier via EMT resulting in ulceration and a non-resolving chronic inflammatory lesion.The project will involve the development of an EMT model system (2D & 3D) using immortal and primary human cells. EMT regulation will subsequently be assessed using molecular and cellular assays including gene & protein expression analysis, cell morphological & histological characterisation, epithelial barrier and bacterial invasion studies which involve relevant interacting immune system cells, e.g. neutrophils. EMT intracellular signalling pathways (e.g. NF-kB, SMAD) will also be dissected in model systems using pharmaceutical modulators. KEY REFERENCES: 1. Milward. MR, Chapple ILC, Wright HJ, Millard JL, Matthews JB, Cooper PR. Differential Activation of NF-kB and gene expression in oral epithelial cells by periodontal pathogens. Clin Exp Immunol. 2007 May;148 (2): 307-324 2. Prime SS, Nixon SV, Crane IJ. The behaviour of human oral squamous cell carcinoma in cell culture. J Pathol 1990; 160 (3): 259-269 3. Khan E, Shelton RM, Cooper PR, Hamburger J, Landini G. Architectural characterization of organotypic cultures of H400 and primary rat keratinocytes. J Biomed Mater Res A. 2012 Jun 26.4. Abdulkareem; Milward; Cooper; Shelton; Landini. Periodontal pathogens promote EMT in oral squamous carcinoma cells in vitro, accepted for publication in Cell Adhesion & Migration 2017.