COBRE: UOK HSC: P1: MICROBIAL MEDIATORS OF TISSUE CELL BEHAVIOR

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 这个拟议的子项目的总体目标是进一步了解口腔细菌及其产物如何直接影响牙周组织内组织细胞的行为，特别是牙龈上皮细胞和结缔组织成纤维细胞。在这个子项目中要测试的中心假设是生物膜细菌对牙周组织内的组织细胞行为产生直接影响。我们的具体目标是：1）分析伴放线放线杆菌（Actinobacillusactinomycemcomitans，Aa）产生的介质的作用（从嗜酸性或生物膜相关生物体释放）和微生物诱导的介质（上皮防御素和来自细菌刺激的细胞培养物的上清液）对牙龈上皮细胞和成纤维细胞的迁移和增殖的影响;和2）确定Aa产生的介质和微生物诱导的宿主组织细胞介质对细胞外基质蛋白合成的影响，细胞因子和其它细胞信号分子。牙龈上皮细胞和成纤维细胞的运动将使用延时视频显微镜和计算机辅助图像分析进行分析。将使用体外伤口模型研究细胞迁移和增殖，其中上皮细胞和成纤维细胞的汇合单层受伤，并随时间测量所创建伤口空间的再增殖。将分析细胞的合成活性，以产生胶原蛋白和非胶原蛋白、胰高血糖素、细胞因子、防御素和防御素样分子。虽然尚未在成纤维细胞中检测到防御素，但在细菌与结缔组织长时间直接接触的情况下，尚未对其进行充分研究。这与牙周病学中的几种临床情况有关，例如暴露的外科材料，以及以细菌侵入牙龈结缔组织为特征的某些牙周疾病，例如侵袭性牙周炎和坏死性溃疡性牙龈炎。总的来说，这组研究将进一步加深我们对牙周病中细菌生物膜对上皮和结缔组织细胞行为和代谢的影响的理解，并为进一步研究这些致病机制如何成为治疗干预的目标铺平道路。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The overall goal of this proposed subproject is to further our understanding of how oral bacteria and their products directly influence the behavior of tissue cells within the periodontium, specifically gingival epithelial cells and connective tissue fibroblasts. The central hypothesis to be tested in this Subproject is that biofilm bacteria exert direct effects on tissue cell behavior within the periodontium. Our specific aims are: 1) to analyze the effects of mediators produced by Actinobacillus actinomycemcomitans (Aa) (released from planktonic or biofilm associated organisms) and microbial-induced mediators (epithelial defensins and supernatants from bacteria stimulated cell cultures) on the migration and proliferation of gingival epithelial cells and fibroblasts; and 2) to determine the effects of Aa-produced mediators and microbial-induced host tissue cell mediators on the synthesis of extracellular matrix proteins, cytokines and other cell signaling molecules by gingival epithelial cells and fibroblasts. Motility of gingival epithelial cells and fibroblasts will be analyzed using time-lapse videomicroscopy and computer-assisted image analysis. Cell migration and proliferation will be studied using an in-vitro wound model, in which a confluent monolayer of epithelial cells and fibroblasts is wounded and repopulation of the created wound space is measured over time. Synthetic activities of cells will be analyzed for production of collagen and non-collagen proteins, prostaglandins, cytokines, defensins, and defensin-like molecules. Although defensins have not been been detected in fibroblasts, this has not been adequately studied in situations where bacteria are in direct contact with connective tissues for extended periods of time. This is relevant to several clinical situations in periodontics, for example, exposed surgical materials, and in certain periodontal diseases characterized by bacterial invasion into gingival connective tissue, such as aggressive periodontitis and necrotizing ulcerative gingivitis. Collectively, this group of studies will further our understanding of the the effects of bacterial biofilms on epithelial and connective tissue cell behavior and metabolism in periodontal diseases and will pave the way for further studies focusing on how these pathogenic mechanisms may become targets for therapeutic intervention.We expect no changes in the gender/minority composition of the subjects.