THROMBIN IN WOUND HEALING--MULTIPLE FUNCTIONS

牙周病是感染特定疾病的病理结果。 具有毒力因子的病原体 或间接导致牙齿支撑结构的破坏。 有压倒性的证据表明，东道主的反应，特别是对 这些病原菌的脂多糖在致病中起主要作用 定义牙周疾病的骨和结缔组织破坏。 大多数研究清楚地表明，多形核白细胞是 对牙周病原体的防御至关重要，细菌 中性粒细胞抗菌功能的逃避和/或中和很常见 成功病原菌的毒力特性。大多数形式的牙周 疾病具有典型的间歇性，代表着一种 组织破坏和伤口愈合的迭代过程 长期接触有毒细菌产品。它是整体的 拟议研究的目标是开始了解相互作用 致病菌和/或其代谢产物之间，炎性 炎症介质和促炎因子，包括急性和慢性炎症 细胞、止血和血栓成分及启动因子 牙周病中的伤口愈合。具体地说，亚细胞 激肽释放酶在中性粒细胞中的定位将被确定。该信号 凝血酶刺激中性粒细胞的转导和第二信使 激肽释放酶合成和释放的上调将是 下定决心。人类中性粒细胞的颗粒含量将被分级 激肽释放酶的分布与 中性粒细胞标志物的特征。激肽释放酶合成， 划分和发布的特征将遵循 凝血酶对中性粒细胞的刺激作用 特征激动剂。颗粒标志物释放，呼吸爆发， 趋化性、细胞形态的改变和粘附性将被表征 凝血酶刺激中性粒细胞和早幼粒细胞 HL60未分化，分化为中性粒细胞或 单核细胞。脉冲追逐实验将用于监测 人中性粒细胞和HL6O细胞合成激肽释放酶。这个 凝血酶对放线杆菌吞噬和杀灭作用的影响 将检测伴生放线菌和牙龈卟啉单胞菌 使用双荧光显微镜分析。促炎症反应的影响 而原嗜中性粒细胞功能的成分将在添加剂中进行检测 使用纯化的细胞因子和炎症介质的实验也是如此 作为血小板、单核细胞和细胞外基质表面。平行 研究将检查细胞类型和细胞类型出现的动力学 选定的生物相关分子的出现和影响 从小鼠皮下腔模型中收集的液体用于 宿主/牙周病原体相互作用的体内评价。这些研究 将深入了解主机之间发生的复杂交互 细胞、致病菌和生物活性分子。是这样的 信息对干预性临床的发展至关重要 细菌介导的组织破坏过程的方法，如 牙周病。

Periodontal diseases are the pathologic outcome of infection with specific pathogens that possess virulence factors that contribute either directly or indirectly to the destruction of the supporting structures of teeth. There is overwhelming evidence that host response especially to the lipopolysaccharide of these pathogens makes a primary contribution to the bone and connective tissue destruction that define periodontal diseases. Most research clearly indicates that the polymorphonuclear leukocyte is central to the defense against periodontopathogens and that the bacterial evasion and/or neutralization of PMN anti-bacterial function are common virulence traits of the successful pathogen. Most forms of periodontal disease are characteristically episodic and represent the result of an iterative process of tissue destruction and wound healing in response to chronic exposure to noxious bacterial products. It is the overall objective of the proposed studies to begin to understand the interactions between pathogenic bacteria, and/or their metabolic products, inflammatory mediators and proinflammatory factors, both acute and chronic inflammatory cells, hemostatic and thrombic components and the initiating factors of wound healing in periodontal diseases. Specifically the subcellular localization of kallikrein in neutrophils will be determined. The signal transduction and second messengers involved in thrombin stimulation of PMN and the up regulation of kallikrein synthesis and release will be determined. The granule content of human neutrophils will be fractionated and the distribution of kallikrein determined relative to that of characterized neutrophil granule markers. Kallikrein synthesis, compartmentalization and release will be characterized following the stimulation of neutrophils with thrombin, as well as, with other well characterized agonists. Release of granule markers, respiratory burst, chemotaxis, change in cell morphology and adherence will be characterized following thrombin stimulation of neutrophils and the promyelocytic cell line, HL60 undifferentiated and differentiated to either neutrophilic or monocytic cells. Pulse-chase experiments will be used to monitor kallikrein synthesis by both human neutrophils and HL6O cells. The influence of thrombin on the phagocytosis and killing of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis will be determined using a dual fluorescent microscopy assay. Influence of proinflammatory and prothombic components on neutrophil function will be examined in admix experiments using purified cytokines and inflammatory mediators, as well as platelets, monocytes, and extracellular matrix surfaces. Parallel studies will examine the kinetics of appearance of cells types and the appearance and influence of selected biologically relevant molecules in fluid collected from a murine subcutaneous chamber model for the evaluation of host/periodontopathogen interaction in vivo. These studies will provide insight into the complex interactions that occur between host cells, pathogenic bacteria, and biologically active molecules. Such information is essential to the development of interventive clinical approaches for bacterial mediated tissue destructive processes such as periodontal diseases.