CELL MOTILITY AND ADHESION IN CORNEAL WOUND HEALING

角膜上皮经常受到机械和化学作用， 创伤，以及一些全身性疾病影响其愈合和粘合 能力的 这些损伤可能导致持续的缺陷或复发 糜烂 长期目标是获得有关角膜的信息， 上皮伤口愈合和粘连，这将导致改善治疗 持续性上皮缺损和复发性角膜糜烂，以及 开发自体上皮移植的程序， 慢性眼表疾病 在过去一年取得的进展的基础上 在供资期间，提出了以下具体目标： 上皮创伤愈合：（1）透射电镜和扫描确定 电子显微镜下复层上皮细胞层移动 进入伤口以形成迁移片的前缘。 顶端或 基底细胞将用颗粒标记， 电镜 (2)通过乳胶球脉冲标记测定 与细胞膜特异性结合，无论细胞膜的数量 每个暴露于撕裂或创伤表面的单个细胞保持恒定， 在愈合过程中膨胀或翻转。 (3)确定是否存在以及如何存在 或缺乏特异性分子（层粘连蛋白、纤连蛋白、硫酸乙酰肝素 蛋白聚糖）对伤口基质的影响愈合率， 细胞表面糖缀合物和角膜上皮合成。 角膜上皮粘连：（1）建立明确的器官培养体系 用于角膜上皮合成和组装基底膜。 将通过TEM、免疫组织化学和SDS-PAGE进行合成试验 电泳 (2)确定的起源，合成，和组成的 锚定纤维，半桥粒（HD）的纤维状基质成分 其功能是将基底膜固定在基质上的复合体。 （三） 表征上存在的新HD形成的成核位点 上皮基底膜。 (4)进行深入的形态测量 分析正常和正常人的HD复合体（包括锚定原纤维）， 糖尿病人角膜，以及如果可用，在持续性 上皮缺损或复发性糜烂史。 上皮移植：（1）确定移植口腔的可行性 粘膜上皮自体移植到兔角膜上的缺损，和（2） 决定他们的命运。

The corneal epithelium is often subjected to mechanical and chemical wounds, and a number of systemic diseases affect its healing and adhesive capabilities. These insults can lead to persistent defects or recurrent erosions. The long-term aims are to obtain information about corneal epithelial wound healing and adhesion that will lead to improved treatment for persistent epithelial defects and recurrent corneal erosions, and to develop procedures for autograft epithelial transplant to humans who have chronic ocular surface disease. Building on progress during the last funding period, the following specific goals are proposed: Epithelial wound healing: (1) Determine by treansmission and scanning electron microscopy which cell layer of the stratified epithelium moves into a wound to form the leading edge of the migrating sheet. Apical or basal cells will be labeled with particles that can be visualized by electron microscopy. (2) Determine, by pulse-labeling with latex spheres that specifically bind to cell membranes, whether the amount of membrane per individual cell exposed to the tear or wound surface remains constant, expands, or turns over during healing. (3) Determine if and how presence or absence of specific molecules (laminin, fibronectin, heparan sulfate proteoglycan) on the wound substrate affect healing rate, expression of cell surface glycoconjugates, and syntheses by the corneal epithelium. Corneal epithelial adhesion: (1) Develop a defined organ culture system for basement membrane synthesis and assembly by the orneal epithelium. Assay for synthesis will be done by TEM, immunohistochemistry, and SDS-PAGE electrophoresis. (2) Determine origin, synthesis, and composition of the anchoring fibril, a fibrillar stromal component of the hemidesmosome (HD) complex whose function is to hold the basement membrane to the stroma. (3) Characterize the nucleation sites for new HD formation that are present on the epithelial basement membrane. (4) Perform in-depth morphometric analysis of the HD complex (including anchoring fibrils) in normal and diabetic human corneas and, if available, in corneas with persistent epithelial defects or a history of recurrent erosion. Epithelial transplant: (1) Determine the feasibility of transplanting oral mucosal epithelial autografts to defects on rabbit corneas, and (2) determine their fate.