METALLOPROTEINASE EXPRESSION IN CORNEAL WOUNDS

细胞外基质重塑，已知由基质介导 金属蛋白酶（MMPs）在正常和病理条件下，可 确定了接受准分子角膜切除术的患者的视力结果。 拟议研究的目的是获得有关 角膜基质金属蛋白酶及其抑制剂的表达 准分子伤口，以确定是否基质内上皮迁移， 与MMP表达相关，并将MMP水平与基质 疤痕和重塑 A.准分子角膜创伤后MMP的表达 角膜切除术，组织学检查显示无白细胞 浸润，以及角膜上皮再形成的相对延迟 基底膜和粘附结构。 MMPs，负责 基底膜和基质降解在其他系统中，是可检测的 在角膜上皮和基质中的表达。 组织抑制剂 金属蛋白酶抑制剂（TIMPs）调节MMP的活化， 对抗细胞外基质降解 体内伤口愈合 将进行实验以将MMP的表达与 酶谱法检测TIMPs与准分子损伤时间、大小和深度的关系 和系列免疫印迹分析。 这些数据将与 手动角膜切除术伤口 免疫组织化学技术也将是 用于MMP和TIMP定位。 B。 体外MMP表达：器官培养伤口愈合系统将 用于确定角膜细胞是否可以在角膜基质中重新合成MMP。 没有白细胞。 此外，TIMPs和合成MMP 将确定对短期MMP表达的抑制剂。 C. 基质内上皮迁移与MMP合成的相关性： MMP介导的细胞外基质降解促进细胞迁移。 我们观察到，在深环形准分子角膜切除术后， 以及上皮细胞增殖进入中央角膜基质。 这种现象似乎是准分子特异性的，并且可能受到以下因素的影响： MMPs 将比较MMP和TIMP在角膜中的免疫定位 与手法环形角膜切除术相比， 上皮细胞进入中央角膜基质。 这种现象 似乎是准分子特异性的，并可能受到MMP的影响。 将比较角膜中MMP和TIMP的免疫定位， 在手动环形角膜切除术的伤口中。 此外， 局部合成MPP抑制剂对间质内上皮迁移的影响 将对vivo进行研究。 D. MMP和TIMP的表达与长期间质增生的关系 准分子角膜切除术后的瘢痕形成和清除： 基质瘢痕形成和随后的清除，临床观察1-6个月 准分子角膜切削术后MMP-2和TIMP的表达可能与角膜切削术后MMP-2和TIMP的表达有关。 疤痕的程度可能取决于这些和其他因素的平衡。 参与基质调节的蛋白质。 基质酶谱分析 MMP将在瘢痕形成之前、期间和之后测定， 清理 这将与角膜光散射以及与 新合成的胶原蛋白 这些实验将增加我们的 了解角膜MMP表达和角膜伤口愈合。

Extracellular matrix remodelling, known to be mediated by matrix metalloproteinases (MMPs) in normal and pathological conditions, may determined the visual outcome of patients undergoing excimer keratectomy. The aims of the proposed research are to obtain information about the expression in corneal tissue of MMPs and their inhibitors following excimer wounds, to determined if intrastromal epithelial migration is associated with MMP expression, and to correlate MMP levels with stromal scarring and remodelling. A. MMP expression following excimer corneal wounds: Following excimer keratectomy, histologic examination shows absence of leukocyte infiltration, and relative delay in the reformation of corneal epithelial basement membrane and adhesion structures. MMPs, held responsible for basement membrane and matrix degradation in other systems, are detectable in corneal epithelium and stroma after excimer wounds. Tissue inhibitors of metalloproteinases (TIMPs) regulate MMP activation and may guard against extracellular matrix degradation. In vivo wound-healing experiments will be performed to correlate the expression of MMPs and TIMPs with the time, size, and depth of excimer wounds using zymography and serial immunoblot assays. These data will be compared to those after manual keratectomy wounds. Immunohistochemical techniques will also be used for MMP and TIMP localization. B. MMP Expression in vitro: An organ culture wound-healing system will be used to determined if corneal cells can synthesize MMP de novo in the absence of leukocytes. In addition, effect of TIMPs and synthetic MMP inhibitors on short-term MMP expression will be determined. C. Correlation of Intrastromal Epithelial Migration with MMP Synthesis: MMP-mediated extracellular matrix degradation facilitated cell migration. We have observer, following deep annular excimer keratectomy, migration and proliferation of epithelial cells into the central corneal stroma. This phenomenon appears to be excimer-specific and may be influence by MMPs. Immunolocalization of MMP and TIMP in the cornea will be compared with that in manual annular keratectomy, migration and proliferation of epithelial cells into the central corneal stroma. This phenomenon appears to be excimer-specific and may be influence by MMPs. Immunolocalization of MMP and TIMP in the cornea will be compared with that in manual annular keratectomy wounds. In addition, the effect of topical synthetic MPP inhibitors on intrastromal epithelial migration in vivo will be studied. D. Correlation of MMP and TIMP expression with long-term stromal scarring and clearing following excimer keratectomy: The process of stromal scarring and subsequent clearing, clinically observer 1-6 months post-excimer keratectomy, may be related to MMP-2 and TIMP expression. The degree of scarring may depend on the balance of these and other proteins involved in matrix regulation. Zymographic analysis of stromal MMP will be determined before, during, and after scar formation and clearing. This will be correlated with corneal light scattering and with newly-synthesized collagen. These experiments will increase our understanding of corneal MMP expression and of corneal wound healing.