Ichijima H, Petroll W M, Barry P A, Andrews P M, Dai M, Cavanagh H D, Jester J V
Department of Ophthalmology, University of Texas, Dallas 75235-9057.
Invest Ophthalmol Vis Sci. 1993 Aug;34(9):2803-12.
To compare and contrast the in vivo mechanism of wound healing after mechanical scrape and transcorneal freeze (TCF) injury in a rabbit eye model by examining changes in the cytoskeletal organization of contractile, filamentous actin (f-actin) microfilaments as relates to differences in cell migration or translocation during endothelial repair.
Endothelial wound healing after mechanical scrape and transcorneal freeze injury was studied in rabbit eyes using laser scanning confocal microscopy (LSCM). Central corneal mechanical scrape injury was made using an olive tip cannula, and TCF injury was made using a 3-mm diameter stainless steel probe cooled with liquid nitrogen. Cytoskeletal changes in f-actin stained with phalloidin-FITC were observed during wound healing using LSCM.
At 6 hours after mechanical scrape, the leading edge of the migrating sheet showed a decrease in the intensity of phalloidin-FITC staining, suggesting a decrease in cortical f-actin. Migrating endothelial cells in vivo did not appear to develop stress fibers after mechanical scrape, which is consistent with an in vitro cell spreading mechanism of endothelial wound healing. By 24 hours, the denuded area was almost fully resurfaced by migrating endothelial cells. On the other hand, TCF injury produced fibroblastic changes in the endothelial cells with extension and elongation of spindle-shaped endothelial cells at the leading edge by 24 hours after injury. Fibroblastic endothelial cells developed prominent actin stress-fibers, which is consistent with an in vitro cell migration mechanism of endothelial wound healing. Three days after TCF, the wounded area was resurfaced with two cell types: rough, fibroblast-like cells forming a retrocorneal fibrous membrane having prominent f-actin bundles or stress fibers with few cell-cell junctions, and smooth, polygonal-shaped endothelial cells having tight cell junctions with a cortical distribution of f-actin. After 28 days the retrocorneal fibrous membrane was posteriorly covered with normal endothelium.
These data support the hypothesis that endothelial wound healing involves two separate, injury-dependent, mechanisms--cell spreading and cell migration.
通过检测收缩性丝状肌动蛋白（f-肌动蛋白）微丝的细胞骨架组织变化，以比较和对比兔眼模型中机械刮伤和经角膜冷冻（TCF）损伤后伤口愈合的体内机制，这些变化与内皮修复过程中的细胞迁移或移位差异有关。
使用激光扫描共聚焦显微镜（LSCM）研究兔眼中机械刮伤和经角膜冷冻损伤后的内皮伤口愈合情况。使用橄榄头插管造成中央角膜机械刮伤，使用直径3毫米的不锈钢探针经液氮冷却造成TCF损伤。在伤口愈合过程中，使用LSCM观察用鬼笔环肽-FITC染色的f-肌动蛋白的细胞骨架变化。
机械刮伤后6小时，迁移细胞片的前沿显示鬼笔环肽-FITC染色强度降低，表明皮质f-肌动蛋白减少。体内迁移的内皮细胞在机械刮伤后似乎未形成应力纤维，这与内皮伤口愈合的体外细胞铺展机制一致。到24小时时，裸露区域几乎完全被迁移的内皮细胞重新覆盖。另一方面，TCF损伤在内皮细胞中产生成纤维细胞样变化，损伤后24小时前沿的梭形内皮细胞延伸和伸长。成纤维细胞样内皮细胞形成突出的肌动蛋白应力纤维，这与内皮伤口愈合的体外细胞迁移机制一致。TCF损伤三天后，伤口区域由两种细胞类型重新覆盖：粗糙的、成纤维细胞样细胞形成后弹力层纤维膜，具有突出的f-肌动蛋白束或应力纤维，细胞间连接较少；光滑的、多边形内皮细胞具有紧密的细胞连接，f-肌动蛋白呈皮质分布。28天后，后弹力层纤维膜后部被正常内皮覆盖。
这些数据支持以下假设，即内皮伤口愈合涉及两种独立的、依赖损伤的机制——细胞铺展和细胞迁移。
