Actin filament organization during endothelial wound healing in the rabbit cornea: comparison between transcorneal freeze and mechanical scrape injuries.
Actin filament organization during endothelial wound healing in the rabbit cornea: comparison between transcorneal freeze and mechanical scrape injuries.
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发表时间：
1993-08
影响因子：
4.4
通讯作者：
H. Ichijima;W. Petroll;P. Barry;P. Andrews;M. Dai;H. D. Cavanagh;J. Jester
中科院分区：
文献类型：
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作者：
H. Ichijima;W. Petroll;P. Barry;P. Andrews;M. Dai;H. D. Cavanagh;J. Jester
目的 通过检查收缩性丝状肌动蛋白（f-肌动蛋白）微丝的细胞骨架组织的变化与内皮修复过程中细胞迁移或易位的差异相关，比较和对比兔眼模型中机械刮擦和经角膜冷冻（TCF）损伤后伤口愈合的体内机制。方法 使用激光扫描共聚焦显微镜 (LSCM) 研究兔眼机械刮擦和经角膜冷冻损伤后的内皮伤口愈合情况。使用橄榄尖插管造成中央角膜机械刮擦损伤，并使用液氮冷却的3毫米直径不锈钢探针造成TCF损伤。使用 LSCM 观察伤口愈合过程中用鬼笔环肽-FITC 染色的 f-肌动蛋白的细胞骨架变化。结果机械刮擦后 6 小时，迁移片的前缘显示鬼笔环肽-FITC 染色强度降低，表明皮质 f-肌动蛋白减少。机械刮擦后，体内迁移的内皮细胞似乎没有产生应力纤维，这与内皮伤口愈合的体外细胞扩散机制一致。 24小时后，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损伤导致内皮细胞发生成纤维细胞变化，损伤后24小时前缘的梭形内皮细胞延伸和伸长。成纤维内皮细胞发育出显着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后三天，受伤区域重新出现两种细胞类型：粗糙的成纤维细胞样细胞，形成角膜后纤维膜，具有突出的f-肌动蛋白束或应力纤维，细胞间连接很少；光滑的多边形内皮细胞，具有紧密的细胞连接，皮质分布有f-肌动蛋白。 28天后，角膜后纤维膜被正常内皮覆盖。结论 这些数据支持这样的假设：内皮伤口愈合涉及两种独立的损伤依赖性机制——细胞扩散和细胞迁移。
PURPOSE 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. METHODS 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. RESULTS 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. CONCLUSIONS These data support the hypothesis that endothelial wound healing involves two separate, injury-dependent, mechanisms--cell spreading and cell migration.