Abstract：
The aim of the study was to investigate the effect of ripasudil on corneal endothelial cell survival and migration after two types of descemetorhexis on a human ex vivo model. Eleven human corneoscleral buttons were incubated in either 50 ml organ culture medium containing 10 μM ripasudil or 50 μl dimethyl sulfoxide (DMSO), the vehicle in ripasudil for 2 days prior to wound creation then for 14 days after. The wound was created with either full trephination scoring or by shallow trephination plus manual peeling. At day 14, immunohistochemistry with vimentin and Na+/K+/ATPase markers was conducted. Tissues were assessed at day 3, 7 and 14 for morphology, cell migration, cell viability and cell density. Full trephination scoring created more damage on tissues compared to shallow trephination with full Descemet membrane peeling. In the full trephination scoring group, no differences in cell viability were noted when ripasudil and DMSO were compared. With the peeling method, Ripasudil could protect the endothelial cell death and maintain the morphology compared to the control. At day 14, no differences in the peripheral cell viability and density were found between ripasudil and DMSO, although the ripasudil group presented significantly increased central cell count and cell viability. Increased cell migration was noted with ripasudil and the initial cell morphology of those migrated cells was similar to that of fibroblasts. In conclusion, ex vivo modelling suggested that peeling resulted in less cell damage than scoring and ripasudil maintained better morphology and promoted migration. These effects might be via transformation of endothelial cells into a more motile spindle-like phenotype.
摘要：
该研究的目的是研究在人离体模型上进行两种类型的异型异型异型异型异型治疗后，rapasudil对角膜内皮细胞存活和迁移的影响。将11个人角膜巩膜纽扣在50ml含有10μMripasudil或50μl二甲基亚砜（DMSO）的器官培养基中孵育，在伤口形成前2天，然后在后14天。伤口是通过完全钻孔划痕或通过浅钻孔加上手动剥离而产生的。在第14天,用波形蛋白和Na+/K+/ATP酶标记进行免疫组织化学。在第3天，第7天和第14天评估组织的形态，细胞迁移，细胞活力和细胞密度。与全Descemet膜剥离的浅钻孔相比，全钻孔评分对组织造成了更多的损伤。在全钻孔评分组中，比较rripasudil和DMSO时，没有发现细胞活力的差异。用剥皮的方法，与对照相比，利帕舒地尔可以保护内皮细胞死亡并保持形态。在第14天，在ricasudil和DMSO之间没有发现外周细胞活力和密度的差异，尽管rapasudil组表现出显着增加的中心细胞计数和细胞活力。用rapasudil观察到增加的细胞迀移,并且那些迀移的细胞的初始细胞形态与成纤维细胞的初始细胞形态相似。总之,离体建模表明，剥离比评分导致的细胞损伤更小，而瑞帕舒地尔保持了更好的形态并促进了迁移。这些作用可能是通过将内皮细胞转化为更活跃的纺锤体样表型。
