Corneal endothelial dysfunction is a major cause of corneal blindness and is mainly treated by corneal transplantation. However, the global shortage of donor cornea hampers its application. Intracameral injection of cultured primary corneal endothelial cells (CECs) was recently confirmed in clinical trials. However, abnormal adhesion of the grafted CECs affects the application of this strategy. In this study, we explored if laminin 511 (LN511) improves the therapeutic function of the intracameral CEC injection for corneal endothelial dysfunction. To mimic the late stage of corneal endothelial diseases, intense scraping was developed to remove CECs and extracellular matrix of the posterior Descemet's membrane (DM) without DM removal in rabbits. Then, Dulbecco's phosphate-buffered saline (DPBS) and LN511 were intracamerally injected as the control and intervention groups, respectively. We found that the injected LN511 could settle and form a coating on the posterior surface of DM. After CEC transplantation, corneal clarity of rabbits in the LN511 group was rapidly recovered within 7 days, whereas the corneal recovery took 14 days in the DPBS group. Corneal thickness of LN511 group decreased to 413.3 ± 20.8 μm 7 days after operation, which was significantly lower than 1086.3 ± 78.6 μm of DPBS group (p < 0.01). Moreover, for the grafted CECs, LN511 promoted the rapid adhesion, tight junction formation, and expression of Na⁺/K⁺-ATPase and ZO-1. In vitro analysis revealed that the functions of LN511 on the cultured human CECs mechanistically depended on the cell density and the nuclear-cytoplasmic translocation of the Yes-associated protein. Our study demonstrated that LN511 precoating promoted the adhesion of the transplanted CECs and enhanced the functional regeneration of the corneal endothelium. Thus, our data suggested that the strategy of LN511 precoating and CECs' intracameral injection could be a potential method for the therapy of corneal endothelial dysfunction.

中文翻译：

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层粘连蛋白 511 预涂层促进移植角膜内皮细胞的功能恢复。

角膜内皮功能障碍是导致角膜失明的主要原因，主要通过角膜移植治疗。然而，供体角膜的全球短缺阻碍了其应用。最近在临床试验中证实了前房内注射培养的原代角膜内皮细胞 (CEC)。然而，嫁接CECs的异常粘附影响了该策略的应用。在这项研究中，我们探讨了层粘连蛋白 511 (LN511) 是否能改善前房内 CEC 注射对角膜内皮功能障碍的治疗功能。为了模拟角膜内皮疾病的晚期阶段，开发了强烈刮除以去除兔的 CEC 和后后旋膜 (DM) 的细胞外基质，而无需去除 DM。然后，杜尔贝科 s 磷酸盐缓冲盐水 (DPBS) 和 LN511 分别作为对照组和干预组进行房内注射。我们发现注入的 LN511 可以沉淀并在 DM 的后表面形成涂层。CEC移植后，LN511组家兔角膜透明度在7天内迅速恢复，而DPBS组角膜恢复需要14天。LN511组术后7天角膜厚度降至413.3±20.8μm，明显低于DPBS组的1086.3±78.6μm（而 DPBS 组的角膜恢复需要 14 天。LN511组术后7天角膜厚度降至413.3±20.8μm，明显低于DPBS组的1086.3±78.6μm（而 DPBS 组的角膜恢复需要 14 天。LN511组术后7天角膜厚度降至413.3±20.8μm，明显低于DPBS组的1086.3±78.6μm（p  < 0.01)。此外，对于嫁接的CECs，LN511促进了Na ⁺ /K ⁺ -ATPase和ZO-1的快速粘附、紧密连接的形成和表达。体外分析表明，LN511 对培养的人类 CEC 的功能在机制上取决于细胞密度和 Yes 相关蛋白的核质易位。我们的研究表明，LN511 预涂层促进了移植 CEC 的粘附并增强了角膜内皮的功能再生。因此，我们的数据表明，LN511 预涂层和 CEC 前房内注射的策略可能是治疗角膜内皮功能障碍的潜在方法。