Laminin 511 precoating promotes the functional recovery of transplanted corneal endothelial cells.
Laminin 511 precoating promotes the functional recovery of transplanted corneal endothelial cells.
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DOI：
10.1089/ten.tea.2020.0047
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发表时间：
2020-06
影响因子：
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通讯作者：
C. Zhao;Qingjun Zhou;H. Duan;Xin Wang;Y. Jia;Yajie Gong;Wenjing Li;Chunxiao Dong;Zongyi Li;W. Shi
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文献类型：
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作者：
C. Zhao;Qingjun Zhou;H. Duan;Xin Wang;Y. Jia;Yajie Gong;Wenjing Li;Chunxiao Dong;Zongyi Li;W. Shi
角膜内皮功能障碍是导致角膜失明的主要原因，目前主要采用角膜移植治疗。然而，全球供体角膜的短缺阻碍了它的应用。培养的原代角膜内皮细胞(CECs)的房内注射最近在临床试验中得到证实。然而，移植的CECs的异常黏附影响了这一策略的应用。在这项研究中，我们探索层粘连蛋白511(LN511)是否能改善房内注射CECs对角膜内皮功能障碍的治疗作用。为模拟晚期角膜内皮疾病，在不去除兔后背膜(DM)的情况下，采用高强度刮削法去除后Descemet膜(DM)中的CEC和细胞外基质。Dulbecco‘s磷酸盐缓冲盐水(DPBS)和LN511分别作为对照组和干预组。我们发现注射的LN511可以在DM的后表面沉降并形成一层涂层。CECs移植后，LN511组兔角膜透明度在7d内迅速恢复，而DPBS组则需14d才能恢复。术后第7天LN511组角膜厚度降至413.3±20.8μm，明显低于DPBS组的1086.3±78.6μm(P<0.01)。此外，LN511还促进了移植的CECs的快速黏附、紧密连接的形成以及Na+/K+-ATPase和ZO-1的表达。体外分析表明，LN511对培养的人CECs的作用机制依赖于细胞密度和Yes相关蛋白的核质转位。我们的研究表明，LN511预涂膜可促进移植的CECs的黏附，促进角膜内皮细胞的功能性再生。因此，我们的数据提示LN511预涂膜和CECs腔内注射的策略可能是治疗角膜内皮功能障碍的一种潜在方法。
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 CECs 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 CECs 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±20.8μm 7 days after operation, which was significantly lower than 1086.3±78.6μm 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.