Department of Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, London, UK.
PLoS One. 2012;7(11):e50993. doi: 10.1371/journal.pone.0050993. Epub 2012 Nov 30.
Current treatments for reversible blindness caused by corneal endothelial cell failure involve replacing the failed endothelium with donor tissue using a one donor-one recipient strategy. Due to the increasing pressure of a worldwide donor cornea shortage there has been considerable interest in developing alternative strategies to treat endothelial disorders using expanded cell replacement therapy. Protocols have been developed which allow successful expansion of endothelial cells in vitro but this approach requires a supporting material that would allow easy transfer of cells to the recipient. We describe the first use of plastic compressed collagen as a highly effective, novel carrier for human corneal endothelial cells. A human corneal endothelial cell line and primary human corneal endothelial cells retained their characteristic cobblestone morphology and expression of tight junction protein ZO-1 and pump protein Na+/K+ ATPase α1 after culture on collagen constructs for up to 14 days. Additionally, ultrastructural analysis suggested a well-integrated endothelial layer with tightly opposed cells and apical microvilli. Plastic compressed collagen is a superior biomaterial in terms of its speed and ease of production and its ability to be manipulated in a clinically relevant manner without breakage. This method provides expanded endothelial cells with a substrate that could be suitable for transplantation allowing one donor cornea to potentially treat multiple patients.
目前,针对由角膜内皮细胞衰竭引起的可逆转失明的治疗方法涉及使用供体组织替换失败的内皮细胞,采用一个供体对应一个受者的策略。由于全球供体角膜短缺的压力不断增加,人们对使用扩展细胞替代疗法治疗内皮紊乱的替代策略产生了浓厚的兴趣。已经制定了允许在体外成功扩展内皮细胞的方案,但这种方法需要一种支持材料,以便将细胞轻松转移到受者身上。我们描述了首次将塑料压缩胶原用作人角膜内皮细胞的高效新型载体。人角膜内皮细胞系和原代人角膜内皮细胞在胶原构建体上培养长达 14 天时,保持了其特征性的鹅卵石形态以及紧密连接蛋白 ZO-1 和泵蛋白 Na+/K+ATPaseα1 的表达。此外,超微结构分析表明,内皮层具有紧密相对的细胞和顶微绒毛,整合良好。在生产速度、生产的简便性以及以临床相关方式进行操作而不会断裂的能力方面,塑料压缩胶原是一种优越的生物材料。这种方法为扩展的内皮细胞提供了一种合适的移植基质,使得一个供体角膜有可能治疗多个患者。