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ORIGINAL ARTICLE
Year : 2018  |  Volume : 131  |  Issue : 14  |  Page : 1710-1714

Constructing a Novel Three-Dimensional Biomimetic Corneal Endothelium Graft by Culturing Corneal Endothelium Cells on Compressed Collagen Gels


Department of Ophthalmology, Peking University Third Hospital; Beijing Key Laboratory of Restoration of Damaged Ocular Nerve, Peking University Third Hospital, Beijing 100191, China

Correspondence Address:
Dr. Yun Feng
Department of Ophthalmology, Peking University Third Hospital, 49 North Garden Road, Haidian District, 100191 Beijing
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0366-6999.235883

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Background: Endothelium allotransplantation is the primary treatment for corneal decompensation. The worldwide shortage of donor corneal tissue has led to increasing pressure to seek an alternative for surgical restoration of corneal endothelium. Compressed collagen (CC) gels have excellent biocompatibility, simple preparation course and easy to be manipulated. This study aimed to form a new biomimetic endothelium graft by CC. Methods: We expanded bovine corneal endothelial cells (B-CECs) on laminin-coated CC to form a biomimetic endothelium graft. Scanning electron microscope was used for ultrastructural analysis and tight junction protein ZO-1 expression was tested by immunohistochemistry. Results: The biomimetic endothelium graft, we conducted had normal cell morphology, ultrastructure and higher cell density (3612.2 ± 43.4 cells/mm2). ZO-1 localization at B-CECs membrane indicated the bioengineered graft possess the basic endothelium function. Conclusions: A biomimetic endothelium graft with B-CECs expanded on CC sheet was constructed, which possessed cells' morphology similar to that of in vivo endothelial cells and specific basic function of endothelium layer. This method provided the possibility of using one donor's cornea to form multiple uniformed endothelium grafts so as to overcome the shortage of cadaveric cornea tissue.

 

 Abstract in Chinese

以压缩胶原为基底构建新型三维仿生角膜内皮植片

摘要

背景:异体角膜内皮移植是治疗角膜内皮失代偿的首选方法。在世界范围内,人角膜供体组织供不应求,急需内皮植片替代材料。压缩胶原具生物相容性佳、易于制备、易于加工的特点,是生物工程研究的重要材料。本研究尝试以压缩胶原为内皮细胞支架构建一种新型角膜内皮植片。

方法:分离牛角膜内皮细胞,经体外扩增后接种于Laminin包被后的压缩胶原薄膜上。观察显微镜下植片内皮细胞结构,并应用扫描电子显微镜观察内皮细胞超微结构。免疫组化染色验证内皮细胞紧密连接蛋白ZO-1的表达情况。

结果:在压缩胶原薄片上制备出单层且具有正常内皮细胞六角形态、超微结构和高细胞密度(3612.2 ± 43.4 cells/mm2)的角膜内皮植片,ZO-1正常表达于植片内皮细胞的细胞膜上。

结论:成功构建以压缩胶原为支架的角膜内皮细胞植片,植片内皮细胞具有正常形态和基础细胞功能。这一方法提供了利用一个供体角膜经细胞体外扩增制备多个角膜内皮移植物的可能性,用以克服人角膜组织供体的不足。



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