Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma
Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma
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DOI：
10.1016/j.biomaterials.2010.05.020
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发表时间：
2010-09-01
期刊：
影响因子：
14
通讯作者：
Soker, Shay
中科院分区：
文献类型：
--
作者：
Choi, Jin San;Williams, James K.;Soker, Shay
角膜移植是一种常见的移植手术，通过用透明的健康供体组织替换不透明或变形的宿主组织来改善视力。然而，由于缺乏高质量的供体角膜，其临床应用受到限制。利用可扩展的人类供体来源的角膜内皮细胞（HCEC）群体创建的生物工程新角膜可以解决目前的短缺问题。本研究的目的是建立HCEC的分离和培养方案，并探讨通过将细胞接种在脱细胞的人角膜基质上来构建生物工程角膜组织的可行性。通过酶消化从眼供体的废弃角膜中取出HCEC。扩增细胞并评价其Na+/K+-ATP酶和闭合蛋白-1（ZO-1）的表达。使用切片机将供体角膜基质切成120-2001 μ m厚的切片，然后脱细胞。测定脱细胞后支架的细胞外基质成分和力学性能。为了工程化新角膜，将130 HCEC/mm（2）接种在脱细胞的人角膜基质上。将所得构建体置于生长培养基中14天，然后使用扫描电子显微镜（SEM）、组织学和免疫细胞化学进行分析。接种的细胞保留了功能标志物Na+/K+-ATP酶和ZO-1的表达，并且构建体具有与正常角膜相似的生物力学性质。这些结果表明，使用来自废弃供体角膜和脱细胞的薄层角膜基质的HCEC构建新角膜，可以产生用于移植的高质量角膜组织的新来源。(C)2010爱思唯尔有限公司保留所有权利。
Corneal transplantation is a common transplant procedure performed to improve visual acuity by replacing the opaque or distorted host tissue by clear healthy donor tissue. However, its clinical utility is limited due to a lack of high quality donor corneas. Bioengineered neo-corneas, created using an expandable population of human donor-derived corneal endothelial cells (HCEC), could address this current shortage. The objectives of this study were to establish HCEC isolation and culture protocols and to investigate the feasibility of bioengineering corneal tissue constructs by seeding the cells on decellularized human corneal stroma. HCECs were removed from the discarded corneas of eye donors by enzymatic digestion. Cells were expanded and evaluated for their expression of Na+/K+-ATPase and zona occludens-1 (ZO-1). Donor corneal stromas were cut to 120-2001 mu m thickness slices using a microtome and then decellularized. Extracellular matrix components and mechanical properties of the scaffolds were measured after decellularization. To engineer neo-corneas, 130 HCEC/mm(2) were seeded on decellularized human corneal stromas. The resulting constructs were placed in growth medium for 14 days and then analyzed using scanning electron microscopy (SEM), histology, and immunocytochemistry. Seeded cells retain expression of the functional markers Na+/K+-ATPase and ZO-1 and constructs have biomechanical properties similar to those of normal corneas. These results indicate that construction of neo-corneas, using HCECs derived from discarded donor corneas and decellularized thin-layer corneal stromas, may create a new source of high quality corneal tissue for transplantation. (C) 2010 Elsevier Ltd. All rights reserved.