Tissue-engineered human corneal endothelial cell sheet transplantation in a rabbit model using functional biomaterials
Tissue-engineered human corneal endothelial cell sheet transplantation in a rabbit model using functional biomaterials
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DOI：
10.1097/01.tp.0000287336.09848.39
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发表时间：
2007-11-27
期刊：
影响因子：
6.2
通讯作者：
Hsiue, Ging-Ho
中科院分区：
文献类型：
--
作者：
Lai, Jul-Yang;Chen, Ko-Hua;Hsiue, Ging-Ho
背景资料。本研究旨在探讨生物工程人角膜内皮细胞(HCEC)片状移植物在兔角膜内皮剥离后能否恢复角膜功能和透明性。用PKH26荧光染料标记成人眼库角膜上皮细胞，在37℃温敏性聚N-异丙基丙烯酰胺(PNIPAAm)接种面上培养3周，20℃孵育45min后收获可移植细胞片，用明胶水凝胶片贴壁，将生物工程细胞单层移植到去除内皮的兔角膜上(HCEC片组)。以创伤兔角膜为对照。术后对角膜进行6个月的临床观察和组织学检查。通过对闭锁带-1蛋白和Na+，K+-三磷酸腺苷酶蛋白的透射电子显微镜和Western印迹分析，表明HCEC膜的结构和功能与天然角膜内皮相似。去除内皮细胞后，各组角膜均出现严重的水肿性混浊。HCEC片剂组角膜透明度逐渐恢复，角膜厚度明显小于对照组(P
Background. This study was performed to investigate whether transplantation of bioengineered human corneal endothelial cell (HCEC) sheet grafts into corneas denuded of endothelium could restore corneal function and clarity in a rabbit model.Methods. After being labeled with PKH26 fluorescent dye, the adult HCECs derived from eye bank corneas were cultivated on the thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) -grafted surfaces for 3 weeks at 37 degrees C, and were harvested as transplantable cell sheets after incubation for 45 min at 20 degrees C. Attached by gelatin hydrogel discs, the bioengineered cell monolayers were transplanted to rabbit corneas denuded of endothelium (HCEC sheet group). Traumatized rabbit corneas were served as controls. Postsurgical corneas under-went clinical observations and histological examinations for 6 months.Results. By transmission electron microscopy and Western blot analysis of zonula occludens- 1 and Na+,K+-adenosine triphosphatase proteins, the structure and function of HCEC sheets resembled those of native corneal endothelium. After endothelial cells were removed, corneas of each group turned severe edematous and opaque. In the HCEC sheet groups, corneal clarity was gradually restored and corneal thickness was significantly less than that in the control groups (P