Influence of substrate on corneal epithelial cell viability within ocular surface models
Influence of substrate on corneal epithelial cell viability within ocular surface models
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DOI：
10.1016/j.exer.2012.05.005
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发表时间：
2012-08-01
影响因子：
3.4
通讯作者：
Connon, Che John
中科院分区：
文献类型：
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作者：
Feng, Yun;Foster, James;Connon, Che John
近年来，角膜组织工程取得了显着进步。现在可以将这些技术进步应用于开发优质的体外眼表模型，以减少动物测试。我们的目的是展示不同基质对扩张的角膜上皮细胞活力的影响，以及那些更准确地模拟基质表面的基质提供针对毒性攻击的最大保护。将压缩胶原凝胶作为人上皮细胞系扩增的基质与两种众所周知的眼表建模基质（聚碳酸酯膜和传统胶原凝胶）进行比较。细胞扩增超过 10 天，此时通过电子显微镜、免疫组织化学和 RT-PCR 评估细胞分层、细胞数量和连接蛋白的表达。通过MTT测定定量增加十二烷基硫酸钠浓度对上皮细胞活力的影响。结果显示，与使用传统胶原凝胶相比，在聚碳酸酯膜或压缩胶原凝胶上扩增的人上皮细胞在分层、细胞数量和紧密连接表达方面有所改善。然而，在压缩胶原凝胶上扩增的细胞的细胞活力显着更高。我们的结论是，压缩胶原凝胶的更自然的成分和机械性能产生了更稳健的角膜模型。 (C) 2012 Elsevier Ltd. 保留所有权利。
Corneal tissue engineering has improved dramatically over recent years. It is now possible to apply these technological advancements to the development of superior in vitro ocular surface models to reduce animal testing. We aim to show the effect different substrates can have on the viability of expanded corneal epithelial cells and that those which more accurately mimic the stromal surface provide the most protection against toxic assault. Compressed collagen gel as a substrate for the expansion of a human epithelial cell line was compared against two well-known substrates for modelling the ocular surface (polycarbonate membrane and conventional collagen gel). Cells were expanded over 10 days at which point cell stratification, cell number and expression of junctional proteins were assessed by electron microscopy, immunohistochemistry and RT-PCR. The effect of increasing concentrations of sodium lauryl sulphate on epithelial cell viability was quantified by MTT assay. Results showed improvement in terms of stratification, cell number and tight junction expression in human epithelial cells expanded upon either the polycarbonate membrane or compressed collagen gel when compared to a the use of a conventional collagen gel. However, cell viability was significantly higher in cells expanded upon the compressed collagen gel. We conclude that the more naturalistic composition and mechanical properties of compressed collagen gels produces a more robust corneal model. (C) 2012 Elsevier Ltd. All rights reserved.