A new 3D reconstituted human corneal epithelium model as an alternative method for the eye irritation test
A new 3D reconstituted human corneal epithelium model as an alternative method for the eye irritation test
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DOI：
10.1016/j.tiv.2010.10.019
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发表时间：
2011-02-01
影响因子：
3.2
通讯作者：
Lim, Kyung-Min
中科院分区：
文献类型：
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作者：
Jung, Kyoung-Mi;Lee, Su-Hyon;Lim, Kyung-Min
人们正在做出许多努力来开发用于眼睛刺激测试的新的替代体外测试方法。在这里，我们报告了一种由原代培养的人角膜缘上皮细胞制备的新的重建人角膜上皮模型（MCTT HCE模型），作为一种新的替代体外眼刺激测试方法。在组织学和免疫组织化学观察中，MCTT HCE模型表现出与完整人角膜相似的形态和生物标志物表达。此外，根据高跨上皮电阻、Triton X-100 的有效时间 50 和角膜厚度测量，屏障功能得到了很好的保存。为了将该模型用作眼睛刺激测试的新替代方法，进行了方案细化并确定了最佳测定条件，包括处理时间、处理量、后孵育时间和冲洗方法。使用改进的方案，对 25 种具有已知眼睛刺激潜力的参考化学品进行了测试。活力截止值为 50%，化学品被分为刺激性或无刺激性。与GHS分类相比，MCTT HCE模型的准确度为88%，敏感性为100%，特异性为77%。这些结果表明 MCTT HCE 模型可能可作为一种新的替代眼刺激测试方法。 (C) 2010 Elsevier Ltd. 保留所有权利。
Many efforts are being made to develop new alternative in vitro test methods for the eye irritation test. Here we report a new reconstructed human corneal epithelial model (MCTT HCE model) prepared from primary-cultured human limbal epithelial cells as a new alternative in vitro eye irritation test method. In histological and immunohistochemical observation, MCTT HCE model displayed a morphology and bio-marker expressions similar to intact human cornea. Moreover, the barrier function was well preserved as measured by high transepithelial electrical resistance, effective time-50 for Triton X-100, and corneal thickness. To employ the model as a new alternative method for eye irritation test, protocol refinement was performed and optimum assay condition was determined including treatment time, treatment volume, post-incubation time and rinsing method. Using the refined protocol, 25 reference chemicals with known eye irritation potentials were tested. With the viability cut-off value at 50%, chemicals were classified to irritant or non-irritant. When compared with GHS classification, the MCTT HCE model showed the accuracy of 88%, sensitivity of 100% and specificity of 77%. These results suggest that the MCTT HCE model might be useful as a new alternative eye irritation test method. (C) 2010 Elsevier Ltd. All rights reserved.