Department Tissue Engineering & Regenerative Medicine (TERM), University Hospital Würzburg, Würzburg, 97070, Germany.
Translational Center for Regenerative Therapies, Fraunhofer Institute for Silicate Research, Würzburg, 97070, Germany.
Sci Rep. 2018 Oct 9;8(1):15049. doi: 10.1038/s41598-018-33118-2.
Highly invasive animal based test procedures for risk assessment such as the Draize eye test are under increasing criticism due to poor transferability for the human organism and animal-welfare concerns. However, besides all efforts, the Draize eye test is still not completely replaced by alternative animal-free methods. To develop an in vitro test to identify all categories of eye irritation, we combined organotypic cornea models based on primary human cells with an electrical readout system that measures the impedance of the test models. First, we showed that employing a primary human cornea epithelial cell based model is advantageous in native marker expression to the primary human epidermal keratinocytes derived models. Secondly, by employing a non-destructive measuring system based on impedance spectroscopy, we could increase the sensitivity of the test system. Thereby, all globally harmonized systems categories of eye irritation could be identified by repeated measurements over a period of 7 days. Based on a novel prediction model we achieved an accuracy of 78% with a reproducibility of 88.9% to determine all three categories of eye irritation in one single test. This could pave the way according to the 3R principle to replace the Draize eye test.
由于对人体的迁移性差和动物福利问题，高度侵入性的基于动物的风险评估测试程序，如 Draize 眼测试，正受到越来越多的批评。然而，尽管付出了所有努力，Draize 眼测试仍然没有完全被无动物替代方法所取代。为了开发一种能够识别所有类别眼部刺激的体外测试方法，我们将基于原代人细胞的器官型角膜模型与一种测量测试模型阻抗的电读取系统相结合。首先，我们表明，采用基于原代人角膜上皮细胞的模型在天然标志物表达方面优于源自原代人表皮角质形成细胞的模型。其次，通过采用基于阻抗谱的非破坏性测量系统，我们可以提高测试系统的灵敏度。因此，可以通过在 7 天的时间内重复测量来识别所有全球协调系统类别（GHS）的眼部刺激。基于一个新的预测模型，我们在单次测试中实现了 78%的准确性和 88.9%的重现性，以确定所有三种眼部刺激类别。这可能符合 3R 原则，以取代 Draize 眼测试。
