College of Pharmacy, Ewha Womans University, Seoul, Republic of Korea.
Amorepacific Corp. R&D Center, Republic of Korea.
Toxicol In Vitro. 2019 Mar;55:173-184. doi: 10.1016/j.tiv.2018.12.003. Epub 2018 Dec 17.
The need for in vitro eye irritation test replacing in vivo is steadily increasing. The MCTT HCE™ eye irritation test (EIT) using 3D reconstructed human cornea-like epithelium, was developed to identify ocular irritants from non-irritants those that are not requiring classification and labelling for eye irritation. Here, we report the results of me-too validation study, which was conducted to evaluate the reliability and relevance of the MCTT HCE EIT, according to performance standards (PS) of OECD TG 492. The optimal cutoff to determine irritation in the prediction model was preliminarily established at 45% with the receiver operation characteristics (ROC) curve for 141 reference substances. To demonstrate the reproducibility of within- and between-laboratory (WLR and BLR), a set of 30 PS reference chemicals were tested in three laboratories three times. The WLR and BLR concordance with the binary decision of whether non-irritant or irritant was estimated to be 90-100% and 90%, respectively, and both met the PS requirements. The predictive capacity of the respective laboratories for the 30 reference chemicals were evaluated based on three different estimation methods, and the results were comparable, with sensitivity ranging from 89.6 to 93.3%, the specificity ranging from 62.2 to 66.7%, and the accuracy ranging from 75.9 to 80.0%. Additional test with the new set of 30 PS substances in the revised OECD GD 216 yielded a performance of sensitivity ranging from 92.6-93.3%, the specificity 62.2-66.7% and the accuracy 77.4-80.0%. 95.0% sensitivity, 67.2% specificity, and 83.0% accuracy were obtained for 141 reference substances in total. Furthermore, separate cutoffs for liquids and solids, 35% and 60%, respectively, produced better predictivity, which was established as a final prediction model. Collectively, our study demonstrated that MCTT HCE EIT meets the reproducibility and predictivity criteria stated in OECD TG 492 PS.
体外眼刺激性试验替代体内试验的需求稳步增加。MCTT HCE™眼刺激性试验（EIT）使用三维重建的人角膜样上皮，旨在识别非刺激性和不需要进行眼刺激性分类和标签的眼部刺激性物质。在这里，我们报告了一项 me-too 验证研究的结果，该研究是根据 OECD TG 492 的性能标准（PS）进行的，旨在评估 MCTT HCE EIT 的可靠性和相关性。根据 141 种参考物质的接收者操作特征（ROC）曲线，初步确定了用于确定刺激的最佳截止值为 45%。为了证明实验室内部和实验室之间（WLR 和 BLR）的重现性，在三个实验室中三次测试了一组 30 种 PS 参考化学品。WLR 和 BLR 对非刺激性或刺激性的二元决策的一致性估计为 90-100%和 90%，均符合 PS 要求。根据三种不同的估计方法，对三个实验室对 30 种参考化学品的预测能力进行了评估，结果具有可比性，敏感性范围为 89.6-93.3%，特异性范围为 62.2-66.7%，准确性范围为 75.9-80.0%。在修订后的 OECD GD 216 中使用新的 30 种 PS 物质进行额外测试，得出的性能为敏感性范围为 92.6-93.3%，特异性为 62.2-66.7%，准确性为 77.4-80.0%。141 种参考物质的总敏感性为 95.0%，特异性为 67.2%，准确性为 83.0%。此外，液体和固体的单独截止值分别为 35%和 60%，产生了更好的预测性，这被确定为最终的预测模型。总的来说，我们的研究表明，MCTT HCE EIT 符合 OECD TG 492 PS 中规定的重现性和预测性标准。
