BASF SE, Experimental Toxicology and Ecology, Ludwigshafen, Germany.
Altern Lab Anim. 2011 Sep;39(4):365-87. doi: 10.1177/026119291103900410.
In 2009, the Bovine Corneal Opacity and Permeability (BCOP) test was accepted by the regulatory bodies for the identification of corrosive and severe ocular irritants (Global Harmonised System [GHS] Category 1). However, no in vitro test is currently accepted for the differentiation of ocular irritants (GHS Category 2) and non-irritants (GHS No Category). Human reconstructed tissue models have been suggested for incorporation into a tiered testing strategy to ultimately replace the Draize rabbit eye irritation test (OECD TG 405). The purpose of this study was to evaluate whether the EpiOcular(TM) reconstructed cornea-like tissue model and the COLIPA pre-validated EpiOcular Eye Irritation Test (EpiOcular-EIT) could be used as suitable components of this testing strategy. The in-house validation of the EpiOcular-EIT was performed by using 60 test substances, including a broad variety of chemicals and formulations for which in vivo data (from the Draize rabbit eye irritation test) were available. The test substances fell into the following categories: 18 severe irritants/corrosives (Category 1), 21 irritants (Category 2), and 21 non-irritants (No Category). Test substances that decreased tissue viability to ≤ 60% (compared to the negative control tissue) were considered to be eye irritants (Category 1/2). Test substances resulting in tissue viability of > 60% were considered to be non-irritants (No Category). For the assessed dataset and the classification cut-off of 60% viability, the EpiOcular-EIT provided 98% and 84% sensitivity, 64% and 90% specificity, and 85% and 86% overall accuracy for the literature reference and BASF proprietary substances, respectively. Applying a 50% tissue viability cut-off to distinguish between irritants and non-irritants resulted in 93% and 82% sensitivity, 68% and 100% specificity, and 84% and 88% accuracy for the literature reference and BASF proprietary substances, respectively. Further, in the EpiOcular-EIT (60% cut-off), 100% of severely irritating substances under-predicted by the BCOP assay were classified as Category 1/2. The results obtained in this study, based on 60 test substances, indicate that the EpiOcular-EIT and the BCOP assay can be combined in a testing strategy to identify strong/severe eye irritants (Category 1), moderate and mild eye irritants (Category 2), and non-irritants (No Category) in routine testing. In particular, when the bottom-up strategy with the 60% viability cut-off was employed, none of the severely irritating substances (Category 1) were under-predicted to be non-irritant. Sensitivity for Category 1/2 substances was 100% for literature reference substances and 89% for BASF SE proprietary substances.
2009 年，牛角膜混浊和通透性（BCOP）试验被监管机构接受，用于鉴定腐蚀性和严重眼部刺激性物质（全球协调系统[GHS]类别 1）。然而，目前还没有用于区分眼部刺激性物质（GHS 类别 2）和非刺激性物质（GHS 无类别）的体外试验。已经提出了人类重建组织模型，以纳入分层测试策略中，最终取代兔眼刺激性试验（OECD TG 405）。本研究的目的是评估 EpiOcular（TM）重建角膜样组织模型和 COLIPA 预先验证的 EpiOcular 眼部刺激性试验（EpiOcular-EIT）是否可作为该测试策略的合适组成部分。通过使用 60 种测试物质对 EpiOcular-EIT 进行了内部验证，其中包括广泛的化学物质和制剂，这些物质具有体内数据（来自兔眼刺激性试验）。测试物质分为以下几类：18 种严重刺激性物质/腐蚀性物质（类别 1）、21 种刺激性物质（类别 2）和 21 种非刺激性物质（无类别）。组织活力降低至≤60%（与阴性对照组织相比）的测试物质被认为是眼部刺激性物质（类别 1/2）。组织活力>60%的测试物质被认为是非刺激性物质（无类别）。对于评估数据集和 60%活力的分类截止值，EpiOcular-EIT 对文献参考物质和巴斯夫专有物质分别提供了 98%和 84%的灵敏度、64%和 90%的特异性和 85%和 86%的总体准确性。应用 50%组织活力截止值来区分刺激性物质和非刺激性物质，对于文献参考物质和巴斯夫专有物质，灵敏度分别为 93%和 82%、特异性分别为 68%和 100%、准确性分别为 84%和 88%。此外，在 EpiOcular-EIT（60%截止值）中，BCOP 测定法预测为严重刺激性的物质全部被分类为类别 1/2。本研究基于 60 种测试物质得出的结果表明，EpiOcular-EIT 和 BCOP 测定法可结合在测试策略中，以在常规测试中识别强/严重眼部刺激性物质（类别 1）、中度和轻度眼部刺激性物质（类别 2）和非刺激性物质（无类别）。特别是，当采用 60%活力截止值的自下而上策略时，没有一种严重刺激性物质（类别 1）被预测为非刺激性物质。文献参考物质的类别 1/2 物质的灵敏度为 100%，巴斯夫 SE 专有物质的灵敏度为 89%。
