Perrot Sébastien, Dutertre-Catella Hélène, Martin Chantal, Rat Patrice, Warnet Jean-Michel
Laboratoire de Toxicologie, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris V, 75270 Paris cedex 06, France.
Toxicol Sci. 2003 Mar;72(1):122-9. doi: 10.1093/toxsci/kfg014.
The main object of our study was to investigate whether the resazurin metabolism assay is a sensitive surfactant and alcohol toxicity test in isolated pig cornea and to compare this recently developed fluorometric assay with the data collected in the eye irritation reference chemical data bank. Resazurin is a substrate that changes color in response to metabolic activity. Isolated pig corneas were immersed for 10 min in surfactants and alcohol irritant solutions. After incubation, resorufin fluorescence was read and corneal viability was assessed. This corneal viability was compared with the maximal modified average score published in the report of ECETOC. This assay highlighted different concentration-dependent irritation potentials of the three surfactants tested, and the same results were obtained with corneas treated with the alcohols. We observed that the degree of surfactant- and alcohol-induced decrease in corneal viability, using the resazurin reduction test, was correlated with the in vivo irritancy measurements as determined by the Draize test and scored with the Modified Maximum Average Score (MMAS). This assay allowed us to classify the ocular irritancy of the tested surfactants and alcohols in the same ranking order as the Draize classification. Corneal viability measurement can be used as a potential alternative for the toxicological assessment of surfactants and alcohols. The nontoxic, nonradioactive resazurin metabolism assay allows rapid assessment of many samples with simple equipment and at reduced cost for continuous monitoring of corneal viability. This assay seems to be suitable as a toxicological screening test for eye irritation determination.
我们研究的主要目的是调查刃天青代谢试验在离体猪角膜中是否是一种灵敏的表面活性剂和酒精毒性测试，并将这种最近开发的荧光测定法与眼刺激参考化学数据库中收集的数据进行比较。刃天青是一种会因代谢活性而改变颜色的底物。将离体猪角膜浸入表面活性剂和酒精刺激溶液中10分钟。孵育后，读取试卤灵荧光并评估角膜活力。将这种角膜活力与欧洲生态毒理学与化学品毒性中心（ECETOC）报告中公布的最大修正平均分进行比较。该试验突出了所测试的三种表面活性剂不同的浓度依赖性刺激潜力，用酒精处理的角膜也得到了相同的结果。我们观察到，使用刃天青还原试验，表面活性剂和酒精诱导的角膜活力下降程度与通过Draize试验测定并以修正最大平均分（MMAS）评分的体内刺激性测量结果相关。该试验使我们能够按照与Draize分类相同的排名顺序对所测试的表面活性剂和酒精的眼刺激性进行分类。角膜活力测量可作为表面活性剂和酒精毒理学评估的一种潜在替代方法。无毒、无放射性的刃天青代谢试验允许使用简单设备快速评估许多样品，并以降低的成本持续监测角膜活力。该试验似乎适合作为一种用于确定眼刺激的毒理学筛选试验。
