Itagaki H, Hagino S, Kato S, Kobayashi T, Umeda M
Shiseido Safety and Analytical Research Center, 1050 Nippa-cho, Kohoku-ku, Yokohama 223, Japan.
Toxicol In Vitro. 1991;5(2):139-43. doi: 10.1016/0887-2333(91)90034-b.
To evaluate a simple in vitro method as an alternative to the Draize eye-irritation test, 12 surfactants were tested by both of the above methods, and the correlation between the results was examined. The in vitro method used was the crystal violet staining method. HeLa cells or SIRC cells from rabbit cornea were inoculated into each well of 96-well microplates which contained serially diluted test chemicals. They were cultured for 72 hr, and then fixed and stained with crystal violet. After spectrophotometric measurement, the concentrations of test materials that inhibited the absorbance to 50% of the control level (IC(50)) were determined. The in vivo data used for comparison were the maximum corneal score and the maximum total score obtained by the eye test when the substances were tested at a concentration of 10%. The correlation coefficients between the maximum corneal scores of the 12 surfactants and their IC(50)s in HeLa cells and in SIRC cells were -0.821 and -0.816, respectively, while those between the maximum total scores and IC(50)s in both cell lines were -0.855 and -0.863, respectively. All of these values are statistically highly significant, indicating that the crystal violet staining method may have value in predicting the eye irritancy of surfactants. Further interlaboratory validation will, however, be needed.
为评估一种简单的体外方法作为替代Draize眼刺激试验的方法，采用上述两种方法对12种表面活性剂进行了测试，并检验了结果之间的相关性。所使用的体外方法是结晶紫染色法。将来自兔角膜的HeLa细胞或SIRC细胞接种到含有系列稀释测试化学品的96孔微孔板的每个孔中。将它们培养72小时，然后固定并用结晶紫染色。经过分光光度测量后，确定将吸光度抑制至对照水平50%的测试材料浓度（IC50）。用于比较的体内数据是当以10%的浓度测试这些物质时，通过眼试验获得的最大角膜评分和最大总分。12种表面活性剂在HeLa细胞和SIRC细胞中的最大角膜评分与其IC50之间的相关系数分别为-0.821和-0.816，而两种细胞系中最大总分与IC50之间的相关系数分别为-0.855和-0.863。所有这些值在统计学上都具有高度显著性，表明结晶紫染色法在预测表面活性剂的眼刺激性方面可能具有价值。然而，还需要进一步的实验室间验证。
