Takahashi Yutaka, Koike Mirei, Honda Hiroshi, Ito Yuichi, Sakaguchi Hitoshi, Suzuki Hiroyuki, Nishiyama Naohiro
Safety Science Research Laboratories, Kao Corporation, 2606 Akabane, Ichikai-Maich, Haga-Gun Tochigi 321-3497, Japan.
Toxicol In Vitro. 2008 Apr;22(3):760-70. doi: 10.1016/j.tiv.2007.11.018. Epub 2007 Dec 7.
Using SIRC (rabbit corneal cell line) cells, we developed an alternative eye irritation test: the short time exposure (STE) test. This STE test is a cytotoxicity test using physiological saline or mineral oil as the test solvent. Evaluation exposure time is short (5 min), which is similar to actual exposure situations, and uses the cell viability (CV) at a constant concentration as the endpoint for irritation potential. First, in order to confirm the usefulness of this STE test in assessing eye irritation potential of chemicals, 51 raw materials were tested and the correlation between CV in the STE test and the eye irritation score in the Draize test was examined. For the undiluted raw materials tested in the Draize test, the 5% test concentration in the STE test gave irritation classes that correlated well with the irritation classes from the Draize test (accuracy: 89.6%). For those materials tested as a 10% solution in the Draize test, STE irritation classes with 0.05% test concentration corresponded well with the Draize irritation classes (accuracy: 80.0%). Next, using the cell viabilities at these two concentrations, the STE prediction model (PM) was developed. A score of 1 or 2 was given for the results from each tested concentration in the STE test and Draize test. The scores from each test were then summed to yield a 3-level (Rank 1: minimally irritant, Rank 2: moderate irritant, Rank 3: severe irritant) eye irritation potential classification. Rank classification in the STE test showed a good correlation mostly to that in the Draize test (irritation class correspondence rate: 70.2%, but after exclusion of data of alcoholic materials, the rate was 91.7%). In most cytotoxicity test, the cytotoxicity of acids and amines is generally underestimated due the use of medium as the solvent. This is the result of the buffering capacity of the media. On the other hand, the STE test could predict the eye irritation potential by evaluating the chemical with a 5% test concentration. Eleven water insoluble materials such as toluene, octanol, and hexanol could be evaluated by using mineral oil as test solvent in the STE test. The STE test demonstrated itself to be simple, promising, have great potential, be of value, and to be an easily standardized alternative eye irritation test.
我们使用SIRC（兔角膜细胞系）细胞开发了一种替代性眼刺激试验：短时间暴露（STE）试验。该STE试验是一种以生理盐水或矿物油作为试验溶剂的细胞毒性试验。评估暴露时间较短（5分钟），这与实际暴露情况相似，并将恒定浓度下的细胞活力（CV）作为刺激潜力的终点指标。首先，为了确认该STE试验在评估化学品眼刺激潜力方面的有效性，对51种原材料进行了测试，并考察了STE试验中的CV与Draize试验中的眼刺激评分之间的相关性。对于在Draize试验中测试的未稀释原材料，STE试验中5%的测试浓度给出的刺激等级与Draize试验中的刺激等级相关性良好（准确率：89.6%）。对于那些在Draize试验中以10%溶液形式测试的材料，STE试验中0.05%测试浓度下的刺激等级与Draize试验中的刺激等级对应良好（准确率：80.0%）。接下来，利用这两种浓度下的细胞活力，开发了STE预测模型（PM）。在STE试验和Draize试验中，对每个测试浓度的结果给出1或2分。然后将每个试验的分数相加，得出一个三级（等级1：轻度刺激，等级2：中度刺激，等级3：重度刺激）的眼刺激潜力分类。STE试验中的等级分类大多与Draize试验中的等级分类具有良好的相关性（刺激等级对应率：70.2%，但排除含醇材料的数据后，该比率为91.7%）。在大多数细胞毒性试验中，由于使用培养基作为溶剂，酸和胺的细胞毒性通常被低估。这是培养基缓冲能力的结果。另一方面，STE试验可以通过评估5%测试浓度的化学品来预测眼刺激潜力。11种水不溶性材料，如甲苯、辛醇和己醇，可以在STE试验中使用矿物油作为试验溶剂进行评估。STE试验证明自身简单、有前景、潜力巨大、有价值，是一种易于标准化的替代性眼刺激试验。
