3T3中性红摄取试验评价皮肤刺激性的初步研究

Preliminary study on neutral red uptake assay as an alternative method for skin irritation test

目的:建立BALB/c-3T3中性红摄取细胞毒性试验并初步探讨其在化妆品原料皮肤刺激毒性评价中的适用性。方法:运用22种已知皮肤刺激性的化妆品原料进行BALB/c-3T3中性红摄取细胞毒性试验,根据判断标准划分皮肤刺激性等级,并与体内试验结果进行比较,分析3T3-中性红摄取细胞毒性实验的预测能力和重现性。结果:3T3-NRU细胞毒性试验与动物实验结果比较呈现较好的等级相关性及分级一致性;三次重复试验分析结果显示体外试验结果间的差别无统计学意义(χ2=0.245,P=0.984),Spearman等级相关系数rs=0.914,P<0.001;说明3T3-NRU细胞毒性试验具有良好的可重复性和稳定性。结论:与Draize试验比较,3T3-NRU试验对参照化学物质具有较好的预测能力和重复性,作为皮肤刺激性化妆品原料评价的筛选方法可进行进一步研究。

Objective：To establish the 3T3-neutral red uptake（NRU） assay and study the applicability of 3T3-NRU assay as an alternative method for skin irritation evaluation of chemicals.Methods： 22 preferential chemicals were selected,of which skin irritation classification were already known.NRU50 values were then determined as criteria for skin irritation classification.Then the test result was compared with that of in vivo test to analyze the predictive ability and reproducibility of 3T3 NRU cytotoxicity test.Results： Rank correlation coefficient（Gamma） and class concordance coefficient（Kappa） between 3T3-NRU assay and animal test were high.The results of triplicate testing of all materials showed that reproducibility and stability of 3T3-NRU assay were good with high Gamma value and Kappa value.Conclusion： It is suggested that 3T3-NRU cytotoxicity assay has good predictive capacity and reproducibility to chemicals when compared with animal test,and it can used as a screening test in safety evaluation of skin.