重组人皮肤模型在化妆品皮肤刺激性评价中的应用研究

Application of the reconstructed human epidermis model in skin irritation evaluation of cosmetics

目的探讨重组人皮肤模型应用于化妆品体外皮肤刺激性评价的可行性。方法以重组人皮肤模型(EpiSkinTM)为受试模型,在对10个已知刺激性分类的标准化学品进行方法验证的基础上,再根据化妆品的使用特性对23个产品进行皮肤刺激性分类评价,其中6个基础型化妆品和10个美容型化妆品暴露18 h、7个清洁型化妆品分别暴露18、4和2 h,检测终点为MTT试验测定细胞活性,并以细胞相对存活率50%为判定依据来分类刺激性和非刺激性物质。分类评价结果同步与本实验室传统Draize兔皮肤刺激性试验历史数据进行一致性比较。结果23个测试产品在暴露时间为18 h的预测性达到73.9%,其中16个基础型和美容型化妆品的预测性可达到100%,而7个清洁型化妆品的预测性仅为14.3%。针对清洁型化妆品缩短上样时间(参照Draize兔皮肤刺激性试验方法),结果该类产品暴露4 h的预测性可提高至42.9%,暴露2 h的预测性可达到100%。结论基于EpiSkin^(TM)的皮肤刺激性体外替代方法(18 h上样)可用于对护肤类和彩妆类化妆品的评价,但全面推广至化妆品的评价尚有局限性,仍需对产品的上样时间进一步研究和摸索。

Objective To investigate the possibility of using the reconstructed human epidermis model in skin irritation test of cosmetics.Methods Firstly ten proficiency substances with known irritation score were selected for the method validation according to OECD TG439(after 15 minutes exposure and 42 hours post-treatment incubation).Next,twenty-three end-products(including six fundamental cosmetics,ten make up and beauty cosmetics and seven cleansing cosmetics)were evaluated by alternative method(18 hours exposure)with EpiSkinTMmodel which produced in China and Draize test.And seven cleansing cosmetics out of 23 end-products were evaluated by the improved method(four and two hours exposure).Then OD values were detected by MTT test at the end of the incubation and the mean tissue viability of 50% was used to identify whether the product is irritating or non-irritating.Results The classification of 10 chemicals was in good accordance with the data provided by validated reference method in OECD TG439.And the result showed that the predictability of 23 products can reach 73.9%.Out of the 23 products,the predictability of 16 products of fundamental and beauty cosmetics can reach 100%,and the predictability of the cleansing cosmetics was only 14.3%.Then the exposure time in Draize test was changed to modify the in vitro method for cleansing cosmetics with bad predictability,and the predictability were only increased to 42.9% with four hours exposure.Although,with two hours exposure,the result showed that the classification of seven cleansing cosmetics was in good accordance with the data provided by Draize test.It could be probably related to the variety and excessive content of surfactants in these products that could inhibit the growth of cells and cause irreversible damage to the cell membrane structure.Conclusion EpiSkin^(TM) skin irritation alternative method with 18 hours exposure could be suitable for the evaluation of the fundamental and beauty cosmetics.Further method development and research are needed to adapt the original test method from OECD guideline for comprehensively application onto cleansing cosmetics and other end-products evaluation.