The ECVAM International Validation Study on in Vitro Tests for Acute Skin Irritation: Report on the Validity of the EPISKIN and EpiDerm Assays and on the Skin Integrity Function Test a
The ECVAM International Validation Study on in Vitro Tests for Acute Skin Irritation: Report on the Validity of the EPISKIN and EpiDerm Assays and on the Skin Integrity Function Test a
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ECVAM 关于急性皮肤刺激体外测试的国际验证研究：EPISKIN 和 EpiDerm 测定以及皮肤完整性功能测试的有效性报告
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发表时间：
2007
期刊：
影响因子：
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通讯作者：
V. Zuang
中科院分区：
文献类型：
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作者：
H. Spielmann;S. Hoffmann;M. Liebsch;P. Botham;J. Fentem;C. Eskes;R. Roguet;J. Cotovio;Thomas Cole;Andrew Paul Worth;J. Heylings;Penny Jones;C. Robles;Helena Kandarova;A. Gamer;M. Remmele;R. Curren;H. Raabe;A. Cockshott;I. Gerner;V. Zuang
ECVAM赞助了一项关于皮肤刺激性的三项体外试验的正式验证研究，其中两项采用重建的人表皮模型（EPISKIN™、EpiDerm™），一项皮肤完整性功能试验（SIFT）采用离体小鼠皮肤。本研究的目的是评估体外试验是否能够根据欧盟分类方案“R38”和“无标签”（即无刺激性）正确预测体内分类。测试了58种化学物质（25种刺激物和33种非刺激物），选择这些化学物质是为了广泛覆盖物理化学性质，并充分分布来自体内兔子皮肤刺激测试的刺激性评分。在第1阶段，由单个牵头实验室对每种方法的20种化学品（9种刺激物和11种非刺激物）进行编码鉴别检测，以确认预验证阶段后引入的方案改进的适用性。当使用细胞活力（通过MTT还原试验评价）作为终点时，认为EpiDerm和EPISKIN的预测能力足以证明其进展至II期，而SIFT的预测能力被判定为不足。由于两种重建皮肤模型在体内分类边界附近提供了错误预测（兔Draize试验评分为2），因此还测定了细胞因子白细胞介素-1 α（IL-1α）的释放。在第二阶段，每个人体皮肤模型在三个实验室进行了测试，使用了58种化学物质。EpiDerm和EPISKIN测量的主要终点是细胞活力。在MTT试验结果高于50%活力阈值的化学品样本中，还测量了IL-1α释放，以确定额外终点是否会提高试验的预测能力。对于EPISKIN，灵敏度为75%，特异性为81%（仅MTT测定）; MTT和IL-1α测定联合使用时，灵敏度增加至91%，特异性为79%。对于EpiDerm，灵敏度为57%，特异性为85%（仅MTT法），而测量IL-1α释放并不能提高EpiDerm的预测能力。经过独立同行评审，ECVAM科学咨询委员会于2007年4月认可了EPISKIN测试作为兔子皮肤刺激方法替代品的科学有效性，以及作为分层测试策略一部分的识别皮肤刺激物的EpiDerm方法的科学有效性。这种新的替代方法可能是在欧洲和国际上首次使用体外毒性试验取代Draize家兔皮肤刺激试验，因为在不久的将来，新的欧盟和经合组织试验指南将被提议用于监管验收。
ECVAM sponsored a formal validation study on three in vitro tests for skin irritation, of which two employ reconstituted human epidermis models (EPISKIN™, EpiDerm™), and one, the skin integrity function test (SIFT), employs ex vivo mouse skin. The goal of the study was to assess whether the in vitro tests would correctly predict in vivo classifications according to the EU classification scheme, “R38” and “no label” (i.e. non-irritant). 58 chemicals (25 irritants and 33 non-irritants) were tested, having been selected to give broad coverage of physico–chemical properties, and an adequate distribution of irritancy scores derived from in vivo rabbit skin irritation tests. In Phase 1, 20 of these chemicals (9 irritants and 11 non-irritants) were tested with coded identities by a single lead laboratory for each of the methods, to confirm the suitability of the protocol improvements introduced after a prevalidation phase. When cell viability (evaluated by the MTT reduction test) was used as the endpoint, the predictive ability of both EpiDerm and EPISKIN was considered sufficient to justify their progression to Phase 2, while the predictive ability of the SIFT was judged to be inadequate. Since both the reconstituted skin models provided false predictions around the in vivo classification border (a rabbit Draize test score of 2), the release of a cytokine, inter-leukin-1α (IL-1α), was also determined. In Phase 2, each human skin model was tested in three laboratories, with 58 chemicals. The main endpoint measured for both EpiDerm and EPISKIN was cell viability. In samples from chemicals which gave MTT assay results above the threshold of 50% viability, IL-1α release was also measured, to determine whether the additional endpoint would improve the predictive ability of the tests. For EPISKIN, the sensitivity was 75% and the specificity was 81% (MTT assay only); with the combination of the MTT and IL-1α assays, the sensitivity increased to 91%, with a specificity of 79%. For EpiDerm, the sensitivity was 57% and the specificity was 85% (MTT assay only), while the predictive capacity of EpiDerm was not improved by the measurement of IL-1α release. Following independent peer review, in April 2007 the ECVAM Scientific Advisory Committee endorsed the scientific validity of the EPISKIN test as a replacement for the rabbit skin irritation method, and of the EpiDerm method for identifying skin irritants as part of a tiered testing strategy. This new alternative approach will probably be the first use of in vitro toxicity testing to replace the Draize rabbit skin irritation test in Europe and internationally, since, in the very near future, new EU and OECD Test Guidelines will be proposed for regulatory acceptance.