表皮皮肤测试1000(EST - 1000)——一种用于体外皮肤腐蚀性测试的新型重建表皮。

Epidermal-skin-test 1,000 (EST-1,000)--a new reconstructed epidermis for in vitro skin corrosivity testing.

作者信息

Hoffmann J, Heisler E, Karpinski S, Losse J, Thomas D, Siefken W, Ahr H-J, Vohr H-W, Fuchs H W

机构信息

Advanced CellSystems GmbH, Mülheimerstr. 26, D-53840 Troisdorf, Germany.

出版信息

Toxicol In Vitro. 2005 Oct;19(7):925-9. doi: 10.1016/j.tiv.2005.06.010. Epub 2005 Aug 2.

Abstract

The determination of a possible corrosive or irritative potential of certain products and ingredients is necessary for their classification and labeling requirements. Reconstructed skin as a model system provides fundamental advantages to single cell culture testing and leads to promising results as shown by different validation studies (for review: Fentem, J.H., Botham, P.A., 2002. ECVAM's activities in validating alternative tests for skin corrosion and irritation. ATLA 30(Suppl. 2), 61-67). In this study we introduce our new reconstructed epidermis "Epidermal-Skin-Test" (EST-1,000). This fully grown epidermis consists of proliferating as well as differentiating keratinocytes. EST-1,000 shows a high comparability to normal human skin as shown by histological and immunohistochemical data. Characteristic markers (KI-67, CK 1/10/5/14, transglutaminase, collagen IV, involucrin, beta 1 integrin) can be identified easily. The main focus of this work was to characterize EST-1,000 especially with respect to its barrier function by testing several substances of known corrosive potential. Skin corrosion was detected by the cytotoxic effect of the substances on a reconstructed epidermis after short-term application to the stratum corneum. The effect was determined by standard MTT assay and accompanying histological analysis. Hence EST-1,000 shows a very high predictive potential and closes the gap between animal testing and the established full-thickness model Advanced-Skin-Test 2,000 (AST-2,000) (Noll, M., Merkle, M.-L., Kandsberger, M., Matthes, T., Fuchs, H., Graeve, T., 1999. Reconstructed human skin (AST-2,000) as a tool for pharmaco-toxicology. ATLA 27, 302).

摘要

确定某些产品和成分可能具有的腐蚀性或刺激性对于其分类和标签要求而言是必要的。作为一种模型系统,重建皮肤相对于单细胞培养测试具有根本优势,并且如不同的验证研究所显示的那样能产生有前景的结果(综述:Fentem, J.H., Botham, P.A., 2002. ECVAM在验证皮肤腐蚀和刺激替代测试方面的活动。《替代与实验室动物》30(增刊2),61 - 67)。在本研究中,我们介绍了我们新的重建表皮“表皮皮肤测试”(EST - 1000)。这种完全成熟的表皮由增殖以及分化的角质形成细胞组成。组织学和免疫组化数据表明,EST - 1000与正常人类皮肤具有高度可比性。特征性标志物(KI - 67、细胞角蛋白1/10/5/14、转谷氨酰胺酶、胶原蛋白IV、内披蛋白、β1整合素)能够被轻易识别出来。这项工作的主要重点是通过测试几种已知具有腐蚀潜力的物质来特别表征EST - 1000的屏障功能。在将物质短期应用于角质层后,通过其对重建表皮的细胞毒性作用来检测皮肤腐蚀情况。该作用通过标准MTT法和附带的组织学分析来确定。因此,EST - 1000显示出非常高的预测潜力,并填补了动物测试与已确立的全层模型高级皮肤测试2000(AST - 2000)之间的差距(Noll, M., Merkle, M.-L., Kandsberger, M., Matthes, T., Fuchs, H., Graeve, T., 1999. 重建的人类皮肤(AST - 2000)作为药物毒理学工具。《替代与实验室动物》27, 302)。

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