Rutten A A, van de Sandt J J
TNO Food and Nutrition Research, Department of Biological Toxicology, PO Box 360, 3700 AJ Zeist, The Netherlands.
Toxicol In Vitro. 1994 Aug;8(4):703-5. doi: 10.1016/0887-2333(94)90047-7.
In order to reduce animal discomfort and to obtain more quantitative endpoints, there is a need for reliable, preferably simple and inexpensive in vitro alternatives to skin toxicity testing. An in vitro model was developed in which full-thickness skin from various species can be cultured (rabbit, pig and human). Subsequent to topically applied test compounds, parameters of dermal toxicity were investigated, including cytotoxicity (MTT assay) and the release of inflammatory mediators (HETE's). Moreover, percutaneous absorption and concurrent biotransformation of compounds was studied. MTT conversion was inhibited in a dose-dependent manner following topical application of a wide range of irritants. Repair of initial damage was observed to some extent. The eicosanoid 12-HETE, which is thought to play an important role in chemotaxis, is released. Interestingly, the anti-inflammatory mediator 15-HETE was released only after a prolonged culture time of 48 hr, possibly indicating repair of the induced damage. The metabolic fate of the pesticide propoxur was investigated. Permeation rates were comparable in human and rabbit skin, while pig skin was found to be twice as permeable. Extensive cutaneous metabolism was observed in all three species.
为了减少动物的不适并获得更具定量性的终点指标，需要一种可靠的、最好是简单且廉价的体外替代方法来进行皮肤毒性测试。开发了一种体外模型，可在其中培养来自不同物种（兔、猪和人）的全层皮肤。在局部应用测试化合物后，研究了皮肤毒性参数，包括细胞毒性（MTT 法）和炎症介质（羟基二十碳四烯酸）的释放。此外，还研究了化合物的经皮吸收和同时进行的生物转化。在局部应用多种刺激物后，MTT 转化呈剂量依赖性抑制。在一定程度上观察到了初始损伤的修复。被认为在趋化作用中起重要作用的类花生酸 12-羟基二十碳四烯酸被释放。有趣的是，抗炎介质 15-羟基二十碳四烯酸仅在延长培养 48 小时后才释放，这可能表明诱导损伤得到了修复。研究了农药残杀威的代谢命运。人皮肤和兔皮肤的渗透速率相当，而猪皮肤的渗透性是其两倍。在所有三个物种中均观察到广泛的皮肤代谢。
