Parnigotto P P, Bernuzzo S, Bruno P, Conconi M T, Montesi F
Dipartimento di Scienze Farmaceutiche, Università di Padova, Italy.
Farmaco. 1998 Feb;53(2):125-31. doi: 10.1016/s0014-827x(97)00020-7.
In recent years in vitro models have been developed to avoid the use of animals in cutaneous toxicological studies. Submerged human keratinocyte cultures in vitro could be so far employed as an alternative to animal testing and a good correlation between skin irritation and cytotoxicity has been demonstrated. Nevertheless, these submerged cultures are lacking in the stratum corneum which acts as a barrier to chemical toxicity, so that this type of culture is far from the in vivo situation. A better alternative method seems to be the use of in vitro reconstructed skin at the air-liquid interface that closely resembles the in vivo situation. In this work, in a first step we have characterized human epidermis reconstructed in vitro on de-epidermized derma (DED) after a two-week air exposure. Human skin reconstituted in vitro on DED was histologically similar to the in vivo skin. A stratified epidermis including the stratum corneum was obtained. The presence of basal lamina as well as of various important markers for epidermal differentiation (involucrin, K10 keratin, and filaggrin) were revealed. In a second step we have tested the cytotoxic and morphological effects of four surfactants on our model. A good rank correlation has been shown to exist between the irritation potency of surfactants on our model and reported ocular irritancy in vivo. From our results, in vitro reconstituted human skin could represent an attractive model for irritancy testing and could be an in vitro replacement for animal testing.
近年来，已开发出体外模型以避免在皮肤毒理学研究中使用动物。迄今为止，体外培养的人角质形成细胞可作为动物试验的替代方法，并且已证明皮肤刺激性与细胞毒性之间具有良好的相关性。然而，这些浸没培养缺乏作为化学毒性屏障的角质层，因此这种培养类型与体内情况相差甚远。一种更好的替代方法似乎是使用气液界面的体外重建皮肤，其与体内情况非常相似。在这项工作中，第一步，我们对在去皮真皮（DED）上体外重建的人表皮进行了为期两周的空气暴露后的表征。在DED上体外重建的人皮肤在组织学上与体内皮肤相似。获得了包括角质层在内的分层表皮。揭示了基底层以及表皮分化的各种重要标志物（内披蛋白、角蛋白K10和丝聚合蛋白）的存在。第二步，我们测试了四种表面活性剂对我们模型的细胞毒性和形态学影响。已表明表面活性剂对我们模型的刺激强度与体内报告的眼刺激性之间存在良好的等级相关性。根据我们的结果，体外重建的人皮肤可能是一种有吸引力的刺激性测试模型，并且可以作为动物测试的体外替代方法。
