cEpiderm, a Canine Skin Analog Suitable for In Vivo Testing Replacement

BioChem Pub Date : 2022-10-20 DOI:10.3390/biochem2040015
Mariana Marques, João Nunes, Bárbara Ustymenko, L. Fialho, Luís Martins, A. Burke, Cesar Filho, Alexandre C. Craveiro, Ana R. Costa, S. Branco, C. Antunes
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Abstract

Skin is one of the organs most tested for toxicity and safety evaluation during the process of drug research and development and in the past has usually been performed in vivo using animals. Over the last few years, non-animal alternatives have been developed and validated epidermis models for human and rat skin are already available. Our goal was to develop a histotypical canine skin analog, suitable for non-animal biocompatibility and biosafety assessment. Canine keratinocytes were seeded in an air-lift culture using an adapted version of the CELLnTEC protocol. Corrosion and irritation protocols were adapted from human EpiSkinTM. For histological analysis, sample biopsies were fixed in neutral-buffered formalin, and paraffin slices were routinely processed and stained with hematoxylin and eosin. A canine multilayer and stratified epidermal-like tissue (cEpiderm), confirmed by histological analysis, was obtained. The cEpiderm tissue exhibited normal morphological and functional characteristics of epidermis, namely impermeability and an adequate response to stressors. The cEpiderm is a promising canine skin model for non-animal safety testing of veterinary pharmaceuticals and/or cosmetics, significantly contributing to reducing undesirable in vivo approaches. cEpiderm is therefore a valid canine skin model and may be made commercially available either as a service or as a product.
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cEpiderm,一种犬类皮肤类似物,适用于体内试验替代品
在药物研发过程中,皮肤是毒性和安全性评价测试最多的器官之一,过去通常在动物体内进行。在过去的几年里,非动物替代品已经被开发出来,并且已经有了人类和大鼠皮肤的表皮模型。我们的目标是开发一种组织典型的犬皮肤类似物,适用于非动物生物相容性和生物安全性评估。采用改良版的CELLnTEC方案,将犬角化细胞植入气举培养中。腐蚀和刺激方案改编自人EpiSkinTM。为了进行组织学分析,活检标本被固定在中性缓冲的福尔马林中,石蜡切片被常规处理,并用苏木精和伊红染色。通过组织学分析证实了犬的多层和分层表皮样组织(cEpiderm)。表皮组织表现出正常的表皮形态和功能特征,即不渗透性和对应激的充分反应。在兽药和/或化妆品的非动物安全性测试中,cEpiderm是一种很有前途的犬皮肤模型,显著有助于减少不良的体内方法。因此,cEpiderm是一种有效的犬皮肤模型,可以作为一种服务或产品进行商业销售。
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