Morales Mariana, Pérez David, Correa Luis, Restrepo Luz
Tissue Engineering and Cell Therapy Group (GITTC), School of Medicine, University of Antioquia, Colombia.
Tissue Engineering and Cell Therapy Group (GITTC), School of Medicine, University of Antioquia, Colombia; Dermatology Department, School of Medicine, University of Antioquia, Colombia.
Toxicol In Vitro. 2016 Oct;36:89-96. doi: 10.1016/j.tiv.2016.07.010. Epub 2016 Jul 21.
Reconstructed human epidermis (RhE) models have been used for in vitro testing of the potential harmful effects of exposure to chemical compounds on health. In the past, skin irritation and corrosion were evaluated in animal models; however, in recent years, due to the bioethics implications of the method and, to minimize the use of experimental animals, alternative procedures have been proposed. The Organisation for Economic Co-operation and Development (OECD) in its test guidelines (TG) 431 and 439 indicates the requirements for validating new methods for the evaluation of skin corrosion and irritation, respectively. Here, we present an in-house human dermal-epidermal model, useful for the performance of these tests. Using the methods described in this work, it was possible to obtain human fibrin-based dermal-epidermal organotypic skin cultures (ORGs) displaying similar histological characteristics to native skin and expressing specific differentiation epithelial proteins. The end points to classify a substance as irritant or corrosive were cell viability evaluated by MTT assay, and cytokine release measured by BD CBA for human inflammatory cytokines. According to the MTT test, the ORGs correctly classified irritating and corrosive substances. Moreover, the cytokine release assay was difficult to interpret in the context of testing chemical hazard classification. Further experiments are needed to validate this new model for the evaluation of surfactants because the fibrin matrix was affected in the presence of these substances.
重建人表皮(RhE)模型已用于体外测试接触化学化合物对健康的潜在有害影响。过去,皮肤刺激性和腐蚀性是在动物模型中评估的;然而,近年来,由于该方法的生物伦理问题,以及为了尽量减少实验动物的使用,人们提出了替代程序。经济合作与发展组织(OECD)在其测试指南(TG)431和439中分别指出了验证皮肤腐蚀性和刺激性评估新方法的要求。在此,我们展示了一种内部人真皮-表皮模型,可用于进行这些测试。使用本文所述方法,可以获得基于人纤维蛋白的真皮-表皮器官型皮肤培养物(ORGs),其显示出与天然皮肤相似的组织学特征,并表达特定的分化上皮蛋白。将一种物质分类为刺激性或腐蚀性物质的终点指标是通过MTT法评估细胞活力,以及通过BD CBA检测人炎症细胞因子来测量细胞因子释放。根据MTT试验,ORGs正确地对刺激性和腐蚀性物质进行了分类。此外,在测试化学危害分类的背景下,细胞因子释放试验难以解释。由于在这些物质存在的情况下纤维蛋白基质会受到影响,因此需要进一步的实验来验证这种用于评估表面活性剂的新模型。
Toxicol In Vitro. 2019-10-20
Toxicol In Vitro. 2007-10
Eur J Pharm Biopharm. 2018-1-6
Toxicol In Vitro. 2012-4-27
Bioengineering (Basel). 2024-10-17
Molecules. 2022-7-14
Curr Top Microbiol Immunol. 2021