Camarena Denisse Esther Mallaupoma, Matsuyama Larissa Satiko Alcântara Sekimoto, Maria-Engler Silvya Stuchi, Catalani Luiz Henrique
Laboratory of Polymeric Biomaterials, Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo 05508-000, Brazil.
Skin Biology Laboratory, Clinical Chemistry & Toxicology Department, School of Pharmaceutical Sciences, University of São Paulo, São Paulo 05508-000, Brazil.
Nanomaterials (Basel). 2020 Dec 16;10(12):2528. doi: 10.3390/nano10122528.
The development of products for topical applications requires analyses of their skin effects before they are destined for the market. At present, the ban on animal use in several tests makes the search for in vitro models (such as artificial skin) necessary to characterize the risks involved. In this work, tissue engineering concepts were used to manufacture collagen-free three-dimensional scaffolds for cell growth and proliferation. Two different human skin models-reconstructed human epidermis and full-thickness skin-were developed from electrospun scaffolds using synthetic polymers such as polyethylene terephthalate, polybutylene terephthalate, and nylon 6/6. After the construction of these models, their histology was analyzed by H&E staining and immunohistochemistry. The results revealed a reconstructed epidermal tissue, duly stratified, obtained from the nylon scaffold. In this model, the presence of proteins involved in the epidermis stratification process (cytokeratin 14, cytokeratin 10, involucrin, and loricrin) was confirmed by immunohistochemistry and Western blot analysis. The nylon reconstructed human epidermis model's applicability was evaluated as a platform to perform irritation and corrosion tests. Our results demonstrated that this model is a promising platform to assess the potential of dermal irritation/corrosion of chemical products.
用于局部应用的产品在投放市场之前需要对其皮肤效应进行分析。目前，多项试验中禁止使用动物，这使得寻找体外模型（如人工皮肤）以确定相关风险成为必要。在这项工作中，利用组织工程概念制造了无胶原蛋白的三维支架，用于细胞生长和增殖。使用聚对苯二甲酸乙二酯、聚对苯二甲酸丁二酯和尼龙6/6等合成聚合物，通过静电纺丝支架开发了两种不同的人类皮肤模型——重建的人类表皮和全层皮肤。构建这些模型后，通过苏木精-伊红染色和免疫组织化学分析其组织学。结果显示，从尼龙支架获得了适当分层的重建表皮组织。在该模型中，通过免疫组织化学和蛋白质印迹分析证实了参与表皮分层过程的蛋白质（细胞角蛋白14、细胞角蛋白10、内披蛋白和兜甲蛋白）的存在。评估了尼龙重建人类表皮模型作为进行刺激和腐蚀试验平台 的适用性。我们的结果表明，该模型是评估化学产品皮肤刺激/腐蚀潜力的一个有前景的平台。
