Rousi Emma, Malheiro Afonso, Harichandan Abhishek, Mohren Ronny, Lourenço Ana Filipa, Mota Carlos, Cillero-Pastor Berta, Wieringa Paul, Moroni Lorenzo
Department of Complex Tissue Regeneration, MERLN Institute for Technology Inspired Regenerative Medicine, Maastricht, The Netherlands.
Institute of Clinical Medicine, Department of Surgery, University of Turku, Turku, Finland.
In Vitro Model. 2022 Jun 10;2(3-4):113-121. doi: 10.1007/s44164-022-00021-0. eCollection 2023 Aug.
A 3D in vitro model of innervated skin would be a useful tool in dermatological research to study the effect of different chemicals and compounds on the sensory properties of skin. Current innervated skin models are limited in composition and often composed of ex vivo skin explants and/or animal-derived material. In this study, our aim was to develop a human innervated skin model with a better biomimicry composition for in vitro research. Fibrin hydrogel and aligned electrospun fibers of poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) were used as a scaffold to generate the 3D in vitro model. The skin component was made of primary human keratinocytes and primary human fibroblasts, while the neuronal component was composed of iPSC-derived sensory neurons. Our results showed that the dermal component consisted of fibroblasts and synthesized collagen. The epidermal component was characterized by the expression of keratins 10 and 14, and involucrin. Finally, sensory neurons extended axons throughout the scaffold and reached the epidermis. Treating the model with a capsaicin solution for 30 min, which was performed as a proof of concept test for sensitization studies, resulted into partial depletion of substance P and tubulin β3. This model could be used for studying skin-neuron interactions and cutaneous toxicity.
The online version contains supplementary material available at 10.1007/s44164-022-00021-0.
具有神经支配的皮肤的三维体外模型将是皮肤病学研究中的一种有用工具，可用于研究不同化学物质和化合物对皮肤感觉特性的影响。当前的神经支配皮肤模型在组成上存在局限性，通常由离体皮肤外植体和/或动物源材料组成。在本研究中，我们的目标是开发一种具有更好仿生组成的人神经支配皮肤模型用于体外研究。使用纤维蛋白水凝胶和聚（对苯二甲酸乙二酯）/聚（对苯二甲酸丁二酯）（PEOT/PBT）的排列电纺纤维作为支架来生成三维体外模型。皮肤成分由原代人角质形成细胞和原代人成纤维细胞制成，而神经成分由诱导多能干细胞衍生的感觉神经元组成。我们的结果表明，真皮成分由成纤维细胞和合成的胶原蛋白组成。表皮成分的特征是角蛋白10、角蛋白14和内披蛋白的表达。最后，感觉神经元在整个支架中延伸轴突并到达表皮。用辣椒素溶液处理该模型30分钟，作为致敏研究的概念验证测试，导致P物质和微管蛋白β3部分耗尽。该模型可用于研究皮肤 - 神经元相互作用和皮肤毒性。
在线版本包含可在10.1007/s44164-022-00021-0获取的补充材料。
