Allergy Research Group, School of Molecular Medical Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, NG7 2UH, UK.
Biofabrication. 2013 Sep;5(3):035011. doi: 10.1088/1758-5082/5/3/035011. Epub 2013 Jul 23.
As the first line of defence, skin is regularly exposed to a variety of biological, physical and chemical insults. Therefore, determining the skin sensitization potential of new chemicals is of paramount importance from the safety assessment and regulatory point of view. Given the questionable biological relevance of animal models to human as well as ethical and regulatory pressure to limit or stop the use of animal models for safety testing, there is a need for developing simple yet physiologically relevant models of human skin. Herein, we describe the construction of a novel immunocompetent 3D human skin model comprising of dendritic cells co-cultured with keratinocytes and fibroblasts. This model culture system is simple to assemble with readily-available components and importantly, can be separated into its constitutive individual layers to allow further insight into cell-cell interactions and detailed studies of the mechanisms of skin sensitization. In this study, using non-degradable microfibre scaffolds and a cell-laden gel, we have engineered a multilayer 3D immunocompetent model comprised of keratinocytes and fibroblasts that are interspersed with dendritic cells. We have characterized this model using a combination of confocal microscopy, immuno-histochemistry and scanning electron microscopy and have shown differentiation of the epidermal layer and formation of an epidermal barrier. Crucially the immune cells in the model are able to migrate and remain responsive to stimulation with skin sensitizers even at low concentrations. We therefore suggest this new biologically relevant skin model will prove valuable in investigating the mechanisms of allergic contact dermatitis and other skin pathologies in human. Once fully optimized, this model can also be used as a platform for testing the allergenic potential of new chemicals and drug leads.
作为第一道防线，皮肤经常暴露于各种生物、物理和化学刺激物中。因此，从安全性评估和监管角度来看，确定新化学物质的皮肤致敏潜力至关重要。鉴于动物模型与人类的生物学相关性存在疑问，以及出于伦理和监管方面的压力，需要限制或停止使用动物模型进行安全性测试，因此需要开发简单但具有生理相关性的人类皮肤模型。在此，我们描述了一种新型免疫活性 3D 人皮肤模型的构建，该模型由与角质形成细胞和成纤维细胞共培养的树突状细胞组成。该模型培养系统易于组装，使用的组件易于获得，重要的是，可以将其分离成其组成的各个层，以进一步深入了解细胞-细胞相互作用，并详细研究皮肤致敏的机制。在这项研究中，我们使用不可降解的微纤维支架和细胞负载凝胶，构建了一种由角质形成细胞和成纤维细胞组成的多层 3D 免疫活性模型，其中散布着树突状细胞。我们使用共聚焦显微镜、免疫组织化学和扫描电子显微镜对该模型进行了表征，并证明了表皮层的分化和表皮屏障的形成。至关重要的是，模型中的免疫细胞能够迁移并对皮肤致敏剂的刺激保持反应性，即使在低浓度下也是如此。因此，我们认为这种新的具有生理相关性的皮肤模型将有助于研究人类过敏性接触性皮炎和其他皮肤病理学的机制。一旦完全优化，该模型还可以用作测试新化学物质和药物先导物致敏潜力的平台。
