1 FORM, Frankfurt Orofacial Regenerative Medicine, Clinic for Maxillofacial and Plastic Surgery, Johann Wolfgang Goethe University , Frankfurt Am Main, Germany .
2 DWI Leibniz-Institute for Interactive Materials , Aachen, Germany .
Tissue Eng Part C Methods. 2018 Sep;24(9):495-503. doi: 10.1089/ten.TEC.2018.0087.
The development of an in vitro model resembling the alveolar-capillary barrier might be a highly beneficial tool to study lung physiology as well as the immune response of the lung to infection or after exposure to nanoparticles. This study is based on an in vitro alveolar barrier developed on a basement membrane mimic, composed of ultrathin nanofiber meshes generated via electrospinning using bioresorbable poly(ɛ-caprolactone). As cellular components, NCI H441, resembling the alveolar epithelial cells, and ISO-HAS-1, an endothelial cell line, were used to perform bipolar coculture experiments for a total cultivation period of 14 days. In addition to immunohistochemical and immunofluorescent studies, transepithelial electrical resistance (TER) and transport capabilities of the in vitro model system were investigated. Alveolar barrier function could be clearly determined for the postulated bipolar coculture system on the basement membrane mimic, since TER increased during the course of bipolar cultivation. Furthermore, to gain first insights into possible lung inflammatory reactions in vitro, this coculture model was further expanded by a human leukemia monocyte cell line (THP-1). This triple-culture system was able to maintain adequately the barrier properties of the bipolar coculture, thus making this in vitro model consisting of epithelial, endothelial, and immune cells on a basement membrane mimic a promising basis for further studies in tissue engineering.
体外模型类似于肺泡毛细血管屏障的发展可能是一个非常有益的工具，可用于研究肺生理学以及肺对感染或暴露于纳米颗粒后的免疫反应。本研究基于基底膜模拟物上开发的体外肺泡屏障，该基底膜模拟物由通过静电纺丝使用可生物吸收的聚（ε-己内酯）生成的超薄纳米纤维网组成。作为细胞成分，NCI H441 类似于肺泡上皮细胞，ISO-HAS-1 是一种内皮细胞系，用于进行双极共培养实验，总培养期为 14 天。除了免疫组织化学和免疫荧光研究外，还研究了体外模型系统的跨上皮电阻 (TER) 和转运能力。由于在双极培养过程中 TER 增加，因此可以清楚地确定假定的双极共培养系统在基底膜模拟物上的肺泡屏障功能。此外，为了初步了解体外可能的肺部炎症反应，进一步通过人白血病单核细胞系 (THP-1) 扩展了这种共培养模型。这种三培养系统能够充分维持双极共培养的屏障特性，从而使该体外模型由上皮细胞、内皮细胞和免疫细胞组成，在基底膜模拟物上成为组织工程进一步研究的有前途的基础。
