Simple design for membrane-free microphysiological systems to model the blood-tissue barriers
Simple design for membrane-free microphysiological systems to model the blood-tissue barriers
复制标题
用于模拟血液组织屏障的无膜微生理系统的简单设计
DOI：
10.1016/j.ooc.2023.100032
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Daniele, Michael
中科院分区：
文献类型：
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作者：
Young, By Ashlyn;Deal, Halston;Rusch, Gabrielle;Pozdin, Vladimir A.;Brown, Ashley C.;Daniele, Michael
微生理系统（MPS）结合了生理相关的显微解剖学，力学和细胞来模拟组织功能。组织屏障（如血液-组织界面（BTI））的可复制和可重复的体外模型对于研究和工业中的下一代MPS应用至关重要。BTI的许多模型受限于需要半透膜、使用同质细胞群或2D培养。这些因素限制了相关的内皮-上皮接触和3D转运，这将最好地模拟BTI。目前的模型也很难组装，需要精确的对齐和组件的分层。本文报道的工作详细介绍了BTI芯片（BTI芯片）的工程设计，该芯片通过展示单层无膜设计来解决当前的缺点。使用层流剖面、可光固化的水凝胶支架和人细胞系来构建BTI芯片，该芯片将内皮与3D工程化组织直接接触并置。一种生物材料复合物，明胶甲基丙烯酰和8臂聚乙二醇硫醇，用于原位制造的Y形微流体装置内的组织结构。为了产生BTI，通过使可光固化的前体溶液与磷酸盐缓冲盐水一起流动来实现层流分布。停止流动后，支架立即通过快速暴露于UV光（<300 mJ/cm 2）进行聚合。在支架形成后，引入血管内皮细胞并使其直接粘附到3D组织支架上，而没有屏障或相位引导器。在上皮组织模型和血脑屏障（BBB）模型中证明了BTI芯片的制造。在上皮模型中，支架与人真皮成纤维细胞接种。对于BBB模型，支架用永生化神经胶质细胞系SVGP 12接种。通过免疫组织化学对BTI芯片的显微解剖结构进行了事后分析，显示了与3D工程组织并列的专利内皮的均匀生产。使用荧光示踪剂分子来表征BTI芯片的渗透性。BTI芯片用外排泵抑制剂环孢菌素A进行激发，以评估生理功能和内皮细胞活化。展示了生理相关BTI芯片的操作和用于高通量MPS生成的新方法，使得未来能够开发候选药物筛选和基础生物学研究。
Microphysiological systems (MPS) incorporate physiologically relevant microanatomy, mechanics, and cells to mimic tissue function. Reproducible and standardizedin vitromodels of tissue barriers, such as the blood-tissue interface (BTI), are critical for next-generation MPS applications in research and industry. Many models of the BTI are limited by the need for semipermeable membranes, use of homogenous cell populations, or 2D culture. These factors limit the relevant endothelial-epithelial contact and 3D transport, which would best mimic the BTI. Current models are also difficult to assemble, requiring precise alignment and layering of components. The work reported herein details the engineering of a BTI-on-a-chip (BTI Chip) that addresses current disadvantages by demonstrating a single layer, membrane-free design. Laminar flow profiles, photocurable hydrogel scaffolds, and human cell lines were used to construct a BTI Chip that juxtaposes an endothelium in direct contact with a 3D engineered tissue. A biomaterial composite, gelatin methacryloyl and 8-arm polyethylene glycol thiol, was used forin situfabrication of a tissue structure within a Y-shaped microfluidic device. To produce the BTI, a laminar flow profile was achieved by flowing a photocurable precursor solution alongside phosphate buffered saline. Immediately after stopping flow, the scaffold underwent polymerization through a rapid exposure to UV light (<300 mJ/cm2). After scaffold formation, blood vessel endothelial cells were introduced and allowed to adhere directly to the 3D tissue scaffold, without barriers or phase guides. Fabrication of the BTI Chip was demonstrated in both an epithelial tissue model and blood-brain barrier (BBB) model. In the epithelial model, scaffolds were seeded with human dermal fibroblasts. For the BBB models, scaffolds were seeded with the immortalized glial cell line, SVGP12. The BTI Chip microanatomy was analyzedpost factoby immunohistochemistry, showing the uniform production of a patent endothelium juxtaposed with a 3D engineered tissue. Fluorescent tracer molecules were used to characterize the permeability of the BTI Chip. The BTI Chips were challenged with an efflux pump inhibitor, cyclosporine A, to assess physiological function and endothelial cell activation. Operation of physiologically relevant BTI Chips and a novel means for high-throughput MPS generation was demonstrated, enabling future development for drug candidate screening and fundamental biological investigations.