Department of General Practice , The First Affiliated Hospital of China Medical University , Shenyang , Liaoning 110001 , China.
College of Textiles & Clothing , Qingdao University , Qingdao 266071 , China.
ACS Appl Mater Interfaces. 2018 Jul 5;10(26):21825-21835. doi: 10.1021/acsami.8b03962. Epub 2018 Jun 22.
The blood-brain barrier (BBB) is an active and complex diffusion barrier that separates the circulating blood from the brain and extracellular fluid, regulates nutrient transportation, and provides protection against various toxic compounds and pathogens. Creating an in vitro microphysiological BBB system, particularly with relevant human cell types, will significantly facilitate the research of neuropharmaceutical drug delivery, screening, and transport, as well as improve our understanding of pathologies that are due to BBB damage. Currently, most of the in vitro BBB models are generated by culturing rodent astrocytes and endothelial cells, using commercially available transwell membranes. Those membranes are made of plastic biopolymers that are nonbiodegradable, porous, and stiff. In addition, distinct from rodent astrocytes, human astrocytes possess unique cell complexity and physiology, which are among the few characteristics that differentiate human brains from rodent brains. In this study, we established a novel human BBB microphysiologocal system, consisting of a three-dimensionally printed holder with a electrospun poly(lactic- co-glycolic) acid (PLGA) nanofibrous mesh, a bilayer coculture of human astrocytes, and endothelial cells, derived from human induced pluripotent stem cells (hiPSCs), on the electrospun PLGA mesh. This human BBB model achieved significant barrier integrity with tight junction protein expression, an effective permeability to sodium fluorescein, and higher transendothelial electrical resistance (TEER) comparing to electrospun mesh-based counterparts. Moreover, the coculture of hiPSC-derived astrocytes and endothielial cells promoted the tight junction protein expression and the TEER value. We further verified the barrier functions of our BBB model with antibrain tumor drugs (paclitaxel and bortezomib) and a neurotoxic peptide (amyloid β 1-42). The human microphysiological system generated in this study will potentially provide a new, powerful tool for research on human BBB physiology and pathology.
血脑屏障（BBB）是一种活跃且复杂的扩散屏障，将循环血液与大脑和细胞外液隔开，调节营养物质的运输，并为各种有毒化合物和病原体提供保护。创建体外微生理 BBB 系统，特别是使用相关的人类细胞类型，将极大地促进神经药物输送、筛选和转运的研究，并增进我们对由于 BBB 损伤导致的病理学的认识。目前，大多数体外 BBB 模型是通过培养啮齿动物星形胶质细胞和内皮细胞，使用市售的 Transwell 膜来构建的。这些膜由不可生物降解的、多孔的和刚性的塑料生物聚合物制成。此外，与啮齿动物星形胶质细胞不同，人类星形胶质细胞具有独特的细胞复杂性和生理学特性，这些特性是人类大脑与啮齿动物大脑的区别之一。在本研究中，我们建立了一种新型的人 BBB 微生理系统，由三维打印支架和电纺聚乳酸-共-羟基乙酸（PLGA）纳米纤维网、由人诱导多能干细胞（hiPSC）衍生的星形胶质细胞和内皮细胞双层共培养组成，在电纺 PLGA 网上。与基于电纺网的对照物相比，该人 BBB 模型具有显著的屏障完整性，紧密连接蛋白表达、对荧光素钠的有效渗透性和更高的跨内皮电阻（TEER）。此外，hiPSC 衍生的星形胶质细胞和内皮细胞的共培养促进了紧密连接蛋白的表达和 TEER 值。我们进一步用抗肿瘤药物（紫杉醇和硼替佐米）和神经毒性肽（淀粉样β 1-42）验证了我们的 BBB 模型的屏障功能。本研究中生成的人微生理系统将为研究人 BBB 的生理学和病理学提供一种新的、强大的工具。
