血脑屏障（BBB）是一种高度选择性的屏障，可确保中枢神经系统（CNS）的稳态环境。血脑屏障功能障碍、炎症和免疫细胞浸润是许多中枢神经系统疾病的标志，包括多发性硬化症和中风。与生理相关的人体血脑屏障体外模型对于提高我们对其在健康和疾病中的功能的理解、识别新的药物靶点以及评估潜在的新疗法至关重要。我们提出了一种 BBB 芯片模型，包含在微流体平台中培养的人脑微血管内皮细胞 (HBMEC)，该平台允许并行培养 40 个芯片。在每个芯片中，灌注的 HBMEC 容器以无膜方式在细胞外基质凝胶上生长。将芯片上的 BBB 暴露于不同浓度的促炎细胞因子肿瘤坏死因子 α (TNFα) 和白细胞介素 1 β (IL-1β) 中以模拟炎症。通过评估芯片上 BBB 的屏障功能、细胞形态和细胞粘附分子的表达来研究炎症条件的影响。原代人类 T 细胞通过芯片上 BBB 的内腔灌注，以研究 T 细胞粘附、外渗和迁移。在炎症条件下，芯片上的 BBB 显示出跨内皮电阻 (TEER) 降低、荧光素钠渗透性增加以及浓度依赖性方式的异常细胞形态。此外，我们观察到细胞粘附分子的表达增加以及伴随的单核细胞粘附。 T 细胞从发炎的血管中渗出，并向 C-X-C 基序趋化因子配体 12 (CXCL12) 梯度迁移。那他珠单抗是一种阻断极晚期抗原 4 (VLA-4) 的抗体药物，用于治疗多发性硬化症，T 细胞粘附显着降低，迁移趋势减少。总之，我们展示了人类血脑屏障的高通量微流体模型，可用于模拟神经炎症并评估抗炎和屏障恢复干预措施以对抗神经系统疾病。版权所有 © 2023 Nair, Groenendijk, Overdevest, Fowke, Annida,莫塞林、德弗里斯和韦弗斯。

The blood-brain barrier (BBB) is a highly selective barrier that ensures a homeostatic environment for the central nervous system (CNS). BBB dysfunction, inflammation, and immune cell infiltration are hallmarks of many CNS disorders, including multiple sclerosis and stroke. Physiologically relevant human in vitro models of the BBB are essential to improve our understanding of its function in health and disease, identify novel drug targets, and assess potential new therapies. We present a BBB-on-a-chip model comprising human brain microvascular endothelial cells (HBMECs) cultured in a microfluidic platform that allows parallel culture of 40 chips. In each chip, a perfused HBMEC vessel was grown against an extracellular matrix gel in a membrane-free manner. BBBs-on-chips were exposed to varying concentrations of pro-inflammatory cytokines tumor necrosis factor alpha (TNFα) and interleukin-1 beta (IL-1β) to mimic inflammation. The effect of the inflammatory conditions was studied by assessing the BBBs-on-chips' barrier function, cell morphology, and expression of cell adhesion molecules. Primary human T cells were perfused through the lumen of the BBBs-on-chips to study T cell adhesion, extravasation, and migration. Under inflammatory conditions, the BBBs-on-chips showed decreased trans-endothelial electrical resistance (TEER), increased permeability to sodium fluorescein, and aberrant cell morphology in a concentration-dependent manner. Moreover, we observed increased expression of cell adhesion molecules and concomitant monocyte adhesion. T cells extravasated from the inflamed blood vessels and migrated towards a C-X-C Motif Chemokine Ligand 12 (CXCL12) gradient. T cell adhesion was significantly reduced and a trend towards decreased migration was observed in presence of Natalizumab, an antibody drug that blocks very late antigen-4 (VLA-4) and is used in the treatment of multiple sclerosis. In conclusion, we demonstrate a high-throughput microfluidic model of the human BBB that can be used to model neuroinflammation and assess anti-inflammatory and barrier-restoring interventions to fight neurological disorders.Copyright © 2023 Nair, Groenendijk, Overdevest, Fowke, Annida, Mocellin, de Vries and Wevers.