Bioelectromechanical Systems Lab, Department of Biomedical Engineering and Mechanics, Virginia Tech, Blacksburg, VA, USA.
Spinneret-Based Tunable Engineering Parameters (STEP) Lab, Department of Mechanical Engineering, Virginia Tech, Blacksburg, VA, USA.
Lab Chip. 2023 Oct 10;23(20):4565-4578. doi: 10.1039/d3lc00304c.
Current basement membrane (BM) mimics used for modeling endothelial and epithelial barriers do not faithfully recapitulate key physiological properties such as BM thickness, porosity, stiffness, and fibrous composition. Here, we use networks of precisely arranged nanofibers to form ultra-thin (∼3 μm thick) and ultra-porous (∼90%) BM mimics for blood-brain barrier modeling. We show that these nanofiber networks enable close contact between endothelial monolayers and pericytes across the membrane, which are known to regulate barrier tightness. Cytoskeletal staining and transendothelial electrical resistance (TEER) measurements reveal barrier formation on nanofiber membranes integrated within microfluidic devices and transwell inserts. Further, significantly higher TEER values indicate a biological benefit for co-cultures formed on the ultra-thin nanofiber membranes. Our BM mimic overcomes critical technological challenges in forming co-cultures that are in proximity and facilitate cell-cell contact, while still being constrained to their respective sides. We anticipate that our nanofiber networks will find applications in drug discovery, cell migration, and barrier dysfunction studies.
目前用于模拟内皮和上皮屏障的基底膜 (BM) 模拟物不能真实地再现 BM 厚度、孔隙率、硬度和纤维组成等关键生理特性。在这里，我们使用精确排列的纳米纤维网络来形成超薄膜（约 3μm 厚）和超高多孔（约 90%）的 BM 模拟物，用于血脑屏障建模。我们表明，这些纳米纤维网络能够使内皮单层细胞和周细胞在膜上紧密接触，这是众所周知的可以调节屏障紧密性的细胞。细胞骨架染色和跨内皮电阻 (TEER) 测量显示，在微流控装置和 Transwell 插入物内集成的纳米纤维膜上形成了屏障。此外，TEER 值显著升高表明在超薄膜上形成的共培养具有生物学益处。我们的 BM 模拟物克服了形成共培养的关键技术挑战，这些共培养物在接近和促进细胞-细胞接触的同时，仍然被限制在各自的一侧。我们预计我们的纳米纤维网络将在药物发现、细胞迁移和屏障功能障碍研究中得到应用。
