(1074-A) Modeling vascular inflammation on the OrganoPlate: A Scalable Platform for Trans-Endothelial Electrical Resistance and Immune Cell Migration

Abstract: Recapitulating the mechanisms of vascular inflammation and the subsequent immune cell response in vitro is critical for the advancement of drug development targeting cardiovascular diseases. To this end, we report on our scalable platform capable of parallel trans endothelial electrical resistance (TEER) measurements across 64 perfused microfluidic human umbilical vein endothelial cell (HUVEC) tubules. . Leveraging the OrganoPlate© 3-lane 64 format, we performed continuous TEER recordings in an incubator environment to monitor endothelial barrier integrity in response to inflammatory stimuli over a 44-hour period. This study presents the largest dataset of TEER measurements to date, with over 250 chips and over 15,000 individual TEER recordings, demonstrating the platform's scalability and potential for high-throughput drug screening.
Our results showcased a significant, dose-dependent impairment of endothelial barrier function upon exposure to pro-inflammatory cytokines TNFα and INF-γ, as well as after interaction with human peripheral blood mononuclear cells (PBMCs). The endothelial response was dynamically captured over a 44-hour period, revealing transient barrier disruptions and subsequent recoveries, particularly in TNFα-treated conditions. The combination of TNFα and INF-γ resulted in the most pronounced and sustained barrier deterioration, with no observed recovery.
Further insights were gained by introducing PBMCs into the endothelial tubules. Unstimulated PBMCs showed no significant impact on barrier integrity; however, stimulated PBMCs induced a barrier disruption akin to that observed with TNFα and INF-γ exposure. This disruption was characterized by a decrease in TEER values, mirroring the inflammatory cytokine effects. Additionally, stimulated PBMCs demonstrated increased adherence to the endothelial wall and, when combined with a chemotactic gradient, exhibited significant transmigration into the ECM, underscoring the platform's utility in studying immune cell behavior in vascular inflammation.
The OrganoPlate 3-lane 64 platform's compatibility with automated liquid handling and high-content imaging systems highlights its suitability for complex disease modeling and drug discovery processes. Our study presents a robust, scalable, and multiplexed approach to investigate vascular inflammation, offering a high-throughput platform for the evaluation of therapeutic agents targeting chronic inflammatory diseases.