Microfluidic Perfusable Pathological Vasculature for Atherosclerosis Drug Screening.

Current in vitro atherosclerosis (AS) models struggle to stimulate blood flow, limiting their ability to replicate endothelial injury and drug transport in AS development. To address this, we developed a 3-dimensional perfusable atherosclerotic vessel-on-a-chip (3D-PAVoC) platform that mimics vascular structure, blood circulating, and disease microenvironment. The system integrates endothelial cells and smooth muscle cells within a flow-enabled arterial construct, exposed to inflammatory (tumor necrosis factor-α and interleukin-1β) and hyperlipidemic stimuli (oxidized low-density lipoprotein) to recreate AS-prone conditions. Flow-dependent endothelial responses including enhanced cell growth and survival were observed, confirming the importance of hemodynamics in disease modeling. Then, rapamycin (RAP) was used as a model drug to evaluate therapeutic effects in the 3D-PAVoC. Compared to static vessel-on-a-chip models and conventional 2-dimensional cultures, 3D-PAVoC exhibited more pronounced AS pathology and higher RAP half-maximal inhibitory concentration, better reflecting in vivo conditions. The effective RAP dose identified in vitro was validated in apolipoprotein E knockout (ApoE-/-) mice, where it partially alleviated AS progression. Transcriptomic analysis revealed RAP-mediated modulation of AS-related gene functions and pathways. Overall, the 3D-PAVoC provides a physiologically relevant platform for anti-AS drug screening, bridging the gap between in vitro testing and in vivo validation, and offering insights into drug action under realistic vascular and pathological conditions.