Abstract
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.