Real-time imaging of platelet dynamics in engineered arterial, venous, and cancer-associated thrombotic microenvironments

Abstract Thrombosis is a leading cause of cardiovascular morbidity and mortality, driven by platelet-mediated mechanisms common across distinct vascular environments. However, the dynamical behavior of platelets during thrombogenesis remains poorly understood due to the lack of a comprehensive analytical framework. Here we present a physiologically relevant, imaging-integrated thrombosis-on-a-chip platform that enables real-time quantitative analysis of platelet dynamics during thrombogenesis under arterial, venous, and cancer-associated conditions. The system incorporates endothelialized 3D-printed vascular geometries into a closed-loop whole-blood perfusion circuit that replicates native hemodynamic and cellular microenvironments. Multimodal imaging captures the spatiotemporal evolution of thrombus formation and shows how hydrodynamic forces, endothelial dysfunction, and tumor-derived factors drive distinct thrombotic signatures. Notably, platelet-endothelium adhesion and circulating platelet aggregation are identified as mechanistically distinct yet closely linked processes, each uniquely modulated by vascular context. This platform offers a robust framework for dissecting thrombogenesis and advancing antithrombotic and cancer-associated thrombosis research.