Sequentially suspended 3D bioprinting of multiple-layered vascular models with tunable geometries for in vitro modeling of arterial disorders initiation

As the main precursor of arterial disorders, endothelial dysfunction preferentially occurs in regions of arteries prone to generating turbulent flow, particularly in branched regions of vasculatures. Although various diseased models have been engineered to investigate arterial pathology, producing a multiple-layered vascular model with branched geometries that can recapitulate the critical physiological environments of human arteries, such as intercellular communications and local turbulent flows, remains challenging. This study develops a sequentially suspended three-dimensional bioprinting (SSB) strategy and a visible-light-curable decellularized extracellular matrix bioink (abbreviated as 'VCD bioink') to construct a biomimetic human arterial model with tunable geometries. The engineered multiple-layered arterial models with compartmentalized vascular cells can exhibit physiological functionality and pathological performance under defined physiological flows specified by computational fluid dynamics simulation. Using different configurations of the vascular models, we investigated the independent and synergetic effects of cellular crosstalk and abnormal hemodynamics on the initiation of endothelial dysfunction, a hallmark event of arterial disorder. The results suggest that the arterial model constructed using the SSB strategy and VCD bioinks has promise in establishing diagnostic/analytic platforms for understanding the pathophysiology of human arterial disorders and relevant abnormalities, such as atherosclerosis, aneurysms, and ischemic diseases.