Year: 2026
Type: preprint
Abstract: Engineering a transplantable artificial liver requires reconstituting the hepatic lobule, whose densely cellular parenchyma is organized around a hierarchically branched vascular network. Achieving high cellular density and hierarchical vascularization within a single fabrication step remains a major challenge. Here, we present an ultrasonic standing-wave (USW)-based strategy that concurrently organizes vascular and parenchymal compartments during a single extrusion. HepG2–endothelial cell aggregates were fabricated as building blocks and co-extruded with single endothelial cells through a glass capillary under USW actuation in a liver-derived decellularized extracellular matrix–alginate hydrogel, enabling rapid, ultraviolet-free crosslinking. Due to their size difference, the single cells were patterned from one supplying line in the acoustic pressure node to four microvascular lines while the aggregates localized away from the acoustic pressure node, reconstituting the basic microvascular–parenchymal unit of the lobule. Aggregate size increased with seeding density while maintaining uniformity and over 90% viability, and the patterned endothelial cells formed perfusable lumina through which 5 µm microparticles flowed freely. Compared with hepatocyte-only cultures, the vascularized constructs showed a 1.7-fold increase in urea production, and 2.7- and 6.2-fold increases in CYP1A2 and CYP3A4 expression, respectively. Following implantation into the mouse liver, the scaffolds showed favorable biocompatibility, sustained proliferation over 21 days, and progressive host integration, with the smallest blood-containing lumina maturing from 26.1 µm to 7.9 µm in diameter and increasing collagen deposition at the graft–host interface. This approach provides a template-free route to hierarchically vascularized hepatic units and a promising step toward implantable liver tissue.
Language: English
Subfield: Surgery
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W7213981317