Hyaluronan Oxime Hydrogels for Bile Duct Engineering

Cholangiopathies account for up to 70% of pediatric and 33% of adult liver transplantations, yet current cholangiocyte organoid systems rely on ill-defined and animal-derived matrices that prevent mechanistic study and clinical translation. Herein, formulations of viscoelastic hyaluronan-oxime (HAO) hydrogels incorporating laminin were developed for three-dimensional cholangiocyte culture. Hydrogels matching the stress relaxation rate of native liver tissue supported robust organoid growth mediated by a FAK-Src-actomyosin-YAP mechanotransduction cascade. Resultant organoids expressed mature biliary markers, demonstrated epithelial polarity, and retained functional transport capabilities. Immobilization of Jagged1 within the viscoelastic hydrogel activated Notch2 signalling and drove unprecedented bile duct-like morphogenesis without feeder cells. This work establishes a defined, tunable platform for advancing biliary disease modelling and liver tissue engineering. ii