Self‐Healing Hydrogel Containing Decellularized Liver Matrix and Endothelial Cell‐Covered Hepatocyte Spheroids for Rescue of Injured Hepatocytes

Abstract Liver fibrosis occurs in many chronic liver diseases, while severe fibrosis can lead to liver failure. A chitosan‐phenol based self‐healing hydrogel (CP) integrated with decellularized liver matrix (DLM) is proposed in this study as a 3D gel matrix to carry hepatocytes for possible therapy of liver fibrosis. To mimic the physiological liver microenvironment, DLM is extracted from pigs and mixed with CP hydrogel to generate DLM‐CP self‐healing hydrogel. Hepatocyte spheroids coated with endothelial cells (ECs) are fabricated using a customized method and embedded in the hydrogel. Hepatocytes injured by exposure to CCl 4 ‐containing medium are used as the in vitro toxin‐mediated liver fibrosis model, where the EC‐covered hepatocyte spheroids embedded in the hydrogel are co‐cultured with the injured hepatocytes. The urea synthesis of the injured hepatocytes reaches 91% of the normal level after 7 days of co‐culture, indicating that the hepatic function of injured hepatocytes is rescued by the hybrid spheroid‐laden DLM‐CP hydrogel. Moreover, the relative lactate dehydrogenase activity of the injured hepatocytes is decreased 49% by the hybrid spheroid‐laden DLM‐CP hydrogel after 7 days of co‐culture, suggesting reduced damage in the injured hepatocytes. The combination of hepatocyte/EC hybrid spheroids and DLM‐CP hydrogel presents a promising therapeutic strategy for hepatic fibrosis.