MicroRNA-Modified DNA Hexahedron-Induced Hepatocyte-Like Cells Integrating 3D Printed Scaffold for Acute Liver Failure Therapy

Acute liver failure (ALF) involves extensive necrosis of liver cells and severe impairment of liver function. Hepatocyte transplantation holds promise for treating ALF by swiftly supporting liver functions and promoting liver regeneration. However, the scarcity of suitable cell sources requires strategies to obtain enough functional hepatocyte-like cells (HLCs) and optimize their in vivo transplantation efficiency. A DNA hexahedral nanostructure (DHN) is developed loaded with microRNA-122 to efficiently induce hepatic differentiation of human adipose-derived mesenchymal stem cells into HLCs. These HLCs can serve as alternative hepatocyte sources, as confirmed by expression of liver-specific genes and proteins, and the restoration of liver functions. To enhance in vivo survival efficiency of HLCs, a versatile scaffold is also created by 3D printing the calcium-cross-linked mixture bioink composed of sodium alginate, gelatin, and silk fibroin with excellent ROS scavenging capabilities. The scaffold is infused with chitosan-DHN hydrogel containing HLCs for tissue engineering orthotopic transplantation in CCl4-induced ALF mice. The transplanted composite scaffold-HLCs successfully repair tissue necrosis in the liver injury area of mice and regulate expressions of genes and proteins associated with inflammation, oxidative stress, and hepatocyte function. Collectively, this study offers a novel approach and strategy for identifying alternative hepatocyte sources and treating ALF. The DNA hexahedral nanostructure-microRNA-122 can induce the differentiation of hADMSC into hepatocyte-like cells. The obtained hepatocyte-like cells, encapsulated in the chitosan hydrogel reintegrated 3D printing scaffold show promising therapeutic efficacy for acute liver failure by restoring hepatocyte functions and providing antioxidative and anti-inflammatory benefits to the damaged liver. image