Microfluidic fabricated cell-laden microgels aggregated into artificial liver microtissue to ameliorate drug-induced liver injury.

Abstract

Acute liver failure (ALF) is a life-threatening disease for which orthotopic liver transplantation remains one of the most effective treatments. Cell therapy, particularly through the transplantation of artificial bioengineered liver grafts, emerges as a promising alternative to the whole liver transplantation in ALF management. However, the limited availability of functional human hepatocytes, the constrained techniques for swiftly creating artificial bioengineered liver grafts, and the poor cellular survival post-transplantation present significant challenges for advancing clinical applications in liver regeneration. In this study, we introduce a novel approach utilizing microfluidics to create cell-laden microgels for fabricating artificial liver microtissues, in which the controlled hepatic differentiation of stem cells and the development of liver microtissues are incorporated. The cell-laden ARGH microgels (GelMA/HAMA microgels containing AuNCs-miRNA) serving as building blocks for rapid engineering tissue construction, demonstrate the capacity for the one-step promotion of hepatic differentiation of stem cells for a short period of time. Additionally, the bioengineered liver microtissues exhibit multiple biological functionalities, including scavenging ROS, modulating inflammatory environment, and promoting angiogenesis, and can maintain long-term stability after transplantation in vivo. Furthermore, our findings reveal that the in vivo transplantation of the bioengineered liver microtissues offers therapeutic benefits in two drug-induced liver failure experimental models. The transcriptome sequencing analysis further elucidates that the microtissue transplantation ameliorated liver failure by improving liver biofunctions, reducing inflammation and oxidative stress, thereby promoting liver regeneration. Overall, this study presents an effective approach leveraging functional biomaterials and microfluidic techniques to fabricate artificial liver microtissues for liver failure treatment.