(610c) Fibrotic Stiffness Promotes Metabolic Dysfunction in Primary Hepatocytes

Metabolic-associated fatty liver disease (MAFLD) is the most common liver disease worldwide, affecting approximately 30% of the population. MAFLD can progress from benign lipid accumulation to nonalcoholic steatohepatitis (NASH) and potentially liver cirrhosis and organ failure. Increased liver stiffness is established as an indicator of disease progression and is linked to poor clinical outcomes. Within liver, hepatocytes are the primary functional cell and accumulate excess lipids in MAFLD. However, the potential role of liver stiffness in potentiating hepatocyte metabolic dysfunction in MAFLD is largely unknown. To explore the role of matrix stiffness in directing hepatocyte function, we cultured primary hepatocytes on our biomimetic platform, BEASTS (Bio-Engineered Adhesive Siloxane substrate with Tunable Stiffness). BEASTS is unique from most commonly used methods for studying matrix stiffness and cell behavior in that it promotes cell attachment without adhesive proteins. With a protein-free surface, the effect of mechanical cues on cells can be isolated while confounding interactions of gel mechanical properties and ligand presentation are avoided. Hepatocytes were cultured on BEASTS tuned to 2 kPa (healthy), 25 kPa, or 55 kPa (advanced fibrosis). In hepatocytes cultured on fibrotic stiffness, we observed elevated intracellular lipids along with changes to lipid metabolic gene expression. Further, hepatocytes cultured on fibrotic stiffness exhibited a shift in bioenergetics demonstrated by increased glycolysis. Decreased glutathione was also measured following culture on fibrotic matrix stiffness, indicating disrupted redox balance. Our data collectively suggest fibrotic liver stiffness is a driver of MAFLD-like dysfunctional changes in hepatocytes. This work contributes to the understanding of hepatocyte response to increased liver stiffness and informs potential approaches for mechano-therapeutic development for treating MAFLD.