Abstract

Abstract In this in vitro study, for the first time, we evaluate the effects of simvastatin‐loaded liposome nanoparticles (SIM‐LipoNPs) treatment on fibrosis‐induced liver microtissues, as simvastatin (SIM) has shown potential benefits in the non‐alcoholic fatty liver disease process. We developed multicellular liver microtissues composed of hepatic stellate cells, hepatoblastoma cells and human umbilical vein endothelial cells. The microtissues were supplemented with a combination of palmitic acid and oleic acid to develop fibrosis models. Subsequently, various groups of microtissues were exposed to SIM and SIM‐LipoNPs at doses of 5 and 10 mg/mL. The effectiveness of the treatments was evaluated by analysing cell viability, production of reactive oxygen species (ROS) and nitric oxide (NO), the expression of Kruppel‐like factor (KLF) 2, and pro‐inflammatory cytokines (interleukin(IL)‐1 α, IL‐1 β, IL‐6 and tumour necrosis factor‐α), and the expression of collagen I. Our results indicated that SIM‐LipoNPs application showed promising results. SIM‐LipoNPs effectively amplified the SIM‐klf2‐NO pathway at a lower dosage compatible with a high dosage of free SIM, which also led to reduced oxidative stress by decreasing ROS levels. SIM‐LipoNPs administration also resulted in a significant reduction in pro‐inflammatory cytokines and Collagen I mRNA levels, as a marker of fibrosis. In conclusion, our study highlights the considerable therapeutic potential of using SIM‐LipoNPs to prevent liver fibrosis progress, underscoring the remarkable properties of SIM‐LipoNPs in activating the KLF2‐NO pathway and anti‐oxidative and anti‐inflammatory response.