Cell mimicry is an emerging concept, which consists of a hierarchical bottom-up assembly strategy aiming to design and engineer the structure and function of nature’s own cells and organelles. Artificial organelles (AOs) are intracellular nanoreactors that equip their host cells with missing function through the loading of natural or artificial enzymes (AEs). Recently, emerging efforts are focused on AC-therapeutics resulting in an increasing number of successful intracellular catalysis using AOs. However, over-simplified cell models have typically been employed. These often lack key features, failing to accurately portray the characteristic of the targeted condition. For instance, fatty hepatic cells cultured in galactose media are a more accurate model for Non-alcoholic Fatty Liver Disease (NAFLD) compared to regularly employed hepatic cells grown in glucose-rich media. Galactose-containing media ensures the dependence on mitochondrial oxidative phosphorylation for energy production. Therefore, we considered lipid enriched HepG2 cells grown in galactose media for the biological evaluation of AOs (Fig.1). The assembled AOs were based on phospholipid-block copolymer hybrid vesicles (HVs) focusing on cell viability, mitochondrial oxygen consumption, uptake efficacy, intracellular fate and intracellular catalytic activity capability. In a first step, we compared the cellular response of two distinct polyanionic homopolymers (poly(cholesteryl methacrylate)-<i>b</i>-poly(2-carboxyethyl acrylate) and poly(cholesteryl methacrylate)-<i>b</i>-poly(methacrylic acid)) in order to select the hydrophilic extension for the amphiphilic block copolymer. Secondly, the successful HVs assembly was confirmed and the non-toxic dose for HepG2 cells was identified. Based on this assessment, the intracellular fate of the HVs was followed focusing on intracellular retention and lysosomal escape. Taken together, we for the first time considered AOs in fatty hepatic cells moving this concept a step closer to be considered in the context of NAFLD.

Polyanionic nanoparticles and their interaction with fatty hepatocytes

ACS Spring Meeting 2021

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.