Cytotoxicity evaluation of functionalized nanocarriers on HepaRG cells

Magnetic mesoporous silica nanoparticles (M-MSN) represent possible tools for cancer diagnostic and therapy. Nevertheless, once functionalized, their biocompatibility must be assessed before to be used as nanocarriers. We... [ view full abstract ]

Magnetic mesoporous silica nanoparticles (M-MSN) represent possible tools for cancer diagnostic and therapy. Nevertheless, once functionalized, their biocompatibility must be assessed before to be used as nanocarriers. We investigated the cellular impact of pristine, PEGylated M-MSN, and lipid bilayer (DMPC) covered M-MSN on HepaRG cells. Based on MTT assay and real-time cell impedance, none of these NPs presented an extensive toxicity on hepatic cells. However, we observed by transmission electron microscopy that the pristine and DMPC M-MSNs were internalized. In comparison, PEG M-MSNs showed a slower cellular uptake. We carried out whole gene expression profiles in a time-and dose-dependent manner and biological data mining with a pathway-driven analysis using Ingenuity Pathways Analysis. The lowest dose tested (1.6 µg/cm²) induced no molecular effect and was defined as ‘No Observed Transcriptional Effect level’. The dose 16 µg/cm² revealed nascent but transient effects. At the highest dose (80 µg/cm²), adverse effects have clearly arisen and increased over time. The limit of biocompatibility for HepaRG cells could be set at 16 µg/cm² for these NPs. At the highest dose, we highlighted the sequence of events that leads to the disruption of hepatobiliary system, elicited by the three types of M-MSNs. The Adverse Outcome Pathway of hepatic cholestasis was implicated. Toxicogenomics applied to cell cultures is an effective tool to characterize and compare the modes of action of many substances. This strategy might be an asset for upstream selection of the safest nanocarriers in the frame of drug safety regulation.