Hepatotoxicity of Copper Sulfide Nanoparticles Towards Hepatocyte Spheroids Using A Novel Multi-Concave Agarose Chip Method

Abstract

<b>Aim:</b> To explore the hepatotoxicity of copper sulfide nanoparticles (CuSNPs) toward hepatocyte spheroids. <b>Materials & methods:</b> Other than the traditional agarose method to generate hepatocyte spheroids, we developed a multi-concave agarose chip (MCAC) method to investigate changes in hepatocyte viability, morphology, mitochondrial membrane potential, reactive oxygen species and hepatobiliary transporter by CuSNPs. <b>Results:</b> The MCAC method allowed a large number of spheroids to be obtained per sample. CuSNPs showed hepatotoxicity <i>in vitro</i> through a decrease in spheroid viability, albumin/urea production and glycogen deposition. CuSNPs also introduced hepatocyte spheroid injury through alteration of mitochondrial membrane potential and reactive oxygen species, that could be reversed by N-acetyl-l-cysteine. CuSNPs significantly decreased the activity of BSEP transporter by downregulating its mRNA and protein levels. Activity of the MRP2 transporter remained unchanged. <b>Conclusion:</b> We observed the hepatotoxicity of CuSNPs <i>in vitro</i> with associated mechanisms in an advanced 3D culture system.