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

Abstract

Aim: To explore the hepatotoxicity of copper sulfide nanoparticles (CuSNPs) toward hepatocyte spheroids. Materials & methods: 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. Results: The MCAC method allowed a large number of spheroids to be obtained per sample. CuSNPs showed hepatotoxicity in vitro 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. Conclusion: We observed the hepatotoxicity of CuSNPs in vitro with associated mechanisms in an advanced 3D culture system.

Keywords::

• copper sulfide nanoparticles
• hepatobiliary transporters
• hepatotoxicity
• reactive oxygen species
• spheroids

Financial & competing interests disclosure

This work was financially supported by the National Natural Science Foundation of China (nos. 31971319, 31771104), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), and the Jiangsu Provincial Key Laboratory of Radiation Medicine and Protection. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations.

Acknowledgments

The authors would like to thank Suzhou Jestar Mold Technology Co., Ltd for the fabrication of the metal mold.

Additional information

Funding

This work was financially supported by the National Natural Science Foundation of China (nos. 31971319, 31771104), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), and the Jiangsu Provincial Key Laboratory of Radiation Medicine and Protection. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.