Investigating the toxic effects induced by iron oxide nanoparticles on neuroblastoma cell line: an integrative study combining cytotoxic, genotoxic and proteomic tools

Abstract

Nanomaterials have gained much attention for their use and benefit in several fields. Iron Oxide Nanoparticles (IONPs) have been used in Biomedicine as contrast agents for imaging cancer cells. However, several studies reported the potential toxicity of those nanoparticles in different models, especially in cells. Therefore, in our present study, we investigated the effects of IONPs on the SH-SY5Y neuroblastoma cell line. We carried out cytotoxic and genotoxic studies to evaluate the phenotypic effects, and proteomic investigation to evaluate the molecular effects and the mechanisms by which this kind of NPs could induce toxicity. Our results showed that the use of three different sizes of IONPs (14, 22 and 30 nm) induced cell detachment, cell morphological changes, size, and concentration-dependent IONP internalization and cell mortality. IONPs induced slight genotoxic damage assayed by modified comet assay without affecting cell cycle, mitochondrial function, membrane integrity, intracellular calcium level, and without inducing ROS generation. All the studies were performed to compare also the effects of IONPs to the ferric iron by incubating cells with equivalent concentration of FeCl₃. In all tests, the NPs exhibited more toxicity than the ferric iron. The proteomic analysis followed by gene ontology and pathway analysis evidenced the effects of IONPs on cytoskeleton, cell apoptosis, and cancer development. Our findings provided more information about IONP effects on human cells and especially on cancer cell line.

Keywords:

• Iron oxide nanoparticles; neuroblastoma; toxicity; proteomics

Acknowledgments

We would like to thank Pr. Lassaad El Mir and his team for providing Iron Oxide Nanoparticles, the EQUIPEX Nano-ID (ANR-10-EQPX-39-01) for the access to the Nano Zetasizer, and Dr. Marie José Stasia and her team for the access to the cell culture laboratory. Thanks also to Dr. Catherine Guette and Dr. Damien Besson for the FASP method adjusted to iTRAQ labeling, and Dr. Sylvie Michelland for her kind assistance in the proteomic study.

Disclosure statement

The authors report no conflicts of interest in this work.

Additional information

Funding

This study was supported by research grants for Dalel Askri from the Tunisian Ministry of Higher Education and Scientific Research, the Auvergne Rhone-Alpes Region (grant No. 16.007278.01) and the French Embassy in Tunisia-Campus, France. The project leading to this publication has also received funding for Dr. Sylvia G. Lehmann from Excellence Initiative of Aix-Marseille University – A*MIDEX, a French ‘Investissements d’Avenir’ program, through its associated Labex SERENADE project.