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Biochemical effects of some CeO2, SiO2, and TiO2 nanomaterials in HepG2 cells

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Abstract

The potential mammalian hepatotoxicity of nanomaterials was explored in dose-response and structure-activity studies in human hepatic HepG2 cells exposed to between 10 and 1000 μg/ml of five different CeO2, three SiO2, and one TiO2-based particles for 3 days. Various biochemical parameters were then evaluated to study cytotoxicity, cell growth, hepatic function, and oxidative stress. Few indications of cytotoxicity were observed between 10 and 30 μg/ml. In the 100 to 300 μg/ml exposure range, a moderate degree of cytotoxicity was often observed. At 1000 μg/ml exposures, all but TiO2 showed a high degree of cytotoxicity. Cytotoxicity per se did not seem to fully explain the observed patterns of biochemical parameters. Four nanomaterials (all three SiO2) decreased glucose 6-phosphate dehydrogenase activity with some significant decreases observed at 30 μg/ml. In the range of 100 to 1000 μg/ml, the activities of glutathione reductase (by all three SiO2) and glutathione peroxidase were decreased by some nanomaterials. Decreased glutathione concentration was also found after exposure to four nanomaterials (all three nano SiO2 particles). In this study, the more responsive and informative assays were glucose 6-phosphate dehydrogenase, glutathione reductase, superoxide dismutase, lactate dehydrogenase, and aspartate transaminase. In this study, there were six factors that contribute to oxidative stress observed in nanomaterials exposed to hepatocytes (decreased glutathione content, reduced glucose 6-phosphate dehydrogenase, glutathione reductase, glutathione peroxidase, superoxide dismutase, and increased catalase activities). With respect to structure-activity, nanomaterials of SiO2 were more effective than CeO2 in reducing glutathione content, glucose 6-phosphate dehydrogenase, glutathione reductase, and superoxide dismutase activities.

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Acknowledgments

We are grateful for the participation of many individuals in this study. Particularly, we thank Drs. Carl Blackman, Michael F. Hughes, and Urmila Kodavanti for reviewing this manuscript as part of EPA clearance procedures. Dr. Xinhua Liang of Missouri University of Science and Technology performed the atomic layer deposition which produced nano SiO2 K1 and SiO2 N2.

Funding

US EPA.

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Authors and Affiliations

  1. Integrated Systems Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, 109 Alexander Drive, B105-03, Durham, NC, 27711, USA

    Kirk T. Kitchin, Brian L. Robinette & Kathleen A. Wallace

  2. Environmental Public Health Division, National Health and Environmental Effects Research Laboratory, 109 Alexander Drive, B105-03, Durham, NC, 27711, USA

    Judy A. Richards & Najwa H. Coates

  3. Institute of Biomedical Studies and Department of Environmental Science, Baylor University, Waco, TX, 76798, USA

    Benjamin T. Castellon

  4. Chemical & Materials Engineering, University of Kentucky, Lexington, KY, 20506-0046, USA

    Eric A. Grulke

Authors
  1. Kirk T. Kitchin
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  2. Judy A. Richards
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  3. Brian L. Robinette
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  4. Kathleen A. Wallace
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  6. Benjamin T. Castellon
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  7. Eric A. Grulke
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Corresponding author

Correspondence to Kirk T. Kitchin.

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The authors declare that they have no conflicts of interest.

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The information in this document has been funded wholly by the U. S. Environmental Protection Agency. It has been subjected to review by the National Health and Environmental Effects Research Laboratory and approved for publication. Approval does not signify that the contents necessarily reflect the views of the Agency, nor does mention of trade names or commercial products that constitute endorsement or recommendation for use.

Electronic supplementary material

Supplementary Table 1

(DOCX 94 kb)

Supplementary Figure 1

% LDH released following metal oxide treatment of HepG2 cells (DOCX 155 kb)

Supplementary Figure 2

% AST released following metal oxide treatment of HepG2 cells (DOCX 165 kb)

Supplementary Figure 3

% ALT released following metal oxide treatment of HepG2 cells (DOCX 215 kb)

Supplementary Figure 4

Effects of nine metal oxides on T BIL concentration (DOCX 172 kb)

Supplementary Figure 5

Effects of nine metal oxides on HepG2 protein content (DOCX 201 kb)

Supplementary Figure 6

Effects of nine metal oxides on GPx activity (DOCX 178 kb)

Supplementary Figure 7

Effects of nine metal oxides on SOD activity (DOCX 199 kb)

Supplementary Figure 8

Effects of nine metal oxides on GSH concentration (DOCX 133 kb)

Supplementary Figure 9

Effects of nine metal oxides on GRD activity (DOCX 204 kb)

Supplementary Figure 10

Effects of nine metal oxides on G6PDH activity (DOCX 189 kb)

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Kitchin, K.T., Richards, J.A., Robinette, B.L. et al. Biochemical effects of some CeO2, SiO2, and TiO2 nanomaterials in HepG2 cells. Cell Biol Toxicol 35, 129–145 (2019). https://doi.org/10.1007/s10565-018-9445-x

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  • DOI: https://doi.org/10.1007/s10565-018-9445-x

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