Assessment of Combined and Comparative Toxicity of Zinc Oxide and Copper Oxide Nanoparticles in the In Vivo Experiment

Introduction: Apart from the targeted production of many metal and oxide nanomaterials with desired properties (socalled engineered nanoparticles) and their wide and diverse use in engineering, science, and medicine, even more important potential health risks to human health may be associated with some old technologies. Non-engineered metal oxide nanoparticles (MeO-NPs) generated spontaneously during arc welding, production of steel and non-ferrous metals, pollute the workplace and ambient air along with submicron particles (> 100 nm) of the same metal oxides. The most important sources of by-production of zinc oxide nanoparticles include primary smelting or re-smelting of brass, an alloy of copper and zinc of various proportions (sometimes with a much lower amount of lead, tin, and other metals). The rationale of the study of the comparative and combined toxicity of zinc oxide nanoparticles (ZnO-NPs) and copper oxide nanoparticles (CuO-NPs) is their simultaneous presence in aerosol emissions from brass metallurgy.

The objective of our study was to estimate the comparative and combined toxicity of ZnO-NPs and CuO-NPs.

Methods: Stable suspensions of MeO-NPs obtained by laser ablation of 99.99 % pure zinc and copper under a layer of deionized water, were injected intraperitoneally 18 times during 6 weeks to outbred male rats separately (in equal mass doses) or in combination for a comparative assessment and analysis of the type of the combined exposure to the studied nanoparticles for a large number of signs (including DNA fragmentation).

Results: We established that, judging by some direct and indirect evidence, the subchronic effect of ZnO-NPs on the body was more detrimental than that of CuO-NPs. The mathematical description of the results using the response surface method showed that, similar to other previously studied binary toxic combinations, the response of the body to the combined exposure to CuO and ZnO nanoparticles was characterized by a complex interaction of various types of combined toxicity, depending on the effect it was evaluated for, the levels of the effect and doses. When analyzing the type of the combined effect of ZnO-NPs and CuO-NPs, we observed both the antagonism and additivity according to some indicators of the state of the body, which makes us evaluate their combined exposure as dangerous.

1. Minigalieva IA, Katsnelson BA, Privalova LI, et al. Combined subchronic toxicity of aluminum (III), titanium (IV) and silicon (IV) oxide nanoparticles and its alleviation with a complex of bioprotectors. Int J Mol Sci. 2018;19(3):837. doi: 10.3390/ijms19030837

2. Nartsissov RP. [Application of n-nitrotetrazole violet for quantitative cytochemistry of human lymphocyte dehydrogenases.] Arkhiv Anatomii, Gistologii i Embriologii. 1969;(5):85–91. (In Russian).

3. Wang S, Ding M, Duan X, et al. Detection of the single nucleotide polymorphism at position rs2735940 in the human telomerase reverse transcriptase gene by the introduction of a new restriction enzyme site for the PCR-RFLP Assay. Ann Clin Lab Sci. 2017;47(5):546–550

4. Sharifiyazdi H, Mirzaei A, Ghanaatian Z. Characterization of polymorphism in the FSH receptor gene and its impact on some reproductive indices in dairy cows. Anim Reprod Sci. 2018;188:45–50. doi: 10.1016/j.anireprosci.2017.11.006

5. Ningombam SS, Chhungi V, Newmei MK, et al. Differential distribution and association of FTO rs9939609 gene polymorphism with obesity: A cross-sectional study among two tribal populations of India with East-Asian ancestry. Gene. 2018;647:198–204. doi: 10.1016/j.gene.2018.01.009

6. Katsnelson BA, Minigaliyeva IA, Panov VG, et al. Some patterns of metallic nanoparticles’ combined subchronic toxicity as exemplified by a combination of nickel and manganese oxide nanoparticles. Food Chem Toxicol. 2015;86:351–64. doi: 10.1016/j.fct.2015.11.012

7. Wang B, Feng WY, Wang TC, et al. Acute toxicity of nanoand micro-scale zinc powder in healthy adult mice. Toxicol Lett. 2006;161(2):115–123. doi: 10.1016/j.toxlet.2005.08.007

8. Cho WS, Duffin R, Howie S, et al. Progressive severe lung injury by zinc oxide nanoparticles; the role of Zn2+ dissolution inside lysosomes. Part Fibre Toxicol. 2011;8:27. doi: 10.1186/1743-8977-8-27

9. Adamcakova-Dodd A, Stebounova LV, Kim JS, et al. Toxicity assessment of zinc oxide nanoparticles using sub-acute and sub-chronic murine inhalation models. Part Fibre Toxicol. 2014;11:15. doi: 10.1186/1743-8977-11-15

10. Filippi C, Pryde A, Cowan P, et al. Toxicology of ZnO and TiO2 nanoparticles on hepatocytes: impact on metabolism and bioenergetics. Nanotoxicology. 2015;9(1):126–34. doi: 10.3109/17435390.2014.895437

11. Choi J, Kim H, Kim P, et al. Toxicity of zinc oxide nanoparticles in rats treated by two different routes: single intravenous injection and single oral administration. J Toxicol Environ Health. 2015;78(4):226–43. doi: 10.1080/15287394.2014.949949

12. Jacobsen NR, Stoeger T, van den Brule S, et al. Acute and subacute pulmonary toxicity and mortality in mice after intratracheal instillation of ZnO nanoparticles in three laboratories. Food Chem Toxicol. 2015;85:84–95. doi: 10.1016/j.fct.2015.08.008

13. Gao F, Ma N, Zhou H, et al. Zinc oxide nanoparticlesinduced epigenetic change and G2/M arrest are associated with apoptosis in human epidermal keratinocytes. Int J Nanomedicine. 2016;11:3859–74. doi: 10.2147/IJN.S107021

14. Wei Y, Li Y, Jia J, Jiang Y, et al. Aggravated hepatotoxicity occurs in aged mice but not in young mice after oral exposure to zinc oxide nanoparticles. NanoImpact. 2016;3–4:1–11. doi: 10.1016/j.impact.2016.09.003

15. Ng CT, Yong LQ, Hande MP, et al. Zinc oxide nanoparticles exhibit cytotoxicity and genotoxicity through oxidative stress responses in human lung fibroblasts and Drosophila melanogaster. Int J Nanomedicine. 2017;12:1621–1637. doi: 10.2147/IJN.S124403

16. Chen Z, Meng H, Xing G, et al. Acute toxicological effects of copper nanoparticles in vivo. Toxicol Lett. 2006;163(2):109–20. doi: 10.1016/j.toxlet.2005.10.003

17. Karlsson H, Cronholm P, Gustafsson J, Möller L. Copper oxide nanoparticles are highly toxic: a comparison between metal oxide nanoparticles and carbon nanotubes. Chem Res Toxicol. 2008;21(9):1726–32. doi: 10.1021/tx800064j

18. Studer AM, Limbach LK, Van Duc L, et al. Nanoparticle cytotoxicity depends on intracellular solubility: comparison of stabilized copper metal and degradable copper oxide nanoparticles. Toxicol Lett. 2010;197(3):169–74. doi: 10.1016/j.toxlet.2010.05.012

19. Bondarenko O, Ivask A, Käkinen A, Kahru A. Sub-toxic effects of CuO nanoparticles on bacteria: kinetics, role of Cu ions and possible mechanisms of action. Environ Pollut. 2012;169:81–9. doi: 10.1016/j.envpol.2012.05.009

20. Magaye R, Zhao J, Bowman L, Ding M. Genotoxicity and carcinogenicity of cobalt-, nickel- and copper-based nanoparticles. Exp Ther Med. 2012;4(4):551–561. doi: 10.3892/etm.2012.656

21. Pang C, Selck H, Misra SK, et al. Effects of sedimentassociated copper to the deposit-feeding snail, Potamopyrgus antipodarum: a comparison of Cu added in aqueous form or as nano- and micro-CuO particles. Aquat Toxicol. 2012;106–107:114–22. doi: 10.1016/j.aquatox.2011.10.005

22. Akhtar MJ, Kumar S, Alhadlaq HA, Alrokayan SA, AbuSalah KM, Ahamed M. Dose-dependent genotoxicity of copper oxide nanoparticles stimulated by reactive oxygen species in human lung epithelial cells. Toxicol Ind Health. 2013;32(5):809–21. doi: 10.1177/0748233713511512

23. Alarifi S, Ali D, Verma A, Alakhtani S, Ali BA. Cytotoxicity and genotoxicity of copper oxide nanoparticles in human skin keratinocytes cells. Int J Toxicol. 2013;32(4):296–307. doi: 10.1177/1091581813487563

24. Cuillel M, Chevallet M, Charbonnier P, et al. Interference of CuO nanoparticles with metal homeostasis in hepatocytes under sub-toxic conditions. Nanoscale. 2014;6(3):1707–15. doi: 10.1039/c3nr05041f

25. Gomes T, Araújo O, Pereira R, Almeida AC, Cravo A, Bebianno MJ. Genotoxicity of copper oxide and silver nanoparticles in the mussel Mytilus galloprovincialis. Mar Environ Res. 2013;84:51–9. doi: 10.1016/j.marenvres.2012.11.009

26. Xu J, Li Z, Xu P, Xiao L, Yang Z. Nanosized copper oxide induces apoptosis through oxidative stress in podocytes. Arch Toxicol. 2013;87(6):1067–73. doi: 10.1007/s00204-012-0925-0

27. Privalova LI, Katsnelson BA, Loginova NV, et al. Some characteristics of free cell population in the airways of rats after intratracheal instillation of copper-containing nanoscale particles. Int J Mol Sci. 2014;15(11):21538–53. doi: 10.3390/ijms151121538

28. Privalova LI, Katsnelson BA, Loginova NV, et al. Subchronic toxicity of copper oxide nanoparticles and its attenuation with the help of a combination of bioprotectors. Int J Mol Sci. 2014;15(7):12379–406. doi: 10.3390/ijms150712379

29. Li Y, Zheng Y, Qian J, et al. Preventive effects of zinc against psychological stress-induced iron dyshomeostasis, erythropoiesis inhibition, and oxidative stress status in rats. Biol Trace Elem Res. 2012;147(1–3):285–91. doi: 10.1007/s12011-011-9319-z

30. Varaksin AN, Katsnelson BA, Panov VG, et al. Some considerations concerning the theory of combined toxicity: a case study of subchronic experimental intoxication with cadmium and lead. Food Chem Toxicol. 2014;64:144–56. doi: 10.1016/j.fct.2013.11.024

31. Panov VG, Katsnelson BA, Varaksin AN, et al. Further development of mathematical description for combined toxicity: A case study of lead–fluoride combination. Toxicol Rep. 2015;2:297–307. doi: 10.1016/j.toxrep.2015.02.002

32. Katsnelson BA, Minigaliyeva IA, Panov VG, et al. Some patterns of metallic nanoparticles’ combined subchronic toxicity as exemplified by a combination of nickel and manganese oxide nanoparticles. Food Chem Toxicol. 2015;86:351–64. doi: 10.1016/j.fct.2015.11.012

33. Katsnelson BA, Tsepilov NA, Panov VG, et al. Applying theoretical premises of binary toxicity mathematical modeling to combined impacts of chemical plus physical agents (A case study of moderate subchronic exposures to fluoride and static magnetic field). Food Chem Toxicol. 2016;95:110–20. doi: 10.1016/j.fct.2016.06.024

34. Minigalieva IA, Katsnelson BA, Panov VG, et al. Experimental study and mathematical modeling of toxic metals combined action as a scientific foundation for occupational and environmental health risk assessment. A summary of results obtained by the Ekaterinburg research team (Russia). Toxicol Rep. 2017;4:194–201. doi: 10.1016/j.toxrep.2017.04.002

35. Bremner I, Beattie JH. Copper and zinc metabolism in health and disease: speciation and interactions. Proc Nutr Soc. 1995;54(2):489–99. doi: 10.1079/PNS19950017