Cytotoxicity of copper- and zinc-containing nanoparticles as health risk factor upon isolated and combined exposure in vivo: Comparative assessmen
М.А. Zemlyanova1,2, М.S. Stepankov1
1Federal Scientific Center for Medical and Preventive Health Risk Management Technologies, 82 Monastyrskaya Str., Perm, 614045, Russian Federation
2Perm State University, 15 Bukireva Str., Perm, 614068, Russian Federation
Provision of healthy and active longevity is among the most significant trends in the scientific and technical development of the Russian Federation. In this respect, it is highly relevant to perform hygienic assessment of health hazards posed by new materials, nano-sized ones included. At present, materials containing zinc and copper nanoparticles (NPs) are being used more and more widely; this goes for both an alloy (Cu-Zn) or using their oxides separately (CuO, ZnO). This results in exposure involving human health risks. Preliminary assessment revealed high potential hazard posed by Cu-Zn NPs due to, among other things, modified cytotoxicity relative to microparticles (MPs) of its chemical analogue because of the nano-sized phase as well as relative to isolated NPs, which is associated with interaction between the two elements upon combined administration into the body. This necessitates investigations with their focus on comparative assessment and analysis of modified cytotoxicity of the Cu-Zn NPs alloy relative to MPs and isolated NPs in vivo.
The study was performed on Wistar rats involving one-time intratracheal exposure to suspensions in the volume of 0.4 cm3, containing Cu-Zn, CuO, ZnO NPs or Cu-Zn MPs in the concentration of 0.013 g/cm3. Cytotoxicity was comparatively assessed 24 hours after the exposure per changes in the cell population in the bronchoalveolar lavage (BAL).
Verification of physical properties confirmed that the tested materials contained NPs. Cu-Zn NPs differed from MPs per their size, specific surface area and total pore volume (up to 2118 times). Cytotoxic effects produced by isolated CuO and ZnO NPs were identified by growing levels of alveolar macrophages (1.9 times higher against the control); declining levels of neutrophil leukocytes, monocytes, lymphocytes and neutrophil to macrophage ratio (up to 3.5 times). Cu-Zn alloy NPs are more cytotoxic relative to MPs (up to 6.1 times) and isolated NPs (up to 29 times) as established per growing levels of neutrophil leukocytes and neutrophil to macrophage ratio and per declining levels of alveolar macrophages, monocytes, and lymphocytes.
Therefore, the study confirms serious health hazard posed by copper- and zinc-containing NPs, which is manifested, among other things, as cytotoxic effects upon intratracheal exposure in vivo. Cytotoxic effects become stronger upon exposure to Cu-Zn alloy NPs relative to MPs due to the nano-sized phase and relative to isolated NPs due to synergy between the two elements. These findings enrich hygienic concepts about cytotoxicity of copper- and zinc-containing NPs upon isolated and combined exposure in vivo, which makes it possible to provide more effective scientific substantiation for preventive measures and mitigating health risks for exposed population.
- Sajid M. Nanomaterials: types, properties, recent advances, and toxicity concerns. Curr. Opin. Environ. Sci. Health, 2022, vol. 25, pp. 100319. DOI: 10.1016/j.coesh.2021.100319
- Molahalli V., Sharma A., Bijapur K., Soman G., Shetty A., Sirichandana B., Maya Patel B.G., Chattham N., Hegde G. Properties, synthesis, and characterization of Cu-based nanomaterials. In book: Copper-based nanomaterials in organic transformations; A. Srivastava, A. Srivastava eds., 2024, vol. 1466, chapter 1, pp. 1–33. DOI: 10.1021/bk-2024-1466.ch001
- Bala N., Saha S., Chakraborty M., Maiti M., Das S., Basu R., Nandy P. Green synthesis of zinc oxide nanoparticles using Hibiscus subdariffa leaf extract: effect of temperature on synthesis, anti-bacterial activity and anti-diabetic activity. RSC Advances, 2015, vol. 5, no. 7, pp. 4993–5003. DOI: 10.1039/C4RA12784F
- Adil M., Bashir S., Bashir S., Aslam Z., Ahmad N., Younas T., Asghar R.M.A., Alkahtani J. [et al.]. Zinc oxide nanoparticles improved chlorophyll contents, physical parameters, and wheat yield under salt stress. Front. Plant Sci., 2022, vol. 13, pp. 932861. DOI: 10.3389/fpls.2022.932861
- Gauba A., Hari S.K., Ramamoorthy V., Vellasamy S., Govindan G., Arasu M.V. The versatility of green synthesized zinc oxide nanoparticles in sustainable agriculture: A review on metal-microbe interaction that rewards agriculture. Physiological and Molecular Plant Pathology, 2023, vol. 125, pp. 102023. DOI: 10.1016/j.pmpp.2023.102023
- Li H., Huang J., Zhang P., Zhang Q. Investigation on tribological behaviors of biodegradable pure Zn and Zn-X (Li, Cu, Ge) binary alloys. J. Mater. Sci. Mater. Med., 2021, vol. 32, no. 12, pp. 149. DOI: 10.1007/s10856-021-06625-4
- Margenot A.J., Rippner D.A., Dumlao M.R., Nezami S., Green P.G., Parikh S.J., McElrone A.J. Copper oxide nanoparticle effects on root growth and hydraulic conductivity of two vegetable crops. Plant Soil, 2018, vol. 431, pp. 333–345. DOI: 10.1007/s11104-018-3741-3
- Agbulut U., Saridemir S., Rajak U., Polat F., Afzal A., Verma T.N. Effects of high-dosage copper oxide nanoparticles addition in diesel fuel on engine characteristics. Energy, 2021, vol. 229, pp. 120611. DOI: 10.1016/j.energy.2021.120611
- Naz S., Gul A., Zia M., Javed R. Synthesis, biomedical applications, and toxicity of
CuO nanoparticles. Appl. Microbiol. Biotechnol., 2023, vol. 107, no. 4, pp. 1039–1061. DOI: 10.1007/s00253-023-12364-z - Rita A., Sivakumar A., Martin Britto Dhas S.A. Influence of shock waves on structural and morphological properties of copper oxide NPs for aerospace applications. J. Nanostruct. Chem., 2019, vol. 9, pp. 225–230. DOI: 10.1007/s40097-019-00313-0
- Morales-Camara S., Choquesillo-Lazarte D., Fernandez B., Rodriguez-Dieguez A., Salcedo-Abraira P., Rojas S. Copper- and zinc-based coordination polymers toward the development of more efficient agrochemicals. ACS Omega, 2025, vol. 10, no. 11, pp. 11274–11281. DOI: 10.1021/acsomega.4c10977
- Zaitseva N.V., Zemlyanova M.A., Stepankov M.S., Ignatova A.M. Copper (II) oxide nano¬par¬ticles toxicity and potential human health hazards. Ekologiya cheloveka, 2021, vol. 28, no. 11, pp. 50–57. DOI: 10.33396/1728-0869-2021-11-50-57 (in Russian).
- Stepankov М.S., Zemlyanova М.А. Assessing potential hazard of zinc oxide nanoparticles to human health. Health Risk Analysis, 2024, no. 4, pp. 123–134. DOI: 10.21668/health.risk/
2024.4.11.eng - Fernandez-Bertolez N., Alba-Gonzalez A., Touzani A., Ramos-Pan L., Mendez J., Reis A.T., Quelle-Regaldie A., Sanchez L. [et al.]. Toxicity of zinc oxide nanoparticles: cellular and behavioral effects. Chemosphere, 2024, vol. 363, pp. 142993. DOI: 10.1016/j.chemosphere.2024.142993
- Singh K.P., Dhasmana A., Rahman Q. Elucidation the toxicity mechanism of zinc oxide nanoparticle using molecular docking approach with proteins. Asian J. Pharm. Clin. Res., 2018, vol. 11, no. 3, pp. 441–446. DOI: 10.22159/ajpcr.2018.v11i3.23384
- Babele P.K. Zinc oxide nanoparticles impose metabolic toxicity by de-regulating proteome and metabolome in Saccharomyces cerevisiae. Toxicol. Rep., 2019, vol. 6, pp. 64–73. DOI: 10.1016/j.toxrep.2018.12.001
- Midander K., Cronholm P., Karlsson H.L., Elihn K., Moller L., Leygraf C., Wallinder I.O. Surface characteristics, copper release, and toxicity of nano- and micrometer-sized copper and copper (II) oxide particles: a cross-disciplinary study. Small, 2009, vol. 5, no. 3, pp. 389–399. DOI: 10.1002/smll.200801220
- Strauch B.M., Hubele W., Hartwig A. Impact of endocytosis and lysosomal acidification on the toxicity of copper oxide nano- and microsized particles: uptake and gene expression related to oxidative stress and the dna damage response. Nanomaterials (Basel), 2020, vol. 10, no. 4, pp. 679. DOI: 10.3390/nano10040679
- Siddiqui M.A., Alhadlaq H.A., Ahmad J., Al-Khedhairy A.A., Musarrat J., Ahamed M. Copper oxide nanoparticles induced mitochondria mediated apoptosis in human hepatocarcinoma cells. PLoS One, 2013, vol. 8, no. 8, pp. e69534. DOI: 10.1371/journal.pone.0069534
- Edelmann M.J., Shack L.A., Naske C.D., Walters K.B., Nanduri B. SILAC-based quantita-tive proteomic analysis of human lung cell response to copper oxide nanoparticles. PLoS One, 2014, vol. 9, no. 12, pp. e114390. DOI: 10.1371/journal.pone.0114390
- Karlsson H.L., Cronholm P., Gustafsson J., Moller L. Copper oxide nanoparticles are highly toxic: a comparison between metal oxide nanoparticles and carbon nanotubes. Chem. Res. Toxicol., 2008, vol. 21, no. 9, pp. 1726–1732. DOI: 10.1021/tx800064j
- Kumbicak U., Cavas T., Cinkilic N., Kumbicak Z., Vatan O., Yilmaz D. Evaluation of in vitro cytotoxicity and genotoxicity of copper–zinc alloy nanoparticles in human lung epithelial cells. Food Chem. Toxicol., 2014, vol. 73, pp. 105–112. DOI: 10.1016/j.fct.2014.07.040
- Onishchenko G.G., Zaitseva N.V., Popova A.Yu., May I.V., Ustinova O.Yu., Trusov P.V., Le T.Kh.Kh., Kleyn S.V. [et al.]. Analiz riska zdorov'yu v strategii gosudarstvennogo sotsial'no-ekonomicheskogo razvitiya [Health risk analysis in the strategy for state socio-economic develop-ment]: monograph in 2 volumes, 2nd ed., rev. and add. In: G.G. Onishchenko, N.V. Zaitseva eds. Moscow; Perm, Perm National Research Polytechnic University Publ., 2024, vol. 1, 580 p. (in Russian).
- Zaitseva N.V., Koldibekova Yu.V., Zemlyanova М.А., Chigvintsev V.М., Ukhabov V.М. Methodical approaches to assessing additional disease risk and loss of life expectancy at birth upon combined exposure to pollutants. Health Risk Analysis, 2025, no. 1, pp. 4–15. DOI: 10.21668/
health.risk/2025.1.01.eng - Griffiths G., Gruenberg J., Marsh M., Wohlmann J., Jones A.T., Parton R.G. Nanoparticle entry into cells; the cell biology weak link. Adv. Drug Deliv. Rev., 2022, vol. 188, pp. 114403. DOI: 10.1016/j.addr.2022.114403
- Lai Y., Chiang P.-C., Blom J.D., Li N., Shevlin K., Brayman T.G., Hu Y., Selbo J.G., Hu L.G. Comparison of in vitro nanoparticles uptake in various cell lines and in vivo pulmonary cellular transport in intratracheally dosed rat model. Nanoscale Res. Lett., 2008, vol. 3, no. 9, pp. 321–329. DOI: 10.1007/s11671-008-9160-2
- Rusakova E., Kosyan D., Sizova E., Miroshnikov S., Sipaylova O. Comparative evaluation of acute toxicity of nanoparticles of zinc, copper and their nanosystems using Stylonychia mytilus. Orient. J. Chem., 2015, vol. 31, no. 1, pp. 105–112. DOI: 10.13005/ojc/31.Special-Issue1.13
- Jain A.K., Singh D., Dubey K., Maurya R., Pandey A.K. Zinc oxide nanoparticles induced gene mutation at the HGPRT locus and cell cycle arrest associated with apoptosis in V-79 cells. J. Appl. Toxicol., 2019, vol. 39, no. 5, pp. 735–750. DOI: 10.1002/jat.3763
- Gupta G., Cappellini F., Farcal L., Gornati R., Bernardini G., Fadeel B. Copper oxide nanoparticles trigger macrophage cell death with misfolding of Cu/Zn superoxide dismutase 1 (SOD1). Part. Fibre Toxicol., 2022, vol. 19, no. 1, pp. 33. DOI: 10.1186/s12989-022-00467-w
- Boyadzhiev A., Halappanavar S. Acute toxicity of metal oxide nanoparticles-role of intracellular localization in vitro in lung epithelial cells. Int. J. Mol. Sci., 2025, vol. 26, no. 17, pp. 8451. DOI: 10.3390/ijms26178451
- Moschini E., Colombo G., Chirico G., Capitani G., Dalle-Donne I., Mantecca P. Biological mechanism of cell oxidative stress and death during short-term exposure to nano CuO. Sci. Rep., 2023, vol. 13, no. 1, pp. 2326. DOI: 10.1038/s41598-023-28958-6
- Niu Y., Tang M. In vitro review of nanoparticles attacking macrophages: Interaction and cell death. Life Sci., 2022, vol. 307, pp. 120840. DOI: 10.1016/j.lfs.2022.120840
- Kaur M., Singh D. Neutrophil chemotaxis caused by chronic obstructive pulmonary disease alveolar macrophages: the role of CXCL8 and the receptors CXCR1/CXCR2. J. Pharmacol. Exp. Ther., 2013, vol. 347, no. 1, pp. 173–180. DOI: 10.1124/jpet.112.201855
- Zadeh F.A., Bokov D.O., Salahdin O.D., Abdelbasset W.K., Jawad M.A., Kadhim M.M., Qasim M.T., Kzar H.H. [et al.]. Cytotoxicity evaluation of environmentally friendly synthesis Cop-per/Zinc bimetallic nanoparticles on MCF-7 cancer cells. Rend. Lincei Sci. Fis., 2022, vol. 33, no. 2, pp. 441–447. DOI: 10.1007/s12210-022-01064-x
- Cao Y., Dhahad H.A., El-Shorbagy M.A., Alijani H.Q., Zakeri M., Heydari A., Bahonar E., Slouf M. [et al.]. Green synthesis of bimetallic ZnO–CuO nanoparticles and their cytotoxicity properties. Sci. Rep., 2021, vol. 11, no. 1, pp. 23479. DOI: 10.1038/s41598-021-02937-1
- Karlsson H.L., Cronholm P., Hedberg Y., Tornberg M., De Battice L., Svedhem S., Wall-inder I.O. Cell membrane damage and protein interaction induced by copper containing nanopar-ticles – Importance of the metal release process. Toxicology, 2013, vol. 313, no. 1, pp. 59–69. DOI: 10.1016/j.tox.2013.07.012
- Zhou B., Chen M., Hao Z., Li L., Zhang Y., Fang B., Shao M., Ren G. [et al.]. Zinc-copper bimetallic nanoplatforms trigger photothermal-amplified cuproptosis and cGAS-STING activation for enhancing triple-negative breast cancer immunotherapy. J. Nanobiotechnology, 2025, vol. 23, no. 1, pp. 137. DOI: 10.1186/s12951-025-03186-4
- Dobrucka R., Kaczmarek M., Lagiedo M., Kielan A., Dlugaszewska J. Evaluation of biologically synthesized Au-CuO and CuO-ZnO nanoparticles against glioma cells and microorganisms. Saudi Pharm. J., 2019, vol. 27, no. 3, pp. 373–383. DOI: 10.1016/j.jsps.2018.12.006

fcrisk.ru

