Zinc ferrite nanoparticle-induced cytotoxicity and oxidative stress in different human cells

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Abstract

Background

Zinc ferrite nanoparticles (NPs) have shown potential to be used in biomedical field such as magnetic resonance imaging and hyperthermia. However, there is limited information concerning the biological response of zinc ferrite NPs. This study was designed to evaluate the cytotoxicity of zinc ferrite NPs in three widely used in vitro cell culture models: human lung epithelial (A549), skin epithelial (A431) and liver (HepG2) cells. Zinc ferrite NPs were characterized by electron microscopy and dynamic light scattering. Cell viability, cell membrane damage, reactive oxygen species (ROS), glutathione (GSH), mitochondrial membrane potential (MMP), transcriptional level of apoptotic genes were determined in zinc ferrite NPs exposed cells.

Results

Zinc ferrite NPs were almost spherical shaped with an average size of 44 nm. Zinc ferrite NPs induced dose-dependent cytotoxicity (MTT and LDH) and oxidative stress (ROS and GSH) in all three types of cells in the dosage range of 10–40 µg/ml. Transcriptional level of tumor suppressor gene p53 and apoptotic genes (bax, caspase-3 and caspase-9) were up-regulated while the anti-apoptotic gene bcl-2 was down-regulated in cells after zinc ferrite NPs exposure. Furthermore, higher activity of caspase-3 and caspase-9 enzymes was also observed in zinc ferrite NPs treated cells. ROS generation, MMP loss and cell death in all three types of cells were abrogated by N-acetyl cysteine (ROS scavenger), which suggests that oxidative stress might be one of the plausible mechanisms of zinc ferrite NPs cytotoxicity. It is worth mentioning that there was marginally difference in the sensitivity of three cell lines against zinc ferrite NPs exposure. Cytotoxicity of zinc ferrite NPs were in following order; A549 > HepG2 > A431.

Conclusion

Altogether, zinc ferrite NPs induced cytotoxicity and oxidative stress in A549, A431 and HepG2 cells, which is likely to be mediated through ROS generation. This study warrants further investigation to explore the potential mechanisms of toxicity of zinc ferrite NPs in normal cells as well as in animal models.

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Most cited references 42

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Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts.

Martin Lundqvist, Johannes Stigler, Giuliano Elia … (2008)

Nanoparticles in a biological fluid (plasma, or otherwise) associate with a range of biopolymers, especially proteins, organized into the "protein corona" that is associated with the nanoparticle and continuously exchanging with the proteins in the environment. Methodologies to determine the corona and to understand its dependence on nanomaterial properties are likely to become important in bionanoscience. Here, we study the long-lived ("hard") protein corona formed from human plasma for a range of nanoparticles that differ in surface properties and size. Six different polystyrene nanoparticles were studied: three different surface chemistries (plain PS, carboxyl-modified, and amine-modified) and two sizes of each (50 and 100 nm), enabling us to perform systematic studies of the effect of surface properties and size on the detailed protein coronas. Proteins in the corona that are conserved and unique across the nanoparticle types were identified and classified according to the protein functional properties. Remarkably, both size and surface properties were found to play a very significant role in determining the nanoparticle coronas on the different particles of identical materials. We comment on the future need for scientific understanding, characterization, and possibly some additional emphasis on standards for the surfaces of nanoparticles.

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Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader.

H. Wang, J. Joseph (1999)

Oxidative stress (OS) has been implicated in various degenerative diseases in aging. In an attempt to quantify OS in a cell model, we examined OS induced by incubating for 30 min with various free radical generators in PC12 cells by using the dichlorofluorescein (DCF) assay, modified for use by a fluorescent microplate reader. The nonfluorescent fluorescin derivatives (dichlorofluorescin, DCFH), after being oxidized by various oxidants, will become DCF and emit fluorescence. By quantifying the fluorescence, we were able to quantify the OS. Our results indicated that the fluorescence varied linearly with increasing concentrations (between 0.1 and 1 mM) of H2O2 and 2,2'-azobios(2-amidinopropane) dihydrochloride (AAPH; a peroxyl radical generator). By contrast, the fluorescence varied as a nonlinear response to increasing concentrations of 3-morpholinosydnonimine hydrochloride (SIN-1; a peroxynitrite generator), sodium nitroprusside (SNP; a nitric oxide generator), and dopamine. Dopamine had a biphasic effect; it decreased the DCF fluorescence, thus acting as an antioxidant, at concentrations <500 microM in cells, but acted as a pro-oxidant by increasing the fluorescence at 1 mM. While SNP was not a strong pro-oxidant, SIN-1 was the most potent pro-oxidant among those tested, inducing a 70 times increase of fluorescence at a concentration of 100 microM compared with control. Collectively, due to its indiscriminate nature to various free radicals, DCF can be very useful in quantifying overall OS in cells, especially when used in conjunction with a fluorescent microplate reader. This method is reliable and efficient for evaluating the potency of pro-oxidants and can be used to evaluate the efficacy of antioxidants against OS in cells.

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Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique.

Richard Murdock, Laura K Braydich-Stolle, Amanda Schrand … (2008)

The need to characterize nanoparticles in solution before assessing the in vitro toxicity is a high priority. Particle size, size distribution, particle morphology, particle composition, surface area, surface chemistry, and particle reactivity in solution are important factors which need to be defined to accurately assess nanoparticle toxicity. Currently, there are no well-defined techniques for characterization of wet nanomaterials in aqueous or biological solutions. Previously reported nanoparticle characterization techniques in aqueous or biological solutions have consisted of the use of ultra-high illumination light microscopy and disc centrifuge sedimentation; however, these techniques are limited by the measurement size range. The current study focuses on characterizing a wide range of nanomaterials using dynamic light scattering (DLS) and transmission electron microscopy, including metals, metal oxides, and carbon-based materials, in water and cell culture media, with and without serum. Cell viability and cell morphology studies were conducted in conjunction with DLS experiments to evaluate toxicological effects from observed agglomeration changes in the presence or absence of serum in cell culture media. Observations of material-specific surface properties were also recorded. It was also necessary to characterize the impact of sonication, which is implemented to aid in particle dispersion and solution mixture. Additionally, a stock solution of nanomaterials used for toxicology studies was analyzed for changes in agglomeration and zeta potential of the material over time. In summary, our results demonstrate that many metal and metal oxide nanomaterials agglomerate in solution and that depending upon the solution particle agglomeration is either agitated or mitigated. Corresponding toxicity data revealed that the addition of serum to cell culture media can, in some cases, have a significant effect on particle toxicity possibly due to changes in agglomeration or surface chemistry. It was also observed that sonication slightly reduces agglomeration and has minimal effect on particle surface charge. Finally, the stock solution experienced significant changes in particle agglomeration and surface charge over time.

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Author and article information

Contributors

Hisham A. Alhadlaq: hhadlaq@ksu.edu.sa

Mohd Javed Akhtar: mohd.j.akhtar@gmail.com

Maqusood Ahamed: +966114698781 , maqusood@gmail.com

Journal

Journal ID (nlm-ta): Cell Biosci

Journal ID (iso-abbrev): Cell Biosci

Title: Cell & Bioscience

Publisher: BioMed Central (London )

ISSN (Electronic): 2045-3701

Publication date (Electronic): 17 September 2015

Publication date PMC-release: 17 September 2015

Publication date Collection: 2015

Volume: 5

Electronic Location Identifier: 55

Affiliations

[ ]Department of Physics and Astronomy, College of Science, King Saud University, Riyadh, 11451 Saudi Arabia

[ ]King Abdullah Institute for Nanotechnology, King Saud University, Riyadh, 11451 Saudi Arabia

Article

Publisher ID: 46

DOI: 10.1186/s13578-015-0046-6

PMC ID: 4574116

SO-VID: 32873921-855b-4fac-ab46-d7a16520397f

License:

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

History

Date received : 30 June 2015

Date accepted : 7 September 2015

Custom metadata

ScienceOpen disciplines: Cell biology

Keywords: zinc ferrite nanoparticles,a549,a431,hepg2,human health,cytotoxicity,reactive oxygen species

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ScienceOpen disciplines: Cell biology

Keywords: zinc ferrite nanoparticles, a549, a431, hepg2, human health, cytotoxicity, reactive oxygen species

Zinc ferrite nanoparticle-induced cytotoxicity and oxidative stress in different human cells

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Abstract

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Stress signalling in stem cells

Most cited references 42

Nanoparticle size and surface properties determine the protein corona with possible implications for biological impacts.

Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader.

Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique.

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