Induction of oxidative stress and cytotoxicity by carbon nanomaterials is dependent on physical properties

Abstract

In this study, we investigated the mechanisms involved in multi-wall carbon particles/nanomaterials (MWCNM) induced cytotoxicity using human embryonic kidney (HEK293) cells and to assess the effect of physicochemical properties on the cytotoxicity and oxidative stress induced by the carbon nanomaterials (CNM). To elucidate the possible mechanisms of CNM-induced cytotoxicity, cell viability (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide [MTT assay]), cell membrane damage (lactate dehydrogenase enzyme [LDH] assay), reduced glutathione (GSH), interleukin-8 (IL-8) and lipid peroxidation levels were quantitatively assessed under carbon nanomaterials exposed (48 h) conditions. Exposure of different sizes of four CNM at dosage levels between 3 and 300 μg/mL decreased cell viability in a concentration- and size-dependent manner. Exposure of CNM (10-100 μg/mL) to HEK cells resulted in size-, surface area- and concentration-dependent cell membrane damage, increased production of IL-8, increased thiobarbituric acid reactive substances (TBARS) and decreased intracellular glutathione levels. In summary, the physical properties of carbon nanoparticles may alter the CNM-induced concentration-dependent cytotoxicity and oxidative stress.

Keywords

Carbon nanoparticles HEK293 cells oxidative stress cytotoxicity in vitro LDH MTT IL-8 MDA glutathione TBARS

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References

Akerboom TPM , Sies H. ( 1981) Assay of glutathione, glutathione disulfide, and glutathione mixed disulfides in biological samples. Methods Enzymology 77: 373-382.

Becker S. , Soukup JM , and Gallagher JE ( 2002) Differential particulate air pollution induced oxidant stress in human granulocytes, monocytes and alveolar macrophages. Toxicology In Vitro 16(3): 209-218.

Berne BJ , Pecora R. ( 1975) Dynamic Light Scattering. New York : John Wiley.

Bradford MM ( 1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254.

Crystal C. , Karin H. , Muller KK , Koziol K. , Jeremy NS , Paul A. , et al. (2009) Toxicity and imaging of multi-walled carbon nanotubes in human macrophage cells. Biomaterials 30(25): 4152-4160.

Cui D. , Tian F. , Ozkan CS , Wang M. , and Gao H. ( 2005) Effect of single wall carbon nanotubes on human HEK293 cells. Toxicology Letters 155(1): 73-85.

Denizot F. , Lang R. ( 1986) Rapid colorimetric assay for cell growth and survival: modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. Journal of Immunological Methods 89(2): 271-277.

Fen W. , Feng G. , Minbo L. , Huihui Y. , Yongping H. , and Jianwen L. ( 2009) Oxidative stress contributes to silica nanoparticle-induced cytotoxicity in human embryonic kidney cells. Toxicology In Vitro 23(5): 808-815.

Hussain SM , Frazier JM ( 2002) Cellular toxicity of hydrazine in primary rat hepatocytes . Toxicology Science 69(2): 424-432.

10.

Ji LL , Chen Y. , and Wang ZT ( 2008) The toxic effect of pyrrolizidine alkaloid clivorine on the human embryonic kidney 293 cells and its primary mechanism. Experimental and Toxicologic Pathology 60(1): 87-93.

11.

Kipen HM , Laskin DL ( 2005) Smaller is not always better: nanotechnology yields nanotoxicology . The American Journal of Physiology - Lung Cellular and Molecular Physiology 289(5): 696-697.

12.

Maynard AD , Baron PA , Foley M. , Shvedova AA , Kisin ER , and Castranova V. ( 2004) Exposure of carbon nanotube material: aerosol release during the handling of unrefined single walled carbon nanotube material. Journal of Toxicology and Environmental Health Part A 67(1): 87-108.

13.

Oberdorster E. ( 2004) Manufactured nanomaterials (Fullerenes, C60) induce oxidative stress in the brain of juvenile largemouth bass. Environmental Health Perspectives 112(10): 1058-1062.

14.

Ohkawa H. , Ohishi N. , and Yagi K. ( 1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95(2): 351-358.

15.

Otwinowski Z. , Minor W. ( 1997) Macromolecular Crystallography, Part A . Vol 276. San Diego, CA: Academic Press Inc, 307-326.

16.

Park EJ , Choi J. , Park Y. , and Park K. ( 2008) Oxidative stress induced by cerium oxide nanoparticles in cultured BEAS-2B cells. Toxicology 1(2): 90-100.

17.

Pulskamp K. , Diabate S. , and Krug HF ( 2007) Carbon nanotubes show no sign of acute toxicity but induce intracellular reactive oxygen species in dependence on contaminants . Toxicology Letters 168(1): 58-74.

18.

Shimida A. , Kawamura N. , Okajima M. , Kaewamatawong T. , Inoue H. , and Morita T. ( 2006) Translocation pathway of the intratracheally instilled ultrafine particles from the lung into blood circulation in the mouse. Toxicology Pathology 34(7): 949-957.

19.

Shvedova AA , Castranova V. , Kisin ER , Schwegler-Berry D. , Murray AR , Gandelsman VZ , et al. (2003) Exposure to carbon nanotube material: assessment of nanotube cytotoxicity using human keratinocyte cells. Journal of Toxicology and Environmental Health A 66(20): 1909-1926.

20.

Stone V. , Johnston H. , and Clift MJ ( 2007) Air pollution, ultrafine and nanoparticles toxicology: cellular and molecular interactions. IEEE Transactions on Nanobioscience 6(4): 331-340.