Skip to main content

Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549

Archives of Toxicology volume 85, pages 743–750 (2011)Cite this article

Abstract

Nanomaterials, especially silver nanoparticles (Ag NPs), are used in a rapidly increasing number of commercial products. Accordingly, the hazards associated with human exposure to nanomaterials should be investigated to facilitate the risk assessment process. A potential route of exposure to NPs is through the respiratory system. In the present study, we investigated the effects of well-characterized PVP-coated Ag NPs and silver ions (Ag+) in the human, alveolar cell line, A549. Dose-dependent cellular toxicity caused by Ag NPs and Ag+ was demonstrated by the MTT and annexin V/propidium iodide assays, and evidence of Ag NP uptake could be measured indirectly by atomic absorption spectroscopy and flow cytometry. The cytotoxicity of both silver compounds was greatly decreased by pretreatment with the antioxidant, N-acetyl-cysteine, and a strong correlation between the levels of reactive oxygen species (ROS) and mitochondrial damage (r s = −0.8810; p = 0.0039) or early apoptosis (r s = 0.8857; p = 0.0188) was observed. DNA damage induced by ROS was detected as an increase in bulky DNA adducts by 32P postlabeling after Ag NP exposure. The level of bulky DNA adducts was strongly correlated with the cellular ROS levels (r s = 0.8810, p = 0.0039) and could be inhibited by antioxidant pretreatment, suggesting Ag NPs as a mediator of ROS-induced genotoxicity.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Arora S, Jain J, Rajwade JM, Paknikar KM (2008) Cellular responses induced by silver nanoparticles: in vitro studies. Toxicol Lett 179(2):93–100

    PubMed  Article  CAS  Google Scholar 

  • AshaRani PV, Low Kah MG, Hande MP, Valiyaveettil S (2009) Cytotoxicity and genotoxicity of silver nanoparticles in human cells. ACS Nano 3(2):279–290

    PubMed  Article  CAS  Google Scholar 

  • Auffan M, Rose J, Bottero JY, Lowry GV, Jolivet JP, Wiesner MR (2009) Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective. Nat Nanotechnol 4(10):634–641

    PubMed  Article  CAS  Google Scholar 

  • Bak H, Autrup H, Thomsen BL, Tjonneland A, Overvad K, Vogel U, Raaschou-Nielsen O, Loft S (2006) Bulky DNA adducts as risk indicator of lung cancer in a Danish case-cohort study. Int J Cancer 118(7):1618–1622

    PubMed  Article  CAS  Google Scholar 

  • Braydich-Stolle L, Hussain S, Schlager JJ, Hofmann MC (2005) In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. Toxicol Sci 88(2):412–419

    PubMed  Article  CAS  Google Scholar 

  • Carlson C, Hussain SM, Schrand AM, Braydich-Stolle LK, Hess KL, Jones RL, Schlager JJ (2008) Unique cellular interaction of silver nanoparticles: size-dependent generation of reactive oxygen species. J Phys Chem B 112(43):13608–13619

    PubMed  Article  CAS  Google Scholar 

  • Foldbjerg R, Olesen P, Hougaard M, Dang DA, Hoffmann HJ, Autrup H (2009) PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. Toxicol Lett 190(2):156–162

    PubMed  Article  CAS  Google Scholar 

  • Hsin YH, Chen CF, Huang S, Shih TS, Lai PS, Chueh PJ (2008) The apoptotic effect of nanosilver is mediated by a ROS- and JNK-dependent mechanism involving the mitochondrial pathway in NIH3T3 cells. Toxicol Lett 179(3):130–139

    PubMed  Article  CAS  Google Scholar 

  • Hussain SM, Hess KL, Gearhart JM, Geiss KT, Schlager JJ (2005) In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicol In Vitro 19(7):975–983

    PubMed  Article  CAS  Google Scholar 

  • Hussain SM, Javorina AK, Schrand AM, Duhart HM, Ali SF, Schlager JJ (2006) The interaction of manganese nanoparticles with PC-12 cells induces dopamine depletion. Toxicol Sci 92(2):456–463

    PubMed  Article  CAS  Google Scholar 

  • Ip M, Lui SL, Poon VKM, Lung I, Burd A (2006) Antimicrobial activities of silver dressings: an in vitro comparison. J Med Microbiol 55(1):59–63

    PubMed  Article  CAS  Google Scholar 

  • Kim S, Choi JE, Choi J, Chung KH, Park K, Yi J, Ryu DY (2009) Oxidative stress-dependent toxicity of silver nanoparticles in human hepatoma cells. Toxicol In Vitro 23(6):1076–1084

    PubMed  Article  CAS  Google Scholar 

  • Kweon MH, Adhami VM, Lee JS, Mukhtar H (2006) Constitutive overexpression of Nrf2-dependent heme oxygenase-1 in A549 cells contributes to resistance to apoptosis induced by epigallocatechin 3-gallate. J Biol Chem 281(44):33761–33772

    PubMed  Article  CAS  Google Scholar 

  • Limbach LK, Wick P, Manser P, Grass RN, Bruinink A, Stark WJ (2007) Exposure of engineered nanoparticles to human lung epithelial cells: influence of chemical composition and catalytic activity on oxidative stress. Environ Sci Technol 41(11):4158–4163

    PubMed  Article  CAS  Google Scholar 

  • Lorenzen A, Kennedy SW (1993) A fluorescence-based protein assay for use with a microplate reader. Anal Biochem 214(1):346–348

    PubMed  Article  CAS  Google Scholar 

  • Melaiye A, Sun Z, Hindi K, Milsted A, Ely D, Reneker DH, Tessier CA, Youngs WJ (2005) Silver(I)-imidazole cyclophane gem-diol complexes encapsulated by electrospun tecophilic nanofibers: formation of nanosilver particles and antimicrobial activity. J Am Chem Soc 127(7):2285–2291

    PubMed  Article  CAS  Google Scholar 

  • Miura N, Shinohara Y (2009) Cytotoxic effect and apoptosis induction by silver nanoparticles in HeLa cells. Biochem Biophys Res Commun 390(3):733–737

    PubMed  Article  CAS  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63

    PubMed  Article  CAS  Google Scholar 

  • Navarro E, Piccapietra F, Wagner B, Marconi F, Kaegi R, Odzak N, Sigg L, Behra R (2008) Toxicity of silver nanoparticles to Chlamydomonas reinhardtii. Environ Sci Technol 42(23):8959–8964

    PubMed  Article  CAS  Google Scholar 

  • Nielsen PS, de Peter N, Okkels H, Autrup H (1996) Environmental air pollution and DNA adducts in Copenhagen bus drivers—effect of GSTM1 and NAT2 genotypes on adduct levels. Carcinogenesis 17(5):1021–1027

    PubMed  Article  CAS  Google Scholar 

  • Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12(5):913–922

    PubMed  Article  CAS  Google Scholar 

  • Pavanello S, Kapka L, Siwinska E, Mielzynska D, Bolognesi C, Clonfero E (2008) Micronuclei related to anti-B[a]PDE-DNA adduct in peripheral blood lymphocytes of heavily polycyclic aromatic hydrocarbon-exposed nonsmoking coke-oven workers and controls. Cancer Epidemiol Biomarkers Prev 17(10):2795–2799

    PubMed  Article  CAS  Google Scholar 

  • Randerath K, Li DH, Randerath E (1990) Age-related DNA modifications (I-compounds): modulation by physiological and pathological processes. Mutat Res 238(3):245–253

    PubMed  CAS  Google Scholar 

  • Schrand AM, Braydich-Stolle L, Schlager JJ, Dai L, Hussain SM (2008) Can silver nanoparticles be useful as potential biological labels? Nanotechnology 19:235104

    Article  Google Scholar 

  • Smith LE, Denissenko MF, Bennett WP, Li H, Amin S, Tang M, Pfeifer GP (2000) Targeting of lung cancer mutational hotspots by polycyclic aromatic hydrocarbons. J Natl Cancer Inst 92(10):803–811

    PubMed  Article  CAS  Google Scholar 

  • Sorensen M, Autrup H, Moller P, Hertel O, Jensen SS, Vinzents P, Knudsen LE, Loft S (2003) Linking exposure to environmental pollutants with biological effects. Mutat Res 544(2–3):255–271

    PubMed  CAS  Google Scholar 

  • Suzuki H, Toyooka T, Ibuki Y (2007) Simple and easy method to evaluate uptake potential of nanoparticles in mammalian cells using a flow cytometric light scatter analysis. Environ Sci Technol 41(8):3018–3024

    PubMed  Article  CAS  Google Scholar 

  • Ueda S, Masutani H, Nakamura H, Tanaka T, Ueno M, Yodoi J (2002) Redox control of cell death. Antioxid Redox Signal 4(3):405–414

    PubMed  Article  CAS  Google Scholar 

  • van Engeland M, Ramaekers FC, Schutte B, Reutelingsperger CP (1996) A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture. Cytometry 24(2):131–139

    PubMed  Article  Google Scholar 

  • Wijnhoven SWP, Peijnenburg WJGM, Herberts CA, Hagens WI, Oomen AG, Heugens EHW, Roszek B, Bisschops J, Gosens I, Van De Meent D, Dekkers S, De Jong WH, van Zijverden M, Sips ANJAM, Geertsma RE (2009) Nano-silver—a review of available data and knowledge gaps in human and environmental risk assessment. Nanotoxicology 3(2):109–138

    Article  CAS  Google Scholar 

  • Xia T, Kovochich M, Liong M, Madler L, Gilbert B, Shi H, Yeh JI, Zink JI, Nel AE (2008) Comparison of the mechanism of toxicity of zinc oxide and cerium oxide nanoparticles based on dissolution and oxidative stress properties. ACS Nano 2(10):2121–2134

    PubMed  Article  CAS  Google Scholar 

  • Xia T, Li N, Nel AE (2009) Potential health impact of nanoparticles. Annu Rev Public Health 30(1):137–150

    PubMed  Article  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Erik Baatrup, Institute of Biology, Aarhus University for conducting AAS measurements. The SUNANO project was granted by the Danish Strategic Research Council, NABIIT2006-06-0015.

Conflict of interest statement

The authors declare that they have no conflict of interest.

Author information

Affiliations

  1. Nanoparticle Safety Group, Department of Environmental and Occupational Medicine, Institute of Public Health, Aarhus University, Bartholins Allé 2, Building 1260, 8000, Aarhus, Denmark

    Rasmus Foldbjerg, Duy Anh Dang & Herman Autrup

Authors
  1. Rasmus Foldbjerg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Duy Anh Dang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Herman Autrup
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rasmus Foldbjerg.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Foldbjerg, R., Dang, D.A. & Autrup, H. Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549. Arch Toxicol 85, 743–750 (2011). https://doi.org/10.1007/s00204-010-0545-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-010-0545-5

Keywords

  • Silver
  • Nanoparticles
  • Reactive oxygen species
  • Apoptosis
  • Bulky DNA adducts