Advertisement

Molecular Biology Reports

, Volume 39, Issue 4, pp 4915–4925 | Cite as

The role of reactive oxygen species in silicon dioxide nanoparticle-induced cytotoxicity and DNA damage in HaCaT cells

  • Chunmei Gong
  • Gonghua Tao
  • Linqing Yang
  • Jianjun Liu
  • Haowei He
  • Zhixiong Zhuang
Article

Abstract

The increasing applications of silicon dioxide (SiO2) nanomaterials have been widely concerned over their biological effects and potential hazard to human health. In this study, we explored the effects of SiO2 nanoparticles (15, 30, and 100 nm) and their micro-sized counterpart on cultured human epidermal Keratinocyte (HaCaT) cells. Cell viability, cell morphology, reactive oxygen species (ROS), DNA damage (8-OHdG, γH2AX and comet assay) and apoptosis were assessed under control and SiO2 nanoparticles exposed conditions. As observed in the Cell Counting Kit-8 (CCK-8) assay, exposure to 15, 30 or 100 nm SiO2 nanoparticles at dosage levels between 0 and 100 μg/ml decreased cell viability in a concentration- and size dependent manner and the IC50 of 24 hour exposure was 19.4 ± 1.3, 27.7 ± 1.5 and 35.9 ± 1.6 μg/ml for 15, 30 and 100 nm SiO2 nanoparticles, respectively. Morphological examination revealed cell shrinkage and cell wall missing after SiO2 nanoparticle exposure. Increase in intracellular ROS level and DNA damage as well as apoptosis were also observed in SiO2 nanoparticle-exposed HaCaT cells. Exposure to SiO2 nanoparticles results in a concentration- and size-dependent cytotoxicity and DNA damage in cultural HaCaT cells which is closely correlated to increased oxidative stress.

Keywords

Reactive oxygen species (ROS) Silicon dioxide nanoparticles Flow cytometry immunofluorescence 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China [30972505, 30700673] and Shenzhen Science Technology Plan Key Project [200901017].

References

  1. 1.
    Stix G (2001) Little big science. Nanotechnology 285:32–37Google Scholar
  2. 2.
    Kipen HM, Laskin DL (2005) Smaller is not always better: nanotechnology yields nanotoxicology. Am J Physiol Lung Cell Mol Physiol 289:L696–L697PubMedCrossRefGoogle Scholar
  3. 3.
    Nel A, Xia T, Mädler L (2006) Toxic potential of materials at the nanolevel. Science 311:622–627PubMedCrossRefGoogle Scholar
  4. 4.
    Borm PJ, Robbins D, Haubold S, Kuhlbusch T, Fissan H, Donaldson K, Schins R, Stone V, Kreyling W, Lademann J, Krutmann J, Warheit D, Oberdorster E (2006) The potential risks of nanomaterials: a review carried out for ECETOC. Part Fibre Toxicol 3:11PubMedCrossRefGoogle Scholar
  5. 5.
    Rothen-Rutishauser BM, Schürch S, Haenni B, Kapp N, Gehr P (2006) Interaction of fine particles and nanopartcles with red blood cells visualized with advanced microscopic technique. Environ Sci Technol 40:353–359CrossRefGoogle Scholar
  6. 6.
    IRAC (1996) Monographs on the evaluation of chemicals to humans. Lyon, IRAC, 10Google Scholar
  7. 7.
    Giraldo AM, Lynn JW, Purpera MN, Godke RA, Bondioli KR (2007) DNA methylation and histone acetylation patterns in cultured bovine fibroblasts for nuclear transfer. Mol Reprod Dev 74:1514–1524PubMedCrossRefGoogle Scholar
  8. 8.
    Espada J, Ballestar E, Esteller M (2001) Qualitative determination of 5-methylcytosine and other components of the DNA methylation machinery: immunofluorescence and chromatin immunoprecipitation. In: Esteller M (ed) DNA methylation: approaches, methods, and applications. CRC press, Boca Raton, pp 122–126Google Scholar
  9. 9.
    Xu B, Kim S, Kastan MB (2001) Involvement of Brca1 in S phase and G2-phase checkpoints after ionizing irradiation. Mol Cell Biol 21:3445–3450PubMedCrossRefGoogle Scholar
  10. 10.
    Tao GH, Yang LQ, Gong CM, Huang HY, Liu JD, Liu JJ, Yuan JH, Chen W, Zhuang ZX (2009) Effect of PARP-1 deficiency on DNA damage and repair in human bronchial epithelial cells exposed to Benzo(a)pyrene. Mol Biol Rep 36(8):2413–2422PubMedCrossRefGoogle Scholar
  11. 11.
    Olive PL, Durand RE (1992) Detection of hypoxic cells in a murine tumor with the use of the comet assay. J Natl Cancer Inst 84:707–711PubMedCrossRefGoogle Scholar
  12. 12.
    Fujiwara K, Suematsu H, Kiyomiya E, Aoki M, Sato M, Moritoki N (2008) Size-dependent toxicity of SiO2 nano-particles to Chlorella kessleri. J Environ Sci Health A Tox Hazard Subst Environ Eng 43:1167–1173PubMedCrossRefGoogle Scholar
  13. 13.
    Rahman IA, Vejayakumaran P, Sipaut SC, Ismail J, Chee KC (2009) Size-dependent physicochemical and optical properties of silica nanoparticles. Mater Chem Phys 114:328–332CrossRefGoogle Scholar
  14. 14.
    Wang F, Gao F, Lan M, Yuan H, Huang Y, Liu J (2009) Oxidative stress contributes to silica nanoparticle-induced cytotoxicity in human embryonic kidney cells. Toxicol in Vitro 23:808–815PubMedCrossRefGoogle Scholar
  15. 15.
    Eom HJ, Choi J (2009) Oxidative stress of silica nanoparticles in human bronchial epithelial cell, Beas-2B. Toxicol In Vitro 23:1326–1332PubMedCrossRefGoogle Scholar
  16. 16.
    Dostert C, Petrilli V, Van Bruggen R, Steele C, Mossman BT, Tschopp J (2008) Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 320:674–677PubMedCrossRefGoogle Scholar
  17. 17.
    Ames BN (1983) Dietary carcinogens, anticarcinogens (oxygen radicals and degenerative diseases). Science 221:1256–1264PubMedCrossRefGoogle Scholar
  18. 18.
    Rojas E, Lopez MC, Valverde M (1999) Single cell gel electrophoresis assay: methodology and applications. J Chromatogr B Biomed Sci Appl 722:225–254PubMedCrossRefGoogle Scholar
  19. 19.
    Motoyama N, Naka K (2004) DNA damage tumor suppressor genes and genomic instability. Curr Opin Genet Dev 14:11–16PubMedCrossRefGoogle Scholar
  20. 20.
    Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622–627PubMedCrossRefGoogle Scholar
  21. 21.
    Nabeshil H, Yoshikawai T, Matsuyamai K (2011) Amorphous nanosilica induce endocytosis dependent ROS generation and DNA damage in human keratinocytes. Part Fibre Toxicol 8:1CrossRefGoogle Scholar
  22. 22.
    Chen M, von Mikecz A (2005) Formation of nucleoplasmic protein aggregates impairs nuclear function in response to SiO2 nanoparticles. Exp Cell Res 305:51–62PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Chunmei Gong
    • 1
    • 2
  • Gonghua Tao
    • 2
  • Linqing Yang
    • 2
  • Jianjun Liu
    • 2
  • Haowei He
    • 3
  • Zhixiong Zhuang
    • 2
  1. 1.School of Public HealthZhengzhou UniversityZhengzhouChina
  2. 2.Shenzhen Center for Disease Control and PreventionShenzhenChina
  3. 3.College of Life ScienceShenzhen UniversityShenzhenChina

Personalised recommendations