Skip to main content
SpringerLink
Log in

Toxicity of amorphous silica nanoparticles on eukaryotic cell model is determined by particle agglomeration and serum protein adsorption effects

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Cell cultures form the basis of most biological assays conducted to assess the cytotoxicity of nanomaterials. Since the molecular environment of nanoparticles exerts influence on their physicochemical properties, it can have an impact on nanotoxicity. Here, toxicity of silica nanoparticles upon delivery by fluid-phase uptake is studied in a 3T3 fibroblast cell line. Based on XTT viability assay, cytotoxicity is shown to be a function of (1) particle concentration and (2) of fetal calf serum (FCS) content in the cell culture medium. Application of dynamic light scattering shows that both parameters affect particle agglomeration. The DLS experiments verify the stability of the nanoparticles in culture medium without FCS over a wide range of particle concentrations. The related toxicity can be mainly accounted for by single silica nanoparticles and small agglomerates. In contrast, agglomeration of silica nanoparticles in all FCS-containing media is observed, resulting in a decrease of the associated toxicity. This result has implications for the evaluation of the cytotoxic potential of silica nanoparticles and possibly also other nanomaterials in standard cell culture.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Oberdörster G, Stone V, Donaldson K (2007) Toxicology of nanoparticles: a historical perspective. Nanotoxicology 1(1):2–25

    Article  Google Scholar 

  2. Brunner TJ, Wick P, Manser P, Spohn P, Grass RN, Limbach LK, Bruinink A, Stark WJ (2006) In vitro cytotoxicity of oxide nanoparticles: comparison to asbestos, silica, and the effect of particle solubility. Environ Sci Technol 40(14):4374–4381

    Article  CAS  Google Scholar 

  3. 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

    Article  CAS  Google Scholar 

  4. Brown SC, Kamal M, Nasreen N, Baumuratov A, Sharma P, Antony VB, Moudgil BM (2007) Influence of shape, adhesion and simulated lung mechanics on amorphous silica nanoparticle toxicity. Adv Powder Technol 18(1):69–79

    Article  CAS  Google Scholar 

  5. Chang JS, Chang KLB, Hwang DF, Kong ZL (2007) In vitro cytotoxicity of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cell line. Environ Sci Technol 41(6):2064–2068

    Article  CAS  Google Scholar 

  6. Di Pasqua AJ, Sharma KK, Shi YL, Toms BB, Ouellette W, Dabrowiak JC, Asefa T (2008) Cytotoxicity of mesoporous silica nanomaterials. J Inorg Biochem 102(7):1416–1423

    Article  Google Scholar 

  7. Yang H, Liu C, Yang DF, Zhang HS, Xi ZG (2009) Comparative study of cytotoxicity, oxidative stress and genotoxicity induced by four typical nanomaterials: the role of particle size, shape and composition. J Appl Toxicol 29(1):69–78

    Article  Google Scholar 

  8. Pumera M (2010) Nanotoxicology: the molecular science point of view. Chem Asian J 6(2):340–348

    Article  Google Scholar 

  9. Lin WS, Huang YW, Zhou XD, Ma YF (2006) In vitro toxicity of silica nanoparticles in human lung cancer cells. Toxicol Appl Pharmacol 217(3):252–259

    Article  CAS  Google Scholar 

  10. Lison D, Thomassen LCJ, Rabolli V, Gonzalez L, Napierska D, Seo JW, Kirsch-Volders M, Hoet P, Kirschhock CEA, Martens JA (2008) Nominal and effective dosimetry of silica nanoparticles in cytotoxicity assays. Toxicol Sci 104(1):155–162

    Article  CAS  Google Scholar 

  11. Napierska D, Thomassen LCJ, Rabolli V, Lison D, Gonzalez L, Kirsch-Volders M, Martens JA, Hoet PH (2009) Size-dependent cytotoxicity of monodisperse silica nanoparticles in human endothelial cells. Small 5(7):846–853

    Article  CAS  Google Scholar 

  12. Lu F, Wu SH, Hung Y, Mou CY (2009) Size effect on cell uptake in well-suspended, uniform mesoporous silica nanoparticles. Small 5(12):1408–1413

    Article  CAS  Google Scholar 

  13. Witasp E, Kupferschmidt N, Bengtsson L, Hultenby K, Smedman C, Paulie S, Garcia-Bennett AE, Fadeel B (2009) Efficient internalization of mesoporous silica particles of different sizes by primary human macrophages without impairment of macrophage clearance of apoptotic or antibody-opsonized target cells. Toxicol Appl Pharmacol 239(3):306–319

    Article  CAS  Google Scholar 

  14. Orts-Gil G, Natte K, Drescher D, Bresch H, Mantion A, Kneipp J, Österle W (2010) Characterisation of silica nanoparticles prior to in vitro studies: from primary particles to agglomerates. J Nanopart Res. doi:10.1007/s11051-010-9910-9

    Google Scholar 

  15. Sager TM, Porter DW, Robinson VA, Lindsley WG, Schwegler-Berry DE, Castranova V (2007) Improved method to disperse nanoparticles for in vitro and in vivo investigation of toxicity. Nanotoxicology 1(2):118–129

    Article  CAS  Google Scholar 

  16. Thomassen LCJ, Aerts A, Rabolli V, Lison D, Gonzalez L, Kirsch-Volders M, Napierska D, Hoet PH, Kirschhock CEA, Martens JA (2010) Synthesis and characterization of stable monodisperse silica nanoparticle sols for in vitro cytotoxicity testing. Langmuir 26(1):328–335

    Article  CAS  Google Scholar 

  17. Murdock RC, Braydich-Stolle L, Schrand AM, Schlager JJ, Hussain SM (2008) Characterization of nanomaterial dispersion in solution prior to In vitro exposure using dynamic light scattering technique. Toxicol Sci 101(2):239–253

    Article  CAS  Google Scholar 

  18. Ding M, Chen F, Shi XL, Yucesoy B, Mossman B, Vallyathan V (2002) Diseases caused by silica: mechanisms of injury and disease development. Int Immunopharmacol 2(2–3):173–182

    Article  CAS  Google Scholar 

  19. Mossman BT, Churg A (1998) Mechanisms in the pathogenesis of asbestosis and silicosis. Am J Respir Crit Care Med 157(5):1666–1680

    CAS  Google Scholar 

  20. Chithrani BD, Ghazani AA, Chan WCW (2006) Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. Nano Lett 6(4):662–668

    Article  CAS  Google Scholar 

  21. Xia T, Kovochich M, Liong M, Zink JI, Nel AE (2008) Cationic polystyrene nanosphere toxicity depends on cell-specific endocytic and mitochondrial injury pathways. ACS Nano 2(1):85–96

    Article  CAS  Google Scholar 

  22. Dutta D, Sundaram SK, Teeguarden JG, Riley BJ, Fifield LS, Jacobs JM, Addleman SR, Kaysen GA, Moudgil BM, Weber TJ (2007) Adsorbed proteins influence the biological activity and molecular targeting of nanomaterials. Toxicol Sci 100(1):303–315

    Article  CAS  Google Scholar 

  23. Lynch I, Dawson KA (2008) Protein-nanoparticle interactions. Nano Today 3(1–2):40–47

    Article  CAS  Google Scholar 

  24. Teeguarden JG, Hinderliter PM, Orr G, Thrall BD, Pounds JG (2007) Particokinetics in vitro: dosimetry considerations for in vitro nanoparticle toxicity assessments. Toxicol Sci 95(2):300–312

    Article  CAS  Google Scholar 

  25. Rezwan K, Studart AR, Voros J, Gauckler LJ (2005) Change of ζ potential of biocompatible colloidal oxide particles upon adsorption of bovine serum albumin and lysozyme. J Phys Chem B 109(30):14469–14474

    Article  CAS  Google Scholar 

  26. Kato H, Fujita K, Horie M, Suzuki M, Nakamura A, Endoh S, Yoshida Y, Iwahashi H, Takahashi K, Kinugasa S (2010) Dispersion characteristics of various metal oxide secondary nanoparticles in culture medium for in vitro toxicology assessment. Toxicol in Vitro 24(3):1009–1018

    Article  CAS  Google Scholar 

  27. Lundqvist M, Sethson I, Jonsson BH (2004) Protein adsorption onto silica nanoparticles: conformational changes depend on the particles' curvature and the protein stability. Langmuir 20(24):10639–10647

    Article  CAS  Google Scholar 

  28. Schulze C, Kroll A, Lehr CM, Schafer UF, Becker K, Schnekenburger J, Isfort CS, Landsiedel R, Wohlleben W (2008) Not ready to use—overcoming pitfalls when dispersing nanoparticles in physiological media. Nanotoxicology 2(2):51–61

    Article  CAS  Google Scholar 

  29. Shang W, Nuffer JH, Dordick JS, Siegel RW (2007) Unfolding of ribonuclease A on silica nanoparticle surfaces. Nano Lett 7(7):1991–1995

    Article  CAS  Google Scholar 

  30. Vertegel AA, Siegel RW, Dordick JS (2004) Silica nanoparticle size influences the structure and enzymatic activity of adsorbed lysozyme. Langmuir 20(16):6800–6807

    Article  CAS  Google Scholar 

  31. Kittler S, Greulich C, Gebauer JS, Diendorf J, Treuel L, Ruiz L, Gonzalez-Calbet JM, Vallet-Regi M, Zellner R, Koller M, Epple M (2010) The influence of proteins on the dispersability and cell-biological activity of silver nanoparticles. J Mater Chem 20(3):512–518

    Article  CAS  Google Scholar 

  32. Díaz B, Sánchez-Espinel C, Arruebo M, Faro J, de Miguel E, Magadán S, Yagüe C, Fernández-Pacheco R, Ibarra MR, Santamaría J, González-Fernández A (2008) Assessing methods for blood cell cytotoxic responses to inorganic nanoparticles and nanoparticle aggregates. Small 4(11):2025–2034

    Article  Google Scholar 

  33. Xing XL, He XX, Peng JF, Wang KM, Tan WH (2005) Uptake of silica-coated nanoparticles by HeLa cells. J Nanosci Nanotechnol 5(10):1688–1693

    Article  CAS  Google Scholar 

  34. Waters KM, Masiello LM, Zangar RC, Tarasevich BJ, Karin NJ, Quesenberry RD, Bandyopadhyay S, Teeguarden JG, Pounds JG, Thrall BD (2009) Macrophage responses to silica nanoparticles are highly conserved across particle sizes. Toxicol Sci 107(2):553–569

    Article  CAS  Google Scholar 

  35. Yu KO, Grabinski CM, Schrand AM, Murdock RC, Wang W, Gu BH, Schlager JJ, Hussain SM (2009) Toxicity of amorphous silica nanoparticles in mouse keratinocytes. J Nanopart Res 11(1):15–24

    Article  CAS  Google Scholar 

  36. Cedervall T, Lynch I, Lindman S, Berggard T, Thulin E, Nilsson H, Dawson KA, Linse S (2007) Understanding the nanoparticle-protein corona using methods to quantify exchange rates and affinities of proteins for nanoparticles. Proc Natl Acad Sci USA 104(7):2050–2055

    Article  CAS  Google Scholar 

  37. Stayton I, Winiarz J, Shannon K, Ma YF (2009) Study of uptake and loss of silica nanoparticles in living human lung epithelial cells at single cell level. Anal Bioanal Chem 394(6):1595–1608

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Michael Weller and Rudolf Schneider of BAM Federal Institute for Materials Research and Testing for use of the cell culture facility. This study has been supported by BAM Federal Institute for Materials Research and Testing within the framework of “BAM Innovationsoffensive” (project NANOTOX 1).

Author information

Authors and Affiliations

  1. BAM Federal Institute for Materials Research and Testing, Richard-Willstätter-Str. 11, 12489, Berlin, Germany

    Daniela Drescher & Janina Kneipp

  2. Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Str. 2, 12489, Berlin, Germany

    Daniela Drescher & Janina Kneipp

  3. BAM Federal Institute for Materials Research and Testing, Unter den Eichen 87, 12205, Berlin, Germany

    Guillermo Orts-Gil, Kishore Natte & Werner Österle

  4. Institute of Integrative Neuroanatomy, Charité - Universitätsmedizin Berlin, Philippstr. 12, 10115, Berlin, Germany

    Gregor Laube & Rüdiger W. Veh

Authors
  1. Daniela Drescher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guillermo Orts-Gil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gregor Laube
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kishore Natte
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rüdiger W. Veh
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Werner Österle
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Janina Kneipp
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Daniela Drescher or Guillermo Orts-Gil.

Electronic supplementary materials

Below is the link to the electronic supplementary material.

Protein adsorption effects

(PDF 222 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Drescher, D., Orts-Gil, G., Laube, G. et al. Toxicity of amorphous silica nanoparticles on eukaryotic cell model is determined by particle agglomeration and serum protein adsorption effects. Anal Bioanal Chem 400, 1367–1373 (2011). https://doi.org/10.1007/s00216-011-4893-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-011-4893-7

Keywords

Search

Navigation