The suitability of different cellular in vitro immunotoxicity and genotoxicity methods for the analysis of nanoparticle-induced events

Abstract Suitable assays and test strategies are needed to analyze potential genotoxic and immunotoxic health effects caused by nanoparticle exposure. The development and validation of such methods is challenging because nanoparticles may show unexpected behavior, like aggregation or interference with optical measurements, when routine in vitro assays are performed. In our interdisciplinary study, the effects of inorganic gold (4.5 nm) and iron oxide (7.3 nm) nanoparticles with a narrow size distribution were tested on human cells using different assay systems. The results show that cytotoxicity as well as immunotoxicity and genotoxicity induced by these two inorganic nanoparticles was low or absent when using a panel of cell-based tests in different laboratories. However, several technical issues had to be tackled that were specific for working with nanoparticles. The methods used, their suitability for nanotoxicity testing, and the technical problems encountered are carefully described and discussed in this paper.

[1]  T. Tetley,et al.  Health effects of nanomaterials. , 2007, Biochemical Society transactions.

[2]  Catherine J. Murphy,et al.  Seeding Growth for Size Control of 5−40 nm Diameter Gold Nanoparticles , 2001 .

[3]  L. Brus,et al.  Magnetite Fe3O4 Nanocrystals: Spectroscopic Observation of Aqueous Oxidation Kinetics† , 2003 .

[4]  Albert Duschl,et al.  SWCNT suppress inflammatory mediator responses in human lung epithelium in vitro. , 2009, Toxicology and applied pharmacology.

[5]  A. Collins,et al.  Oxidative damage to DNA: do we have a reliable biomarker? , 1996, Environmental health perspectives.

[6]  H. Byrne,et al.  Spectroscopic analysis confirms the interactions between single walled carbon nanotubes and various dyes commonly used to assess cytotoxicity , 2007 .

[7]  P. Vandenabeele,et al.  Generation of dendritic cells from bone marrow progenitors using GM‐CSF, TNF‐α, and additional cytokines: antagonistic effects of IL‐4 and IFN‐γ and selective involvement of TNF‐α receptor‐1 , 1997, Immunology.

[8]  P. Jain,et al.  Au nanoparticles target cancer , 2007 .

[9]  C. Borrebaeck,et al.  Global reprogramming of dendritic cells in response to a concerted action of inflammatory mediators. , 2002, International immunology.

[10]  R. Service.  Can High-Speed Tests Sort Out Which Nanomaterials Are Safe? , 2008, Science.

[11]  P. Bossù,et al.  Balance between autocrine interleukin-1beta and caspases defines life versus death of polymorphonuclear cells. , 2001, European cytokine network.

[12]  J. Hooyberghs,et al.  Microarray analyses in dendritic cells reveal potential biomarkers for chemical-induced skin sensitization. , 2007, Molecular immunology.

[13]  Maureen R. Gwinn,et al.  Nanoparticles: Health Effects—Pros and Cons , 2006, Environmental health perspectives.

[14]  H. Norppa,et al.  Induction of micronuclei by five pyrethroid insecticides in whole-blood and isolated human lymphocyte cultures. , 1995, Mutation research.

[15]  Minnamari Vippola,et al.  Genotoxicity of nanomaterials: DNA damage and micronuclei induced by carbon nanotubes and graphite nanofibres in human bronchial epithelial cells in vitro. , 2009, Toxicology letters.

[16]  Jinhee Choi,et al.  Oxidative stress induced by cerium oxide nanoparticles in cultured BEAS-2B cells. , 2008, Toxicology.

[17]  Ignacy Gryczynski,et al.  Metal-enhanced fluorescence: an emerging tool in biotechnology. , 2005, Current opinion in biotechnology.

[18]  T. Faunce.  Integrated Research into the Nanoparticle-Protein Corona: A New Focus for Safe, Sustainable and Equitable Development of Nanomedicines , 2008, Nanomedicine.

[19]  L. Verschaeve,et al.  Genotoxic effects of carbon black particles, diesel exhaust particles, and urban air particulates and their extracts on a human alveolar epithelial cell line (A549) and a human monocytic cell line (THP‐1) , 2001, Environmental and molecular mutagenesis.

[20]  M. Fenech.  Cytokinesis-block micronucleus cytome assay , 2007, Nature Protocols.

[21]  N. Gu,et al.  Effects of Proteins from Culture Medium on Surface Property of Silanes- Functionalized Magnetic Nanoparticles , 2008, Nanoscale research letters.

[22]  Julie W. Fitzpatrick,et al.  Principles for characterizing the potential human health effects from exposure to nanomaterials: elements of a screening strategy , 2005, Particle and Fibre Toxicology.

[23]  David B Warheit,et al.  How meaningful are the results of nanotoxicity studies in the absence of adequate material characterization? , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

[24]  R. Pieters,et al.  Lung dendritic cells are stimulated by ultrafine particles and play a key role in particle adjuvant activity. , 2008, The Journal of allergy and clinical immunology.

[25]  L. Migliore,et al.  Analysis of cobalt ferrite nanoparticles induced genotoxicity on human peripheral lymphocytes: comparison of size and organic grafting-dependent effects , 2007 .

[26]  J. Joseph,et al.  Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. , 1999, Free radical biology & medicine.

[27]  Jian Qin,et al.  The importance of an endotoxin-free environment during the production of nanoparticles used in medical applications. , 2006, Nano letters.

[28]  K. Wittmaack.  In Search of the Most Relevant Parameter for Quantifying Lung Inflammatory Response to Nanoparticle Exposure: Particle Number, Surface Area, or What? , 2006, Environmental health perspectives.

[29]  George C. Schatz,et al.  The Optical Properties of Metal Nanoparticles: The Influence of Size, Shape, and Dielectric Environment , 2003 .

[30]  F. Coppedè,et al.  Genotoxicity induced by arsenic compounds in peripheral human lymphocytes analysed by cytokinesis-block micronucleus assay. , 2007, Mutagenesis.

[31]  O. Herbarth,et al.  An experimental model for the determination of immunomodulating effects by volatile compounds. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[32]  Albert Duschl,et al.  Dispersion medium modulates oxidative stress response of human lung epithelial cells upon exposure to carbon nanomaterial samples. , 2009, Toxicology and applied pharmacology.

[33]  S. Gibbs,et al.  Progress on the development of human in vitro dendritic cell based assays for assessment of the sensitizing potential of a compound. , 2009, Toxicology and applied pharmacology.

[34]  Z. Berneman,et al.  Capacity of CD34+ progenitor-derived dendritic cells to distinguish between sensitizers and irritants. , 2005, Toxicology letters.

[35]  Neus G Bastús,et al.  Peptides conjugated to gold nanoparticles induce macrophage activation. , 2009, Molecular immunology.

[36]  M. Kandlikar,et al.  The impact of toxicity testing costs on nanomaterial regulation. , 2009, Environmental science & technology.

[37]  Thomas Kuhlbusch,et al.  Particle and Fibre Toxicology BioMed Central Review The potential risks of nanomaterials: a review carried out for ECETOC , 2006 .

[38]  S. Georas,et al.  Diesel-enriched particulate matter functionally activates human dendritic cells. , 2007, American journal of respiratory cell and molecular biology.

[39]  J. Gearhart,et al.  In vitro toxicity of nanoparticles in BRL 3A rat liver cells. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[40]  G. Oostingh,et al.  A high-throughput screening method based on stably transformed human cells was used to determine the immunotoxic effects of fluoranthene and other PAHs. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[41]  V. Puntes,et al.  Shuttling Gold Nanoparticles into Tumoral Cells with an Amphipathic Proline‐Rich Peptide , 2009, Chembiochem : a European journal of chemical biology.

[42]  Alberto Mantovani,et al.  Macrophage activation and polarization. , 2008, Frontiers in bioscience : a journal and virtual library.

[43]  A. Alivisatos,et al.  Synthesis of hcp-Co Nanodisks. , 2002, Journal of the American Chemical Society.

[44]  Robert N Grass,et al.  Exposure of engineered nanoparticles to human lung epithelial cells: influence of chemical composition and catalytic activity on oxidative stress. , 2007, Environmental science & technology.

[45]  J. B. Hall,et al.  Characterization of nanoparticles for therapeutics. , 2007, Nanomedicine.

[46]  K. Batarseh.  A novel silver (I) chelate: cytotoxicity and apoptotic mechanism of action on human hematopoietic malignancies , 2008, Leukemia.

[47]  A. Dhillon,et al.  Characterisation of inorganic microparticles in pigment cells of human gut associated lymphoid tissue. , 1996, Gut.

[48]  T. Yoshikawa,et al.  Effects of Diesel Exhaust Particles on Antigen-Presenting Cells and Antigen-Specific Th Immunity in Mice , 2009, Experimental biology and medicine.

[49]  M. A. Curotto de Lafaille,et al.  Diesel Exhaust Particle-Exposed Human Bronchial Epithelial Cells Induce Dendritic Cell Maturation1 , 2006, The Journal of Immunology.

[50]  Kenneth A. Dawson,et al.  Nanotoxicology: nanoparticles reconstruct lipids. , 2009, Nature nanotechnology.

[51]  E Sabbioni,et al.  Comparative genotoxicity of cobalt nanoparticles and ions on human peripheral leukocytes in vitro. , 2008, Mutagenesis.

[52]  V. Puntes,et al.  In vitro investigation of immunomodulatory effects caused by engineered inorganic nanoparticles – the impact of experimental design and cell choice , 2009 .

[53]  J. Schnekenburger,et al.  Not ready to use – overcoming pitfalls when dispersing nanoparticles in physiological media , 2008 .

[54]  M. Fenech,et al.  HUman MicroNucleus project: international database comparison for results with the cytokinesis‐block micronucleus assay in human lymphocytes: I. Effect of laboratory protocol, scoring criteria, and host factors on the frequency of micronuclei , 2001, Environmental and molecular mutagenesis.

[55]  H. Takano,et al.  Carbon black nanoparticles promote the maturation and function of mouse bone marrow-derived dendritic cells. , 2008, Chemosphere.

[56]  N. Monteiro-Riviere,et al.  Limitations and relative utility of screening assays to assess engineered nanoparticle toxicity in a human cell line. , 2009, Toxicology and applied pharmacology.

[57]  T Hartung,et al.  In vitro tests to evaluate immunotoxicity: a preliminary study. , 2007, Toxicology.

[58]  Parag Aggarwal,et al.  Preclinical studies to understand nanoparticle interaction with the immune system and its potential effects on nanoparticle biodistribution. , 2008, Molecular pharmaceutics.

[59]  H. Krug,et al.  Oops they did it again! Carbon nanotubes hoax scientists in viability assays. , 2006, Nano letters.

[60]  A. Stammati,et al.  The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics , 2005, Cell Biology and Toxicology.

[61]  I. Nelissen,et al.  Flow cytometric characterisation of antigen presenting dendritic cells after in vitro exposure to diesel exhaust particles. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[62]  P. Krammer,et al.  Involvement of Jun and Rel Proteins in Up-regulation of Interleukin-4 Gene Activity by the T Cell Accessory Molecule CD28* , 1998, The Journal of Biological Chemistry.

[63]  Roel P F Schins,et al.  Genotoxicity of Poorly Soluble Particles , 2007, Inhalation toxicology.

[64]  Tian Xia,et al.  The role of oxidative stress in ambient particulate matter-induced lung diseases and its implications in the toxicity of engineered nanoparticles. , 2008, Free radical biology & medicine.

[65]  Sara Linse,et al.  The nanoparticle-protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. , 2007, Advances in colloid and interface science.

[66]  Nastassja A. Lewinski,et al.  Cytotoxicity of nanoparticles. , 2008, Small.