Exposure of silver-nanoparticles and silver-ions to lung cells in vitro at the air-liquid interface

BackgroundDue to its antibacterial properties, silver (Ag) has been used in more consumer products than any other nanomaterial so far. Despite the promising advantages posed by using Ag-nanoparticles (NPs), their interaction with mammalian systems is currently not fully understood. An exposure route via inhalation is of primary concern for humans in an occupational setting. Aim of this study was therefore to investigate the potential adverse effects of aerosolised Ag-NPs using a human epithelial airway barrier model composed of A549, monocyte derived macrophage and dendritic cells cultured in vitro at the air-liquid interface. Cell cultures were exposed to 20 nm citrate-coated Ag-NPs with a deposition of 30 and 278 ng/cm2 respectively and incubated for 4 h and 24 h. To elucidate whether any effects of Ag-NPs are due to ionic effects, Ag-Nitrate (AgNO3) solutions were aerosolised at the same molecular mass concentrations.ResultsAgglomerates of Ag-NPs were detected at 24 h post exposure in vesicular structures inside cells but the cellular integrity was not impaired upon Ag-NP exposures. Minimal cytotoxicity, by measuring the release of lactate dehydrogenase, could only be detected following a higher concentrated AgNO3-solution. A release of pro-inflammatory markers TNF-α and IL-8 was neither observed upon Ag-NP and AgNO3 exposures as well as was not affected when cells were pre-stimulated with lipopolysaccharide (LPS). Also, an induction of mRNA expression of TNF-α and IL-8, could only be observed for the highest AgNO3 concentration alone or even significantly increased when pre-stimulated with LPS after 4 h. However, this effect disappeared after 24 h. Furthermore, oxidative stress markers (HMOX-1, SOD-1) were expressed after 4 h in a concentration dependent manner following AgNO3 exposures only.ConclusionsWith an experimental setup reflecting physiological exposure conditions in the human lung more realistic, the present study indicates that Ag-NPs do not cause adverse effects and cells were only sensitive to high Ag-ion concentrations. Chronic exposure scenarios however, are needed to reveal further insight into the fate of Ag-NPs after deposition and cell interactions.

[1]  Seung-Heon Shin,et al.  The effects of nano-silver on the proliferation and cytokine expression by peripheral blood mononuclear cells. , 2007, International immunopharmacology.

[2]  D. Dix,et al.  Informing Selection of Nanomaterial Concentrations for ToxCast in Vitro Testing Based on Occupational Exposure Potential , 2011, Environmental health perspectives.

[3]  W. D. de Jong,et al.  Nano-silver – a review of available data and knowledge gaps in human and environmental risk assessment , 2009 .

[4]  Kyunghee Choi,et al.  Induction of inflammatory responses and gene expression by intratracheal instillation of silver nanoparticles in mice , 2011, Archives of pharmacal research.

[5]  Stella M. Marinakos,et al.  Mechanism of silver nanoparticle toxicity is dependent on dissolved silver and surface coating in Caenorhabditis elegans. , 2012, Environmental science & technology.

[6]  Steffen Foss Hansen,et al.  Survey on basic knowledge about exposure and potential environmental and health risks for selected nanomaterials , 2011 .

[7]  W. Stark,et al.  Effects of flame made zinc oxide particles in human lung cells - a comparison of aerosol and suspension exposures , 2012, Particle and Fibre Toxicology.

[8]  S. Okabe,et al.  In vitro toxicity of silver nanoparticles at noncytotoxic doses to HepG2 human hepatoma cells. , 2009, Environmental science & technology.

[9]  Mustafa Culha,et al.  Interaction of multi-functional silver nanoparticles with living cells , 2010, Nanotechnology.

[10]  Wolfgang J Parak,et al.  Fluorescent-magnetic hybrid nanoparticles induce a dose-dependent increase in proinflammatory response in lung cells in vitro correlated with intracellular localization. , 2010, Small.

[11]  Peter Gehr,et al.  Dendritic cells and macrophages form a transepithelial network against foreign particulate antigens. , 2007, American journal of respiratory cell and molecular biology.

[12]  Peter Gehr,et al.  A three-dimensional cellular model of the human respiratory tract to study the interaction with particles. , 2005, American journal of respiratory cell and molecular biology.

[13]  G. Oberdörster,et al.  Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles , 2005, Environmental health perspectives.

[14]  O. Schmid,et al.  Effects and uptake of gold nanoparticles deposited at the air-liquid interface of a human epithelial airway model. , 2010, Toxicology and applied pharmacology.

[15]  H. White,et al.  Electrochemistry of Sulfur Adlayers on the Low-Index Faces of Silver , 1996 .

[16]  W. D. de Jong,et al.  The effect of particle size on the cytotoxicity, inflammation, developmental toxicity and genotoxicity of silver nanoparticles. , 2011, Biomaterials.

[17]  Christian Mühlfeld,et al.  Quantitative evaluation of cellular uptake and trafficking of plain and polyethylene glycol-coated gold nanoparticles. , 2010, Small.

[18]  Albert Duschl,et al.  Shape matters: effects of silver nanospheres and wires on human alveolar epithelial cells , 2011, Particle and Fibre Toxicology.

[19]  I. Yu,et al.  Subchronic inhalation toxicity of silver nanoparticles. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[20]  Enrique Navarro,et al.  Toxicity of silver nanoparticles to Chlamydomonas reinhardtii. , 2008, Environmental science & technology.

[21]  Seoyoung Park,et al.  Cellular Toxicity of Various Inhalable Metal Nanoparticles on Human Alveolar Epithelial Cells , 2007, Inhalation toxicology.

[22]  J. Lead,et al.  Silver nanoparticles: behaviour and effects in the aquatic environment. , 2011, Environment international.

[23]  M. Rai,et al.  Silver nanoparticles as a new generation of antimicrobials. , 2009, Biotechnology advances.

[24]  M. Hande,et al.  Cytotoxicity and genotoxicity of silver nanoparticles in human cells. , 2009, ACS nano.

[25]  M. Ahamed,et al.  Silver nanoparticle applications and human health. , 2010, Clinica chimica acta; international journal of clinical chemistry.

[26]  Peter Wick,et al.  Pulmonary surfactant coating of multi-walled carbon nanotubes (MWCNTs) influences their oxidative and pro-inflammatory potential in vitro , 2012, Particle and Fibre Toxicology.

[27]  B. Rothen‐Rutishauser,et al.  A newly developed in vitro model of the human epithelial airway barrier to study the toxic potential of nanoparticles. , 2008, ALTEX.

[28]  J. Yi,et al.  Oxidative stress-dependent toxicity of silver nanoparticles in human hepatoma cells. , 2009, Toxicology in vitro : an international journal published in association with BIBRA.

[29]  Sophie Lanone,et al.  Comparative toxicity of 24 manufactured nanoparticles in human alveolar epithelial and macrophage cell lines , 2009, Particle and Fibre Toxicology.

[30]  J. Schlager,et al.  DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells. , 2008, Toxicology and applied pharmacology.

[31]  H J Klasen,et al.  Historical review of the use of silver in the treatment of burns. I. Early uses. , 2000, Burns : journal of the International Society for Burn Injuries.

[32]  R. L. Jones,et al.  Unique cellular interaction of silver nanoparticles: size-dependent generation of reactive oxygen species. , 2008, The journal of physical chemistry. B.

[33]  Jeffrey M. Perkel LIFE SCIENCE TECHNOLOGIES: Animal-Free Toxicology: Sometimes, in Vitro is Better , 2012 .

[34]  Dae Hong Jeong,et al.  Antimicrobial effects of silver nanoparticles. , 2007, Nanomedicine : nanotechnology, biology, and medicine.

[35]  Jinhee Choi,et al.  Oxidative stress‐related PMK‐1 P38 MAPK activation as a mechanism for toxicity of silver nanoparticles to reproduction in the nematode Caenorhabditis elegans , 2012, Environmental toxicology and chemistry.

[36]  Il Je Yu,et al.  Lung function changes in Sprague-Dawley rats after prolonged inhalation exposure to silver nanoparticles. , 2008, Inhalation toxicology.

[37]  R. Hurt,et al.  Ion release kinetics and particle persistence in aqueous nano-silver colloids. , 2010, Environmental science & technology.

[38]  Christian Mühlfeld,et al.  In vitro models of the human epithelial airway barrier to study the toxic potential of particulate matter , 2008, Expert opinion on drug metabolism & toxicology.

[39]  Revista Mundo Nano Survey on basic knowledge about exposure and potential environmental and health risks for selected nanomaterials. Mikkelsen, Sonja H., Hansen, Erik.; Christensen, Trine B.; Baun, Anders; Hansen, Steffen F., Binderup, Mona-Lise... , 2014 .

[40]  H. Autrup,et al.  PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes. , 2009, Toxicology letters.

[41]  Kyunghee Choi,et al.  Silver nanoparticles induce cytotoxicity by a Trojan-horse type mechanism. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[42]  Vicki Stone,et al.  Intracellular imaging of nanoparticles: Is it an elemental mistake to believe what you see? , 2010, Particle and Fibre Toxicology.

[43]  Ke Karlovu,et al.  The bactericidal effect of silver nanoparticles , 2010 .

[44]  V. Edwards-Jones The benefits of silver in hygiene, personal care and healthcare , 2009, Letters in applied microbiology.

[45]  Naomi Lubick,et al.  Nanosilver toxicity: ions, nanoparticles--or both? , 2008, Environmental science & technology.

[46]  B. Rothen‐Rutishauser,et al.  An optimized in vitro model of the respiratory tract wall to study particle cell interactions. , 2006, Journal of aerosol medicine : the official journal of the International Society for Aerosols in Medicine.

[47]  M. Ochs,et al.  Visualization and quantitative analysis of nanoparticles in the respiratory tract by transmission electron microscopy , 2007, Particle and Fibre Toxicology.

[48]  Martin Mohr,et al.  Oxidative stress and inflammation response after nanoparticle exposure: differences between human lung cell monocultures and an advanced three-dimensional model of the human epithelial airways , 2010, Journal of The Royal Society Interface.

[49]  E. J. Foster,et al.  Investigating the interaction of cellulose nanofibers derived from cotton with a sophisticated 3D human lung cell coculture. , 2011, Biomacromolecules.

[50]  B. Rothen‐Rutishauser,et al.  Laser scanning microscopy combined with image restoration to analyse a 3D model of the human epithelial airway barrier. , 2010, Swiss medical weekly.

[51]  M. Hande,et al.  Anti-proliferative activity of silver nanoparticles , 2009, BMC Cell Biology.

[52]  Barbara Rothen-Rutishauser,et al.  A dose-controlled system for air-liquid interface cell exposure and application to zinc oxide nanoparticles , 2009, Particle and Fibre Toxicology.

[53]  Herman Autrup,et al.  Toxicity of silver nanoparticles - nanoparticle or silver ion? , 2012, Toxicology letters.

[54]  Vicki Stone,et al.  Toxicology of nanoparticles: A historical perspective , 2007 .

[55]  A. Lund,et al.  Conduction electron spin resonance of small silver particles. , 2006, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[56]  H. Autrup,et al.  Cytotoxicity and genotoxicity of silver nanoparticles in the human lung cancer cell line, A549 , 2011, Archives of Toxicology.

[57]  Larissa V Stebounova,et al.  Nanosilver induces minimal lung toxicity or inflammation in a subacute murine inhalation model , 2011, Particle and Fibre Toxicology.

[58]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[59]  Pedro J J Alvarez,et al.  Negligible particle-specific antibacterial activity of silver nanoparticles. , 2012, Nano letters.

[60]  L. Marr,et al.  Toxicity of Silver Nanoparticles at the Air-Liquid Interface , 2012, BioMed research international.