The oxidative potential of differently charged silver and gold nanoparticles on three human lung epithelial cell types

BackgroundNanoparticle (NPs) functionalization has been shown to affect their cellular toxicity. To study this, differently functionalized silver (Ag) and gold (Au) NPs were synthesised, characterised and tested using lung epithelial cell systems.MethodsMonodispersed Ag and Au NPs with a size range of 7 to 10 nm were coated with either sodium citrate or chitosan resulting in surface charges from −50 mV to +70 mV. NP-induced cytotoxicity and oxidative stress were determined using A549 cells, BEAS-2B cells and primary lung epithelial cells (NHBE cells). TEER measurements and immunofluorescence staining of tight junctions were performed to test the growth characteristics of the cells. Cytotoxicity was measured by means of the CellTiter-Blue ® and the lactate dehydrogenase assay and cellular and cell-free reactive oxygen species (ROS) production was measured using the DCFH-DA assay.ResultsDifferent growth characteristics were shown in the three cell types used. A549 cells grew into a confluent mono-layer, BEAS-2B cells grew into a multilayer and NHBE cells did not form a confluent layer. A549 cells were least susceptible towards NPs, irrespective of the NP functionalization. Cytotoxicity in BEAS-2B cells increased when exposed to high positive charged (+65-75 mV) Au NPs. The greatest cytotoxicity was observed in NHBE cells, where both Ag and Au NPs with a charge above +40 mV induced cytotoxicity. ROS production was most prominent in A549 cells where Au NPs (+65-75 mV) induced the highest amount of ROS. In addition, cell-free ROS measurements showed a significant increase in ROS production with an increase in chitosan coating.ConclusionsChitosan functionalization of NPs, with resultant high surface charges plays an important role in NP-toxicity. Au NPs, which have been shown to be inert and often non-cytotoxic, can become toxic upon coating with certain charged molecules. Notably, these effects are dependent on the core material of the particle, the cell type used for testing and the growth characteristics of these cell culture model systems.

[1]  Weiping Qian,et al.  Facile synthesis of Ag and Au nanoparticles utilizing chitosan as a mediator agent. , 2008, Colloids and surfaces. B, Biointerfaces.

[2]  T. Minko,et al.  Biodegradable Janus Nanoparticles for Local Pulmonary Delivery of Hydrophilic and Hydrophobic Molecules to the Lungs , 2014, Langmuir : the ACS journal of surfaces and colloids.

[3]  David A Groneberg,et al.  Gold nanoparticles: recent aspects for human toxicology , 2013, Journal of Occupational Medicine and Toxicology.

[4]  W. Tiyaboonchai.  CHITOSAN NANOPARTICLES: A PROMISING SYSTEM FOR DRUG DELIVERY , 2003 .

[5]  S. Barcikowski,et al.  Reprotoxicity of gold, silver, and gold-silver alloy nanoparticles on mammalian gametes. , 2014, The Analyst.

[6]  J. Lechner,et al.  A serum-free method for culturing normal human bronchial epithelial cells at clonal density , 1985 .

[7]  E. Bourdon,et al.  The antioxidant properties of serum albumin , 2008, FEBS letters.

[8]  L. Qin,et al.  Inhibition effects of gold nanoparticles on proliferation and migration in hepatic carcinoma-conditioned HUVECs. , 2014, Bioorganic & medicinal chemistry letters.

[9]  Iseult Lynch,et al.  Physical-chemical aspects of protein corona: relevance to in vitro and in vivo biological impacts of nanoparticles. , 2011, Journal of the American Chemical Society.

[10]  Christine K Payne,et al.  Nanoparticle surface charge mediates the cellular receptors used by protein-nanoparticle complexes. , 2012, The journal of physical chemistry. B.

[11]  M. Kruszewski,et al.  Silver nanoparticles -- allies or adversaries? , 2013, Annals of agricultural and environmental medicine : AAEM.

[12]  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.

[13]  E. Hudson,et al.  Transformed human bronchial epithelial cells (beas-2b) alter the growth and morphology of normal human bronchial epithelial cells in vitro , 1990, Cell Biology and Toxicology.

[14]  B. Rothen‐Rutishauser,et al.  In vitro human lung cell culture models to study the toxic potential of nanoparticles , 2009 .

[15]  C. Instanes,et al.  Toxicity of Silver Nanomaterials in Higher Eukaryotes , 2011 .

[16]  V. Puntes,et al.  Altered characteristics of silica nanoparticles in bovine and human serum: the importance of nanomaterial characterization prior to its toxicological evaluation , 2013, Particle and Fibre Toxicology.

[17]  Albert Duschl,et al.  Interaction of nanoparticles with proteins: relation to bio-reactivity of the nanoparticle , 2013, Journal of Nanobiotechnology.

[18]  K. Siegmann,et al.  Physical and chemical properties of airborne nanoscale particles and how to measure the impact on human health , 1998 .

[19]  K. Leong,et al.  Chitosan nanoparticles for oral drug and gene delivery , 2006, International journal of nanomedicine.

[20]  V. Sharma,et al.  Silver nanoparticles: green synthesis and their antimicrobial activities. , 2009, Advances in colloid and interface science.

[21]  Rudolf Hagen,et al.  Silver nanoparticles: evaluation of DNA damage, toxicity and functional impairment in human mesenchymal stem cells. , 2011, Toxicology letters.

[22]  Barry H. Smith,et al.  A continuous tumor‐cell line from a human lung carcinoma with properties of type II alveolar epithelial cells , 1976, International journal of cancer.

[23]  David M. Brown,et al.  Increased inflammation and intracellular calcium caused by ultrafine carbon black is independent of transition metals or other soluble components , 2000, Occupational and environmental medicine.

[24]  Iseult Lynch,et al.  Formation and characterization of the nanoparticle-protein corona. , 2013, Methods in molecular biology.

[25]  Iseult Lynch,et al.  Minimal analytical characterization of engineered nanomaterials needed for hazard assessment in biological matrices , 2011, Nanotoxicology.

[26]  B. Smith.  Cell line A549: a model system for the study of alveolar type II cell function. , 1977, The American review of respiratory disease.

[27]  Dong Yang,et al.  Hypermethylation of CpG island in O6-methylguanine-DNA methyltransferase gene was associated with K-ras G to A mutation in colorectal tumor. , 2005, World journal of gastroenterology.

[28]  Tit Meng Lim,et al.  Pharmacological activity of peroral chitosan-insulin nanoparticles in diabetic rats. , 2005, International journal of pharmaceutics.

[29]  Biju Jacob,et al.  Toxicity and antibacterial assessment of chitosancoated silver nanoparticles on human pathogens and macrophage cells , 2012, International journal of nanomedicine.

[30]  P A Ward,et al.  Measurement of Intracellular Fluorescence of Human Monocytes Relative to Oxidative Metabolism , 1988, Journal of leukocyte biology.

[31]  W. Hennink,et al.  Chitosan-based formulations of drugs, imaging agents and biotherapeutics. Preface. , 2010, Advanced drug delivery reviews.

[32]  Maren Schneider,et al.  Reactive oxygen species as mediators of membrane-dependent signaling induced by ultrafine particles. , 2010, Free radical biology & medicine.

[33]  Jinbin Liu,et al.  Chitosan-capped gold nanoparticles for selective and colorimetric sensing of heparin , 2013, Journal of Nanoparticle Research.

[34]  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.

[35]  Helinor J Johnston,et al.  Review of carbon nanotubes toxicity and exposure—Appraisal of human health risk assessment based on open literature , 2010, Critical reviews in toxicology.

[36]  J. Pourová,et al.  Reactive oxygen and nitrogen species in normal physiological processes , 2010, Acta physiologica.

[37]  Ling Ye,et al.  Inhalable microspheres embedding chitosan-coated PLGA nanoparticles for 2-methoxyestradiol , 2014, Journal of drug targeting.

[38]  Jerold A. Last,et al.  TOXIC RESPONSES OF THE RESPIRATORY SYSTEM , 2001 .

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

[40]  Oliver Eickelberg,et al.  Efficient bioactive delivery of aerosolized drugs to human pulmonary epithelial cells cultured in air-liquid interface conditions. , 2014, American journal of respiratory cell and molecular biology.

[41]  P. Friedl,et al.  Impaired induction of adhesion molecule expression in immortalized endothelial cells leads to functional defects in dynamic interactions with lymphocytes. , 2007, The Journal of investigative dermatology.

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

[43]  David M. Brown,et al.  Measurement of reactive species production by nanoparticles prepared in biologically relevant media. , 2007, Toxicology letters.

[44]  E. Pinto,et al.  Differential elemental distribution of retained particles along the respiratory tract , 2011, Inhalation toxicology.

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

[46]  K. Donaldson,et al.  Signs of stress , 2006, Nature nanotechnology.

[47]  Robert Rallo,et al.  Differential expression of syndecan-1 mediates cationic nanoparticle toxicity in undifferentiated versus differentiated normal human bronchial epithelial cells. , 2011, ACS nano.

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

[49]  L. Låstbom,et al.  Human alveolar macrophage phagocytic function is impaired by aggregates of ultrafine carbon particles. , 2001, Environmental research.

[50]  Albert Duschl,et al.  Time evolution of the nanoparticle protein corona. , 2010, ACS nano.

[51]  M. Gaisberger,et al.  Effects of Ionized Waterfall Aerosol on Pediatric Allergic Asthma , 2012, The Journal of asthma : official journal of the Association for the Care of Asthma.

[52]  Ranbir Singh,et al.  J. Mol. Struct. (Theochem) , 1996 .

[53]  Lei Wang,et al.  Preparation and electrochemical properties of LiFePO4/C nanoparticles using different organic carbon sources , 2013, Journal of Nanoparticle Research.

[54]  B. Ekstrand-Hammarström,et al.  Human primary bronchial epithelial cells respond differently to titanium dioxide nanoparticles than the lung epithelial cell lines A549 and BEAS-2B , 2012, Nanotoxicology.

[55]  J. Hillier,et al.  A study of the nucleation and growth processes in the synthesis of colloidal gold , 1951 .

[56]  Jian-Zhong Shao,et al.  Toxicity evaluation of biodegradable chitosan nanoparticles using a zebrafish embryo model , 2011, International journal of nanomedicine.

[57]  S. Barcikowski,et al.  Toxicity of gold nanoparticles on somatic and reproductive cells. , 2012, Advances in experimental medicine and biology.

[58]  Christian Mühlfeld,et al.  Translocation and cellular entering mechanisms of nanoparticles in the respiratory tract. , 2008, Swiss medical weekly.

[59]  M. Hande,et al.  Toxicological profile of small airway epithelial cells exposed to gold nanoparticles , 2013, Experimental biology and medicine.

[60]  Wei Wang,et al.  Cytotoxicity induced by engineered silver nanocrystallites is dependent on surface coatings and cell types. , 2012, Langmuir : the ACS journal of surfaces and colloids.

[61]  Albert Duschl,et al.  Hardening of the nanoparticle-protein corona in metal (Au, Ag) and oxide (Fe3O4, CoO, and CeO2) nanoparticles. , 2011, Small.

[62]  E. Hoek,et al.  A review of the antibacterial effects of silver nanomaterials and potential implications for human health and the environment , 2010 .

[63]  Zi-rong Xu,et al.  In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells. , 2005, World journal of gastroenterology.

[64]  T. Xia,et al.  Toxic Potential of Materials at the Nanolevel , 2006, Science.

[65]  G. Bachand,et al.  Cytotoxicity and inflammation in human alveolar epithelial cells following exposure to occupational levels of gold and silver nanoparticles , 2012, Journal of Nanoparticle Research.

[66]  V. Puntes,et al.  Facile preparation of cationic gold nanoparticle-bioconjugates for cell penetration and nuclear targeting. , 2012, ACS nano.

[67]  M. Politis,et al.  ULTRAFINE PARTICLES (UFP) AND HEALTH EFFECTS. DANGEROUS. LIKE NO OTHER PM? REVIEW AND ANALYSIS , 2008 .

[68]  Saber M Hussain,et al.  Metal-based nanoparticles and their toxicity assessment. , 2010, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[69]  Stephan Barcikowski,et al.  Cytotoxicity and ion release of alloy nanoparticles , 2012, Journal of Nanoparticle Research.

[70]  Erik Swietlicki,et al.  Measurement techniques for respiratory tract deposition of airborne nanoparticles: a critical review. , 2014, Journal of aerosol medicine and pulmonary drug delivery.

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

[72]  Min-Ki Lee,et al.  Air-liquid interface (ALI) culture of human bronchial epithelial cell monolayers as an in vitro model for airway drug transport studies. , 2007, Journal of pharmaceutical sciences.

[73]  Amanda Hayes,et al.  Nanoparticles: a review of particle toxicology following inhalation exposure , 2012, Inhalation toxicology.

[74]  Y. Ikada,et al.  Macrophage phagocytosis of biodegradable microspheres composed of L-lactic acid/glycolic acid homo- and copolymers. , 1988, Journal of biomedical materials research.

[75]  S. Aaronson,et al.  In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. , 1973, Journal of the National Cancer Institute.