Silver nanoparticle-induced cytotoxicity in rat brain endothelial cell culture.

Silver nanoparticles (AgNPs) are among the most widely commercialised engineered nanomaterials, because of their antimicrobial properties. They are already commonly used in medical devices, household products and industry. Concerns have been raised about potential adverse health effects due to increasing dispersion of AgNPs in the environment. The present study examined the cytotoxic effects of spherical, citrate-coated AgNPs (10, 50 and 100 nm) in rat brain endothelial (RBE4) cells and investigated whether the observed effects can be explained by the intrinsic toxicity of the particles or the silver ions released from the particles. The results indicated that exposure of RBE4 cells to AgNPs lead to significant reduction in dye uptake as measured with the Neutral red (NR) assay. The effect was found to be related to particle size, surface area, dose and exposure time. In contrast, silver ions increased NR uptake (ca. 10%) in RBE4 cells after 1h, while a reduction in NR uptake was observed after 24h exposure at high concentrations (20-30 μM). Colony formation, as an indicator of proliferation ability, was completely inhibited by AgNPs at concentrations higher than 1 μg/ml. Silver ions had less effect on the colony formation of RBE4 cells than AgNPs.

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

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

[3]  Peter Wick,et al.  Barrier Capacity of Human Placenta for Nanosized Materials , 2009, Environmental health perspectives.

[4]  T. Xi,et al.  Influence of silver nanoparticles on neurons and blood-brain barrier via subcutaneous injection in rats , 2008 .

[5]  Saber M Hussain,et al.  Assessment of the toxicity of silver nanoparticles in vitro: a mitochondrial perspective. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[6]  T. Syversen,et al.  The colony formation assay as an indicator of carbon nanotube toxicity examined in three cell lines , 2009 .

[7]  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.

[8]  J. Didziapetriene,et al.  Transport of nanoparticles through the placental barrier. , 2011, The Tohoku journal of experimental medicine.

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

[10]  Zhiqiang Hu,et al.  Size dependent and reactive oxygen species related nanosilver toxicity to nitrifying bacteria. , 2008, Environmental science & technology.

[11]  Wolfgang Kreyling,et al.  Toxicological hazards of inhaled nanoparticles--potential implications for drug delivery. , 2004, Journal of nanoscience and nanotechnology.

[12]  Tobias Wang,et al.  Silver nanoparticles and silver nitrate cause respiratory stress in Eurasian perch (Perca fluviatilis). , 2010, Aquatic toxicology.

[13]  Robert Gelein,et al.  EXTRAPULMONARY TRANSLOCATION OF ULTRAFINE CARBON PARTICLES FOLLOWING WHOLE-BODY INHALATION EXPOSURE OF RATS , 2002, Journal of toxicology and environmental health. Part A.

[14]  Robert J. Allaway,et al.  Gold nanoparticle trafficking of typically excluded compounds across the cell membrane in JB6 Cl 41-5a cells causes assay interference , 2011, Nanotoxicology.

[15]  Christina M. Powers,et al.  Silver Nanoparticles Compromise Neurodevelopment in PC12 Cells: Critical Contributions of Silver Ion, Particle Size, Coating, and Composition , 2010, Environmental health perspectives.

[16]  Karluss Thomas,et al.  Research strategies for safety evaluation of nanomaterials, part VII: evaluating consumer exposure to nanoscale materials. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[17]  Clinton F Jones,et al.  In vitro assessments of nanomaterial toxicity. , 2009, Advanced drug delivery reviews.

[18]  G. Oberdörster,et al.  Safety assessment for nanotechnology and nanomedicine: concepts of nanotoxicology , 2010, Journal of internal medicine.

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

[20]  Jong Hoon Park,et al.  Comparison of acute responses of mice livers to short-term exposure to nano-sized or micro-sized silver particles , 2008, Biotechnology Letters.

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

[22]  T. Nilsen,et al.  Trace elements in serum from patients with Parkinson's disease — a prospective case-control study The Nord-Trøndelag Health Study (HUNT) , 2008, Brain Research.

[23]  A. Lansdown Silver in health care: antimicrobial effects and safety in use. , 2006, Current problems in dermatology.

[24]  S. Oldenburg,et al.  Evaluation of Silver Nanoparticle Toxicity in Skin in Vivo and Keratinocytes in Vitro , 2009, Environmental health perspectives.

[25]  Gordon Chambers,et al.  Comparative in vitro cytotoxicity study of silver nanoparticle on two mammalian cell lines. , 2012, Toxicology in vitro : an international journal published in association with BIBRA.

[26]  W. Kreyling,et al.  Translocation of Inhaled Ultrafine Particles to the Brain , 2004, Inhalation toxicology.

[27]  Matthias Epple,et al.  TOXICITY OF SILVER NANOPARTICLES INCREASES DURING STORAGE BECAUSE OF SLOW DISSOLUTION UNDER RELEASE OF SILVER IONS , 2010 .

[28]  R. Mumper,et al.  Paclitaxel nanoparticles for the potential treatment of brain tumors. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

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

[30]  Merle G Paule,et al.  Silver nanoparticle induced blood-brain barrier inflammation and increased permeability in primary rat brain microvessel endothelial cells. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[31]  R. Dietz,et al.  The pharmacological relevance of vital staining with neutral red , 1979, Experientia.

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

[33]  Andrew D Maynard,et al.  The new toxicology of sophisticated materials: nanotoxicology and beyond. , 2011, Toxicological sciences : an official journal of the Society of Toxicology.

[34]  X. Chen,et al.  Nanosilver: a nanoproduct in medical application. , 2008, Toxicology letters.

[35]  Kirk G Scheckel,et al.  Surface charge-dependent toxicity of silver nanoparticles. , 2011, Environmental science & technology.

[36]  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.

[37]  E Borenfreund,et al.  Toxicity determined in vitro by morphological alterations and neutral red absorption. , 1985, Toxicology letters.

[38]  Saber M Hussain,et al.  Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique. , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

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

[40]  N. Miura,et al.  Cytotoxic effect and apoptosis induction by silver nanoparticles in HeLa cells. , 2009, Biochemical and biophysical research communications.

[41]  Gordon Chambers,et al.  A new approach to the toxicity testing of carbon-based nanomaterials--the clonogenic assay. , 2007, Toxicology letters.

[42]  I. Rodushkin,et al.  Improved multi-elemental analyses by inductively coupled plasma-sector field mass spectrometry through methane addition to the plasma , 2005 .

[43]  R. Sinha,et al.  Interaction and nanotoxic effect of ZnO and Ag nanoparticles on mesophilic and halophilic bacterial cells. , 2011, Bioresource technology.

[44]  S. Franzen,et al.  Probing BSA binding to citrate-coated gold nanoparticles and surfaces. , 2005, Langmuir : the ACS journal of surfaces and colloids.

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

[46]  O. Salata,et al.  Applications of nanoparticles in biology and medicine , 2004, Journal of nanobiotechnology.

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

[48]  J. Schlager,et al.  In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[49]  K. Paknikar,et al.  Interactions of silver nanoparticles with primary mouse fibroblasts and liver cells. , 2009, Toxicology and applied pharmacology.

[50]  Holger Moch,et al.  Nanoparticle cytotoxicity depends on intracellular solubility: comparison of stabilized copper metal and degradable copper oxide nanoparticles. , 2010, Toxicology letters.

[51]  J. Yeh,et al.  Induction of cytotoxicity and apoptosis in mouse blastocysts by silver nanoparticles. , 2010, Toxicology letters.

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

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