Uptake of gold nanoparticles in murine macrophage cells without cytotoxicity or production of pro-inflammatory mediators

Abstract More information characterizing the biological responses to nanoparticles is needed to allow the U.S. Food and Drug Administration to evaluate the safety and effectiveness of products with nano-scale components. The potential cytotoxicity and inflammatory responses of Au NPs (60 nm, NIST standard reference materials) were investigated in murine macrophages. Cytotoxicity was evaluated by MTT and LDH assays. Cytokines (IL-6, TNF-α), nitric oxide, and ROS were assayed to assess inflammatory responses. Morphological appearance and localization of particles were examined by high resolution illumination microscopy, transmission electron microscopy (TEM), and scanning TEM coupled with EDX spectroscopy. Results showed no cytotoxicity and no elevated production of proinflammatory mediators; however, imaging analyses demonstrated cellular uptake of Au NPs and localization within intracellular vacuoles. These results suggest that 60 nm Au NPs, under the exposure conditions tested, are not cytotoxic, nor elicit pro-inflammatory responses. The localization of Au NPs in intracellular vacuoles suggests endosomal containment and an uptake mechanism involving endocytosis.

[1]  P. Goering,et al.  Effects of particulate and soluble cadmium species on biochemical and functional parameters in cultured murine macrophages. , 2000, In vitro & molecular toxicology.

[2]  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.

[3]  Sabine Neuss,et al.  Size-dependent cytotoxicity of gold nanoparticles. , 2007, Small.

[4]  Vincent M Rotello,et al.  Functionalized gold nanoparticles for drug delivery. , 2007, Nanomedicine.

[5]  P. Jain,et al.  Gold nanoparticles: interesting optical properties and recent applications in cancer diagnostics and therapy. , 2007, Nanomedicine.

[6]  Sara Linse,et al.  Understanding the nanoparticle–protein corona using methods to quantify exchange rates and affinities of proteins for nanoparticles , 2007, Proceedings of the National Academy of Sciences.

[7]  L. Dai,et al.  Can silver nanoparticles be useful as potential biological labels? , 2008, Nanotechnology.

[8]  Marina A Dobrovolskaia,et al.  Evaluation of nanoparticle immunotoxicity. , 2009, Nature nanotechnology.

[9]  Saber M Hussain,et al.  Are diamond nanoparticles cytotoxic? , 2007, The journal of physical chemistry. B.

[10]  Rebecca S. Shawgo,et al.  Biocompatibility and biofouling of MEMS drug delivery devices. , 2003, Biomaterials.

[11]  Anil K Patri,et al.  Method for analysis of nanoparticle hemolytic properties in vitro. , 2008, Nano letters.

[12]  Paresh Chandra Ray,et al.  Challenge in Understanding Size and Shape Dependent Toxicity of Gold Nanomaterials in Human Skin Keratinocytes. , 2008, Chemical physics letters.

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

[14]  D. Astruc,et al.  Gold nanoparticles: assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology. , 2004, Chemical reviews.

[15]  C. Mirkin,et al.  Optically and Chemically Encoded Nanoparticle Materials for DNA and Protein Detection , 2005 .

[16]  C. Murphy,et al.  Gold nanoparticles are taken up by human cells but do not cause acute cytotoxicity. , 2005, Small.

[17]  P. Quincey,et al.  Characterisation of the de-agglomeration effects of bovine serum albumin on nanoparticles in aqueous suspension. , 2010, Colloids and surfaces. B, Biointerfaces.

[18]  D. Tsai,et al.  Biosensing, Cytotoxicity, and Cellular Uptake Studies of Surface-Modified Gold Nanorods , 2009 .

[19]  Chad A. Mirkin,et al.  Programmed Materials Synthesis with DNA. , 1999, Chemical reviews.

[20]  Huabei Jiang,et al.  Gold-Speckled Multimodal Nanoparticles for Noninvasive Bioimaging , 2008, Chemistry of materials : a publication of the American Chemical Society.

[21]  M. Lipscomb.  Lung defenses against opportunistic infections. , 1989, Chest.

[22]  L. Dai,et al.  Preparation of cells for assessing ultrastructural localization of nanoparticles with transmission electron microscopy , 2010, Nature Protocols.

[23]  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.

[24]  H. Krug,et al.  Carbon nanotubes show no sign of acute toxicity but induce intracellular reactive oxygen species in dependence on contaminants. , 2007, Toxicology letters.

[25]  Vincent M Rotello,et al.  Toxicity of gold nanoparticles functionalized with cationic and anionic side chains. , 2004, Bioconjugate chemistry.

[26]  Alexandra Kroll,et al.  Current in vitro methods in nanoparticle risk assessment: limitations and challenges. , 2009, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[27]  N. Wang,et al.  Comparison of cytotoxic and inflammatory responses of photoluminescent silicon nanoparticles with silicon micron‐sized particles in RAW 264.7 macrophages , 2009, Journal of applied toxicology : JAT.

[28]  Michael S. Strano,et al.  Size-dependent cellular uptake and expulsion of single-walled carbon nanotubes: single particle tracking and a generic uptake model for nanoparticles. , 2009, ACS nano.

[29]  D. Jaffray,et al.  Intracellular uptake, transport, and processing of nanostructures in cancer cells. , 2009, Nanomedicine : nanotechnology, biology, and medicine.

[30]  Petra Krystek,et al.  Particle size-dependent organ distribution of gold nanoparticles after intravenous administration. , 2008, Biomaterials.

[31]  S. Hersch,et al.  A Novel Procedure for Pre-embedding Double Immunogold–Silver Labeling at the Ultrastructural Level , 2001, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[32]  Warren C W Chan,et al.  Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. , 2007, Nano letters.

[33]  R. Shukla,et al.  Biocompatibility of gold nanoparticles and their endocytotic fate inside the cellular compartment: a microscopic overview. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[34]  D. B. Lyle,et al.  Screening biomaterials for stimulation of nitric oxide-mediated inflammation. , 2009, Journal of biomedical materials research. Part A.

[35]  R. Shukla,et al.  Gastrin releasing protein receptor specific gold nanorods: breast and prostate tumor avid nanovectors for molecular imaging. , 2009, Nano letters.

[36]  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.

[37]  S. Hsu,et al.  Cytotoxicity and immunological response of gold and silver nanoparticles of different sizes. , 2009, Small.

[38]  Xiaohua Huang,et al.  Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer. , 2005, Nano letters.

[39]  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.