Non-specific cellular uptake of surface-functionalized quantum dots.

We report a systematic empirical study of nanoparticle internalization into cells via non-specific pathways. The nanoparticles were comprised of commercial quantum dots (QDs) that were highly visible under a fluorescence confocal microscope. Surface-modified QDs with basic biologically significant moieties, e.g. carboxyl, amino, and streptavidin, were used, in combination with surface derivatization with polyethylene glycol (PEG) for a range of immortalized cell lines. Internalization rates were derived from image analysis and a detailed discussion about the effect of nanoparticle size, charge and surface groups is presented. We find that PEG derivatization dramatically suppresses the non-specific uptake while PEG-free carboxyl and amine functional groups promote QD internalization. These uptake variations displayed a remarkable consistency across different cell types. The reported results are important for experiments concerned with cellular uptake of surface-functionalized nanomaterials, both when non-specific internalization is undesirable and when it is intended for material to be internalized as efficiently as possible.

[1]  Jasmina Lovrić,et al.  Fate of micelles and quantum dots in cells. , 2007, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[2]  S. Carotta,et al.  DNA/polyethylenimine transfection particles: Influence of ligands, polymer size, and PEGylation on internalization and gene expression , 2001, AAPS PharmSci.

[3]  W. Webb,et al.  Water-Soluble Quantum Dots for Multiphoton Fluorescence Imaging in Vivo , 2003, Science.

[4]  Stephanie E. A. Gratton,et al.  The effect of particle design on cellular internalization pathways , 2008, Proceedings of the National Academy of Sciences.

[5]  S. Zigmond,et al.  Inhibition of receptor-mediated but not fluid-phase endocytosis in polymorphonuclear leukocytes , 1985, The Journal of cell biology.

[6]  Shuming Nie,et al.  Cell-penetrating quantum dots based on multivalent and endosome-disrupting surface coatings. , 2007, Journal of the American Chemical Society.

[7]  Moungi G Bawendi,et al.  Compact biocompatible quantum dots functionalized for cellular imaging. , 2008, Journal of the American Chemical Society.

[8]  Joseph L. Goldstein,et al.  Coated pits, coated vesicles, and receptor-mediated endocytosis , 1979, Nature.

[9]  Jean-Pierre Benoit,et al.  Parameters influencing the stealthiness of colloidal drug delivery systems. , 2006, Biomaterials.

[10]  Masato Yasuhara,et al.  Use of fluorescent quantum dot bioconjugates for cellular imaging of immune cells, cell organelle labeling, and nanomedicine: surface modification regulates biological function, including cytotoxicity , 2007, Journal of Artificial Organs.

[11]  Nancy A Monteiro-Riviere,et al.  Mechanisms of quantum dot nanoparticle cellular uptake. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[12]  Yuliang Zhang,et al.  Transmembrane delivery of the cell-penetrating peptide conjugated semiconductor quantum dots. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[13]  K Dane Wittrup,et al.  Monovalent, reduced-size quantum dots for imaging receptors on living cells , 2008, Nature Methods.

[14]  M. Howarth,et al.  Targeting quantum dots to surface proteins in living cells with biotin ligase. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Hanning Xiao,et al.  Investigation of PEG adsorption on the surface of zinc oxide nanoparticles , 2004 .

[16]  Michael A. Schreuder,et al.  Controlling the reactivity of ampiphilic quantum dots in biological assays through hydrophobic assembly of custom PEG derivatives. , 2008, Bioconjugate chemistry.

[17]  Arezou A Ghazani,et al.  Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. , 2006, Nano letters.

[18]  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.

[19]  Ron C. Hardman.  A Toxicologic Review of Quantum Dots: Toxicity Depends on Physicochemical and Environmental Factors , 2005, Environmental health perspectives.

[20]  M. Belting.  Heparan sulfate proteoglycan as a plasma membrane carrier. , 2003, Trends in biochemical sciences.

[21]  Kaushal Rege,et al.  Cancer-cell-phenotype-dependent differential intracellular trafficking of unconjugated quantum dots. , 2009, Small.

[22]  Chi‐Hwa Wang,et al.  Effect of PEG conformation and particle size on the cellular uptake efficiency of nanoparticles with the HepG2 cells. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[23]  Igor L. Medintz,et al.  Quantum dot bioconjugates for imaging, labelling and sensing , 2005, Nature materials.

[24]  K. Audus,et al.  Microparticulate uptake mechanisms of in‐vitro cell culture models of the respiratory epithelium , 2001, The Journal of pharmacy and pharmacology.

[25]  Martin C Garnett,et al.  The effect of poly(ethylene glycol) molecular architecture on cellular interaction and uptake of DNA complexes. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[26]  D. Scherman,et al.  Lipospermine-based gene transfer into the newborn mouse brain is optimized by a low lipospermine/DNA charge ratio. , 1995, Human gene therapy.

[27]  G. Jiang,et al.  Progress in the toxicological researches for quantum dots , 2008 .

[28]  Feng Xu,et al.  Influence of PEG chain on the complement activation suppression and longevity in vivo prolongation of the PCL biomedical nanoparticles , 2009, Biomedical microdevices.

[29]  K. Higashitani,et al.  Effect of the physicochemical properties of poly(ethylene glycol) brushes on their binding to cells. , 2007, Biophysical journal.

[30]  L. Rajendran,et al.  Subcellular targeting strategies for drug design and delivery , 2010, Nature Reviews Drug Discovery.

[31]  Elizabeth L. Bentzen,et al.  Surface modification to reduce nonspecific binding of quantum dots in live cell assays. , 2005, Bioconjugate chemistry.

[32]  F. Brodsky.  Living with clathrin: its role in intracellular membrane traffic. , 1988, Science.

[33]  Brad A. Kairdolf,et al.  Minimizing nonspecific cellular binding of quantum dots with hydroxyl-derivatized surface coatings. , 2008, Analytical chemistry.

[34]  Igor L. Medintz,et al.  Delivering quantum dots into cells: strategies, progress and remaining issues , 2009, Analytical and bioanalytical chemistry.

[35]  Subra Suresh,et al.  Size‐Dependent Endocytosis of Nanoparticles , 2009, Advanced materials.

[36]  I. Zuhorn,et al.  Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. , 2004, The Biochemical journal.

[37]  O. Bourdon,et al.  Relationship between complement activation, cellular uptake and surface physicochemical aspects of novel PEG-modified nanocapsules. , 2001, Biomaterials.

[38]  Philippe Rostaing,et al.  Diffusion Dynamics of Glycine Receptors Revealed by Single-Quantum Dot Tracking , 2003, Science.

[39]  M. Wilchek,et al.  Streptavidin contains an RYD sequence which mimics the RGD receptor domain of fibronectin. , 1990, Biochemical and biophysical research communications.