Abstract
In vitro experiments typically measure the uptake of nanoparticles by exposing cells at the bottom of a culture plate to a suspension of nanoparticles, and it is generally assumed that this suspension is well-dispersed. However, nanoparticles can sediment, which means that the concentration of nanoparticles on the cell surface may be higher than the initial bulk concentration, and this could lead to increased uptake by cells. Here, we use upright and inverted cell culture configurations to show that cellular uptake of gold nanoparticles depends on the sedimentation and diffusion velocities of the nanoparticles and is independent of size, shape, density, surface coating and initial concentration of the nanoparticles. Generally, more nanoparticles are taken up in the upright configuration than in the inverted one, and nanoparticles with faster sedimentation rates showed greater differences in uptake between the two configurations. Our results suggest that sedimentation needs to be considered when performing in vitro studies for large and/or heavy nanoparticles.
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References
Ghosh, P., Han, G., De, M., Kim, C. K. & Rotello, V. M. Gold nanoparticles in delivery applications. Adv. Drug Deliv. Rev. 60, 1307–1315 (2008).
Maeda, H., Wu, J., Sawa, T., Matsumura, Y. & Hori, K. Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J. Control. Rel. 65, 271–284 (2000).
Sawant, R. M. et al. ‘SMART’ drug delivery systems: double-targeted pH-responsive pharmaceutical nanocarriers. Bioconj. Chem. 17, 943–949 (2006).
Rosler, A., Vandermeulen, G. W. M. & Klok, H-A. Advanced drug delivery devices via self-assembly of amphiphilic block copolymers. Adv. Drug Deliv. Rev. 53, 95–108 (2001).
Jain, P. K., Huang, X., El-Sayed, I. H. & El-Sayed, M. A. Noble metals on the nanoscale: optical and photothermal properties and some applications in imaging, sensing, biology, and medicine. Acc. Chem. Res. 41, 1578–1586 (2008).
Skrabalak, S. E. et al. Gold nanocages: synthesis, properties, and applications. Acc. Chem. Res. 41, 1587–1595 (2008).
Lal, S., Clare, S. E. & Halas, N. J. Nanoshell-enabled photothermal cancer therapy: impending clinical impact. Acc. Chem. Res. 41, 1842–1851 (2008).
Murphy, C. J. et al. Gold nanoparticles in biology: beyond toxicity to cellular imaging. Acc. Chem. Res. 41, 1721–1730 (2008).
Zerda, A. D. L. et al. Carbon nanotubes as photoacoustic molecular imaging agents in living mice. Nature Nanotech. 3, 557–562 (2008).
Gao, X., Cui, Y., Levenson, R. M., Chung, L. W. K. & Nie, S. In vivo cancer targeting and imaging with semiconductor quantum dots. Nature Biotechnol. 22, 969–976 (2004).
Sun, C., Lee, J. S. H. & Zhang, M. Magnetic nanoparticles in MR imaging and drug delivery. Adv. Drug Deliv. Rev. 60, 1252–1265 (2008).
Nel, A., Xia, T., Madler, L. & Li, N. Toxic potential of materials at the nanolevel. Science 311, 622–627 (2006).
Teeguarden, J. G., Hinderliter, P. M., Orr, G., Thrall, B. D. & Pounds, J. G. Particokinetics in vitro: dosimetry considerations for in vitro nanoparticle toxicity assessments. Toxicol. Sci. 95, 300–312 (2007).
Lison, D. et al. Nominal and effective dosimetry of silica nanoparticles in cytotoxicity assays. Toxicol. Sci. 104, 155–162 (2008).
Jiang, W., Kim, B. Y. S., Rutka, J. T. & Chan, W. C. W. Nanoparticle-mediated cellular response is size-dependent. Nature Nanotech. 3, 145–150 (2008).
Alivisatos, A. P., Gu, W. & Larabell, C. Quantum dots as cellular probes. Annu. Rev. Biomed. Eng. 7, 55–76 (2005).
Rejman, J., Oberle, V., Zuhorn, I. S. & Hoekstra, D. Size-dependent internalization of particles via the pathways of clathrinand caveolae-mediated endocytosis. Biochem. J. 377, 159–169 (2004).
Panyam, J. & Labhasetwar, V. Biodegradable nanoparticles for drug and gene delivery to cells and tissue. Adv. Drug Deliv. Rev. 55, 329–347 (2003).
Prabha, S., Zhou, W-Z., Panyam, J. & Labhasetwar, V. Size-dependency of nanoparticle-mediated gene transfection: studies with fractionated nanoparticles. Int. J. Pharm. 244, 105–115 (2002).
Cho, E. C., Au, L., Zhang, Q. & Xia, Y. The effects of size, shape, and surface functional group old nanoparticles on their adsorption and internalization by cells. Small 6, 517–522 (2010).
Chithrani, B. D. & Chan, W. C. W. Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. Nano Lett. 7, 1542–1550 (2007).
Verma, A. & Stellacci, F. Effect of surface properties on nanoparticle–cell interactions. Small 6, 12–21 (2010).
Verma, A. et al. Surface-structure-regulated cell-membrane penetration by monolayer-protected nanoparticles. Nature Mater. 7, 588–595 (2008).
Leroueil, P. R. et al. Nanoparticle interaction with biological membranes: does nanotechnology present a Janus face? Acc. Chem. Res. 40, 335–342 (2007).
Cho, E. C., Xie, J., Wurm, P. A. & Xia, Y. Understanding the role of surface charges in cellular adsorption versus internalization by selectively removing gold nanoparticles on the cell surface with a I2/KI etchant. Nano Lett. 9, 1080–1084 (2009).
Zorko, M. & Langel, U. Cell-penetrating peptides: mechanism and kinetics of cargo delivery. Adv. Drug Deliv. Rev. 57, 529–545 (2005).
Sudimack, J. & Lee, R. J. Targeted drug delivery via the folate receptor. Adv. Drug Deliv. Rev. 41, 147–162 (2000).
Sager, T. M. et al. Improved method to disperse nanoparticles for in vitro and in vivo investigation of toxicity. Nanotoxicology 1, 118–129 (2007).
Xu, C., Tung, G. A. & Sun, S. Size and concentration effect of gold nanoparticles on X-ray attenuation as measured on computed tomography. Chem. Mater. 20, 4167–4169 (2008).
Hinderliter, P. M. et al. ISDD: a computational model of particle sedimentation, diffusion and target cell dosimetry for in vitro toxicity studies. Particle Fibre Toxicol. 7, 36–54 (2010).
Kong, H. J. et al. Non-viral gene delivery regulated by stiffness of cell adhesion substrates. Nature Mater. 4, 460–464 (2005).
Alkilany, A. M. et al. Cellular uptake and cytotoxicity of gold nanorods: molecular origin of cytotoxicity and surface effects. Small 5, 701–708 (2009).
Lynch, I. et al. The nanoparticle–protein complex as a biological entity: a complex fluids and surface science challenge for the 21st century. Adv. Colloid Interface Sci. 134–135, 167–174 (2007).
Conner, S. D. & Schmid, S. L. Regulated portals of entry into the cell. Nature 422, 37–44 (2003).
Feldman, K., Ha1hner, G., Spencer, N. D., Harder, P. & Grunze, M. Probing resistance to protein adsorption of oligo(ethylene glycol)-terminated self-assembled monolayers by scanning force microscopy. J. Am. Chem. Soc. 121, 10134–10141 (1999).
Sigal, G. B., Mrksich, M. & Whitesides, G. M. Effect of surface wettability on the adsorption of proteins and detergents. J. Am. Chem. Soc. 120, 3464–3473 (1998).
Horbett, T. A. & Brash, J. L. (eds) Proteins at Interfaces II 396 (American Chemical Society, 1995).
Cho, E. C., Liu, Y. & Xia, Y. A simple spectroscopic method for differentiating cellular uptakes of gold nanospheres and nanorods from their mixtures. Angew. Chem. Int. Ed. 49, 1976–1980 (2010).
Leckband, D. Nanomechanics of adhesion proteins. Curr. Opin. Struct. Biol. 14, 524–530 (2004).
Dammer, U. et al. Specific antigen/antibody interactions measured by force microscopy. Biophy. J. 70, 2437–2441 (1996).
Allen, S. et al. Detection of antigen–antibody binding events with the atomic force microscope. Biochemistry 36, 7457–7463 (1997).
Tha, S. P., Shuster, J. & Goldsmith, H. L. Interaction forces between red cells agglutinated by antibody. II. Measurement of hydrodynamic force of breakup. Biophys. J. 50, 1117–1126 (1986).
Vasir, J. K. & Labhasetwar, V. Quantification of the force of nanoparticle–cell membrane interactions and its influence on intracellular trafficking of nanoparticles. Biomaterials 29, 4244–4252 (2008).
Hiemenz, P. C. & Rajagopalan, R. Principles of Colloid and Surface Chemistry 94 (Marcel Dekker, 1997).
Brown, P. H. & Schuck, P. Macromolecular size-and-shape distributions by sedimentation velocity analytical ultracentrifugation. Biophys. J. 90, 4651–4661 (2006).
Sperling, R. A. et al. Size determination of (bio)conjugated water-soluble colloidal nanoparticles: a comparison of different techniques. J. Phys. Chem. 111, 11552–11559 (2007).
Shim, J. et. al. Transdermal delivery of mixnoxidil with block copolymer nanoparticles. J. Control. Rel. 97, 477–484 (2004).
Sonavane, G. et al. In vitro permeation of gold nanoparticles through rat skin and rat intestine: effect of particle size. Colloids Sur. B 65, 1–10 (2008).
Acknowledgements
Y.X. thanks the National Institutes of Health (NIH) for a 2006 Director's Pioneer Award (DP1 OD000798) and a grant (1R01 CA138527). E.C.C. was also partially supported by a fellowship from the Korea Research Foundation (KRF-2007-357-D00070). Part of the work was performed at the Nano Research Facility, a member of the National Nanotechnology Infrastructure Network (NNIN), supported by the National Science Foundation (NSF) under award ECS-0335765.
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E.C.C. and Y.X. conceived and designed the experiments. E.C.C. performed the experiments, analysed the data and prepared the manuscript. Y.X. revised the manuscript. Q.Z. synthesized the nanocages.
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Cho, E., Zhang, Q. & Xia, Y. The effect of sedimentation and diffusion on cellular uptake of gold nanoparticles. Nature Nanotech 6, 385–391 (2011). https://doi.org/10.1038/nnano.2011.58
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DOI: https://doi.org/10.1038/nnano.2011.58
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