Nanoparticle-Cell Interactions and Intracellular Fate; Roles of Particle Surface Properties
EMSL Project ID
30433
Abstract
The growing commerce in micro- and nanotechnology is expected to increase human exposure to submicrometer and nanoscale particles, including certain forms of gold and amorphous silica. Adverse effects of these particles on living systems have been demonstrated, but the underlying mechanism is far from being understood. When inhaled, these particles are likely to reach the alveoli, where alveolar type II epithelial cells are found. These cells play critical roles in the function of the alveoli and participate in the immune response to certain particles by releasing chemokines. The cellular interactions of the particles, which drive the cellular response, are still unclear. Furthermore, the role of particle surface properties in the cellular response is largely unknown. Here we propose to investigate the cellular interactions, internalization pathways and fate of individual fluorescent nanoparticles with well defined surface properties, and correlate these processes with the response and survival of the cell. By focusing on cultured alveolar type II epithelial cells, these studies will identify properties of inhaled nanoparticles that govern their toxicity or biocompatibility, and will aid in formulating preventative approaches. These studies will directly contribute to EMSL mission in supporting environmental research, and target EMSL Science Theme by focusing on the Impact of Nanotechnology on Living Systems. Accumulating observations suggest that nanoscale particles exert harmful effects on human health to a greater extent than larger particles, and these effects have been linked to the surface properties of nanomaterial. Although large aggregates of nanoparticles have been found within cells, it is thought that such agglomerations occur as the result of experimental conditions. It is currently thought that inhaled nanoparticles are able to escape the alveolar macrophages and might directly enter the circulatory system through the epithelial wall. Our studies will therefore be guided by the working hypothesis that the initial interaction of nanoparticles with the living cell in vivo occurs at the level of individual or small nanoparticle aggregates (<100 nm), and that the surface properties of the individual nanoparticle dictate its mechanisms of interaction with the cell, and ultimately govern the level of toxicity. The investigation of dynamic processes that underlie the cellular interactions of individual or small clusters of nanoparticles, as they are likely to be presented to cells in vivo, requires tools that gain insights into living cells with high spatial and temporal resolutions. We will take advantage of EMSLs unique capabilities in single-molecule fluorescence imaging to identify and track individual nanoparticles with well defined surface properties. Using time-lapse single-molecule fluorescence imaging techniques, combined with total internal reflection fluorescence (TIRF) and DIC microscopy, it will be possible to follow individual nanoparticles and delineate their interactions and pathways within subcellular structures in real time. Using multi-channel fluorescence imaging, we will identify the endocytic pathways and final destination of the particles. By correlating the cellular interactions and fate of the particles with the cellular response and survival it will be possible to understand the mechanisms that underlie nanoparticle potential toxicity and delineate desired surface properties for designing safer nanomaterials and exposure guidelines.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2008-08-14
End Date
2011-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
6 Orr G, Panther DJ, Cassens KJ, Phillips JL, Tarasevich BJ, Pounds JG. Syndecan-1 mediates the coupling of positively charged submicrometer amorphous silica particles with actin filaments across the alveolar epithelial cell membrane. Toxicology and Applied Pharmacology. 2009, 236, 210-220.
6 Orr G, Panther DJ, Cassens KJ, Phillips JL, Tarasevich BJ, Pounds JG. Syndecan-1 mediates the coupling of positively charged submicrometer amorphous silica particles with actin filaments across the alveolar epithelial cell membrane. Toxicology and Applied Pharmacology. 2009, 236, 210?220.
de la Iglesia, Diana , Harper, Stacey , Hoover, Mark D. , Klaessig, Fred, Lippell, Phil, Maddux, Bettye, Morse, Jeffrey, Nel, Andre, Rajan, Krishna, Reznik-Zellen, Rebecca, Tuominen, Mark T.
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Harper S, JL Carriere, JM Miller, JE Hutchison, BL Maddux, and R Tanguay. 2011. "Systematic Evaluation of Nanomaterial Toxicity: Utility of Standardized Materials and Rapid Assays." ACS Nano 5(6):4688–4697. doi:10.1021/nn200546k
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DOI: 10.1021/cr050943k
Orr G, WB Chrisler, KJ Cassens, R Tan, BJ Tarasevich, LM Markillie, RC Zangar, and BD Thrall. 2010. "Cellular Recognition and Trafficking of Amorphous Silica Naoparticles by Macrophage Scavenger Receptor A." Nanotoxicology. [In Press]
Spying on chameleon nanoparticles inside living cells - (outreach article): 2014, February: http://sustainable-nano.com/2014/02/04/spying-on-chameleon-nanoparticles-inside-living-cells/#more-1728
Waters KM, LM Masiello, RC Zangar, BJ Tarasevich, NJ Karin, RD Quesenberry, S Bandyopadhyay, JG Teeguarden, JG Pounds, and BD Thrall. 2009. "Macrophage Responses to Silica Nanoparticles are Highly Conserved Across Particle Sizes." Toxicological Sciences 107(2):553-569.
Xie Y, NG Williams, A Tolic, WB Chrisler, JG Teeguarden, BL Maddux, JG Pounds, A Laskin, and G Orr. 2012. "Aerosolized ZnO nanoparticles induce toxicity in alveolar type II epithelial cells at the air-liquid interface." Toxicological Sciences 125(2):450-461. doi:10.1093/toxsci/kfr251