Cellular Interactions and Fate of Engineered Nanomaterials in Alveolar Epithelial Cells at the Air-Liquid Interface
EMSL Project ID
44627
Abstract
The growing commerce in micro- and nanotechnology is expected to increase human exposure to submicrometer and nanoscale particles. 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 cells, grown and exposed at the air-liquid interface (ALI). 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 molecular research, and target EMSL Science of Interfacial Phenomena Theme by investigating the interactions of nanoparticles with biological 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 EMSL's unique capabilities in single-molecule fluorescence imaging to identify and track individual nanoparticles with well defined surface properties. We will also take advantage of EMSL's new multi-photon confocal fluorescence and FLIM microscope to study particle interactions and pathways within subcellular structures, and the new flowcytometer to quantify particle uptake and cellular response. 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
2011-10-01
End Date
2014-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
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