Spectroscopic study of the bioactivity of nanoparticles by exposure to human lung cells. (Summer Research Institute, PNNL Scope #16248)
EMSL Project ID
18413
Abstract
Recent investigations of inhalation hazards have begun to indicate possible correlations between ultrafines (particles with an aerodynamic radius of 100 nm or less) and cardiovascular damage that may be the result of increased production of cytokines by macrophages resident in the lung. Such effects are strongly composition dependent. The complexity of issues relating PM composition, particle size, deposition in the lung, and cellular responses necessitates monitoring the health impact of inhaled PM hazards and developing methods, which can capture the multifaceted nature of this problem. The goal is to develop a new airborne testing capability with complementary infrared (IR) [e.g., attenuated total reflectance (ATR) spectroscopy] and fluorescence spectroscopic techniques that will be useful for real-time, in situ, deployable, high throughput screening of cellular responses to PM. Under this project, we will use ATR spectroscopy for monitoring in situ biological activity induced by nanoparticles (eg carbon nanotubes, quantum dots) in real time. An IR transparent crystal (ZnSe) surface shall be used as both a substrate/waveguide and sensing element, to permits remote collection of IR spectra. The principle of measurement is based on the general concept of the ATR spectroscopy. Under appropriate conditions, light can be forced to travel near or at the surface of a transparent medium, and in its course of travel it will interact with the environment in the near-surface region of that medium. Various absorptions occur based on the resonant structures of the molecules during this interaction. These absorptive losses can be measured and quantified using an IR spectrometer. The objective is to coat the surface of the IR transparent medium with macrophages or epithelial cells so that cell response to controlled PM or LPS exposures can be monitored evanescently.Macrophages or epithelial cells will be grown on ZnSe surfaces and exposed to the nanoparticles. IR observable changes will be defined as a function of time and samples will be analyzed for biomarker expression using fluorescence microscopy in parallel. This approach will enable the determination of whether 1) the cells are exhibiting the expected inflammatory response when seeded on IR fibers and 2) whether IR observable changes in the cells directly correlates with the appearance of biomarkers of the inflammatory response. The goal is to identify biomarkers of response that are both detected by the IR system and are predictive of an inflammatory response to enable real time monitoring of cells and potentially deployment of cells as sensors. The surface of the carbon nanotubes and quantum dot shall be functionalized to study the specificity of the nanoparticles. TEM, SEM/EDS shall be used to study the surface morphology and chemistry.
Project Details
Project type
Exploratory Research
Start Date
2006-05-08
End Date
2007-03-22
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members