Characterizing Nanoparticle Dosimetry In Vitro

Elizabeth Jaye Brown

Abstract


Nanoparticles, which are less than 100 nm in at least one dimension, are used in a variety of fields from engineering to health care. However, adverse effects of nanoparticle exposure are still under investigation. In vitro studies generally use high bolus doses that fail to emulate realistic effects on the human lung. The Air Liquid Interface (ALI) system models the effects of airborne nanoparticles on the lung by delivering aerosols to cells grown at the air-liquid interface. This may be superior to conventional in vitro exposure methods because it creates a more realistic nanoparticle deposition on the cell surface by minimizing overlying fluid and delivering the dose as an aerosol rather than a bolus. Initial experiments with the ALI system were performed using only RPMI culture medium in the wells to analyze gold nanoparticle (25 - 200 nm) deposition on the transwell membrane, and to test well-to-well variability. Once even deposition was confirmed, type I alveolar epithelial cells of the spontaneously immortalized R3/1 line were grown on transwells and utilized to determine effects following deposition of the nanoparticles. This cell type was used due to its usefulness as a target for nanoparticles, as type I squamous epithelium of the lung constitutes about 95-98% of the total available surface area. Scanning electron microscopy was used to visualize the deposited nanoparticles following exposure. LDH assays were conducted to measure cell response (cytotoxicity). The goal of this project is to compare the ALI nanoparticle delivery method to conventional exposure methods to determine if the latter overestimate or underestimate response. Future studies may include comparing the response from a constant dose of nanoparticles using the ALI system to the response generated using conventional exposure methods. These could lead to the development of highly accurate in vitro studies, which would exclude the use of animals.

Keywords


Nanoparticles, Dosimetry, In Vitro

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