10th International Aerosol Conference September 2 - September 7, 2018 America's Center Convention Complex St. Louis, Missouri, USA
Abstract View
Development of an in Vitro Exposure System Enabled to Track Biological Impacts of Aerosol on Human Lung Cells in Real-Time
NIMA AFSHAR-MOHAJER, Lakshmana Chandrala, Kristine Nishida, Venkataramana Sidhaye, Joseph Katz, Kirsten Koehler, Johns Hopkins School of Public Health
Abstract Number: 408 Working Group: Instrumentation
Abstract In vitro exposure systems for studying aerosol-lung cells interactions have the limitations of inability of tracking the effects in real-time, non-uniform dose delivery onto the cell layer and being expensive. With the goal of addressing these shortcomings, we developed a novel in vitro exposure system, which simulates exposure of human bronchial epithelial cells to aerosols. We assessed the physical and biological performances of the system in comparison to a 3-well-insert stainless steel VitroCell® exposure system (VitroCell Inc., Germany) using cigarette smoke as a challenge aerosol. Then, a microscopy system was incorporated to allow visualization of the lung cells when exposed to crude oil and polystyrene latex droplets, which possess fluorescent properties. The imaging system consisted of a 20× long-distance objective, a tube lens, and a camera illustrated alterations in the ciliary beat frequency and spatial distribution of the depositing particles throughout the aerosol exposure. During exposure, live fluorescent stains were used to tag the live cells to determine cell viability. Based on the gravimetric analysis results, the average mass of the collected particles using our system versus VitroCell® were 0.177±0.027 vs 0.172±0.025 µg when exposed to 32 puffs, with a total volume of 35 mL at an incoming flow rate of 0.2 L/min per insert. Our exposure system provided a more uniform cellular exposure when exposed to the cigarette smoke. Toxicity assessment demonstrated 30% reduction in trans-epithelial electrical resistance of the exposed cells in our system compared to 8% reduction when VitroCell® was used. The percentage change in permeability of the cells monolayer after exposure to the cigarette smoke was 195% vs 175%.
We observed a more uniform delivery in our proposed exposure system at a much lower cost. Our system also facilitates the live visualization of cells during the exposure and allows an investigation of the changes within the epithelium to disrupt tissue integrity in response to inhalation of aerosols.