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Development and validation of a semiautomatic system for generation and deposition of sprays on isolated cells of the respiratory tract

: Knebel, J.W.; Ritter, D.; Hoffmann, K.; Lödding, H.; Windt, H.; Koch, W.; Aufderheide, M.


Toxicology methods 11 (2001), Nr.3, S.161-171
ISSN: 1051-7235
Fraunhofer ITA ( ITEM) ()
liquid interface; air interface; aerosol; in vitro; human; epithelial cell

A computer-controlled spray-generating and spray-exposure unit called RHINOCON was designed and constructed to offer the possibility of depositing, for example, environmental or pharmaceutical aerosols at the air/liquid interface under realistic exposure conditions. Here, isolated human respiratory cells are chosen to simulate, as far as possible, the in vivo situation. The exposure unit is equipped to hold up to 6 commercially available pump-spray dispensers, which are prepared mechanically for exposure as described in the manufacturer's users' guide. A transwell insert containing the cells is placed opposite each dispenser in a special plate head first. After starting the program, the spray dispensers are shaken and actuated automatically. The spray is released into an air flow passing over the cells, where the spray droplets are deposited due to their inertia. After exposure, the inserts can easily be removed and processed to measure a variety of cell toxicity parameters, such as cell viability or cell metabolism. Validation of the experimental setup was performed in an initial set of experiments: (1) by determination of the deposited amount of nebulized test substance on transwell membranes, and (2) by investigation of the cellular reactions of a human respiratory epithelial cell line (HFBE 21) exposed to nebulized hydrogen peroxide or to standard medium as a reference control.