(9�C11) Although there is immense literature about the interaction of NPs with normal airways and airway epithelial cells, investigations involving diseased airways are unknown. There is also a gap in understanding of as to how atmospheric Belinostat ptcl pollutants such as ozone modify the particulates by itself or the interaction with cells of the airways and the response of the airways. In this study we have used engineered NPs, that is, polystyrene NPs to investigate their deposition and interaction with CF airway epithelial cells in absence or presence of ozone and compared them with those of non-CF. Materials and Methods Airway epithelial cell culture Normal and CF cell lines CF41o-, CF45o-, and a wild-type (healthy/non-CF) airway epithelial cell line, 16HBEo-, were provided by Prof. D.

Gruenert (California Pacific Medical Center Research Institute, University of California at San Francisco). They were cultured in Eagle’s minimal essential medium (Invitrogen, Carlsbad, CA), supplemented with 10% fetal bovine serum (FBS), L-glutamine, and penicillin/streptomycin at 37��C under 5% CO2. 16HBEo- cells with stable expression of sense (16HBE-S or S-1) and antisense CFTR (16HBE-AS or AS-3) oligonucleotides were cultured as described previously.(12) Polarized cultures growing on fibronectin-collagen coated Snapwells? (0.4-��M pore size, Corning, Corning, NY). These Snapwells? are inserts that are suspended on a frame that hangs in a six-well plate. Primary healthy (non-CF) and CF airway epithelial cells Primary human bronchial epithelial cell harvest and culture was performed using established procedures previously described in detail(13) under National Jewish institutional review board (NJIRB)-approved protocols.

Primary epithelial cells (passage 2; 5��105 cells) were seeded onto 12-mm diameter type VI collagen (Sigma, St. Louis, MO)-coated Snapwells? (0.4-��M pore size, Corning, Corning, NY). Following confluence on days 4�C5 the cultures were maintained at an air�Cliquid interface (ALI). All experiments comparing primary non-CF and CF cells were performed with ALI cultures grown simultaneously and matched for passage number, number of cells plated, and days in culture. Undifferentiated cultures of primary airway epithelial cells obtained from three non-CF (normal) and three CF subjects were used (detailed description in Ahmad et al.(14)). Trans-epithelial electrical resistance (TEER) was measured as described previously.(15,16) For TEER measurement medium was added to the upper chamber 30min before TEER was measured and again removed after the measurements. GSK-3 Addition of media apically provides solution on the upper surface to dip one arm of the chopstick electrode and is necessary for TEER measurement.