Hyperoxia-induced changes in human airway epithelial cells: the protective effect of perflubron.
OBJECTIVE
To determine the protective effect of perflubron (PFB), a type of perfluorochemical liquid, in hyperoxia-induced cellular injury in the human airway epithelial cells.


METHODS
A controlled, in vitro laboratory study.


METHODS
Tertiary-care children's hospital.


METHODS
Human airway epithelial cells.


METHODS
Human airway epithelial cells, Calu-3 cells, grown on polycarbonate porous filters at an air-liquid interface culture were exposed to normoxic (Fico(2) = 5%, balance air) or hyperoxic (Fio(2) = 95%, balance CO(2)) conditions. Hyperoxia-induced cellular changes were monitored by measuring transepithelial resistance (TER) of monolayers, histology of cells, total protein, and interleukin-8 (IL-8) secretion in apical surface fluid (ASF) washings. Under hyperoxic conditions, the protective effect of PFB was assessed by directly adding PFB liquid to the apical surface of monolayers.


RESULTS
During hyperoxic gas-liquid interface culture, Calu-3 monolayers exhibited a loss of cellular integrity morphologically, decreased protein concentration, and IL-8 level in ASF washings. During hyperoxic PFB-liquid interface culture, there was an overall increase in TER value of monolayers, improved histology, decreased total protein secretion in ASF washings, and unaltered IL-8 secretion. Cytomorphologic observations of PFB-treated Calu-3 cells indicated the presence of varying numbers of differently sized intracellular vacuoles during both normoxic and hyperoxic conditions.


CONCLUSIONS
We conclude that the air-liquid interface culture of Calu-3 may be helpful in understanding mechanisms of lung injuries caused in clinical practice, and PFB protects against hyperoxia-induced airway epithelial cell injury by promoting cellular integrity as well as cytologic modifications. PFB-liquid interface culture of Calu-3 may be a useful in vitro model for studying the cytoprotective role of liquid ventilation.
Babu
Polani B Ramesh
PB
Nemours Research Lung Center, Alfred I. duPont Hospital for Children, Wilmington, DE 19803, USA.
Chidekel
Aaron
A
Shaffer
Thomas H
TH
D018929:Cell Culture Techniques; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology*; D005466:Fluorocarbons / Q000627:therapeutic use*; D006801:Humans; D006842:Hydrocarbons, Brominated; D018496:Hyperoxia / Q000150:complications / Q000517:prevention & control*; D016209:Interleukin-8 / Q000378:metabolism; D021061:Liquid Ventilation; D008168:Lung / Q000378:metabolism / Q000473:pathology*; D012131:Respiratory Insufficiency / Q000378:metabolism / Q000628:therapy
D005466:Fluorocarbons; D006842:Hydrocarbons, Brominated; D016209:Interleukin-8; C003072:perflubron