Hyperoxia-induced changes in human airway epithelial cells: the protective effect of perflubron

Abstract

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.

Design: A controlled, in vitro laboratory study.

Setting: Tertiary-care children's hospital.

Subjects: Human airway epithelial cells.

Interventions: 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.

Measurements and main 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.