Effects of oxygen concentration and exposure time on cultured human airway epithelial cells

Abstract

Objectives: To distinguish the direct effects of oxygen dose and exposure time on human airway epithelial cells. We hypothesized that progressive oxygen exposure would induce cell dysfunction and inflammation in a dose-dependent manner.

Design: Interventional laboratory study.

Setting: An academic medical research facility in the northeastern United States.

Subjects: Calu-3 human airway epithelial cell culture.

Interventions: Cells were cultured at a gas-liquid interface with the cells fed basolaterally with medium and grown to full confluence. The apical surfaces were then exposed to gas containing 21%, 40%, 60%, or 80% oxygen, 5% CO2, and balance nitrogen for 24 or 72 hrs.

Measurements and main results: The effects of oxygen concentration and time-induced cellular change were examined by measuring transepithelial resistance of monolayers, cell viability by trypan blue exclusion, basolateral lactate concentration, histology of monolayer cross-sections, and cytospin slides, plus interleukin (IL)-6 and IL-8 secretion in apical surface fluid. Transepithelial resistance decreased in a dose- and time-dependent manner (p < .001), whereas cell viability was reduced only at 72 hrs and in all hyperoxic groups (p < .05). IL-6 secretion was elevated in all hyperoxic groups at 24 hrs (p < .001), and both IL-6 and IL-8 levels were greater in the 40% FiO2 group compared with all other groups at 72 hrs (p < .01).

Conclusions: In this model, airway epithelial cells demonstrate profound concentration and time-dependent responses to hyperoxic exposure with respect to cell physiology, viability, histology, and secretion of inflammatory mediators. This model might be a valuable tool for preliminary analysis of potentially protective therapies against hyperoxia-induced airway epithelial injury.