Although elevated oxygen fraction is used in intensive care units around the world, pathological changes in pulmonary tissue have been shown to occur with prolonged exposure to hyperoxia. In this work a bovine bronchus culture model has been successfully used to evaluate the effects of hyperoxia on ciliated epithelium in vitro. Samples were cultured using an air interface method and exposed to normoxia, 21% O-2 or hyperoxia, 95% O-2. Cilial coverage was assessed using scanning electron microscopy (SEM). Tissue damage (lactate dehydrogenase, LDH, in the medium), lipid peroxidation (thiobarbituric acid reactive substances, TEARS), DNA damage (comet assay), protein oxidation (OxyBlot kit) and antioxidant status (total glutathione) were used to assess whether the hyperoxia caused significant oxidative stress. Hyperoxia caused a time-dependent decline (t(1/2) = 3.4d compared to 37.1 d under normoxia) in cilial coverage (P < 0.0001). This was associated with a significant increase in the number of cells (2.80 +/- 0.27 x 10(6) compared to 1.97 +/- 0.23 x 10(6) ml(-1) after 6 d), many apparently intact, in the medium (P < 0.05): LDH release (1.06 +/- 0.29 compared to 0.83 +/- 0.36 mu mol min(-1) g(-1) after 6 d; P < 0.001); lipid peroxidation (352 +/- 16 versus 247 +/- 11 mu mol MDA g(-1) for hyperoxia and normoxia, respectively); % tail DNA (18.7 +/- 2.2 versus 11.1 +/- 1.5); protein carbonyls (P < 0.05); and total glutathione (229 +/- 20 mu mol g(-1) versus 189 +/- 15 mu mol g(-1)). Vitamins E (10(-7) M) and C (10(-6) or 10(-7) M) alone or in combination (10(-7) M and 10(-6)M, respectively) had a significant protective effect on the hyperoxia-induced reduction in percentage cilial coverage (P < 9.05). In conclusion, hyperoxia caused damage to cultured bovine bronchial epithelium and denudation of cilia. The antioxidant vitamins E and C significantly protected against hyperoxia-induced cilia loss. (C) 2012 Elsevier Inc. All rights reserved.