Objective To investigate the effect of endotracheal tube cuff inflation pressure on the occurrence of liquid aspiration and tracheal wall damage.Study design Prospective, randomized experimental study.Animals Ten healthy horses, weighing 535 ± 55 kg.Methods Horses were anesthetized, orotracheally intubated, placed in dorsal recumbency, and maintained on isoflurane in oxygen with controlled ventilation for 175 ± 15 minutes. The horses were randomly assigned to an endotracheal cuff pressure of 80–100 or 120 cm H2O. The cuff pressure was continuously monitored and maintained at a constant pressure. Methylene blue in saline was instilled proximal to the cuff. After euthanasia, the trachea was opened distal to the endotracheal tube tip to check for evidence of dye leaking past the cuff. The cervical trachea was then resected and opened longitudinally for gross and histologic examinations.Results No blue staining was found distal to the cuff in any horse. Visual examination of the tracheal mucosa revealed hyperemic and hemorrhagic lesions at the site of the cuff contact. Histologic changes included epithelium attenuation or erosion, submucosal neutrophilic infiltration, and submucosal hemorrhages. Lesions were absent or less extensive in the lower cuff pressure group as compared to the high cuff pressure group.Conclusions The endotracheal tube cuff produced a seal sufficient to prevent leakage in both groups. Tracheal wall damage was more severe and occurred more frequently in the higher cuff pressure group.Clinical relevance Tracheal mucosal damage induced by cuff inflation is pressure-dependent. Cuff pressure monitoring is recommended.