OBJECTIVE To investigate the effect of continuous positive airway pressure (CPAP) on middle ear pressure in patients with obstructive sleep apnea syndrome (OSAS). METHODS Prospective clinical case-control study. METHODS Seventy-eight patients with moderate and severe OSAS (the mean apnea-hypopnea indexes were 22.70 ± 3.59 and 48.59 ± 12.50, respectively) using CPAP (study group) and 60 patients with no sleep apnea syndrome (control group) were included in the study. Forty-two of the patients were female and 96 were male. The mean age in the study group was 47.38 ± 6.23, whereas it was 46.10 ± 11.96 in the control group (P > .05). The middle ear pressure of all study subjects was evaluated with tympanometry at the beginning of the study and 6 months later. Tympanometric measurements of both groups were compared. Furthermore, the same data for the study group were also evaluated according to CPAP pressure levels. RESULTS The middle ear peak pressure values of the patients in the study group were significantly increased from -63.04 ± 55.82 daPa to -39.6 ± 27.72 daPa after 6 months (P < .01). The middle ear pressure in the control group was found to be -13.26 ± 22.60 daPa at the beginning of the study and -13.60 ± 38.82 daPa after 6 months (P > .05). The mean middle ear pressure level was significantly higher in patients using CPAP at 12 to 14 cm H2 O pressure than in those using CPAP at 8 to 10 cm H2 O pressure (P < .05). CONCLUSIONS There was a significant increase in the middle ear pressure of patients using CPAP regularly for 6 months. This increase was proportional to the pressure level of the CPAP device. METHODS 3b.