Dexmedetomidine is a preferred anesthetic agent in otological surgery because it provides controlled hypotension and good surgical field visibility. The aim of this study was to evaluate the influence of this novel agent on middle ear pressure. This prospective clinical trial was performed in 60 patients who were scheduled for elective surgery. They received dexmedetomidine or saline infusion for 20 minutes before induction of anesthesia. Tympanometric measurements were recorded for both ears at preoperative, intraoperative, and postoperative states. Mean difference of tympanometric peak pressure from baseline was statistically significant between dexmedetomidine and control group at the 30th minute of operation (24.8 daPa, P = 0.003 for right ear; 20.5 daPa, P = 0.02 for left ear) and at the end of the operation (25.8 daPa, P = 0.01 for right ear; 28.1 daPa, P = 0.004 for left ear). Dexmedetomidine anesthesia raises the tympanometric parameters, but they never exceed the limits of normal.