The aim of the study is to evaluate the Eustachian tube functionality in a group of aviators to underline a subclinical dysfunction and the related risk of ear fullness or barotitis. Hypobaric chamber allows to simulate the pressure variation of the flight. This prospective study enrolled 42 aviation pilots, members of the Italian Air Force, to whom were evaluated Eustachian tube functionality by Tubomanometry, patients subjective assessments concerning feasibility of Valsalva's and Toynbee's clinical symptoms, tympanometry, and objective Valsalva before and after exposure to hypobaric chamber. The new Eustachian tube score (ETS-7) was also calculated for each pilot before and after exposure to hypobaric chamber. Results of our examination showed that: before chamber exposition, in 92.8 % of pilots, an ETS-7 of eight or more was found bilaterally. In three (7.2 %) cases, a unilateral ETS-7 ≤ 7 was found and two of those had a positive history. After undergoing the hypobaric chamber session, the evaluation of ETS-7 showed only 19 % of pilots with an ETD score ≤7, in particular three bilateral and five unilateral cases. Three of those pilots were clinically positive: two airmen reported persistent fullness, while the other one had a barotitis. Therefore, the combined use of TMM and ETS-7 before and after hypobaric chamber exposure appears to be a reliable method for assessing the functional capacity of the Eustachian tube in aviators, the stressful effect of flight on it, to exclude subjects at increased risk of ear pain, fullness, or barotrauma