Neck pain associated with helmet-wear is an occupational health problem often observed in helicopter pilots and aircrew. Whether aircrew helmet wearing is associated with physiological and biomechanical differences between sexes is currently unknown. This study investigated neuromuscular activation patterns during different helmet-wearing conditions. The helmet load was manipulated through a novel Helmet Balancing System (HBS) in healthy, non-pilot male and female participants (n = 10 each, age 19-45 years) in two phases. Phase A assessed the acute effects of helmet-wear on neck muscles activation during head movements. Phase B examined changes in muscle activity and cervical disc height after wearing a helmet for 45 min. In Phase A, muscle activity was similar between sexes in many movements, but it was higher in female participants when wearing a helmet than in males. The HBS reduced muscle activity in both sexes. In Phase B, female participants exhibited a greater level of muscular fatigue, and male participants' cervical disc height was significantly decreased [5.7 (1.4) vs. 4.4 (1.5) mm, P