Research has proven that mechanical deformations (MD) caused by hand flexing affect the structure of disposable protective gloves. However, these studies do not consider the presence of a physiological solution such as sweat. The combination of mechanical constraints and sweat might modify the mechanical and physical properties of the protective gloves and therefor their effectiveness. The main objective of this work is to evaluate the combined effects of human sweat and MDs on certain physical and mechanical phenomena that could affect the structure of nitrile rubber protective gloves. The strain energy, crystallinity, swelling and glove surface profile were investigated. Combined with MDs, the physiological solution significantly affects the structure of the gloves. Contact of the glove sample with the physiological solution modified the strain energy and induced a swelling that modified mechanical properties. Also, the deformation frequency greatly affected the strain energy and the number of deformations changed the degree of crystallinity. A qualitative analysis by SEM showed the deterioration of the surface of the gloves in contact with skin. Based on these results, further investigation is needed on the overall effect of sweat on the effectiveness of protective gloves against mechanical and chemical hazards.