[Purpose] This study investigated how the toe spreader affects muscle activities during gait in order to enhance the clinical decision-making process regarding the different characteristics of toe spreader materials. [Subjects] Twenty healthy participants were recruited. [Methods] Repeated measures of tibialis anterior (TA) and peroneus longus (PL) EMG activities (%RVC) were evaluated with and without toe separators made of two different materials (i.e., soft and hard). [Results] Compared to walking without a toe spreader, TA activity was significantly increased by the soft toe spreader and was decreased by the hard one. PL activity was significantly decreased by the soft toe spreader. [Conclusion] The altered TA and PL activities observed in this study may be the result of the toe spreader providing medio-lateral foot support during walking.