While the need for caregivers' brushing have increased, the appropriate brushing motion is still unclear. The aim of this study was to clarify the gender difference of brushing motions between self-brushing and caregivers' brushing.Tooth brushing motions of 15 male and 20 female dental professionals were analyzed using a three-dimensional accelerometer and a strain tension gage. They brushed the buccal surfaces of the right lower molars using two brushing situations. Multilevel model analysis was applied in this study.Concerning self-brushing, gender difference was significant in cycle duration, buccal-lingual motion and brushing force, whereas for caregivers' brushing, a significant difference was observed only in the brushing force. In addition, the brushing force during caregivers' brushing was significantly greater than that of self-brushing on both genders.This study clearly demonstrated gender difference of brushing motion and force. Furthermore, a significant difference between self-brushing and caregivers' brushing was also evident. These results might contribute to improving the caregivers' brushing.