Introduction: Gait evaluation and training using treadmill will be increasingly used in near future. However it is con¬troversial whether the treadmill replicates the overground environment. Goal: Aim of this study was to compare overground and treadmill ambulation for possible differences in gait tempo¬ral variables and leg joint kinematics. Materials and Methods: A total of 10 participants walked on overground and treadmill. Participants walked at their preferred velocity on overground. The treadmill velocity was adjusted average velocity obtained in overground walking. Walking in two conditions was captured by high speed camera and analyzed by motion analyses software. Results: The maximum hip flexion angle (P=0.046), maximum hip extension (P=0.0001), maximum knee extension (P=0.0001) and maximum ankle dorsiflexion (P=0.022) were significantly different in the two conditions. Conclusions: The present study suggest that statistically significant differences exist between overground and treadmill walking in healthy subjects for some joint kinematic and temporal variables.