Asymmetrical stiff knee gait is a mechanical pathology that can disrupt lower extremity muscle coordination. A better understanding of this condition can help identify potential complications. This study proposes the use of dynamic musculoskeletal modelling simulation to investigate the effect of induced mechanical perturbation on the kneeand to examine the muscle behaviour without invasive technique. Thirty-eight healthy participants were recruited. Asymmetrical gait was simulated using knee brace. Knee joint angle, joint moment and knee flexor and extensor muscle forces were computed using OpenSim. Differences inmuscle force between normal and abnormal conditions were investigated using ANOVA and Tukey-Kramer multiple comparison test.The results revealed that braced knee experienced limited range of motion with smaller flexion moment occuring at late swing phase. Significant differences were found in all flexormuscle forces and in several extensor muscle forces (p<0.05). Normal knee produced larger flexor muscle force than braced knee. Braced knee generated the largest extensor muscle force at early swing phase. In summary, musculoskeletal modelling simulation can be a computational tool to map and detect the differences between normal and asymmetrical gaits.