© 2020 Cardiff Metropolitan University.Biofeedback is used for enhancing performance through providing real-time stimulus to the individual during physical activities. This randomised controlled trial aimed investigating the changes in physiological and kinematic variables of treadmill running in response to biofeedback. 24 age-matched, healthy, and recreationally active participants were randomly assigned to biofeedback and no-biofeedback groups. During 15-min treadmill running, group-based knee ROM, COM and heart rate were collected and analysed. Heart rate change was statistically significant for no-biofeedback group (156.60 ± 5.18, 160.20 ± 5.22 and 163.60 ± 5.93 F (2, 9) = 13.878, p < 0.001). No such change was found for biofeedback group (152.20 ± 6.63, 152.10 ± 6.12 and 155.30 ± 6.22 F (2, 9) = 2.787, p = 0.088). According to the Pearson Correlation Coefficient and RMSE techniques knee movement was highly correlated (initial-mid R2 = 0.995, RMSE = 1.558; initial-final R2 = 0.994, RMSE = 1.407) for biofeedback group compared to no biofeedback group (initial-mid R2 = 0.989, RMSE = 1.988; initial-final R2 = 0.985, RMSE = 4.978). Heart rate and COM variability were minimised; correspondingly, consistent knee movement was obtained in biofeedback group. Future research should determine if muscle involvement in motion and fatigue are regulated with biofeedback.