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Background The equivalency of treadmill and overground walking has been investigated in a large number of studies. However, no systematic review has been performed on this topic. Research question The aim of this study was to compare the biomechanical, electromyographical and energy consumption outcomes of motorized treadmill and overground walking. Methods Five databases, ScienceDirect, SpringerLink, Web of Science, PubMed, and Scopus, were searched until January 13, 2021. Studies written in English comparing lower limb biomechanics, electromyography and energy consumption during treadmill and overground walking in healthy young adults (20-40 years) were included. Results Twenty-two studies (n=409 participants) were included and evaluated via the Cochrane Collaboration’s tool. These 22 studies showed that some kinematic (reduced pelvic ROM, maximum hip flexion angle for females, maximum knee flexion angle for males and cautious gait pattern), kinetic (sagittal plane joint moments: dorsiflexor moments, knee extensor moments and hip extensor moments and sagittal plane joint powers at the knee and hip joints, peak backwards, lateral and medial COP velocities and propulsive forces during late stance) and electromyographic (lower limbs muscles activities) outcome measures were significantly different for motorized treadmill and overground walking. Significance Spatiotemporal, kinematic, kinetic, electromyographic and energy consumption outcome measures were largely comparable for motorized treadmill and overground walking. However, the differences in kinematic, kinetic and electromyographic parameters should be taken into consideration by clinicians, trainers, and researchers when working on new protocols related to patient rehabilitation, fitness rooms or research as to be as close as possible to the outcome measures of overground walking. The protocol registration number is CRD42021236335 (PROSPERO International Prospective Register of Systematic Reviews).