This systematic review evaluated the available evidence for the effects of Masai Barefoot Technology (MBT) footwear on lower limb biomechanics during gait. Electronic databases (MEDLINE, EMBASE, CINAHL, SPORTDiscus, and PubMed) were searched in January 2015. Methodological quality of included studies was evaluated using the Quality Index. Standardised mean differences and 95% confidence intervals were calculated, and meta-analysis was conducted where possible. 17 studies satisfied the inclusion criteria; 16 cross-sectional studies and one randomised control trial (RCT). Quality Index scores ranged from 7 to 12 (out of 15). All 17 studies investigated walking gait only. Evidence showed that MBT footwear caused asymptomatic individuals to walk with a shorter stride length, reduced peak hip flexion, increased peak knee extension, and reduced hip and knee range of motion throughout gait. All kinematic effects occurred in the sagittal plane. There was a trend towards a decrease in internal and external joint moments and power, except for the foot, where increases in force were observed. There were only a small number of changes to lower limb muscle amplitude and timing. No statistically significant effects were observed in symptomatic individuals with knee osteoarthritis or following total knee replacement, but there was an increase in cadence and a decrease in step length in individuals following tibiotalar arthrodesis. These findings suggest that MBT footwear does change lower limb biomechanics in both asymptomatic and symptomatic individuals during gait. However, further clinical trials need to be undertaken to determine whether these changes are therapeutically beneficial.