BackgroundThe nonlinear variability present during steady-state gait may provide a signature of health and showcase one’s walking adaptability. Although treadmills can capture vast amounts of walking data required for estimating variability within a small space, gait patterns may be misrepresented compared to an overground setting. Smartphones may provide a low-cost and user-friendly estimate of gait patterns among a variety of walking settings. However, no study has investigated differences in gait patterns derived from a smartphone between treadmill walking (TW) and overground walking (OW). Research questionThis study implemented a smartphone accelerometer to compare differences in temporal gait variability and gait dynamics between TW and OW. MethodsSixteen healthy adults (8F; 24.7 ± 3.8 years) visited the laboratory on three separate days and completed three 8-minute OW and three TW trials, at their preferred speed, during each visit. The inter-stride interval was calculated as the time difference between right heel contact events located within the vertical accelerometery signals recorded from a smartphone while placed in participants front right pant pocket during walking trials. The inter-stride interval series was used to calculate stride time standard deviation (SD) and coefficient of variation (COV), statistical persistence (fractal scaling index), and statistical regularity (sample entropy). Two-way analysis of variance compared walking condition and laboratory visits for each measure. ResultsCompared to TW, OW displayed significantly (p < 0.01) greater stride time SD (0.014 s, 0.020 s), COV (1.26 %, 1.82 %), fractal scaling index (0.70, 0.79) and sample entropy (1.43, 1.63). No differences were found between visits for all measures. SignificanceSmartphone-based assessment of gait provides the ability to distinguish between OW and TW conditions, similar to previously established methodologies. Furthermore, smartphones may be a low-cost and user-friendly tool to estimate gait patterns outside the laboratory to improve ecological validity, with implications for free-living monitoring of gait among various populations.