The study aims to determine the validity and reproducibility of step duration and step length parameters measured during walking in healthy participants using an accelerometer embedded in smart eyeglasses. Twenty young volunteers participated in two identical sessions comprising a 30 s gait assessment performed at three different treadmill speeds under two conditions (i.e., with and without a cervical collar). Spatiotemporal parameters (i.e., step duration and step length normalized by the lower limb length) were obtained with both the accelerometer embedded in smart eyeglasses and an optoelectronic system. The relative intra- and inter-session reliability of step duration and step length computed from the vertical acceleration data were excellent for all experimental conditions. An excellent absolute reliability was observed for the eyeglasses for all conditions and concurrent validity between systems was observed. An accelerometer incorporated in smart eyeglasses is accurate to measure step duration and step length during gait.