The topic of potential long-term neurological consequences from having multiple concussions during a career in collision sports is controversial. We sought to investigate white matter microstructure using diffusion tensor imaging (DTI) in retired professional Australian National Rugby League (NRL) players (  = 11) with a history of multiple self-reported concussions compared with age- and education-matched controls (  = 13) who have had no history of brain trauma. Diffusion-weighted images were acquired with a Siemens 3T scanner. All participants completed a clinical interview. There were no significant differences between groups on measures of depression, anxiety, stress, or post-concussion symptoms; however, NRL players scored significantly higher on the alcohol use disorder identification test (AUDIT). Voxelwise analyses of DTI measures were performed using tract-based spatial statistics (TBSS) with age and AUDIT scores included as covariates. TBSS revealed significantly reduced fractional anisotropy (FA), and increased radial diffusivity (RD), axial diffusivity (AD), and trace (TR) in white matter regions of recently retired NRL players compared with controls. FA was significantly reduced in the right superior longitudinal fasciculus and right corticospinal tract while TR, RD, and AD were increased in these regions, as well as the corpus callosum, forceps major, right uncinate fasciculus, and left corticospinal tract. In summary, DTI in a small cohort of recently retired professional NRL players with a history of multiple concussions showed differences in white matter microstructure compared with age- and education-matched controls with no history of brain trauma.