AbstractThe glymphatic system is a perivascular fluid clearance system, most active during sleep, considered important for clearing the brain of waste products and toxins. Glymphatic failure is hypothesized to underlie brain protein deposition in neurodegenerative disorders like Alzheimer’s disease. Preclinical evidence suggests that a functioning glymphatic system is also essential for recovery from traumatic brain injury, which involves release of debris and toxic proteins that need to be cleared from the brain. In a cross-sectional observational study, we estimated glymphatic clearance using diffusion tensor imaging along perivascular spaces, an MRI-derived measure of water diffusivity surrounding veins in the periventricular region, in 13 non-injured controls and 37 subjects who had experienced traumatic brain injury ∼5 months previously. We additionally measured the volume of the perivascular space usingT2-weighted MRI. We measured plasma concentrations of neurofilament light chain, a biomarker of injury severity, in a subset of subjects. Diffusion tensor imaging along perivascular spaces index was modestly though significantly lower in subjects with traumatic brain injury compared with controls when covarying for age. Diffusion tensor imaging along perivascular spaces index was significantly, negatively correlated with blood levels of neurofilament light chain. Perivascular space volume did not differ in subjects with traumatic brain injury as compared with controls and did not correlate with blood levels of neurofilament light chain, suggesting it may be a less sensitive measure for injury-related perivascular clearance changes. Glymphatic impairment after traumatic brain injury could be due to mechanisms such as mislocalization of glymphatic water channels, inflammation, proteinopathy and/or sleep disruption. Diffusion tensor imaging along perivascular spaces is a promising method for estimating glymphatic clearance, though additional work is needed to confirm results and assess associations with outcome. Understanding changes in glymphatic functioning following traumatic brain injury could inform novel therapies to improve short-term recovery and reduce later risk of neurodegeneration.