BackgroundKnowledge of joint biomechanics is crucial for understanding the pathophysiology of osteoarthritis (OA). Biomechanical changes, especially in early-stage OA, are subtle and therefore require highly accurate and precise ways of measuring joint loading (i.e., forces) and kinematics (i.e., motion). Most OA research investigating kinematics to date using conventional measurement methods, has been restricted either to measures of functional, weight-bearing activities with limited precision, or to highly precise measures of non-functional activities. X-ray fluoroscopy, however, is a medical imaging modality that can be used to precisely measure high-speed dynamic human movement in vivo under weight-bearing conditions. ObjectiveTo illustrate the potential of fluoroscopy to measure human movement and study joint biomechanics in OA by highlighting several exemplary applications in studies on knee OA. DesignWe searched PubMed to identify a selection of relevant studies with fluoroscopic evaluation of joint mechanics in individuals with, or at risk of developing, knee OA. ResultsFluoroscopy has been utilized in biomechanical studies in populations with early stages of knee OA or at risk of developing knee OA, as well as in studies evaluating conservative treatment strategies for knee OA. Due to its ability to capture highly accurate and precise kinematic measures, fluoroscopy holds great promise in providing new insights in the role of biomechanics in OA pathogenesis. ConclusionsThe use of fluoroscopy in larger cohorts and longitudinal (interventional) studies may lead to more robust and generalizable results and could clarify the mediation pathways of joint biomechanics in the onset and progression of OA.