BACKGROUND: The movement coordination in patients with knee osteoarthritis may be impaired and the identification of the deficits in lower limb inter-segmental coordination is crucial to understand the effect of knee osteoarthritis on knee function. RESEARCH QUESTION: This study utilizes continuous relative phase to investigate the pattern and variability of lower limb inter-segmental coordination in patients with knee osteoarthritis and in healthy subjects during walking, and to evaluate inter-segmental coordination alterations in patients. METHODS: Gait was measured by a three-dimensional motion capture system for 44 patients with late-stage knee osteoarthritis and 22 healthy subjects. Segmental kinematic parameters, continuous relative phase and its variability were calculated. Independent samples t-tests were used to detect differences between patients and healthy subjects. RESULTS: Thigh-shank continuous relative phase of patients is significantly decreased by 16.04° and 16.18° during late stance and swing phase as compared with healthy subjects (P < 0.05). Shank-foot continuous relative phase of patients is significantly decreased by 6.89° during early stance and significantly increased by 5.49° and 6.39° during late stance and swing phase (P < 0.05). Patients also exhibit increased variability of thigh-shank continuous relative phase during late stance and swing phase by 1.90° and 1.65° respectively, and increased variability of shank-foot continuous relative phase during early stance and swing phase by 0.83° and 0.88° respectively as compared to healthy subjects (P < 0.05). SIGNIFICANCE: Patients with knee osteoarthritis exhibit altered coordination patterns and increased coordination variability of thigh-shank and shank-foot. Knee dysfunction results in altered lower limbs coordination and unstable motor control during walking. Investigation of inter-segmental coordination could therefore provide insights into changes in neuromuscular control of gait in patients with knee osteoarthritis.