- OBJECTIVES: When measuring changes in knee cartilage thickness in vivo after loading, mean values may not reflect local changes. The objectives of this investigation were: (1) use statistical parametric mapping (SPM) to determine regional deformation patterns of tibiofemoral cartilage in response to running; (2) quantify regional differences in cartilage thickness between males and females; and (3) explore the influence of sex on deformation. MATERIALS AND METHODS: Asymptomatic males (n = 15) and females (n = 15) had MRI imaging of their right knee before and after 15 min of treadmill running. Medial and lateral tibial, and medial and lateral weight-bearing femoral cartilage were segmented. SPM was completed on cartilage thickness maps to test the main effects of Running and Sex, and their interaction. F-statistic maps were thresholded; clusters above this threshold indicated significant differences. RESULTS: Deformation was observed in all four compartments; the lateral tibia had the largest area of deformation (p < 0.0001). Thickness differences between sexes were observed in all four compartments, showing females have thinner cartilage (p ≤ 0.009). The lateral tibia had small clusters indicating an interaction of sex on deformation (p ≤ 0.012). DISCUSSION: SPM identified detailed spatial information on tibiofemoral cartilage thickness differences observed after running, and between sexes and their interaction.