This study examined the effects of obesity on cartilage mechanics and longitudinal failure probability at the medial tibiofemoral compartment, using combined musculoskeletal simulation and probabilistic failure modelling approaches. The current investigation examined twenty obese females (BMI > 30.0 kg/m2) and 20 healthy weight (BMI < 25.0 kg/m2) females. Walking kinematics were obtained via an 8-camera optoelectric system, and a force plate was used to collect ground reaction forces. Musculoskeletal simulation and probabilistic failure modelling were utilized to explore medial tibiofemoral forces and cartilage probability. Comparisons between groups were undertaken using linear mixed-effects models. Net peak cartilage forces, stress and strain were significantly larger in the obese group (force = 2013.92 N, stress = 3.03 MPa & strain = 0.25), compared to health weight (force = 1493.21 N, stress 2.26 MPa & strain = 0.19). In addition, medial tibiofemoral cartilage failure probability was also significantly larger in the obese group (42.98%) compared to healthy weight (11.63%). The findings from the current investigation show that obesity has a profoundly negative influence on longitudinal medial knee cartilage health and strongly advocates for the implementation of effective weight management programs into long-term musculoskeletal management strategies.