Background: The objective of this study was to determine the influence of loading frequency on the failure of articular cartilage-on-bone specimens under three-point bending. Methods: In this study, cyclic three-point bending was used to introduce failure into cartilage-on-bone specimens at varying loading frequencies. Sinusiodally varying maximum compressive loads in the range 40-130 N were applied to beam-shaped cartilage-on-bone specimens at frequencies of 1, 10, 50 and 100 Hz. Results: The number of cycles to failure decreased when loading frequency increased from normal and above gait (1 and 10 Hz) to impulsive loading frequencies (50 and 100 Hz). It was found that 67 and 27% of the specimens reached run-out at loading of 10,000 cycles at frequencies of 1 and 10 Hz, respectively. However, 0% of the specimens reached run-out at loading frequencies of 50 and 100 Hz. Conclusion: The results indicate that increasing the loading frequency reduces the ability of specimens to resist fracture during bending. The findings underline the importance of the loading frequency concerning the failure of articular cartilage-on-bone and it may have implications in the early onset of osteoarthritis.