Soft biological polymers, such as articular cartilage, possess exceptional fracture and fatigue resistance, offering inspiration for the development of novel materials. However, we lack a detailed understanding of changes in cartilage material behavior and of crack propagation following cyclic compressive loading. We investigated the structure and mechanical behavior of cartilage as a function of loading frequency and number of cycles. Microcracks were initiated in cartilage samples using microindentation, then cracks were extended under cyclic compression. Thickness, apparent stiffness, energy dissipation, phase angle, and crack length were measured to determine the effects of cyclic loading at two frequencies (1 Hz and 5 Hz). To capture the fatigue-induced material response (thickness, stiffness, energy dissipation, and phase angle), material properties were compared between pre-and-post diagnostic tests. The findings indicate that irreversible structural damage (reduced thickness), cartilage softening (reduced apparent stiffness), and reduced energy dissipation (including phase angle) increased with an increase in the number of cycles. Higher frequency loading resulted in less reduction in energy dissipation, phase angle, and thickness change. Crack lengths, quantified through brightfield imaging, increased with number of cycles and frequency. This study sheds light on the complex response of cartilage under cyclic loading resulting in softening, structural damage, and altered dynamic behavior. The findings provide better understanding of failure mechanisms in cartilage and thus may help in diagnosis and treatment of osteoarthritis.