The multimodal approach combining AFM micro-indentation and MALDI-FTICR mass spectrometry imaging was applied to investigate the local biomechanical and biochemical properties of healthy and osteoarthritic human menisci. Healthy menisci exhibited a distinct zonal stiffness gradient and coherent spatial organization of extracellular-matrix components. Osteoarthritic tissues showed reduced mechanical heterogeneity, disrupted molecular gradients, and localized ECM remodelling. Spatial integration of both datasets revealed associations between mechanical softening and altered matrix composition, providing new insight into meniscal structure–property relationships and their disruption during osteoarthritis.