This study integrated AFM micro-indentation with MALDI-FTICR mass spectrometry imaging to spatially correlate the mechanical and molecular properties of healthy and osteoarthritic human menisci. Healthy menisci exhibited a distinct zonal stiffness gradient and coherent molecular organization, whereas osteoarthritic tissues showed reduced zonal mechanical differences, disrupted molecular gradients, and localized matrix remodelling. The correlative workflow revealed spatial associations between mechanical softening and altered ECM organization, providing a platform for characterizing meniscal degeneration at the microscale.