The mechanical property of articular cartilage determines to a great extent the functionality of diarthrodial joints. Consequently, the early detection of mechanical and, thus, functional changes of cartilage is crucial for preventive measures to maintain the mobility and the quality of life of individuals. An alternative to conventional mechanical testing is the inverse finite element approach, enabling non-destructive testing of the tissue. We evaluated a method for the assessment of the equilibrium material properties of the patellar cartilage based on magnetic resonance imaging during patellofemoral compression. We performed ex vivo testing of two equine patellas with healthy cartilage, one with superficial defects, and one with synthetically degenerated cartilage to simulate a pre-osteoarthritic stage. Static compression with 400 N for 2 h resulted in morphological changes comparable to physiological in vivo deformations in humans. We observed a decrease of the equilibrium Young’s modulus of the degenerated cartilage by -59%, which was in the range of the results from indentation (-74%) and confined compression tests (-58%). With the reported accuracy of magnetic resonance imaging and its reproducibility, the results indicate the potential to measure differences in Young’s modulus with regard to cartilage degeneration and consequently to distinguish between healthy and pre-osteoarthritic cartilage.