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Abstract Clinical Magnetic Resonance Imaging (MRI) of joints is limited to mere morphologic evaluation and fails to directly visualize joint or ligament function. In this controlled laboratory study, we show that knee joint functionality may be quantified in situ and as a function of graded posterior cruciate ligament (PCL)-deficiency by combining MRI and standardized loading. 11 human knee joints underwent MRI under standardized posterior loading in the unloaded and loaded (147 N) configurations and in the intact, partially, and completely PCL-injured conditions. For each specimen, configuration, and condition, 3D joint models were implemented to analyse joint kinematics based on 3D Euclidean vectors and their projections on the Cartesian planes. Manual 2D measurements served as reference. With increasing PCL deficiency, vector projections increased significantly in the anteroposterior dimension under loading and manual measurements demonstrated similar patterns of change. Consequently, if combined with advanced image post-processing, stress MRI is a powerful diagnostic adjunct to evaluate ligament functionality and joint laxity in multiple dimensions and may have a role in differentiating PCL injury patterns, therapeutic decision-making, and treatment monitoring.