Knee ligaments guide and restrain joint motion, and their properties influence joint mechanics. Inverse modeling schemes have been used to estimate specimen-specific ligament properties, where external joint forces are assumed to balance with internal ligament and contact forces. This study simplifies this assumption by adjusting experimental loads to remove internal contact forces. The purpose of this study was to use novel experimental loading in an inverse modeling scheme to estimate ligament slack lengths, perform validation using additional loading scenarios, and evaluate sensitivity to the applied loading. Joint kinematics and kinetics were experimentally measured for a set of load cases. An optimization scheme used a specimen-specific forward kinematics model to estimate ligament slack lengths by minimizing the residual between model and experimentally measured kinetics. The calibrated model was used for a form of validation by evaluating non-optimized load cases. Additionally, uncertainty analysis related kinetic errors to previously reported kinematic errors. The six DOF tibial reactions realized RMS errors less than 23 N and 0.75 Nm for optimized load cases, and 33 N and 2.25 Nm for the non-optimized load cases. The uncertainty analysis, which was performed using the optimized load cases, showed average kinetic RMS errors less than 26 N and 0.45 Nm. The model’s recruitment patterns were similar to those found in clinical and cadaveric studies. This study demonstrated that experimental distraction loading can be used in an inverse modeling scheme to estimate ligament slack lengths with a forward kinematics model
