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A key requirement in both native knee joints and total knee arthroplasty is a stable capsular ligament complex. However, knee stability is highly individual and ranges from clinically loose to tight. So far, hardly any in vivo data on the intrinsic mechanical of the knee are available. This study investigated if stiffness of the native ligament complex may be determined in vivo using a standard knee balancer. Measurements were obtained with a commercially available knee balancer, which was initially calibrated in vitro. 5 patients underwent reconstruction of the force-displacement curves of the ligament complex. Stiffness of the medial and lateral compartments were calculated to measure the stability of the capsular ligament complex. All force-displacement curves consisted of a non-linear section at the beginning and of a linear section from about 80 N onwards. The medial compartment showed values of 28.4 ± 1.2 N/mm for minimum stiffness and of 39.9 ± 1.1 N/mm for maximum stiffness; the respective values for the lateral compartment were 19.9 ± 0.9 N/mm and 46.6 ± 0.8 N/mm. A commercially available knee balancer may be calibrated for measuring stiffness of knee ligament complex in vivo, which may contribute to a better understanding of the intrinsic mechanical behaviour of knee joints.