BackgroundGeneric musculoskeletal models based on literature data are often used to estimate joint reaction forces (JRFs) that otherwise could only be measured invasively. Estimated JRFs are sensitive to changes in maximum isometric force (Fiso) of the muscles, but these are normally simply scaled using a multiplicative coefficient. The impact of varying Fiso, or strength, of muscles crossing each lower limb joint on estimated JRFs has not been systematically explored in musculoskeletal models of the lower limb. Research QuestionHow do alterations in the strength of joint-crossing muscles influence the lower limb JRF magnitudes computed through a generic musculoskeletal model? MethodsBy modifying Fiso of muscles crossing hip, knee, ankle, or all joints at once up to ± 40% in 10% increments, thirty-two models were created to simulate the gait of a patient with an instrumented tibial prosthesis (5th Grand Challenge dataset). A standard workflow (inverse kinematics, static optimization, joint reaction analysis) was utilized to calculate JRFs. Both alterations in JRF magnitudes due to joint crossing muscles’ strength modifications and their accuracy against in vivo knee loading measurements were quantified. ResultsThe knee JRF was the most sensitive force to changes in the joint-crossing muscles’ strength (variations ranging from −37.9 ± 0.5% to +37.9 ± 3.2%), while the hip and ankle JRFs were almost unaffected (maximum variation: +6.1%). Reducing the strength of knee and ankle-crossing muscles and intensifying the strength of hip-crossing muscles lowered the knee JRF. The knee JRF was best estimated (peak error: 0.42 ± 0.15 body weight, root mean squared error: 0.37 ± 0.06 body weight, coefficient of determination: 0.76 ± 0.10) by the model with -40% weakened knee-crossing muscles. SignificanceAltering strengths mainly affects knee JRF estimated with generic musculoskeletal models, suggesting that personalization of strength of joint-crossing muscles is required for accurate knee JRF estimations. Rehabilitation regimes meant to strengthen muscles crossing a joint should be carefully designed to avoid undesired effects on the other joints.