| Summary: | This cross-sectional study analyzed the lower limb (LL) behavior in terms of gait asymmetry and joints’ kinematic parameters, comparing people with multiple sclerosis (pwMS) and unaffected individuals. Methods: Data from 15 patients, EDSS ≤ 4.5, and 15 healthy control volunteers were gathered. The VICON Motion Capture System (14 infrared cameras), NEXUS software, Plug-in–Gait skeleton model and reflective markers were used to collect data for each subject during five gait cycles on a plane surface. Biomechanical analysis included evaluation of LL joints’ range of motion (ROM) bilaterally, as well as movement symmetry. Results: Comparative biomechanical analysis revealed a hierarchy of vulnerability between the groups: the ankle is the most affected joint in pwMS (<i>p</i> = 0.008–0.014), the knee is moderately affected (<i>p</i> = 0.015 in swing phase), and the hip is the least affected (<i>p</i> > 0.05 in all phases). The swing phase showed the most significant left–right asymmetry impairment, as reflected by root mean square error (RMSE) values: swing-phase RMSE = 9.306 ± 4.635 (higher and more variable) versus stance-phase RMSE = 6.363 ± 2.306 (lower and more consistent). Conclusions: MS does not affect the joints structurally; rather, it eliminates the ability to differentiate the fine-tuning control between them. The absence of significant left–right joint asymmetry differences during complete gait cycle indicates dysfunction in the global motor control.
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