Background <p>Patellofemoral pain syndrome (PFPS) is one of the most common conditions affecting the knee joint, yet its pathomechanics remain unclear. The aim of this study was to investigate changes in muscle activation and gait patterns and to analyze the relationship between muscle activation and kinetic gait patterns in patients with PFPS.</p> Methods <p>This study included 31 patients with PFPS and 28 healthy volunteers without any symptoms. The sagittal plane motion of the knee joint, representing primary movement of the knee joint, was evaluated to identify changes in gait patterns. Electromyography (EMG) was used to measure muscle activation of vastus medialis (VM), vastus lateralis (VL), semitendinosus (ST), and gastrocnemius (GCM) muscles during gait analysis. Biomechanical features were analyzed during the three phases of the gait cycle; weight acceptance (WA), single limb support (SLS), and swing limb advancement (SLA) (0 ~ 12%, 13 ~ 50%, and 51 ~ 100% of the gait cycle, respectively).</p> Results <p>The average knee extension moment (KEM) during WA was lower in the patient group and no significant differences were observed in the knee flexion angle (KFA). With respect to muscle activation, the patient group showed significantly higher muscle activation of the ST muscle in all phases. As the absolute value of the moment increased, the activation of the VM, VL, and ST muscles increased more rapidly in the patient group, especially when KEM was under −1% body weight × height (Bw × Ht) or over 5% Bw × Ht.</p> Conclusions <p>Patients with PFPS exhibit elevated muscle activation, particularly in response to changes in the knee extension moment, which is likely a compensatory mechanism to manage knee joint loading during gait. These results highlight altered neuromuscular adaptations in PFPS, suggesting targeted therapies may help improve functional outcomes.</p> <p><i>Level of evidence</i> III, cross-sectional study</p>