Insufficient quadriceps force production and altered knee joint biomechanics after anterior cruciate ligament reconstruction (ACLR) may contribute to a heightened risk of osteoarthritis. Quadriceps muscle lengthening dynamics affect force production and knee joint loading; however, no study to our knowledge has quantified in vivo quadriceps dynamics during walking in individuals with ACLR or examined correlations with joint biomechanics. Our purpose was to quantify bilateral vastus lateralis (VL) fascicle length change and the association thereof with gait biomechanics during weight acceptance in individuals with ACLR. The authors hypothesized that ACLR limbs would exhibit more fascicle lengthening than contralateral limbs. The authors also hypothesized that ACLR limbs would exhibit positive correlations between VL fascicle lengthening and knee joint biomechanics during weight acceptance in walking. The authors quantified VL contractile dynamics via cine B-mode ultrasound imaging in 18 individuals with ACLR walking on an instrumented treadmill. In partial support of our hypothesis, ACLR limb VL fascicles activated without length change on average during weight acceptance while fascicle length on the contralateral limb decreased on average. The authors found a positive association between fascicle lengthening and increase in knee extensor moments in both limbs. Our results suggest that examining quadriceps muscle dynamics may elucidate underlying mechanisms relevant to osteoarthritis.