This study investigated whether the onset of electromyographic (EMG) activity of vastus medialis oblique (VMO) was delayed relative to that of vastus lateralis (VL) in patients with osteoarthritis (OA) of the knee compared to asymptomatic participants during open kinetic chain activities. An exploratory observational cross sectional study was carried out. Two groups were tested, symptomatic OA knee patients, diagnosed by an orthopaedic surgeon, (n = 17), mean (SD) age 66.0 (7.6) years, and asymptomatic participants (n = 17), 56.7 (8.6) years. Surface EMG activity of VMO and VL was measured, during concentric contractions extending the knee from 90° flexion, and during maximal voluntary isometric contractions at 60° knee flexion. The EMG onset times of VMO and VL were determined visually and by algorithm. The onset timing difference (OTD) between the two muscles was calculated for each subject, by subtracting the onset time of VL from VMO. Mann-Whitney U-tests revealed that the OTD between VMO and VL was not significantly different between the groups during either contraction type (both p>0.05). The results of this exploratory study may have implications for rehabilitation programmes aimed at developing preferential activation of VMO compared to VL in OA knee patients.