Abstract Background Osteoarthritis (OA) is a chronic disease, usually characterized by pain, which is associated with reduced muscle strength, disability and progressive loss of function. However, the pain influence over proprioception and motor behaviour remains unclear. Thus, the purpose of the study was to identify the levels of pain, the proprioceptive acuity and the pattern of muscle recruitment during stair ascent and descent in elderly patients with mild and moderate osteoarthritis (OA) compared to healthy subjects. Methods The study participants included 11 healthy elderly subjects (7 women and 4 men) and 31 elderly patients with knee OA (19 women and 12 men). The functional capacity was assessed by the Western Ontario and McMaster Universities (WOMAC) osteoarthritis index; the pain was evaluated by Wong-Baker faces pain rating scale (WBS) and pressure pain threshold (PPT); the proprioceptive acuity was based on the joint position sense evaluated by electrogoniometer; and the electromyographic (EMG) activity of the major muscles of the lower limb were evaluated during a task of stair ascent and descent of 15 cm. For statistical analysis it was used Statistic for Windows software (StatSoft Inc., version 5.0). Data from the WOMAC index, WBS, the proprioceptive acuity and IEMG (for each muscle in each phase) were analyzed using the Mann–Whitney U test and data from PPT was used Kruskal-Wallis test. Results Higher scores were found in the WOMAC index and WBS whereas lower scores were seen in PPT in patients with knee OA compared to healthy subjects. In contrast, there were no significant differences in the proprioceptive acuity and EMG results of most muscles analyzed between the groups. Conclusion The presence of pain does not influence the proprioception and the motor behavior of the thigh muscles during stair ascent and descent in subjects with mild and moderate knee OA.