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Background: Acupuncture has been used as a treatment for pain associated with osteoarthritis (OA) for thousands of years; however, there is a lack of definitive evidence for this indication in humans or animals. The aim of this study was to prospectively evaluate the efficacy of acupuncture on lameness and clinical function in dogs affected by naturally-occurring OA using objective outcome measures. A total of 32 client-owned dogs completed this prospective, randomized, placebo-controlled, blinded clinical trial, using a cross-over design. Participants were assigned to receive placebo or acupuncture treatment once weekly for 4 weeks in random order with a two-week wash-out period in between treatment phases. Outcome measures included ground reaction forces (GRF), subjective orthopedic scoring (SOS), activity counts (AC), and owner-completed clinical metrology instruments (CMI; Canine Brief Pain Inventory [CBPI] and Client Specific Outcome Measures [CSOM]). For statistical comparison, baseline GRF, SOS, and CMI data were compared to data obtained 1 week after each treatment phase. Similarly, total weekly AC of the final week of each treatment phase were compared to the baseline week. Results: Evidence of differences between baseline versus acupuncture and placebo treatments was not identified for the following outcome measures: GRF, AC, or SOS. However, evidence of differences was identified for some of the CMI scores, including the CSOM questionnaire which showed evidence of improvement when comparing baseline versus acupuncture (p = 0.0002) as well as between placebo versus acupuncture treatments (p = 0.035) but not between baseline versus placebo treatments (p = 0.221). Conclusions: The applied acupuncture protocol did not show improvement in function when using objective outcome measures for OA in dogs; however, certain CMI measurements recorded some degree of treatment response. [ABSTRACT FROM AUTHOR]