Intramuscular administration of polysulfated glycosaminoglycan (PSGAG) reduces joint inflammation and pain, and has demonstrated efficacy in the treatment of canine osteoarthritis. This study sought to determine whether injection of PSGAG using aqua-acupuncture (aqua-AP) technique is more effective than standard intramuscular (IM) treatment. Arthritic canines were randomly assigned to Control Group (IM) or Test Group (aqua-AP) injections. All subjects received a commercial FDA-approved PSGAG dosed at 2 mg/lb (4.4 mg/kg) twice weekly for four weeks. Outcomes were compared between groups using scores from Canine Brief Pain Inventory (CBPI) and Quality of Life (QoL) completed by the owners (blinded to treatment group). Twenty-eight patients completed the study (12 controls, 16 test dogs). Evaluation of CBPI pain severity score improvement showed that test dogs had significant improvement (p=0.0004) achieved after 1 week of treatment and the improvement remained significant throughout the course of the trial. Controls also improved with significance attained at Week 3 (p=0.047). For CBPI pain interference score, the Test Group again had significant improvement (p=0.002) after 1 week of treatment, and improvement remained significant throughout the 4-week trial. Controls also had significant improvement, starting slightly later at Week 2 (p=0.025) and continuing through Week 4. Both groups demonstrated improvement for QoL score, with statistical significance starting from Week 2 for aqua-AP test dogs (p=0.011) and from Week 3 for controls (p=0.004). The results from this study suggest PSGAG administration using aqua-AP injection technique is non-inferior and potentially more effective and efficient than standard IM injection in mitigating pain severity for canine osteoarthritis patients.