The purpose of this study was to compare the chondroprotective effects of chondroitin sulfate (CS)-hyaluronic acid (HA) (CS-HA) injection and HA injection in an experimental model of osteoarthritis. After induction of osteoarthritis in rabbits, 28 rabbits were randomized into four groups: control group, 'HA' group, 'CS' group, and 'CS-HA' group. After 7 days, rabbits in the control group, 'HA' group, 'CS' group and 'CS-HA' group were respectively treated with normal saline, HA, CS, or CS-HA injection in the knees. All animals were treated once weekly. The animals were treated continuously for 5 weeks. Histological and biochemical evaluations were performed. As shown by histological observation, CS-HA injection treatment showed a chondroprotective effect on osteoarthritis. However, the histological scores of 'HA' group and 'CS-HA' group were not significantly different (p > 0.05). The results of biochemical evaluation showed that the expression levels of IL-1β, TNF-α, TIMP-1 and NO in synovial fluid of treated groups were all different from the control group (p < 0.05). However, the expression levels of these biochemical molecules in three treated groups were not significantly different (p > 0.05). In conclusion, CS-HA injection showed no obvious advantage over HA injection in osteoarthritis treatment.