Background: Osteoarthritis (OA) is the most common form of arthritis, which cause pain and disability that can affect the quality of life. The inflammation in OA is chronic and characterized by the infiltration of immune cells and the secretion of cytokines. Previous OA treatment used hyaluronic acid intraarticular injection, but several studies show no satisfactory clinical outcome. This study aims to determine and compare the anti-inflammatory and anti-catabolism effects of stem cell-derived secretomes versus hyaluronic acid as the gold standard for the treatment of OA. Methods: This research was conducted using an experimental randomized post-test-only control group design. According to veterinarians, the population in this study were local rabbits, aged 6-8 months, weighing > 1,500 grams, and healthy. Data were analyzed using SPSS version 22.0 for Windows. Results: Twenty-two subjects were included in each treatment and control group. From the descriptive analysis, it was found that MMP-13 levels were 7.55 ± 0.23 (Treatment) (ng/mL) and 8.36 ± 1.04 (Control) (ng/mL), ADAMTS-5 levels were 0.83 ± 0.08 (Treatment) (ng/mL) and 1.14 ± 0.33 (Control) (ng/mL), and levels of IL-1β 23.0 5± 2.48 (Treatment) (ng/mL) and 26.81 ± 5.57 (Control) (ng/mL). From the inferential test, it was found that all research results were statistically significant (MMP-13= 0.81; p = 0.021; 95%CI =-1.48 – (-0.13)) (Agrekanase 2 = 0.32; p = 0.006; 95% CI =-0.53 – (-0.09)) (IL-1β = 3.76; p = 0.023; 95% CI =-7.59 – (-0.08)). Conclusion: Administration of mesenchymal stem cell-derived secretomes resulted in significantly lower levels of MMP-13, Agrekanase 2, and IL-1β compared to administration of hyaluronic acid in rabbit knee osteoarthritis.