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Objective: As a progressive chronic condition, osteoarthritis (OA) causes substantial pain and impairment. Secrete proinflammatory cytokines are essential mediators involved in the pathophysiology of OA. In this regard, the clinical effectiveness of autologous conditioned serum (ASC) has been shown through its injection into OA tissues. This study aimed to assess the effectiveness and concentration level of ACS components produced by Nano-carbon glass beads. Intravenous whole blood was obtained from each New Zealand male rabbit by 10-ml syringes, comprising 33 medical-grade Nano carbon-coated glass beads. Serum retrieving was performed after 6–8 h incubation (37 C, 5% Co2), and then centrifuged. The ACS was then injected into OA rabbits to assess its function. Results: Glass beads-prepared ACS coated with Nano-carbon, induced a huge amount of cytokines and growth factors production. The concentration level of anti-inflammatory cytokines and proinflammatory cytokines was improved throughout Nano-carbon coated glass beads stimulation. ACS also shortened the recovery time and improved the function and mobility of OA rabbits. We showed that ACS improved the function and mobility of OA rabbits, as well as shortened the recovery time. It is suggested that further studies evaluate this effectiveness. [ABSTRACT FROM AUTHOR]