A retrospective cohort study reviewed the records of 46 patients with osteoarthritis (OA) treated with warm acupuncture with moxibustion from March 2022 to December 2024, along with 46 age- and sex-matched healthy controls. The treatment was applied three times a week for 8 weeks to the acupuncture points Dubi (ST35), Zusanli (ST36), Yinlingquan (SP9), Yanglingquan (GB34) and Heding. Lysholm knee score (LKSS) increased after treatment, while VAS and WOMAC scores decreased (P < 0.05). Levels of osteoprotegerin (OPG) and bone gla protein (BGP) increased, while levels of MMP-1, MMP-3, prostaglandin E2, interleukin-1β, and TNF-α decreased (P < 0.05). Cx26 mRNA expression decreased, whereas Cx43 and Cx45 expression increased (P < 0.05). Connexin levels after treatment were comparable to the control group (P > 0.05). Warm acupuncture with moxibustion improved joint function, reduced pain and inflammation, and normalized connexin expression in OA patients.