The objective of the clinical case is to report the symptomatic effect and, for the first time, the disease-modifying effect of ozone in a patient with osteoarthritis of the knee. A 67-year-old woman has been infiltrated with ozone 1 time/week for 4 weeks, using an oxygen-ozone volume of 20 ml, at a 20-µg/ml concentration. Another cycle of ozone treatment was boosted at 6 months follow-up. Clinical outcomes such as pain, function, and quality of life were measured by the Western Ontario and McMaster University Index for Osteoarthritis (WOMAC) scale. Biochemical markers of inflammation such as C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and uric acid were performed at the beginning and at the end of ozone (O2-O3) treatment protocol. Radiological outcome was evaluated as the change in minimal joint space at the medial compartment of joint space, at the beginning and at 12 months follow-up in both knees. In the first and second ozone cycles, clinical (pain, function, rigidity) and biochemical variables (CRP, ESR, uric acid) improved in our case report. Minimal joint space at medial compartment improved in both knees after 12 months follow-up. Intra-articular ozone (O2-O3) has shown a symptomatic effect and especially a disease-modifying effect in a patient with knee osteoarthritis, improving pain, function, and rigidity; decreasing inflammation biomarkers (CRP, ESR, and uric acid); and increasing the joint space of the medial component evidenced radiologically. The structural effect is reported for the first time in literature. No adverse effects have been observed after intra-articular ozone infiltrations.