. Background. Knee osteoarthritis (KOA) is a common chronic degenerative joint disease. 3D printing technology has become one of the important directions of medical development along with individualized precision treatment in orthopedics. Objective. To investigate the effect of 3D printing technology-assisted total knee arthroplasty (TKA) on cartilage in rabbits with KOA. Methods. A rabbit model of KOA was established and treated by TKA or 3D printing-assisted TKA. Four weeks after treatment, radiological evaluation of rabbit knees was performed by X-ray examination, in order to observe the severity of osteoarthritic lesions. Then the knee joints of rabbits were collected for Hematoxylineosin, Toluidine blue, and Safranin O-Fast green staining. The expressions of cartilage matrix metabolism-related and apoptosis-related genes were scrutinized by real-time quantitative reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry. The levels of inflammatory- related factors in the cartilage tissues of rabbits were tested by enzyme-linked immunosorbent assay. Results. In rabbits with KOA, 3D printing technology-assisted TKA alleviated the inflammation and bone remodeling of the knee joint, relieved synovial hyperplasia and inflammatory cell infiltration in the articular cartilage, reduced articular cartilage degradation, suppressed cartilage matrix metabolism, and mitigated the inflammatory response and apoptosis of cartilage cells. Conclusion. 3D printing technology-assisted TKA exhibits a good treatment effect in rabbit KOA. This study provides an important basis for the clinical application of 3D printing technology-assisted TKA in KOA treatment.