Objective This study aims to investigate the effect of glucosamine intra-articular injection in temporomandibular joint(TMJ) osteoarthritis(OA) treatment of rabbits. Methods Intra-articular injection of type Ⅱ collagenase was performed bilaterally in six rabbits to induce OA(3 mg each on Days 1 and 4). On Week 6, the right joint(experimental group) was injected with glucosamine(once a week for 5 weeks), and the left joint(control group) was injected with physiological saline. On Week 11, six rabbits were sacrificed, and the TMJs were dissected for histological comparison. Results All joints showed osteoarthritic-like change, but the degree in the experimental group was significantly less severe than that in the control group. Moreover, the scale of the experiment group was significantly less than that of the control group. Conclusion Intra-articular injection of glucosamine can significantly relieve TMJ OA in rabbits.