Background: Elastic ear cartilage is a good source of tissue for support or augmentation in plastic and reconstructive surgery. However, the amount of ear cartilage is limited and excessive use of cartilage can cause deformation of the auricular framework. This animal study investigated the potential of periosteal chondrogenesis in an ear cartilage defect model. Methods: Twelve New Zealand white rabbits were used in the present study. Four ear cartilage defects were created in both ears of each rabbit, between the central artery and marginal veins. The defects were covered with perichondrium (group 1), periosteum taken from the calvarium (group 2), or periosteum taken from the tibia (group 3). No coverage was performed in a control group (group 4). All animals were sacrificed 6 weeks later, and the ratio of neo-cartilage to defect size was measured. Results: Significant chondrogenesis occurred only in group 1 (cartilage regeneration ratio: mean± standard deviation, 0.97± 0.60), whereas the cartilage regeneration ratio was substantially lower in group 2 (0.10± 0.11), group 3 (0.08± 0.09), and group 4 (0.08± 0.14) (p= 0.004). Instead of chondrogenesis, osteogenesis was observed in the periosteal graft groups. No statistically significant differences were found in the amount of osteogenesis or chondrogenesis between groups 2 and 3. Group 4 showed fibrous tissue accumulation in the defect area. Conclusion Periosteal grafts showed weak chondrogenic potential in an ear cartilage defect model of rabbits; instead, they exhibited osteogenesis, irrespective of their embryological origin.