Objective To investigate the feasibility of repairing thyroid cartilage defects by implantation of chondrocyte-allogenous acellular cartilaginousmatrix(chondrocyte-ACM) composite in rabbits. Methods The thyroid chondrocyteswere isolated and co-cultured in vitro with allogenous acellular cartilaginousmatrix(ACM) to form the chondrocyte-ACM composite. The composite was analyzed histologically and was used to repair defects of thyroid cartilage. Eighteen New Zealand adult rabbits were made the defect models of thyroid cartilage at the two sides and divided into three groups. The defects were repaired with chondrocyte-ACM composite in the experimental group(n=6), with simple ACM in the ACM group (n=6)and without any material in the control group(n=6). The animals were sacrificed at 8 weeks after operation. The specimens were evaluated histologically. Results In vitro, the growth of chondrocytes was observed on the surface of allogenous acellular cartilaginous matrix and no chondrocytes grew inside the matrix. The defect filled with muscle and connective tissues in control group; the lymphocyte infiltration was observed in the matrix and no new cartilage formationoccurred at 8 weeks after operation in simple ACM group and experimental group.So the defect repair of rabbits thyroid cartilage failed. Conclusion The allogenous acellular cartilaginous matrixfailed to serve as a scaffold for chondrocytes both in vitro and in vivo. The allogenous acellular cartilaginous matrixshould be improved.