A Novel Method to Acquire Decellularized Tracheal Matrix from Neonatal Rabbit

Tissue engineering of tracheal with decellularized tracheal matrix has been proposed as a therapeutic alternative to tracheal replacement. At present, the method to acquire decellularized tracheal matrix was not efficient enough, which should be improved. The aim of our study is to explore a novel method to acquire decellularized tracheal matrix efficiently. Here, the tracheas were taken from New Zealand white rabbits and randomly divided into three groups: Control group (C group), stored in sterile D-Hanks solution; Traditional method group (T group), digested by the detergentenzymatic method; Novel method group (N group), digested by the Trypsin-EDTA comprised with Detergent-enzymatic method. After the same time of decellularization, the efficacy of decellularization, the histoarchitecture and mechanical properties of decellularized tracheal matrix were detected in each group. Bone marrow mesenchymal stem cells (MSCs) from neonatal rabbit were seeded in the decellularized tracheal matrix, and SEM was used to detect the growth of MSCs. After the same time of decellularization, most of the cellular components and nuclear material were removed in N group, compared with C group and T group. SEM and TEM analysis revealed that both N group and T group could retain the hierarchical structure of the native trachea. Manual compression suggested the trachea of T group was much weaker than N group. Tensile test result showed that tensile breaking load, tensile strength, elongation at break, modulus of elasticity of T group and N group were similar to C group. In addition, 1 days and 3 days after seeded, much more MSCs could adherent in N group than T group. This study suggested that the novel method can generate a decellularized tracheal matrix superior to the traditional method.