Evaluation of an immune-privileged scaffold for In vivo implantation of tissue-engineered trachea

Forty tracheas were harvested from donor New Zealand rabbits. Thirty of the tracheas were randomly divided into four treatment groups corresponding to 4, 5, 6, or 7% NaClO₄ and one untreated group (n = 6 each group). Scanning electron microscopy distinctly revealed the cilium of epithelial cells in the fresh trachea. The internal surface of the trachea was rough in the 4% treatment group and smooth in the 5% treatment group, whereas the matrix was fractured in the 6% treatment group and highly fractured in the 7% treatment group. We observed that the number of nuclei in the cells of the 4, 5, 6, and 7% treatment groups decreased compared to the cells of the untreated group (p < 0.05). Although there was a significant decrease in maximum tensile strength, tensile strain at fracture and the elastic modulus (p < 0.05) with increasing concentrations of NaClO₄, the content of glycosaminoglycans (GAGs) did not significantly decline (p > 0.05) in the 5% treatment group. In addition, histopathological analysis showed that the fiber component and basement membrane of the matrix in the 5% treatment group were retained after optimal decellularization. Despite the preserved cartilage, in vitro immunohistochemical analysis revealed that the matrix did not show the presence of major histocompatibility complex (MHC) antigens. The remaining ten donor tracheas, which were divided into a positive control group and an optimal decellularized group, were used for allogeneic transplantation. Blood samples were taken regularly, and histologic examinations were performed at 30 days postimplantation, which showed no significant immune rejection. In conclusion, we surveyed the structural integrity through morphological observation and compared the biomechanical and immunogenic changes in the tracheal matrix under the different treatments. The optimal decellularized tracheal matrix with preserved cartilage, which was acquired via 5% NaClO4 treatment, exhibited structural integrity, antigen cell removal and immune privilege and would be suitable for use as a tissue-engineered trachea for in vivo transplantation in rabbit models.