Structural integrity, ECM components and immunogenicity of decellularized laryngeal scaffold with preserved cartilage

Decellularization techniques have been widely used as an alternative strategy for organ reconstruction. However, the compliance of tracheal or laryngeal tissues can be increased during the decellularization process, which might cause postoperative stenosis due to elimination of chondrocytes. The purpose of our study was to construct a decellularized, whole, laryngeal scaffold with preserved chondrocytes using perfusion techniques and to evaluate the immunogenicity of the decellularized scaffold in vitro and in vivo in a rat model. The cellular components and immunogenicity of the scaffold were decellularized after 14 h of perfusion with detergent and 48 h of perfusion with phosphate buffered saline. However, the cartilage was well preserved via histological analysis and a chondrocyte viability test. Compared to the fresh larynx, the decellularized larynx did not show the presence of major histocompatibility complex antigens via immunohistochemical analysis in vitro and no significant immune rejection occurred 12 weeks post-implantation. In conclusion, decellularization via perfusion can achieve a decellularized, whole-laryngeal scaffold with the cell components removed and the cartilage and extra-cellular matrix well preserved. T cell-mediated immune rejection was significantly reduced in decellularized laryngeal cartilaginous scaffolds in vivo.