Development and Characterization of Decellularized Rabbit Tracheal Cartilage Matrix for Use in Tissue Engineering

Abstract

Reconstruction of large upper airway defects requires replacement tissue. Unfortunately, there is no widely available tracheal tissue substitute that can be used at this time. To overcome this limitation, tissue engineering approaches have been used to generate tracheal grafts. The objective of this study was to decellularize and characterize rabbit tracheal tissue with the aim of generating an extracellular matrix scaffold for tracheal tissue engineering. Rabbit tracheal tissue was subjected to novel decellularization treatments. The decellularization processes involved cycles of modified enzymatic-detergent treatments. For characterization, decellularized and fresh specimens underwent histological, biochemical, and mechanical analyses. Scanning electron microscopy and biocompatibility assay were also performed. The decellularization treatments resulted in significant reduction of genetic/cellular material. The glycosaminoglycan content of the extracellular matrix was not significantly altered in most cases. The effectiveness of decellularization was also confirmed on histological, immunohistochemical, and scanning electron microscopic analyses. Mechanical testing results showed that the tensile parameters were largely maintained after decellularization. Contact cytotoxicity assay showed that the decellularized extracellular matrix was biocompatible. Overall, the decellularization treatments resulted in significant reduction of genetic/cellular material while preserving the underlying extracellular matrix structure. The tracheal extracellular matrix generated by decellularization has potential to be a viable scaffold material in tracheal tissue engineering.