Background: A tracheal matrix scaffold decellularized by detergent-enzymatic treatment has been shown as a promising scaffold in tracheal tissue engineering. The objectives of this study were to evaluate the impact of this technique on tracheal extracellular matrix integrity and characterize the matrix environment for recellularization.
Methods: Brown Norway rat tracheae were decellularized using a modified detergent-enzymatic treatment. Antigenicity and cellularity were monitored during processing. Glycosaminoglycan content, histoarchitecture, and mechanical properties were also evaluated. Matrix compatibility was determined by cytotoxicity assay. Surface ultrastructure of the matrix and its interaction with seeded bone marrow stem cell–derived chondrocytes and tracheal epithelial cells were examined by scanning electron microscopy.
Results: Rat trachea treated with five detergent-enzymatic treatment cycles demonstrated complete elimination of antigenicity. Although there was a significant loss of glycosaminoglycan (t test, p < 0.01), histoarchitecture of tracheal cartilage and basement membrane was retained after decellularization. Stiffness decreased, but sufficient compressive strength was preserved to maintain lumen patency. The decellularized matrix showed good cell compatibility and favored adhesion and growth of chondrocytes and respiratory epithelial cells, as demonstrated by scanning electron microscopy.
Conclusions: At the point of complete antigen removal, detergent-enzymatic treatment altered tracheal extracellular matrix composition but preserved the major structure and adequate mechanical strength. The matrix provided a compatible and supportive environment for recellularization.