Efficient decellularization of rabbit trachea to generate a tissue engineering scaffold biomatrix

Objectives: Most tracheal decellularization protocols are lengthy and can lead to reduced biomechanical stability. The objectives of this study were: 1) to generate a tracheal extracellular matrix scaffold using an efficient decellularization process and 2) to characterize the decellularized scaffold to assess its suitability for tissue engineering applications. Methods: Twelve rabbit tracheae underwent a decellularization process that involved enzymatic-detergent treatments. For characterization, fresh (control) and decellularized tissues underwent histological, immunohistochemical, and biochemical analyses. Tensile testing, scanning electron microscopy, and biocompatibility assay were also conducted. Results: Post-decellularization, the tracheal tissue had significantly less genetic material while the structural integrity was maintained. Specifically, the deoxyribonucleic acid content was significantly reduced and the glycosaminoglycan content was unchanged. Cell and cellular components were largely removed; at the same time the tensile properties and surface ultrastructural characteristics were unaltered. Biocompatibility was confirmed by contact cytotoxicity assay. Conclusions: Overall, an efficient decellularization process was used to treat rabbit tracheal tissue. The effectiveness of the decellularization process was demonstrated and at the same time there was preservation of the underlying extracellular matrix structure. This decellularized material may serve as a potential scaffold for tracheal tissue engineering.