Optimization and Characterization of a Physicochemical Decellularization Protocol for Porcine Tracheal Scaffolds

Reconstruction of extensive tracheal defects remains a challenge, and the development of decellularized scaffolds represents a promising strategy for tissue engineering. This study aimed to optimize a porcine tracheal decellularization protocol using sodium deoxycholate (SDC), reducing detergent exposure time without compromising extracellular matrix (ECM) preservation or biomechanical properties. Ten decellularization cycles were evaluated by residual DNA quantification, histological analysis, and residual SDC quantification, and cycles 1, 2, and 10 were selected for ECM characterization, cytocompatibility assessment, and biomechanical analysis. The first cycle was effective for submucosal decellularization, although chondrocytes remained within the tracheal cartilage. ECM architecture and its biomechanical properties were preserved. Additional cycles provided no further benefit to decellularization and were associated with alterations in longitudinal biomechanical parameters. Residual SDC was detected in the scaffolds, and undiluted extracts showed cell viability below 70%. These results indicate that 24 h of SDC exposure achieved effective submucosal decellularization while chondrocytes remained in the cartilage regardless of the number of decellularization cycles. ECM characteristics and tissue biomechanical properties were preserved.