Comparative study of two perfusion routes with different flow in decellularization to harvest an optimal pulmonary scaffold for recellularization

Decellularization processes may variably distort or degrade extracellular matrix (ECM) structure. In this study, two perfusion routes (PR) were tested on SD rat lung samples. One decellularization protocol, PR1, was perfused through the pulmonary artery. The other decellularization protocol, PR2, was perfused through the trachea. Both decellularization protocols were used by the same detergent-based (sodium dodecyl sulphate and Triton X-100) with different flow continuous perfusion. There was no visible difference in vessel architecture between PR1- and PR2-decellularized scaffold. However, the airway structure and alveoli architecture of pulmonary decellularized scaffolds generated through PR2 at a flow rate of 8 mL/min were destroyed partly when compared to that in native lung and PR1-decellularized scaffold. Ultramicroscopic assessment of scaffolds was similar in both protocols and showed filamentous ECM with preserved fiber disposition and structure. Histological analysis and immunostaining showed no detectable cells remaining in the pulmonary scaffolds compare with native lung. The DNA concentration was significantly reduced in the decellularized scaffolds compared to the native lungs. A549 cells reseeded onto decellularized pulmonary scaffolds were no significant difference between PR1 and PR2 in cell viability, p > 0.05. We conclude that under the same high flow velocity status, perfusion decellularization through the pulmonary artery may be an optimal pathway to obtain decellularized scaffolds for pulmonary regeneration. This article is protected by copyright. All rights reserved. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2567-2575, 2016.