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

Abstract

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.

Effects of two different decellularization routes on the mechanical properties of decellularized lungs

“…Both protocols demonstrated preserved alveolar septum and visceral pleura, which are important pulmonary structures that determine optimum decellularized lungs, similar to the previous results. Wang et al, 2016 [ 9 ], recently compared the same routes for lung decellularization, the trachea and pulmonary artery, and demonstrated preserved ECM, but the airway structure and alveoli architecture of the pulmonary decellularized lung was partially destroyed. This probably occurred because of the high flow applied in the pulmonary artery during the decellularization process, unlike the constant physiological pressure applied in our protocol, which maintained the lung structures.…”

“…The agents for lung decellularization can be applied by using two different infusion routes: the pulmonary vasculature and the airway tree [ 9 ]. A previous study from our group demonstrated that decellularization process by both routes did not induce any significant differences in the micro-scale local stiffness of the decellularized lung [ 10 ].…”

Lung Microvascular Niche, Repair, and Engineering

“…The lung, however, has an additional possible route for decellularization since the detergents can also be infused through the trachea. In fact, both routes (trachea and pulmonary artery) alone or combined have been used to effectively decellularize lungs (Price et al, 2010;Maghsoudlou et al, 2013;Tsuchiya et al, 2016;Wang et al, 2016). By using an intermittent intra-tracheal flow of detergent for decellularization, the same combination of SDC/Triton X-100 yielded an acellular scaffold in a shorter time FIGURE 2 | ECM decomposition and reconstruction after recellularization.…”

Rabbit thyroid extracellular matrix as a 3D bioscaffold for thyroid bioengineering: a preliminary in vitro study

“…Both SDS and Triton X-100 have been commonly used as detergents for cell lysis applications in TE. Notably, studies have demonstrated that perfusion decellularization protocols have been commonly performed on relatively large organs with distinct blood vessels or catheters, such as kidneys and lungs [ 14 , 16 ]. More so, in tissues where blood vessels hardly separate like in small intestine submucosa (SIS) and testis tissues, immersion/agitation decellularization could be an alternative option [ 17 , 18 ].…”