Ionic detergent under pressure-vacuum as an innovative strategy to generate canine tracheal scaffold for organ engineering

Abstract

Decellularized scaffolds applied in tissue engineering offer improvements, supplying the elevated necessity for organs and tissues for replacement. However, obtaining a functional trachea for autotransplantation or allotransplantation is tricky due to the organ anatomical and structural complexity. Most tracheal decellularization protocols are lengthy, expensive and could damage the tracheal extracellular matrix (ECM) architecture and functionality. Here we aim to evaluate the effectiveness of three different decellularization protocols combined with chemical and physical methods to obtain acellular canine tracheal scaffolds. Six adult dog tracheas were incised (tracheal segments) resulting in twenty-eight rings for control tissue and eighty-four rings for decellularization (5-7 mm thick). Following, decellularized tracheal scaffolds were microscopic/macroscopic characterized by histological analysis (Hematoxylin-Eosin, Masson Trichrome, Picrosirius red, Alcian Blue and Safranin O), immunohistochemistry for ECM components, scanning electron microscopy and genomic DNA quantification. After decellularization, the tracheal tissue revealed reduced genomic DNA, and maintenance of ECM components preserved (structural proteins, adhesive glycoproteins, glycosaminoglycans and proteoglycans) suggesting ECM integrity and functionality. Comparatively, the combined ionic detergent with high vacuum pressure decellularization protocol revealed superior genomic DNA decrease (13.5 ng/mg) and improvement on glycosaminoglycans (GAGs) and proteoglycans (PGs) preservation regarding the other decellularized trachea scaffolds and native tissue. Our results indicate that the three chemical/physical protocols reduce the decellularization time, without ECM proteins damage. Notwithstanding, the use of ionic detergent under vacuum pressure was able to generate an innovative strategy to obtain acellular canine tracheal scaffolds with the highest levels of adhesive proteins that support its potentiality for recellularization and future tissue engineering application.