Tergitol Based Decellularization Protocol Improves the Prerequisites for Pulmonary Xenografts: Characterization and Biocompatibility Assessment

Abstract

Right ventricle outflow tract obstruction (RVOTO) is a congenital pathological condition that contributes to about 15% of congenital heart diseases. In most cases, the replacement of the right ventricle outflow in pediatric age requires subsequent pulmonary valve replacement in adulthood. The aim of this study was to investigate the extracellular matrix scaffold obtained by decellularization of the porcine pulmonary valve using a new detergent (Tergitol) instead of Triton X-100. The decellularized scaffold was evaluated for the integrity of its extracellular matrix (ECM) structure by testing for its biochemical and mechanical properties, and the cytotoxicity/cytocompatibility of decellularized tissue was assessed using bone marrow-derived mesenchymal stem cells. We concluded that Tergitol could remove the nuclear material efficiently while preserving the structural proteins of the matrix, but without an efficient removal of the alpha-gal antigenic epitope. Therefore, Tergitol can be used as an alternative detergent to replace the Triton X-100.

Optimization of Enzymatic and Chemical Decellularization of Native Porcine Heart Valves for the Generation of Decellularized Xenografts

“…Our results show clearly that while the structure of the ECM and its protein content stayed intact and preserved, the nuclear component was efficiently removed by using Tergitol (Figure 4). Our results are similar to the results from Tondato et al [37], where they showed that Tergitol can be a reliable replacement for Triton X-100.…”

“…In addition, in decellularization, Triton X-100, a non-ionic detergent, is commonly used to remove remaining ionic detergent [35], and also due to its efficient removal of cellular content by disrupting lipid-lipid and protein-lipid bonds [36]. Since maintaining GAGs and lipids is crucial to preserve the ECM structure and integrity [5], Triton X-100 is replaced here by Tergitol, which is a mild and more environmentally friendly detergent [37]. Our results show clearly that while the structure of the ECM and its protein content stayed intact and preserved, the nuclear component was efficiently removed by using Tergitol (Figure 4).…”