Preparation and characterization of small-diameter decellularized scaffolds for vascular tissue engineering in an animal model

Abstract

BackgroundThe development of a suitable extracellular matrix (ECM) scaffold is the first step in vascular tissue engineering (VTE). Synthetic vascular grafts are available as an alternative to autologous vessels in large-diameter arteries (>8 mm) and medium-diameter arteries (6–8 mm). In small-diameter vessels (<6 mm), synthetic vascular grafts are of limited use due to poor patency rates. Compared with a vascular prosthesis, natural tissue ECM has valuable advantages. Despite considerable progress in recent years, identifying an optimal protocol to create a scaffold for use in small-diameter (<6 mm) fully natural tissue-engineered vascular grafts (TEVG), remains elusive. Although reports on different decellularization techniques have been numerous, combination of and comparison between these methods are scarce; therefore, we have compared five different decellularization protocols for making small-diameter (<6 mm) ECM scaffolds and evaluated their characteristics relative to those of fresh vascular controls.ResultsThe protocols differed in the choice of enzymatic digestion solvent, the use of non-ionic detergent, the durations of the individual steps, and UV crosslinking. Due to their small diameter and ready availability, rabbit arteria carotis were used as the source of the ECM scaffolds. The scaffolds were subcutaneously implanted in rats and the results were evaluated using various microscopy and immunostaining techniques.ConclusionsOur findings showed that a 2 h digestion time with 1× EDTA, replacing non-ionic detergent with double-distilled water for rinsing and the application of UV crosslinking gave rise to an ECM scaffold with the highest biocompatibility, lowest cytotoxicity and best mechanical properties for use in vivo or in situ pre-clinical research in VTE in comparison.Electronic supplementary materialThe online version of this article (doi:10.1186/s12938-017-0344-9) contains supplementary material, which is available to authorized users.

Implantation of VEGF‐functionalized cell‐free vascular grafts: regenerative and immunological response

“…Over the past decade, several groups have demonstrated the feasibility of completely acellular, off-the-shelf grafts in various animal models as well as human clinical trials. Decellularized and devitalized tissue-engineered constructs have been utilized with increasing frequency and have demonstrated improved patency and regeneration potential (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13). Devitalized constructs are based on bioengineered tissue constructs that are stripped of the cellular components while leaving extracellular matrix (ECM) components intact and then implanted into the animal model of choice (14).…”

Sequential adaptation of perfusion and transport conditions significantly improves vascular construct recellularization and biomechanics

“…For the scaffold choice, among the various possible approaches, ex vivo ‐ derived materials such as decellularized vessels have the structural and mechanical advantage of being composed of a native extracellular matrix (ECM) and possess bioactive molecules that aid tissue homeostasis and regeneration (Bejleri & Davis, ; Chen & Liu, ; M. C. Moore et al, ). Indeed, positive cell interactions with a structurally appropriate ECM aid and guide the regenerative response (Dahan et al, ; Uzarski, Van De Walle, & McFetridge, ; Xu et al, ). Clinical success of decellularized tissues has been reported in a range of applications, including trachea (Gonfiotti et al, ; Hamilton et al, ), heart valve (Dohmen, Lembcke, Holinski, Pruss, & Konertz, ; Tudorache et al, ), right ventricular outflow tract (Bibevski et al, ; Konertz et al, ) as well as vascular grafts (Olausson et al, , ).…”

“…Indeed, positive cell interactions with a structurally appropriate ECM aid and guide the regenerative response (Dahan et al, 2017;Uzarski, Van De Walle, & McFetridge, 2013;Xu et al, 2017). Clinical success of decellularized tissues has been reported in a range of applications, including trachea (Gonfiotti et al, 2014;Hamilton et al, 2015), heart valve (Dohmen, Lembcke, Holinski, Pruss, & Konertz, 2011;Tudorache et al, 2016), right ventricular outflow tract (Bibevski et al, 2017;Konertz et al, 2011) as well as vascular grafts (Olausson et al, 2014(Olausson et al, , 2012.…”

Investigation of survival and migration potential of differentiated cardiomyocytes transplanted with decellularized heart scaffold

“…The decellularization method has been employed to simulate this complex structure effectively. To date, many groups have reported that transplanted decellularized ECM biomaterials loaded with different cell types improved myocardial function . However, the efficiency and long‐term viability still remains as a major challenge.…”

“…To date, many groups have reported that transplanted decellularized ECM biomaterials loaded with different cell types improved myocardial function. 9,10 However, the efficiency and long-term viability still remains as a major challenge. Therefore, the first step toward understanding the biology and physiology of transplanted cells is through documenting survival, homing, differentiation, and migration potential of these cells.…”