title: Functional Heart Valve Scaffolds Obtained by Complete Decellularization of Porcine Aortic Roots in a Novel Differential Pressure Gradient Perfusion System.
abstract: There is a great need for living valve replacements for patients of all ages. Such constructs could be built by tissue engineering, with perspective of the unique structure and biology of the aortic root. The aortic valve root is composed of several different tissues, and careful structural and functional consideration has to be given to each segment and component. Previous work has shown that immersion techniques are inadequate for whole-root decellularization, with the aortic wall segment being particularly resistant to decellularization. The aim of this study was to develop a differential pressure gradient perfusion system capable of being rigorous enough to decellularize the aortic root wall while gentle enough to preserve the integrity of the cusps. Fresh porcine aortic roots have been subjected to various regimens of perfusion decellularization using detergents and enzymes and results compared to immersion decellularized roots. Success criteria for evaluation of each root segment (cusp, muscle, sinus, wall) for decellularization completeness, tissue integrity, and valve functionality were defined using complementary methods of cell analysis (histology with nuclear and matrix stains and DNA analysis), biomechanics (biaxial and bending tests), and physiologic heart valve bioreactor testing (with advanced image analysis of open-close cycles and geometric orifice area measurement). Fully acellular porcine roots treated with the optimized method exhibited preserved macroscopic structures and microscopic matrix components, which translated into conserved anisotropic mechanical properties, including bending and excellent valve functionality when tested in aortic flow and pressure conditions. This study highlighted the importance of (1) adapting decellularization methods to specific target tissues, (2) combining several methods of cell analysis compared to relying solely on histology, (3) developing relevant valve-specific mechanical tests, and (4) in vitro testing of valve functionality.
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Leslie Neil
authors.initials: LN
authors.lastname: Sierad
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Eliza Laine
authors.initials: EL
authors.lastname: Shaw
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Alexander
authors.initials: A
authors.lastname: Bina
authors.affiliation: 2 Tissue Bioengineering Laboratory, Department of Agricultural and Biological Engineering, Mississippi State University , Starkville, Mississippi.
authors.forename: Bryn
authors.initials: B
authors.lastname: Brazile
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Rierson
authors.affiliation: 2 Tissue Bioengineering Laboratory, Department of Agricultural and Biological Engineering, Mississippi State University , Starkville, Mississippi.
authors.forename: Sourav S
authors.initials: SS
authors.lastname: Patnaik
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Allison
authors.initials: A
authors.lastname: Kennamer
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Rebekah
authors.initials: R
authors.lastname: Odum
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Ovidiu
authors.initials: O
authors.lastname: Cotoi
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Preda
authors.initials: P
authors.lastname: Terezia
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Klara
authors.initials: K
authors.lastname: Branzaniuc
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Harrison
authors.initials: H
authors.lastname: Smallwood
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Radu
authors.initials: R
authors.lastname: Deac
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Imre
authors.initials: I
authors.lastname: Egyed
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Pavai
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Annamaria
authors.initials: A
authors.lastname: Szanto
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Lucian
authors.initials: L
authors.lastname: Harceaga
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Horatiu
authors.initials: H
authors.lastname: Suciu
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Victor
authors.initials: V
authors.lastname: Raicea
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Peter
authors.initials: P
authors.lastname: Olah
authors.affiliation: 4 Cardiovascular Tissue Engineering and Regenerative Medicine Laboratory, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Agneta
authors.initials: A
authors.lastname: Simionescu
authors.affiliation: 2 Tissue Bioengineering Laboratory, Department of Agricultural and Biological Engineering, Mississippi State University , Starkville, Mississippi.
authors.forename: Jun
authors.initials: J
authors.lastname: Liao
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Ionela
authors.initials: I
authors.lastname: Movileanu
authors.affiliation: 3 Tissue Engineering and Regenerative Medicine Laboratory, Department of Anatomy, University of Medicine and Pharmacy , Targu Mures, Romania .
authors.forename: Marius
authors.initials: M
authors.lastname: Harpa
authors.affiliation: 1 Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University , Clemson, South Carolina.
authors.forename: Dan Teodor
authors.initials: DT
authors.lastname: Simionescu
mesh_terms: D000818:Animals; D001011:Aorta / Q000737:chemistry*; D006351:Heart Valves / Q000737:chemistry*; D013552:Swine; D054457:Tissue Scaffolds / Q000737:chemistry*