title: A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix.
abstract: A combined physical-chemical protocol for whole full-thickness bladder decellularization is proposed, based on organ cyclic distention through repeated infusion/withdrawal of the decellularization agents through the urethra. The dynamic decellularization was intended to enhance cell removal efficiency, facilitating the delivery of detergents within the inner layers of the tissue and the removal of cell debris. The use of mild chemical detergents (hypotonic solution and non-ionic detergent) was employed to limit adverse effects upon matrix 3D ultrastructure. Inspection of the presence of residual DNA and RNA was carried out on decellularized matrices to verify effective cell removal. Histological investigation was focused on assessing the retention of adequate structural and functional components that regulate the biomechanical behaviour of the acellular tissue. Biomechanical properties were evaluated through uniaxial tensile loading tests of tissue strips and through ex vivo filling cystometry to evaluate the whole-organ mechanical response to a physiological-like loading state. According to our results, a dynamic decellularization protocol of 17 h duration with a 5 ml/min detergent infusion flow rate revealed higher DNA removal efficiency than standard static decellularization, resulting in residual DNA content < 50 ng/mg dry tissue weight. Furthermore, the collagen network and elastic fibres distribution were preserved in the acellular ECM, which exhibited suitable biomechanical properties in the perspective of its future use as an implant for bladder augmentation.
authors.affiliation: Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Consolo
authors.affiliation: Department of Health, Animal Science and Food Safety, Surgery Unit, Faculty of Veterinary Medicine, Università delgi Studi di Milano, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Brizzola
authors.affiliation: Department of Health, Animal Science and Food Safety, Surgery Unit, Faculty of Veterinary Medicine, Università delgi Studi di Milano, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Tremolada
authors.affiliation: Department of Veterinary Science and Public Health, Faculty of Veterinary Medicine, Università degli Studi di Milano, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Grieco
authors.affiliation: Department of Veterinary Science and Public Health, Faculty of Veterinary Medicine, Università degli Studi di Milano, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Riva
authors.affiliation: Department of Health, Animal Science and Food Safety, Surgery Unit, Faculty of Veterinary Medicine, Università delgi Studi di Milano, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Acocella
authors.affiliation: Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Italy.
authors.forename: G B
authors.initials: GB
authors.lastname: Fiore
authors.affiliation: Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Soncini
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D012313:RNA / Q000378:metabolism; D011817:Rabbits; D013194:Staining and Labeling; D023822:Tissue Engineering / Q000379:methods*; D001743:Urinary Bladder / Q000166:cytology* / Q000502:physiology*; D016474:Weight-Bearing
chemical_list: D012313:RNA; D004247:DNA
keywords: biological scaffold; bladder acellular matrix; bladder tissue engineering; whole-organ decellularization