title: Effect of three decellularisation protocols on the mechanical behaviour and structural properties of sheep aortic valve conduits.
abstract: OBJECTIVE
  To determine the best method for decellularisation of aortic valve conduits (AVCs) that efficiently removes the cells while preserving the extracellular matrix (ECM) by examining the valvular and conduit sections separately.
  
  
  METHODS
  Sheep AVCs were decellularised by using three different protocols: detergent-based (1% SDS+1% SDC), detergent and enzyme-based (Triton+EDTA+RNase and DNase), and enzyme-based (Trypsin+RNase and DNase) methods. The efficacy of the decellularisation methods to completely remove the cells while preserving the ECM was evaluated by histological evaluation, scanning electron microscopy (SEM), hydroxyproline analysis, tensile test, and DAPI staining.
  
  
  RESULTS
  The detergent-based method completely removed the cells and left the ECM and collagen content in the valve and conduit sections relatively well preserved. The detergent and enzyme-based protocol did not completely remove the cells, but left the collagen content in both sections well preserved. ECM deterioration was observed in the aortic valves (AVs), but the ultrastructure of the conduits was well preserved, with no media distortion. The enzyme-based protocol removed the cells relatively well; however, mild structural distortion and poor collagen content was observed in the AVs. Incomplete cell removal (better than that observed with the detergent and enzyme-based protocol), poor collagen preservation, and mild structural distortion were observed in conduits treated with the enzyme-based method.
  
  
  CONCLUSIONS
  The results suggested that the detergent-based methods are the most effective protocols for cell removal and ECM preservation of AVCs. The AVCs treated with this detergent-based method may be excellent scaffolds for recellularisation.
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Khorramirouz
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Shabnam
authors.initials: S
authors.lastname: Sabetkish
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Aram
authors.initials: A
authors.lastname: Akbarzadeh
authors.affiliation: Cancer Research Center, Iranian Cancer Institute, Department of Pathology, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Ahad
authors.initials: A
authors.lastname: Muhammadnejad
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Heidari
authors.affiliation: Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children's Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: kajbafzd@sina.tums.ac.ir.
authors.forename: Abdol-Mohammad
authors.initials: AM
authors.lastname: Kajbafzadeh
mesh_terms: D000818:Animals; D001021:Aortic Valve / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D055598:Chemical Phenomena; D003902:Detergents / Q000737:chemistry; D005109:Extracellular Matrix / Q000737:chemistry* / Q000648:ultrastructure; D007492:Iran; D055595:Mechanical Phenomena; D034561:Sheep, Domestic; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D003902:Detergents
keywords: Acellular; Aorta; Aortic conduit; Heart valve; Sheep