title: In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization.
abstract: BACKGROUND
  Selection of a biomaterial-based scaffold that mimics native myocardial extracellular matrix (ECM) architecture can facilitate functional cell attachment and differentiation. Although decellularized myocardial ECM accomplishes these premises, decellularization processes may variably distort or degrade ECM structure.
  
  
  METHODS
  Two decellularization protocols (DP) were tested on porcine heart samples (epicardium, mid myocardium and endocardium). One protocol, DP1, was detergent-based (SDS and Triton X-100), followed by DNase I treatment. The other protocol, DP2, was focused in trypsin and acid with Triton X-100 treatments. Decellularized myocardial scaffolds were reseeded by embedding them in RAD16-I peptidic hydrogel with adipose tissue-derived progenitor cells (ATDPCs).
  
  
  RESULTS
  Both protocols yielded acellular myocardial scaffolds (~82% and ~94% DNA reduction for DP1 and DP2, respectively). Ultramicroscopic assessment of scaffolds was similar for both protocols and showed filamentous ECM with preserved fiber disposition and structure. DP1 resulted in more biodegradable scaffolds (P = 0.04). Atomic force microscopy revealed no substantial ECM stiffness changes post-decellularization compared to native tissue. The Young's modulus did not differ between heart layers (P = 0.69) or decellularization protocols (P = 0.15). After one week, recellularized DP1 scaffolds contained higher cell density (236 ± 106 and 98 ± 56 cells/mm(2) for recellularized DP1 and DP2 scaffolds, respectively; P = 0.04). ATDPCs in both DP1 and DP2 scaffolds expressed the endothelial marker isolectin B4, but only in the DP1 scaffold ATDPCs expressed the cardiac markers GATA4, connexin43 and cardiac troponin T.
  
  
  CONCLUSIONS
  In our hands, DP1 produced myocardial scaffolds with higher cell repopulation and promotes ATDPCs expression of endothelial and cardiomyogenic markers.
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Isaac
authors.initials: I
authors.lastname: Perea-Gil
authors.affiliation: Biophysics and Bioengineering Unit, Faculty of Medicine, University of Barcelona Barcelona, Spain ; CIBER Enfermedades Respiratorias Madrid, Spain.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Uriarte
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Prat-Vidal
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Gálvez-Montón
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Roura
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Aida
authors.initials: A
authors.lastname: Llucià-Valldeperas
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Soler-Botija
authors.affiliation: Biophysics and Bioengineering Unit, Faculty of Medicine, University of Barcelona Barcelona, Spain ; CIBER Enfermedades Respiratorias Madrid, Spain ; Institut d'Investigacions Biomèdiques August Pi i Sunyer Barcelona, Spain.
authors.forename: Ramon
authors.initials: R
authors.lastname: Farré
authors.affiliation: Biophysics and Bioengineering Unit, Faculty of Medicine, University of Barcelona Barcelona, Spain ; CIBER Enfermedades Respiratorias Madrid, Spain ; Institute for Bioengineering of Catalonia Barcelona, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Navajas
authors.affiliation: ICREC (Heart Failure and Cardiac Regeneration) Research Lab, Health Sciences Research Institute Germans Trias i Pujol (IGTP). Cardiology Service, Hospital Universitari Germans Trias i Pujol Badalona, Barcelona, Spain ; Department of Medicine, Autonomous University of Barcelona (UAB) Barcelona, Spain.
authors.forename: Antoni
authors.initials: A
authors.lastname: Bayes-Genis
keywords: Myocardial infarction; acellular myocardial scaffold; adipose tissue-derived progenitor cells; decellularization protocols; extracellular matrix; recellularization