In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization

Isaac Perea-Gil, Juan J. Uriarte, Cristina Prat-Vidal, Carolina Gálvez-Montón, Santiago Roura, Aida Llucià-Valldeperas, Carolina Soler-Botija, Ramon Farré, Daniel Navajas, Antoni Bayes-Genis

Research output: Contribution to journalArticleResearchpeer-review

34 Citations (Scopus)

Abstract

© 2015, E-Century Publishing Corporation. All right reserved. Introduction. 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. Materials and 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/mm2 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.
Original languageEnglish
Pages (from-to)558-573
JournalAmerican Journal of Translational Research
Volume7
Issue number3
Publication statusPublished - 1 Jan 2015

Keywords

  • Acellular myocardial scaffold
  • Adipose tissue-derived progenitor cells
  • Decellularization protocols
  • Extracellular matrix
  • Myocardial infarction
  • Recellularization

Fingerprint Dive into the research topics of 'In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization'. Together they form a unique fingerprint.

  • Cite this

    Perea-Gil, I., Uriarte, J. J., Prat-Vidal, C., Gálvez-Montón, C., Roura, S., Llucià-Valldeperas, A., Soler-Botija, C., Farré, R., Navajas, D., & Bayes-Genis, A. (2015). In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization. American Journal of Translational Research, 7(3), 558-573.