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Publications

Journal Article Research Support, Non-U.S. Gov't Epublish 11 citations

PubMed: 36230907 DOI: 10.3390/cells11192945

Bio-Engineered Scaffolds Derived from Decellularized Human Esophagus for Functional Organ Reconstruction.

Cells 11(19)

Authors: Silvia Barbon, Andrea Biccari, Elena Stocco, Giovanni Capovilla, Edoardo D'Angelo, Martina Todesco, Deborah Sandrin, Andrea Bagno, Filippo Romanato, Veronica Macchi, Raffaele De Caro, Marco Agostini, Stefano Merigliano, Michele Valmasoni, Andrea Porzionato

Affiliations

  • Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy.
  • L.i.f.e.L.a.b. Program, Consorzio per la Ricerca Sanitaria, 35128 Padova, Italy.
  • Foundation for Biology and Regenerative Medicine, Tissue Engineering and Signaling-TES, Onlus, 35136 Padova, Italy.
  • Department of Surgical Oncological and Gastroenterological Sciences, University of Padova, 35128 Padova, Italy.
  • Department of Industrial Engineering, University of Padova, 35131 Padova, Italy.
  • Department of Physics and Astronomy "G. Galilei", University of Padova, 35131 Padova, Italy.

Abstract

Esophageal reconstruction through bio-engineered allografts that highly resemble the peculiar properties of the tissue extracellular matrix (ECM) is a prospective strategy to overcome the limitations of current surgical approaches. In this work, human esophagus was decellularized for the first time in the literature by comparing three detergent-enzymatic protocols. After decellularization, residual DNA quantification and histological analyses showed that all protocols efficiently removed cells, DNA (<50 ng/mg of tissue) and muscle fibers, preserving collagen/elastin components. The glycosaminoglycan fraction was maintained (70-98%) in the decellularized versus native tissues, while immunohistochemistry showed unchanged expression of specific ECM markers (collagen IV, laminin). The proteomic signature of acellular esophagi corroborated the retention of structural collagens, basement membrane and matrix-cell interaction proteins. Conversely, decellularization led to the loss of HLA-DR expression, producing non-immunogenic allografts. According to hydroxyproline quantification, matrix collagen was preserved (2-6 µg/mg of tissue) after decellularization, while Second-Harmonic Generation imaging highlighted a decrease in collagen intensity. Based on uniaxial tensile tests, decellularization affected tissue stiffness, but sample integrity/manipulability was still maintained. Finally, the cytotoxicity test revealed that no harmful remnants/contaminants were present on acellular esophageal matrices, suggesting allograft biosafety. Despite the different outcomes showed by the three decellularization methods (regarding, for example, tissue manipulability, DNA removal, and glycosaminoglycans/hydroxyproline contents) the ultimate validation should be provided by future repopulation tests and in vivo orthotopic implant of esophageal scaffolds.

MeSH

Glycosaminoglycans, Collagen, Esophagus, Dna, Cells, Extracellular Matrix, Tissues, Tissue Engineering, Allografts, Hydroxyproline

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