Jérôme Duisit1, Hadrien Amiel2, Tsering Wüthrich3, Adriano Taddeo4, Adeline Dedriche5, Vincent Destoop6, Thomas Pardoen7, Caroline Bouzin8, Virginie Joris9, Derek Magee10, Esther Vögelin11, David Harriman12, Chantal Dessy13, Giuseppe Orlando14, Catherine Behets15, Robert Rieben16, Pierre Gianello17, Benoît Lengelé18. 1. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium; Department of Plastic and Reconstructive Surgery, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10, B-1200 Brussels, Belgium. Electronic address: jerome.duisit@uclouvain.be. 2. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: hadrien.amiel@student.uclouvain.be. 3. Department for BioMedical Research, University of Bern, Murtenstrasse 50, CH-3008 Bern, Switzerland. Electronic address: tsering.w@students.unibe.ch. 4. Department for BioMedical Research, University of Bern, Murtenstrasse 50, CH-3008 Bern, Switzerland; Department of Plastic, Reconstructive and Hand Surgery, Inselspital, University Hospital, CH-3010 Bern, Switzerland. Electronic address: adriano.taddeo@dbmr.unibe.ch. 5. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: adeline.dedriche@uclouvain.be. 6. Institute of Mechanics, Materials and Civil Engineering, Materials and Process Engineering, Place Sainte Barbe 2/L5.02.02, B-1348 Louvain-la-Neuve, Belgium. Electronic address: vincent.destoop@uclouvain.be. 7. Institute of Mechanics, Materials and Civil Engineering, Materials and Process Engineering, Place Sainte Barbe 2/L5.02.02, B-1348 Louvain-la-Neuve, Belgium. Electronic address: thomas.pardoen@uclouvain.be. 8. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: caroline.bouzin@uclouvain.be. 9. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: virginie.joris@uclouvain.be. 10. School of Computing, University of Leeds, Leeds LS2 9JT, UK; HeteroGenius Limited, 21 Parkland Crescent, LS6 4PR Leeds, UK. Electronic address: D.R.Magee@leeds.ac.uk. 11. Department of Plastic, Reconstructive and Hand Surgery, Inselspital, University Hospital, CH-3010 Bern, Switzerland. Electronic address: esther.voegelin@insel.ch. 12. Department of Surgery, Section of Transplantation, Wake Forest School of Medicine, 1 Medical Center Blvd, Winston-Salem, NC 27157, USA. 13. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: chantal.dessy@uclouvain.be. 14. Department of Surgery, Section of Transplantation, Wake Forest School of Medicine, 1 Medical Center Blvd, Winston-Salem, NC 27157, USA. Electronic address: gorlando@wakehealth.edu. 15. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: catherine.behets@uclouvain.be. 16. Department for BioMedical Research, University of Bern, Murtenstrasse 50, CH-3008 Bern, Switzerland. Electronic address: robert.rieben@dbmr.unibe.ch. 17. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium. Electronic address: pierre.gianello@uclouvain.be. 18. Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, Avenue Hippocrate 55/B1.55.04, B-1200 Brussels, Belgium; Department of Plastic and Reconstructive Surgery, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10, B-1200 Brussels, Belgium. Electronic address: benoit.lengele@uclouvain.be.
Abstract
INTRODUCTION: Human ear reconstruction is recognized as the emblematic enterprise in tissue engineering. Up to now, it has failed to reach human applications requiring appropriate tissue complexity along with an accessible vascular tree. We hereby propose a new method to process human auricles in order to provide a poorly immunogenic, complex and vascularized ear graft scaffold. METHODS: 12 human ears with their vascular pedicles were procured. Perfusion-decellularization was applied using a SDS/polar solvent protocol. Cell and antigen removal was examined by histology and DNA was quantified. Preservation of the extracellular matrix (ECM) was assessed by conventional and 3D-histology, proteins and cytokines quantifications. Biocompatibility was assessed by implantation in rats for up to 60 days. Adipose-derived stem cells seeding was conducted on scaffold samples and with human aortic endothelial cells whole graft seeding in a perfusion-bioreactor. RESULTS: Histology confirmed cell and antigen clearance. DNA reduction was 97.3%. ECM structure and composition were preserved. Implanted scaffolds were tolerated in vivo, with acceptable inflammation, remodeling, and anti-donor antibody formation. Seeding experiments demonstrated cell engraftment and viability. CONCLUSIONS: Vascularized and complex auricular scaffolds can be obtained from human source to provide a platform for further functional auricular tissue engineered constructs, hence providing an ideal road to the vascularized composite tissue engineering approach. STATEMENT OF SIGNIFICANCE: The ear is emblematic in the biofabrication of tissues and organs. Current regenerative medicine strategies, with matrix from donor tissues or 3D-printed, didn't reach any application for reconstruction, because critically missing a vascular tree for perfusion and transplantation. We previously described the production of vascularized and cell-compatible scaffolds, from porcine ear grafts. In this study, we ---- applied findings directly to human auricles harvested from postmortem donors, providing a perfusable matrix that retains the ear's original complexity and hosts new viable cells after seeding. This approach unlocks the ability to achieve an auricular tissue engineering approach, associated with possible clinical translation.
INTRODUCTION:Human ear reconstruction is recognized as the emblematic enterprise in tissue engineering. Up to now, it has failed to reach human applications requiring appropriate tissue complexity along with an accessible vascular tree. We hereby propose a new method to process human auricles in order to provide a poorly immunogenic, complex and vascularized ear graft scaffold. METHODS: 12 human ears with their vascular pedicles were procured. Perfusion-decellularization was applied using a SDS/polar solvent protocol. Cell and antigen removal was examined by histology and DNA was quantified. Preservation of the extracellular matrix (ECM) was assessed by conventional and 3D-histology, proteins and cytokines quantifications. Biocompatibility was assessed by implantation in rats for up to 60 days. Adipose-derived stem cells seeding was conducted on scaffold samples and with human aortic endothelial cells whole graft seeding in a perfusion-bioreactor. RESULTS: Histology confirmed cell and antigen clearance. DNA reduction was 97.3%. ECM structure and composition were preserved. Implanted scaffolds were tolerated in vivo, with acceptable inflammation, remodeling, and anti-donor antibody formation. Seeding experiments demonstrated cell engraftment and viability. CONCLUSIONS: Vascularized and complex auricular scaffolds can be obtained from human source to provide a platform for further functional auricular tissue engineered constructs, hence providing an ideal road to the vascularized composite tissue engineering approach. STATEMENT OF SIGNIFICANCE: The ear is emblematic in the biofabrication of tissues and organs. Current regenerative medicine strategies, with matrix from donor tissues or 3D-printed, didn't reach any application for reconstruction, because critically missing a vascular tree for perfusion and transplantation. We previously described the production of vascularized and cell-compatible scaffolds, from porcine ear grafts. In this study, we ---- applied findings directly to human auricles harvested from postmortem donors, providing a perfusable matrix that retains the ear's original complexity and hosts new viable cells after seeding. This approach unlocks the ability to achieve an auricular tissue engineering approach, associated with possible clinical translation.
Authors: Paul Girard; Joelle Dulong; Jerome Duisit; Camille Mocquard; Simon Le Gallou; Benoit Chaput; Elise Lupon; Eric Watier; Audrey Varin; Karin Tarte; Nicolas Bertheuil Journal: Front Bioeng Biotechnol Date: 2022-09-13