Development and In Vitro/In Vivo Comparative Characterization of Cryopreserved and Decellularized Tracheal Grafts
Elena Stocco,
Silvia Barbon,
Marco Mammana,
Diletta Trojan,
Alice Bianchin,
Francesca Favaretto,
Martina Contran,
Giovanni Zambello,
Andrea Vogliardi,
Marta Confalonieri,
Silvia Todros,
Piero G. Pavan,
Filippo Romanato,
Maria Teresa Conconi,
Veronica Macchi,
Raffaele De Caro,
Federico Rea,
Andrea Porzionato
Affiliations
Elena Stocco
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Silvia Barbon
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Marco Mammana
L.i.f.e.L.a.b. Program, Consorzio per la Ricerca Sanitaria (CORIS), Veneto Region, 35128 Padova, Italy
Diletta Trojan
Tissue Bank, Fondazione Banca dei Tessuti del Veneto ETS, 31100 Treviso, Italy
Alice Bianchin
Tissue Bank, Fondazione Banca dei Tessuti del Veneto ETS, 31100 Treviso, Italy
Francesca Favaretto
Tissue Bank, Fondazione Banca dei Tessuti del Veneto ETS, 31100 Treviso, Italy
Martina Contran
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Giovanni Zambello
Thoracic Surgery Division, Department of Cardiac, Thoracic, Vascular Sciences and Public Health, Padova University Hospital, Via Giustiniani, 2, 35128 Padova, Italy
Andrea Vogliardi
Department of Physics and Astronomy ‘G. Galilei’, University of Padova, 35131 Padova, Italy
Marta Confalonieri
Department of Industrial Engineering, University of Padova, 35131 Padova, Italy
Silvia Todros
Department of Industrial Engineering, University of Padova, 35131 Padova, Italy
Piero G. Pavan
Department of Industrial Engineering, University of Padova, 35131 Padova, Italy
Filippo Romanato
L.i.f.e.L.a.b. Program, Consorzio per la Ricerca Sanitaria (CORIS), Veneto Region, 35128 Padova, Italy
Maria Teresa Conconi
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, 35131 Padova, Italy
Veronica Macchi
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Raffaele De Caro
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Federico Rea
L.i.f.e.L.a.b. Program, Consorzio per la Ricerca Sanitaria (CORIS), Veneto Region, 35128 Padova, Italy
Andrea Porzionato
Section of Human Anatomy, Department of Neuroscience, University of Padova, 35121 Padova, Italy
Tracheal reconstruction represents a challenge when primary anastomosis is not feasible. Within this scenario, the study aim was to develop a new pig-derived decellularized trachea (DecellT) to be compared with the cryopreserved counterpart (CryoT) for a close predictive analysis. Tracheal segments underwent decellularization by a physical + enzymatic + chemical method (12 cycles); in parallel, cryopreserved samples were also prepared. Once decellularized (histology/DNA quantification), the two groups were characterized for Alpha-Gal epitopes/structural proteins (immunohistochemistry/histology/biochemical assays/second harmonic generation microscopy)/ultrastructure (Scanning Electron Microscopy (SEM))/mechanical behaviour. Cytotoxicity absence was assessed in vitro (extract-test assay/direct seeding, HM1SV40 cell line) while biocompatibility was verified in BALB/c mice, followed by histological/immunohistochemical analyses and SEM (14 days). Decellularization effectively removed Alpha-Gal epitopes; cartilage histoarchitecture was retained in both groups, showing chondrocytes only in the CryoT. Cryopreservation maintained few respiratory epithelium sparse cilia, not detectable in DecellT. Focusing on ECM, preserved structural/ultrastructural organization and collagen content were observed in the cartilage of both; conversely, the GAGs were significantly reduced in DecellT, as confirmed by mechanical study results. No cytotoxicity was highlighted by CryoT/DecellT in vitro, as they were also corroborated by a biocompatibility assay. Despite some limitations (cells presence/GAGs reduction), CryoT/DecellT are both appealing options, which warrant further investigation in comparative in vivo studies.