title: Immunogenicity of intensively decellularized equine carotid arteries is conferred by the extracellular matrix protein collagen type VI.
abstract: The limited biocompatibility of decellularized scaffolds is an ongoing challenge in tissue engineering. Here, we demonstrate the residual immunogenicity of an extensively decellularized equine carotid artery (dEAC(intens)) and identify the involved immunogenic components. EAC were submitted to an elaborated intensified decellularization protocol with SDS/sodium desoxycholate for 72 h using increased processing volumes (dEAC(intens)), and compared to dEAC(ord) prepared by an ordinary protocol (40 h, normal volumes). Matrix integrity was checked via correlative volumetric visualization which revealed only minor structural changes in the arterial wall. In dEAC(intens), a substantial depletion of cellular components was obvious for smooth muscle actin (100%), MHC I complexes (97.8%), alphaGal epitops (98.4% and 91.3%) and for DNA (final concentration of 0.34 ± 0.16 ng/mg tissue). However, dEAC(intens) still evoked antibody formation in mice after immunization with dEAC(intens) extracts, although to a lower extent than dEAC(ord). Mouse plasma antibodies recognized a 140 kDa band which was revealed to contain collagen VI alpha1 and alpha2 chains via mass spectrometry of both 2D electrophoretically separated and immunoprecipitated proteins. Thus, even the complete removal of cellular proteins did not yield non-immunogenic dEAC as the extracellular matrix still conferred immunogenicity by collagen VI. However, as lower antibody levels were achieved by the intensified decellularization protocol, this seems to be a promising basis for further development.
authors.affiliation: GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Boeer
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Falk F R
authors.initials: FF
authors.lastname: Buettner
authors.affiliation: GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Klingenberg
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e.V., Hannover, Germany.
authors.forename: Georgios C
authors.initials: GC
authors.lastname: Antonopoulos
authors.affiliation: Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany; Biomedical Optics Department, Laser Zentrum Hannover e.V., Hannover, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Meyer
authors.affiliation: GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Axel
authors.initials: A
authors.lastname: Haverich
authors.affiliation: GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Mathias
authors.initials: M
authors.lastname: Wilhelmi
mesh_terms: D000818:Animals; D002339:Carotid Arteries / Q000276:immunology* / Q000637:transplantation; D024141:Collagen Type IV / Q000276:immunology*; D005260:Female; D064593:Heterografts; D006648:Histocompatibility; D006736:Horses; D051379:Mice; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D024141:Collagen Type IV