Immunogenicity of intensively decellularized equine carotid arteries is conferred by the extracellular matrix protein collagen type VI
Creators: name: Boeer, Ulrike
name: Buettner, Falk F.R.
name: Klingenberg, Melanie
name: Antonopoulos, Georgios C.
name: Meyer, Heiko
name: Haverich, Axel
name: Wilhelmi, Mathias
Publisher: name: San Francisco, CA : Public Library of Science (PLoS)
Year: 2014
Subjects: subject: cell protein
subject: collagen type 4
subject: collagen type 6
subject: deoxycholate sodium
subject: DNA
subject: epitope
subject: major histocompatibility antigen class 1
subject: scleroprotein
subject: smooth muscle actin
subject: animal tissue
subject: antibody production
subject: artery wall
subject: article
subject: biocompatibility
subject: carotid artery
subject: clinical protocol
subject: controlled study
subject: DNA content
subject: Equidae
subject: extracellular matrix
subject: immunization
subject: immunogenicity
subject: immunoprecipitation
subject: liquid chromatography
subject: mass spectrometry
subject: mouse
subject: nonhuman
subject: protein analysis
subject: two dimensional gel electrophoresis
subject: volumetry
subject: animal
subject: female
subject: histocompatibility
subject: horse
subject: immunology
subject: tissue engineering
subject: tissue scaffold
subject: transplantation
subject: xenograft
subject: Animals
subject: Carotid Arteries
subject: Collagen Type IV
subject: Female
subject: Heterografts
subject: Histocompatibility
subject: Horses
subject: Mice
subject: Tissue Engineering
subject: Tissue Scaffolds
subject: Dewey Decimal Classification::500 | Naturwissenschaften
subject: Dewey Decimal Classification::600 | Technik::610 | Medizin, Gesundheit

description: 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 dEACord 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 dEACintens, 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, dEACintens still evoked antibody formation in mice after immunization with dEACintens extracts, although to a lower extent than dEACord. 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.