Zusammenfassung: | |
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 <inf>intens</inf>) 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 <inf>intens</inf>), and compared to dEAC<inf>ord</inf> 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<inf>intens</inf>, 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<inf>intens</inf> still evoked antibody formation in mice after immunization with dEAC<inf>intens</inf> extracts, although to a lower extent than dEAC<inf>ord</inf>. 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.
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Lizenzbestimmungen: | CC BY 4.0 Unported - https://creativecommons.org/licenses/by/4.0/ |
Publikationstyp: | Article |
Publikationsstatus: | publishedVersion |
Erstveröffentlichung: | 2014 |
Schlagwörter (englisch): | cell protein, collagen type 4, collagen type 6, deoxycholate sodium, DNA, epitope, major histocompatibility antigen class 1, scleroprotein, smooth muscle actin, collagen type 4, animal tissue, antibody production, artery wall, article, biocompatibility, carotid artery, clinical protocol, controlled study, DNA content, Equidae, extracellular matrix, immunization, immunogenicity, immunoprecipitation, liquid chromatography, mass spectrometry, mouse, nonhuman, protein analysis, two dimensional gel electrophoresis, volumetry, animal, carotid artery, female, histocompatibility, horse, immunology, tissue engineering, tissue scaffold, transplantation, xenograft, Animals, Carotid Arteries, Collagen Type IV, Female, Heterografts, Histocompatibility, Horses, Mice, Tissue Engineering, Tissue Scaffolds |
Fachliche Zuordnung (DDC): | 500 | Naturwissenschaften, 610 | Medizin, Gesundheit |
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