Cargando…

Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI

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 el...

Descripción completa

Detalles Bibliográficos
Autores principales: Boeer, Ulrike, Buettner, Falk F. R., Klingenberg, Melanie, Antonopoulos, Georgios C., Meyer, Heiko, Haverich, Axel, Wilhelmi, Mathias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4144968/
https://www.ncbi.nlm.nih.gov/pubmed/25157402
http://dx.doi.org/10.1371/journal.pone.0105964
_version_ 1782332110280851456
author Boeer, Ulrike
Buettner, Falk F. R.
Klingenberg, Melanie
Antonopoulos, Georgios C.
Meyer, Heiko
Haverich, Axel
Wilhelmi, Mathias
author_facet Boeer, Ulrike
Buettner, Falk F. R.
Klingenberg, Melanie
Antonopoulos, Georgios C.
Meyer, Heiko
Haverich, Axel
Wilhelmi, Mathias
author_sort Boeer, Ulrike
collection PubMed
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 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.
format Online
Article
Text
id pubmed-4144968
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-41449682014-08-29 Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI Boeer, Ulrike Buettner, Falk F. R. Klingenberg, Melanie Antonopoulos, Georgios C. Meyer, Heiko Haverich, Axel Wilhelmi, Mathias PLoS One Research Article 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. Public Library of Science 2014-08-26 /pmc/articles/PMC4144968/ /pubmed/25157402 http://dx.doi.org/10.1371/journal.pone.0105964 Text en © 2014 Boeer et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Boeer, Ulrike
Buettner, Falk F. R.
Klingenberg, Melanie
Antonopoulos, Georgios C.
Meyer, Heiko
Haverich, Axel
Wilhelmi, Mathias
Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title_full Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title_fullStr Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title_full_unstemmed Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title_short Immunogenicity of Intensively Decellularized Equine Carotid Arteries Is Conferred by the Extracellular Matrix Protein Collagen Type VI
title_sort immunogenicity of intensively decellularized equine carotid arteries is conferred by the extracellular matrix protein collagen type vi
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4144968/
https://www.ncbi.nlm.nih.gov/pubmed/25157402
http://dx.doi.org/10.1371/journal.pone.0105964
work_keys_str_mv AT boeerulrike immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT buettnerfalkfr immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT klingenbergmelanie immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT antonopoulosgeorgiosc immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT meyerheiko immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT haverichaxel immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi
AT wilhelmimathias immunogenicityofintensivelydecellularizedequinecarotidarteriesisconferredbytheextracellularmatrixproteincollagentypevi