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Home / Equine Research Bank / PLoS One / Immunogenicity of intensively decellularized equine carotid ...
PloS one2014; 9(8); e105964; doi: 10.1371/journal.pone.0105964

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.
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Publication Date: 2014-08-26 PubMed ID: 25157402PubMed Central: PMC4144968DOI: 10.1371/journal.pone.0105964Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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The study investigates the remaining immunity-triggering components in extremely decellularized horse carotid artery samples, identifying collagen type VI as the primary cause. Despite efforts to remove all cellular proteins, the extracellular matrix retained enough immunogenicity to provoke an immune response.

Decellularization and Objective of the Study

  • The study seeks to solve the problem of low biocompatibility in decellularized scaffolds, a significant concern in tissue engineering.
  • To understand the source of remaining immunogenicity in extensively cleaned equine carotid artery (dEAC) samples, researchers subjected them to a more intensive decellularization process. This involved using higher volumes of SDS/sodium desoxycholate for an extended period.

Evaluation of Decellularization Process and Finding

  • The structural integrity of the treated arterial wall was preserved, significant as structural changes typically arise in tissue engineering.
  • A high percentage of cellular components were removed in this process, including 100% of smooth muscle actin, nearly all MHC 1 complexes and alphaGal epitops, and a significant reduction in DNA.
  • However, even after this radical treatment, the material still generated an immune response when presented to the mice’s immune system, though less than the ordinarily treated samples.

Role of Collagen VI in Immunogenic Response

  • The immune responses were traced back to a 140 kDa band of mouse plasma antibodies.
  • Meticulous testing identified this as being composed of collagen VI alpha1 and alpha2 chains.
  • Despite the removal of cellular proteins, collagen VI in the extracellular matrix remained, triggering an immune response.
  • The study suggests a connection between the immune response and collagen type VI, a critical finding that may help improve decellularization protocols in future tissue engineering experiments.

Conclusion of the Study

  • This study provides robust evidence that even extensive decellularization might not entirely prevent an immune response, with extracellular matrix elements like collagen VI identified as potential triggers.
  • However, the lower levels of immune response witnessed under the intensive decellularization protocol suggest it might be a promising foundation for future development in tissue engineering.

Cite This Article

APA
Boeer U, Buettner FF, Klingenberg M, Antonopoulos GC, Meyer H, Haverich A, Wilhelmi M. (2014). Immunogenicity of intensively decellularized equine carotid arteries is conferred by the extracellular matrix protein collagen type VI. PLoS One, 9(8), e105964. https://doi.org/10.1371/journal.pone.0105964

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 9
Issue: 8
Pages: e105964
PII: e105964

Researcher Affiliations

Boeer, Ulrike
  • GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
Buettner, Falk F R
  • Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
Klingenberg, Melanie
  • GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
Antonopoulos, Georgios C
  • Biomedical Optics Department, Laser Zentrum Hannover e.V., Hannover, Germany.
Meyer, Heiko
  • Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany; Biomedical Optics Department, Laser Zentrum Hannover e.V., Hannover, Germany.
Haverich, Axel
  • GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.
Wilhelmi, Mathias
  • GMP- model laboratory for tissue engineering, Hannover, Germany; Division for Cardiac-, Thoracic-, Transplantation- and Vascular Surgery, Hannover Medical School, Hannover, Germany.

MeSH Terms

  • Animals
  • Carotid Arteries / immunology
  • Carotid Arteries / transplantation
  • Collagen Type IV / immunology
  • Female
  • Heterografts
  • Histocompatibility
  • Horses
  • Mice
  • Tissue Engineering
  • Tissue Scaffolds

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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Citations

This article has been cited 12 times.
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