The effect of detergent-based decellularization procedures on cellular proteins and immunogenicity in equine carotid artery grafts.
Abstract: Decellularized equine carotid arteries (dEAC) may represent a reasonable alternative to alloplastic materials in vascular replacement therapy. Acellularity of the matrix is standardly evaluated by DNA quantification what however may not record sufficiently the degree of matrix immunogenicity. Thus, our aim was to analyze dEAC with a low DNA content for residual cellular proteins. A detergent-based decellularization protocol including endonuclease treatment resulted in dEAC with 0.6 ± 0.15 ng DNA/mg dry weight representing 0.33 ± 0.14% of native tissue DNA content. In contrast, when matrices were homogenized and extracted by high detergent concentrations westernblot analyses revealed cytosolic and cytosceleton proteins like GAPDH and smooth muscle actin which were depleted to 4.1 ± 1.9% and 13.8 ± 0.55%, resp. Also putative immunogenic MHC I complexes and the alpha-Gal epitop were reduced to only 14.8 ± 1.2% and 15.1 ± 2.05%. Mass spectrometry of matrix extracts identified 306 proteins belonging to cytosol, organelles, nucleus and cell membrane. Moreover, aqueous matrix extracts evoked a pronounced antibody formation when administered in mice and thus display high immunogenic potential. Our data indicate that an established decellularization protocol which results in acellular matrices evaluated by low DNA content reduces but not eliminates cellular components which may contribute to its immunogenic potential in vivo.
Copyright © 2011 Elsevier Ltd. All rights reserved.
Publication Date: 2011-09-23 PubMed ID: 21944468DOI: 10.1016/j.biomaterials.2011.09.015Google Scholar: Lookup
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Summary
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The research article explores the effect of detergent-based decellularization procedures on cellular proteins and potential immune reactions in decellularized equine carotid artery grafts (dEAC). Despite these grafts showing low DNA content, implying successful decellularization, the study found that they still contain residual cellular proteins which may induce an immune response in the recipient.
Decellularization Procedures and their Impact on Cellular Proteins
- The researchers analyzed the dEAC for residual cellular proteins, despite the grafts showing a low DNA content (0.6 ± 0.15 ng DNA/mg dry weight), which represented only 0.33 ± 0.14% of the native tissue DNA content.
- A decellularization protocol based on detergent, which included endonuclease treatment, was used attempting total elimination of cellular components.
- When the researchers homogenized and extracted the matrices using a high concentration of detergent, their analysis revealed the presence of cytosolic and cytoskeleton proteins, GAPDH and smooth muscle actin.
- These proteins were only partially depleted (to 4.1 ± 1.9% and 13.8 ± 0.55% respectively), despite the rigorous decellularization protocol.
Potential Immunogenicity in the Decellularized Matrices
- The research also explored the potential immune response these grafts could trigger. MHC I complexes and the alpha-Gal epitop, both recognized to induce an immune response, were reduced to only 14.8 ± 1.2% and 15.1 ± 2.05% respectively.
- Through mass spectrometry, 306 proteins belonging to different cellular components such as the cytosol, organelles, nucleus, and cell membrane were detected in the matrix extracts.
- Further, when administered in mice, these aqueous matrix extracts provoked a significant antibody formation, suggesting a high immunogenic potential.
Implications of the Study
- The study implies that a decellularization protocol, known to result in acellular matrices evaluated by low DNA content, may reduce but not eliminate cellular components entirely. These residual cellular proteins may contribute to the graft’s potential to induce an immune response when transplanted.
- This underscores the need for more refined decellularization protocols to ensure the graft’s biocompatibility and the success of vascular replacement therapy.
Cite This Article
APA
Böer U, Lohrenz A, Klingenberg M, Pich A, Haverich A, Wilhelmi M.
(2011).
The effect of detergent-based decellularization procedures on cellular proteins and immunogenicity in equine carotid artery grafts.
Biomaterials, 32(36), 9730-9737.
https://doi.org/10.1016/j.biomaterials.2011.09.015 Publication
Researcher Affiliations
- GMP Model Laboratory for Tissue Engineering, Feodor-Lynen-Str. 31, 30625 Hannover, Germany. boeer.ulrike@mh-hannover.de
MeSH Terms
- Actins / metabolism
- Animals
- Antibodies / blood
- Antibody Specificity / immunology
- Blood Vessel Prosthesis
- Carotid Arteries / cytology
- Carotid Arteries / drug effects
- Carotid Arteries / immunology
- Carotid Arteries / transplantation
- Cell Extracts
- Detergents / pharmacology
- Epitopes / immunology
- Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism
- Histocompatibility Antigens Class I / immunology
- Horses
- Immunization
- Mass Spectrometry
- Mice
- Muscle, Smooth / drug effects
- Muscle, Smooth / metabolism
- Proteins / metabolism
- Tissue Engineering / methods
- alpha-Galactosidase / immunology
Citations
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