A Glutaraldehyde-Free Crosslinking Method for the Treatment of Collagen-Based Biomaterials for Clinical Application
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Published:2023-10-25
Issue:11
Volume:10
Page:1247
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ISSN:2306-5354
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Container-title:Bioengineering
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language:en
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Short-container-title:Bioengineering
Author:
Steitz Marvin123ORCID, Zouhair Sabra2ORCID, Khan Mahamuda Badhon2ORCID, Breitenstein-Attach Alexander123ORCID, Fritsch Katharina4ORCID, Tuladhar Sugat Ratna5ORCID, Wulsten Dag6ORCID, Wolkers Willem-Frederik5ORCID, Sun Xiaolin13ORCID, Hao Yimeng2, Emeis Jasper2, Lange Hans-E.2ORCID, Berger Felix123ORCID, Schmitt Boris123ORCID
Affiliation:
1. Department of Pediatric Cardiology and Congenital Heart Disease, German Heart Center Berlin (Charité), D-13353 Berlin, Germany 2. Department of Pediatric Cardiology and Congenital Heart Disease, Charité University Medicine Berlin, D-13353 Berlin, Germany 3. German Centre for Cardiovascular Research, D-10785 Berlin, Germany 4. Department Dynamics and Transport in Quantum Materials, Helmholtz-Zentrum Berlin für Materialien und Energie GmbH, D-14109 Berlin, Germany 5. Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hanover, D-30625 Hannover, Germany 6. Julius Wolff Institute—Center for Musculoskeletal Biomechanics and Regeneration, D-13353 Berlin, Germany
Abstract
Biological bioprostheses such as grafts, patches, and heart valves are often derived from biological tissue like the pericardium. These bioprostheses can be of xenogenic, allogeneic, or autologous origin. Irrespective of their origin, all types are pre-treated via crosslinking to render the tissue non-antigenic and mechanically strong or to minimize degradation. The most widely used crosslinking agent is glutaraldehyde. However, glutaraldehyde-treated tissue is prone to calcification, inflammatory degradation, and mechanical injury, and it is incapable of matrix regeneration, leading to structural degeneration over time. In this work, we are investigating an alternative crosslinking method for an intraoperative application. The treated tissue‘s crosslinking degree was evaluated by differential scanning calorimetry. To confirm the findings, a collagenase assay was conducted. Uniaxial tensile testing was used to assess the tissue’s mechanical properties. To support the findings, the treated tissue was visualized using two-photon microscopy. Additionally, fourier transform infrared spectroscopy was performed to study the overall protein secondary structure. Finally, a crosslinking procedure was identified for intraoperative processing. The samples showed a significant increase in thermal and enzymatic stability after treatment compared to the control, with a difference of up to 22.2 °C and 100%, respectively. Also, the tissue showed similar biomechanics to glutaraldehyde-treated tissue, showing greater extensibility, a higher failure strain, and a lower ultimate tensile strength than the control. The significant difference in the structure band ratio after treatment is proof of the introduction of additional crosslinks compared to the untreated control with regard to differences in the amide-I region. The microscopic images support these findings, showing an alteration of the fiber orientation after treatment. For collagen-based biomaterials, such as pericardial tissue, the novel phenolic crosslinking agent proved to be an equivalent alternative to glutaraldehyde regarding tissue characteristics. Although long-term studies must be performed to investigate superiority in terms of longevity and calcification, our novel crosslinking agent can be applied in concentrations of 1.5% or 2.0% for the treatment of biomaterials.
Funder
EXIST—Forschungstransfer Federal Ministry of Education and Research—Kleine Patienten, großer Bedarf—Medizintechnische Lösungen für eine kindgerechte Gesundheitsversorgung
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