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Stepwise solubilization-based antigen removal for xenogeneic scaffold generation in tissue engineering.

Maelene L Wong1, Janelle L Wong, Kyriacos A Athanasiou

  • 1Department of Veterinary Medicine: Medicine and Epidemiology, University of California, Davis, Davis, CA 95616, USA.

Acta Biomaterialia
|January 17, 2013
PubMed
Summary

A new two-step antigen removal (AR) process effectively reduces both hydrophilic and lipophilic antigens in bovine pericardium scaffolds. This method enhances xenogeneic tissue suitability for heart valve engineering by minimizing immune rejection risks.

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Immunology

Background:

  • Residual antigens in decellularized xenogeneic scaffolds can trigger immune responses.
  • Current antigen removal (AR) methods primarily target hydrophilic proteins, leaving lipophilic antigens as a challenge.
  • Bovine pericardium (BP) is a common xenogeneic material requiring improved AR for tissue engineering.

Purpose of the Study:

  • To develop and evaluate a sequential, two-step AR strategy for bovine pericardium (BP).
  • To investigate methods for solubilizing lipophilic proteins as a second AR step.
  • To assess the impact of the AR process on antigenicity, xenoantigen removal, and material properties.

Main Methods:

  • A sequential AR strategy was employed: first, hydrophilic AR using dithiothreitol and potassium chloride, followed by lipophilic AR using amidosulfobetaine-14 (ASB-14).

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  • Bovine pericardium after AR (BP-AR) was analyzed for residual hydrophilic and lipophilic antigenicity.
  • Removal of key xenoantigens (Galactose-α(1,3)-galactose and MHC I) and assessment of tensile properties and ECM structure were performed.

    • Main Results:

    • The sequential AR strategy significantly reduced both hydrophilic and lipophilic antigenicity compared to single-step methods or SDS decellularization.
    • The 1% ASB-14 lipophilic AR step effectively eliminated galactose-α(1,3)-galactose and major histocompatibility complex I (MHC I).
    • The process preserved the tensile properties and extracellular matrix structure of the bovine pericardium.

    Conclusions:

    • A sequential, differential protein solubilization approach is crucial for comprehensive antigen removal in xenogeneic scaffolds.
    • This enhanced AR method effectively reduces key barriers to xenotransplantation in bovine pericardium.
    • The developed AR process yields a biomaterial suitable for heart valve tissue engineering with reduced immunogenicity.