Korean J Thorac Cardiovasc Surg.  2010 Dec;43(6):576-587.

Study on Effective Preservation of Bovine Pericardium Using Decellularization and alpha-galactosidase for Eliminating Xenoreactive Antigen

• Kim MS
• Park CJ
• Kim SH
• Lim HG
• Kim YJ

Abstract

BACKGROUND
Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with alpha-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. MATERIAL AND METHOD: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with alpha-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and alpha-galactose staining were carried out to each groups. RESULT: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups. EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups.
CONCLUSION
We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.

Keyword

Tissue engineering; Glutaraldehyde; Pericardium; Xenograft

MeSH Terms

alpha-Galactosidase
Compliance
DNA
Glutaral
Pericardium
Permeability
Physical Examination
Pronase
Sodium Dodecyl Sulfate
Sulfhydryl Compounds
Tensile Strength
Tissue Engineering
Tissue Preservation
Transplantation, Heterologous
DNA
Glutaral
Pronase
Sodium Dodecyl Sulfate
Sulfhydryl Compounds
alpha-Galactosidase