Study on Effective Preservation of Bovine Pericardium Using Decellularization and alpha-galactosidase for Eliminating Xenoreactive Antigen.
- Author:
Min Seok KIM
1
;
Cham Jin PARK
;
Soo Hwan KIM
;
Hong Gook LIM
;
Yong Jin KIM
Author Information
1. Department of Thoracic and Cardiovascular Surgery, Seoul National University Hospital, Seoul National University College of Medicine, Korea. kyj@plaza.snu.ac.kr
- Publication Type:Original Article
- Keywords:
Tissue engineering;
Glutaraldehyde;
Pericardium;
Xenograft
- MeSH:
alpha-Galactosidase;
Compliance;
DNA;
Glutaral;
Pericardium;
Permeability;
Physical Examination;
Pronase;
Sodium Dodecyl Sulfate;
Sulfhydryl Compounds;
Tensile Strength;
Tissue Engineering;
Tissue Preservation;
Transplantation, Heterologous
- From:The Korean Journal of Thoracic and Cardiovascular Surgery
2010;43(6):576-587
- CountryRepublic of Korea
- Language:Korean
-
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.
