High-concentration glutaraldehyde fixation of bovine pericardium in organic solvent and post-fixation glycine treatment: in vitro material assessment and in vivo anticalcification effect.
OBJECTIVE
Glutaraldehdye (GA)-fixed xenografts are widely used in cardiovascular surgery. The objective of this study was to evaluate the anticalcification effect of glycine treatment and high-concentration GA fixation in organic solvent on GA-fixed bovine pericardium, and to evaluate the possible synergistic effect of combined treatment.


METHODS
Bovine pericardial tissues were divided into four groups according to the methods of treatment. Group 1 consisted of tissues fixed with 0.5% GA (control), group 2 fixed with 0.5% GA and post-treated with glycine, group 3 fixed with 2% GA in organic solvent (65% ethanol+5% octanol), and group 4 fixed with 2% GA in organic solvent and post-treated with glycine. The material characteristics of the treated tissues were assessed by amino acid analysis, thermal stability test, uniaxial mechanical test and light microscopy. The tissues were subcutaneously implanted into 4-week-old rats for 8 weeks, and the calcium contents of the explanted tissues were measured.


RESULTS
Differently treated tissues resulted in no significant alterations in material characteristics and morphology as assessed by amino acid analysis, thermal stability test, uniaxial mechanical test, and light microscopy. Median calcium contents of groups 1, 2, 3, and 4 were 80.5 μg mg(-1), 1.0 μg mg(-1), 0.5 μg mg(-1) and 1.7 μg mg(-1), respectively. The calcium contents of groups 2, 3 and 4 were all significantly lower than that of group 1 (p < 0.05).


CONCLUSIONS
Post-fixation treatment with glycine, high-concentration GA fixation in organic solvent and combined treatment of these all strongly prevented calcification of GA-fixed bovine pericardium in rat subcutaneous implantation model.
Lee
Cheul
C
Department of Thoracic and Cardiovascular Surgery, Cardiovascular Center, Sejong General Hospital, Bucheon, Republic of Korea.
Kim
Soo Hwan
SH
Choi
Seung-Hwa
SH
Kim
Yong Jin
YJ
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001705:Bioprosthesis; D002114:Calcinosis / Q000517:prevention & control*; D002118:Calcium / Q000032:analysis; D002417:Cattle; D003094:Collagen / Q000032:analysis; D004357:Drug Synergism; D005260:Female; D005404:Fixatives / Q000494:pharmacology*; D005976:Glutaral / Q000494:pharmacology*; D005998:Glycine / Q000494:pharmacology; D006358:Hot Temperature; D010496:Pericardium / Q000737:chemistry / Q000187:drug effects* / Q000637:transplantation* / Q000648:ultrastructure; D011183:Postoperative Complications / Q000517:prevention & control; D051381:Rats; D017207:Rats, Sprague-Dawley; D012997:Solvents / Q000494:pharmacology; D013718:Tensile Strength; D016707:Tissue Fixation / Q000379:methods; D014183:Transplantation, Heterologous
D000596:Amino Acids; D005404:Fixatives; D012997:Solvents; D003094:Collagen; D002118:Calcium; D005976:Glutaral; D005998:Glycine