Mitigated calcification of glutaraldehyde-fixed bovine pericardium by tannic acid in rats

Abstract

Bioprosthetic heart valves derived from glutaraldehyde (Glut)-fixed xenografts have been widely used to replace diseased cardiac valves. However, calcification and degeneration are common following their implantation. Inflammation is closely associated with calcification of Glut-fixed xenografts via macrophage infiltration. Tannic acid (TA) possesses anti-inflammatory effects. This study was designed to investigate the anti-calcification of TA treatment on the Glut-fixed bovine pericardium (BP) in a rat subdermal model. Fresh BP was divided into two groups (10 in each group) and separately subjected to different fixation procedures as follows: (1) Glut group: fixation with 0.6% Glut alone; (2) Glut/TA group: fixation with 0.6% Glut and subsequent 0.3% TA. Then the BP samples were subdermally implanted in juvenile male Sprague-Dawley rats and explanted 21 days after implantation. Each explanted BP sample was divided into three parts for calcium content analysis, von Kossa's staining and immunohistochemical staining, and reverse transcription-polymerase chain reaction study. The data from quantitative calcium analysis and von Kossa's staining showed that Glut-fixed BP developed significantly more calcification than Glut/TA-fixed BP ((90.3 +/- 32.5) mg/g dry weight vs (6.4 +/- 1.3) mg/g dry tissue, P < 0.01). Immunostaining demonstrated lower matrix metalloproteinase-9 and tenascin-C expression as well as macrophage infiltration into Glut/TA-fixed BP than in its Glut-fixed counterpart (P < 0.01 for all). Additionally, the reverse transcription-polymerase chain reaction study showed that higher levels of expression of matrix metalloproteinase-9 and tenascin-C mRNA occurred within Glut-fixed BP than within the Glut/TA-fixed ones (P < 0.01 for all). TA exerts excellent anti-calcification effects on Glut-fixed BP via inhibiting macrophage infiltration and expression of matrix metalloproteinase-9 and tenascin-C.