Calcium mitigation in bioprosthetic tissues by iron pretreatment: the challenge of iron leaching

Preliminary studies in our laboratory have shown that iron pretreatment of glutaraldehyde-preserved tissues inhibited calcification. The present study was designed to further investigate this finding and to optimize the technique. Porcine valve tissue and bovine pericardium preserved in glutaraldehyde and pretreated by iron at different concentrations and incubation times were implanted either subcutaneously in rats or as a mitral valve substitute in sheep. Compared with control groups, calcification was markedly inhibited in the groups pretreated with Fe3+. There was a direct correlation between the degree of calcification and the iron content within the tissue. A minimal iron content within the tissue of 0.5% for porcine valve tissue and 0.2% for pericardial tissue was found to be necessary for calcium mitigation. A progressive leaching of Fe3+ was observed that could be stabilized by using a higher concentration of iron in the solution and or by additional pretreatment in glutaraldehyde at high temperature, but only for subcutaneously implanted tissues. By contrast, leaching was faster and more complete in the iron-pretreated tissue valves placed in the circulating blood environment, therefore resulting in calcified valves. Pretreatment in glutaraldehyde at high temperature (50 degrees C) alone was able to mitigate calcification in both subcutaneous and circulating blood environments, a new finding that may lead to an improved method for glutaraldehyde preservation of tissues.