Improved spontaneous endothelialization by postfixation treatment of bovine pericardium

The longstanding release of locally cytotoxic glutaraldehyde concentrations from fixed biological materials is one reason for the lack of spontaneous endothelialization in vivo and in vitro of biological grafts (and valves). Preliminary studies have shown that bovine pericardium postfixation-treated with aminoacids may be endothelialized in vitro. To test whether such treatment improves spontaneous endothelialization in vivo 8 cm long segments grafts (6 mm I.D.) were interposed into the carotid arteries of 10 sheep. Ten grafts were sewn from postfixation treated pericardium (PTP), 4 from standard glutaraldehyde fixed pericardium (GAP) and 6 polytetrafluoroethylane (PTFE) grafts were implanted to study the degree of spontaneous endothelialization. In two pregnant animals, all grafts occluded (2 PTP, 1 GAP, 1 PTFE). In the remaining animals 1 additional PTFE graft occluded and 2 PTFE and 1 GA grafts showed significant thrombotic obstruction. The patent grafts were harvested at 3 months and the surface covered with red thrombus was determined by planimetry. PTP grafts had significantly less (12.2% +/- 15%) thrombotic appositions than glutaraldehyde treated grafts (49% +/- 20%; P = 0.01) and PTFE grafts (40.5% +/- 13%; P = 0.01). In the central areas of the PTP grafts, endothelial cells spread directly on the collagenous matrix and produced a basal lamina. In GAP, endothelial cells spread on amorphous material or macrophages and in PTFE grafts, only a neointima composed of myofibroblasts was endothelialized. Neutralization of glutaraldehyde concentrations by postfixation with aminoacids improves spontaneous endothelialization in vivo in biological materials.