Successful endothelialization of porcine glutaraldehyde-fixed aortic valves in a heterotopic sheep model

The purpose of our study was to evaluate the stability of an artificially seeded endothelial cell layer on porcine aortic prostheses under in vivo conditions in the arterial system.

Ten female sheep were divided into two groups. Animals of the study group (n = 7) had dissection of their right external jugular vein for cell harvesting. Myofibroblasts and endothelial cells were labelled with PKH-26, seeded onto pretreated (10% citric acid) porcine glutaraldehyde-fixed aortic valves (Freestyle, Medtronic Inc, Duesseldorf, Germany), and the valves were implanted into the descending aorta. Controls (n = 3) received pretreated but unseeded valves. A shunt between the aortic arch and the left atrial appendage ensured systolic or diastolic leaflet motions, or both, that were documented by sonography. After 3 months the valves were explanted. Specimens for scanning electron microscopy and immunohistochemical staining were taken prior to implantation and after explantation.

A neointimal proliferation was detected in the control group. No endothelial cells were found on the leaflets and the sinuses, but erythrocytes and thrombocytes were seen entrapped within the collagen fibers. Thrombus formation was documented macroscopically and histologically on the leaflets and the sinuses. In the study group a confluent endothelial cell layer was documented on the walls and leaflets. Neither neointimal proliferation nor any clots were seen. Some cells were still labelled positively indicating their origin from the initial cell seeding. No dilatation of any prosthesis was observed, but all valves showed slight thickening of the leaflets.

The artificially seeded endothelial cell layers remained stable under in vivo conditions in the arterial system. Biocompatibility of the prostheses seemed to be improved by reduction of thrombogenicity.