In the swine abdominal aorta, most atherosclerotic lesions arise from naturally occuring collections of intimal cells called intimal cell masses. The main objective of this study was to determine if the tritiated thymidine labeling index of endothelial cells lying over intimal cell masses was greater than that of endothelial cells not over intimal cell masses. A higher endothelial cell labeling index over intimal cell masses would indicate possibly a greater turnover in the area. Such a finding would suggest possible transient but repeated breaks in the endothelial cell barrier that might contribute to the initiation of the atherosclerotic lesion in intimal cell masses.

This study shows that the endothelial cell turnover over the early porcine atherosclerotic lesion is more or less uniform in distribution. This is contrary to the findings in White Carneau pigeons where the increased endothelial cell turnover has a zonal pattern, being much greater at the lesion edge. The difference in the endothelial cell pattern of turnover in the two lesions probably is a reflection of their different manner of development. The porcine abdominal aortic lesion is derived from naturally occurring preexisting collections of intimal smooth-muscle cells called intimal cell masses. After 90 days of a hyperlipidemic diet these lesions are composed predominantly of smooth-muscle cells, with very few monocytes. The White Carneau pigeon lesion develops from circulating monocytes that have been shown to adhere to the edge of the lesion.

Intravenously (IV) injected Evans blue dye consistently accumulates at particular areas in aortas of normal swine. Theses areas are also where IV injected tracer cholesterol accumulates. We studied the ultrastructural characteristics of blue vs nonblue areas in the abdominal aortas of both control and atherosclerotic swine. In blue areas, the basement membrane was largely missing beneath the endothelial cells in both normal swine and swine with raised atherosclerotic lesions; the basement membrane that was present was often discontinuous. If the basement membrane helps regulate the transport of Evans blue dye across the arterial wall, its absence may play a part in the accumulation of the dye. In raised atherosclerotic lesions, those portions that accumulated Evans blue dye had a larger amount of amorphous extracellular matrix than did nonblue areas. No consistent difference between blue and nonblue areas were seen in the endothelial cell junctions of abdominal aortas in either control or atherosclerotic swine. Similarly, no differences were demonstrable in the number of Golgi complexes, mitochondria, or profiles of smooth or rough endoplasmic reticulum per cell.

Young male Yorkshire swine underwent denudation of the abdominal aorta by transfemoral intraarterial ballooning followed by the intravenous injection of Evans blue dye at intervals up to 28 days after ballooning. A small number of swine were also sacrificed 30 min after ballooning. Approximately half the swine were fed a hypercholesterolemic diet, while the others received a mash diet. Quantitation by light microscopy showed that ballooning had removed 99% of the endothelial cells and approximately 50% of the native intimal smooth muscle cells. Three hours after ballooning, the entire luminal surface of the abdominal aorta was stained with Evans blue. By 3 days, a continuous cellular lining had been reformed over at least one-fifth of the denuded surface and the area stained with Evans blue had sharply decreased. The number of intimal smooth muscle cells had returned to nearly normal values, and continued to increase steadily from that day onward. By 8 days, the continuous lining cells had further increased in number and most could be identified as endothelial cells by electron microscopy. Proliferation of the intimal smooth muscle cells and endothelial cells appeared independent, and not influenced by each other. From 8 to 28 days, the percentage of reendothelialized aortic surface increased and Evans blue staining decreased so that by 28 days, values for both were close to those in nonballooned controls. The rate of return of endothelial cells with consequent decrease in Evans blue staining did not appear to be influenced by the hypercholesterolemic diet in any consistent manner. However, at Day 28, the number of intimal smooth muscle cells was approximately four times greater in the hypercholesterolemic group than in the mash-fed and ballooned group. Thirty minutes postballooning, layers of platelets covering most of the surface were adherent to the denuded abdominal aorta. By 3 hr, they were found only as scattered clumps and became even scarcer at later times. If adherent platelets were a factor in the intimal smooth muscle cell proliferation, they appear to have been operative only in the immediate postballooing period.

A two-part study concerned the accumulation of intravenously injected Evans blue dye in the abdominal aorta of swine. In normolipidemic swine weighing up to 90 kg each, Evans blue accumulated predominantly in areas of the aorta that were the site of intimal smooth muscle cell collections. The second part of the study dealt with Evans blue accumulation in swine with advanced atherosclerosis. The extent of blue staining in these animals was not significantly different than in normolipidemic, nonatherosclerotic swine of the same size. The dye showed a predilection for raised atherosclerotic lesions, but only one quarter of the surface of these lesions was stained. Histologic studies of the uppermost portion of blue and nonblue lesion areas showed that the dye preferentially accumulated in regions with lower concentrations of foam cells. Injected isotopic cholesterol tended to accumulate in the blue rather than the nonblue areas.