Reendothelialization and maintenance of endothelial integrity in longitudinal denuded tracks in the thoracic aorta of rats

Abstract

Endothelial repair was studied during, and up to 26 weeks following reendothelialization of longitudinal tracks denuded of endothelium. Deendothelialized tracks were produced on the ventral aspect of the thoracic aortas of rats. A standardized denudation procedure was used in conjunction with continuous intravenous infusion of [ 3H]thymidine to obtain quantitative evaluations of replication, migration, and cell density. The method of denudation was relatively gentle, selective, and reproducible. Reendothelialization was completed in < 66 h. During reendothelialization, cell migration and replication proceeded simultaneously until confluence was reached. Following confluence, as migration ceased, cell density returned toward control levels. It continued to rise, associated with continued, though attenuated, nuclear incorporation of [ 3H]thymidine, until it reached 1 1 2 –2 1 2 times that of adjacent uninjured control endothelium. The boundaries, or margins, of the tracks were well demarcated as judged by the pattern of labeled cells within and unlabeled cells adjacent to reendothelialized tracks. Widths of deendothelialized and reendotheliazed tracks were similar. Thus, endothelial cells in uninjured regions surrounding denuded tracks were not observed to contribute to reendothelialization by proliferation. During reendothelialization, unlabeled cells were observed within reendothelializing tracks. Thus, some cells migrated relatively long distances (0.3 – 0.4 mm) before either replicating or sloughing from the luminal surface. Endothelial cell density within reendothelialized tracks remained elevated at 12 and 26 weeks following denudation. At 12 weeks there was little intermingling of cells inside tracks (labeled during reendothelialization) with unlabeled cells in adjacent uninjured areas. At 26 weeks, after a terminal 12-day continuous labeling period in previously unlabeled animals, replication of cells inside tracks was far less than that in adjacent uninjured endothelium. Thus, the newly regenerated cell population in tracks remained synchronously quiescent and physically segregated for long periods of time, despite normally occurring hemodynamic and remodeling factors which might be expected to favor continuous cell turnover and migration. Our findings relating to endothelial cell migration and replication during reendothelialization vary somewhat from those reported using other methods of deendothelialization.