THE ROLE OF THE ADVENTITIA IN THE ARTERIAL RESPONSE TO ANGIOPLASTY: THE EFFECT OF INTRAVASCULAR RADIATION

Abstract Purpose: In the current series of experiments we have characterized cell proliferation leading to vascular lesion formation in a porcine model for post-angioplasty restenosis adn examined the mechanism of action intravascular beta irradiation in the prevention of lesion formation in this model. Methods and Materials: Juvenile male pigs were subjected to balloon overstretch injury of the left anterior descending and circumflex coronary arteries using clinical angioplasty catheters. Proliferating cells were labelled by injections of 50 mg/kg of bromo-deoxyuridine (BrDU) 24, 16 and 8 hrs prior to sacrifice adn were detected by immunohistochemistry using a specific antibody to BrDU. In some cases. BrDI was given as a pulse 3 days after angioplasty and the animals sacrificed on day 14 to follow the migration of the cells which had proliferated earlier. Characterization of the proliferating cells was performed by immunohistochemistry using antibodies to specific cytoskeletal proteins specific for smooth muscle cells and myofibroblasts. Some vessels were treated at the time of angioplasty with 14 or 28 Gy (to a depth of 2 mm) intravascular irradiation using a flexible catheter with a pure beta emitter 90 SR/Y and the effect on cell proliferation and terminal transferase-mediated UTP nick-end labelling (TUNEL) examined 3 or 7 days later. Results: The first major site of cell proliferation between 2–3 days after angioplasty is the adventitia and not the medial wall. Seven days after angiosplasty cell proliferation is predominant is predominant in the neontima and is reduced in the media and adventitia. Differential staining with antibodies directed against smooth muscle alpha actin and other cytoskeletal proteins indicates that the proliferating adventitial cells are myofibroblasts. Pulse label studies with BrDU indicates that the proliferating adventitial myofibroblasts migrate into the neointima and contribute to the mass of the restenosis lesion. Fourteen days after angioplasty the myofibroblasts in the neointima and the adventitia express alpha smooth muscle actin and form a fibrotic scar in the adventitia surrounding the injury site. Endovascular irradiation appears to inhibit development of the restenosis lesion by significantly reducing cell proliferation in the media and adventitia at early time points after injury. There were no significant differences in the percent of TUNEL labelled cells in the irradiated vessels compared to controls. Alpha actin staining of myofibroblasts in the adventitia was reduced in the irradiated vessels suggesting a positive effect of intravascular irradiation on vascular remodeling. Conclusions: These studies have shown that adventitial myofibroblasts contribute to the problem of post-angioplasty restenosis by proliferating, forming a fibrotic scar surrounding the injury site, and migrating into the neointima. We hypothesize that the adventitial fibrosis which develops at the injury site contributes to negative vascular remodeling associated with clinical restenosis. Experiments in which vessels were exposed to intravascular irradiation at the time of angiosplasty indicate that this treatment reduces post-angioplasty restenosis by inhibiting early cell proliferation in the media and adventitia and by preventing the fibrotic changes in the adventitia without a corresponding increase in cellular death or apoptosis in these tissues.