Vascular smooth muscle cell apoptosis induced by "supercooling" and rewarming.
OBJECTIVE
The underlying mechanisms for the reduction in restenosis caused by cryoplasty for peripheral atherosclerotic lesions are not well understood. Because vascular smooth muscle cells (SMCs) are known to play a critical role in restenosis and neointimal hyperplasia, the aim of this study was to determine SMC survival under conditions of "supercooling" and/or rewarming.


METHODS
Bovine aortic SMCs were supercooled to -10 degrees C for 0, 60, or 120 seconds with a custom-designed conduction cooling stage and then rewarmed to 37 degrees C in an incubator for 0, 12, or 24 hours. A terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was used to measure the degree of apoptosis. Activation of Akt (ie, protein kinase B), a key signal protein involved in cell survival, was assessed by Western blot analysis.


RESULTS
An increase in apoptotic SMCs was observed with increasing supercooling and rewarming time. Akt was significantly activated at only the most severe condition (120 seconds of supercooling and 24 hours of rewarming), which showed a 2.03-fold increase compared with the group without rewarming.


CONCLUSIONS
The data suggest that SMC apoptosis occurs with supercooling and rewarming. Protective cell survival mechanisms were activated only late in the rewarming phase. This may partially explain the long-term patency observed with cryoplasty of atherosclerotic peripheral lesions.
Yiu
Wai-ki
WK
Department of Vascular Surgery, Yale University School of Medicine, 333 Cedar Street, New Haven, Connecticut 06520-8062, USA, and Department of Vascular Surgery, Queen Mary Hospital, Hong Kong.
Cheng
Stephen W K
SW
Sumpio
Bauer E
BE
D000704:Analysis of Variance; D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology*; D017209:Apoptosis; D015153:Blotting, Western; D002417:Cattle; D003080:Cold Temperature; D020287:In Situ Nick-End Labeling; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D010766:Phosphorylation; D017694:Rewarming