In vitro evaluation of vascular endothelial and smooth muscle cell survival and apoptosis in response to hypothermia and freezing

The success of endovascular techniques such as balloon angioplasty and stenting in the treatment of atherosclerotic vascular disease has been limited by an aggressive proliferative response leading to neointimal hyperplasia and re-stenosis. A new endovascular therapy combining cold treatment with balloon dilation has been proposed to prevent arterial restenosis. In order to evaluate the potential of this application, studies were conducted investigating the effects of hypothermia and freezing on human arteries at the cellular level. Cultured arterial endothelial cells and smooth muscle cells were chilled or frozen under controlled thermal conditions. The viability response of the cells was measured with a variety of assays quantifying necrosis, apoptosis, and cell proliferation. These data establish correlations between thermal conditions and the extent and nature of arterial freezing injury. Arterial smooth muscle cells were found to be susceptible to freeze-induced apoptosis in a temperature range of -5 to -15 degrees C. Endovascular cryotherapy designed to induce apoptosis in arterial smooth muscle cells may limit neointimal formation and thereby improve the durability of conventional angioplasty.