Currently, one of our laboratories is exploring additional assays to assess monocyte binding under conditions that mimic flow types found in healthy and diseased arteries ( Cockcroft et al., 2009), which in essence re-creates vascular physiology in an in vitro system. Development of the first grossly visible atherosclerotic lesion (fatty streak lesion) is characterised by the presence of macrophage foam cells. The initial recruitment of blood monocytes, their differentiation to macrophages and their subsequent progression to foam cells is well described (Ross, 1999). The presence

of free radicals in cigarette smoke is thought to contribute to the modification of lipids and lipoproteins, which results in their increased recognition and uptake by macrophages. Studies by Yokode et al., 1988 and Yokode et al., 1994 have shown that low density lipoprotein (LDL) exposure to an aqueous extract ABT-888 solubility dmso of cigarette smoke significantly increases lipid droplets (as measured with oil red O staining) and the concentration of cholesteryl ester in cultured macrophages. The lipoprotein particles were shown to be extensively modified while control selleck inhibitor particles were protected

by superoxide dismutase; however, thiobarbituric acid-reactive substances (TBARS) were similar with and without exposure to aqueous extracts of cigarette smoke. Expanding on these early studies is necessary to understand the role of reactive oxygen species on lipoprotein modification resulting in lipoprotein uptake by macrophages. Furthermore, this type of in vitro assay is anticipated to have the capacity to differentiate between tobacco extracts prepared from traditional cigarettes, PREPs and smokeless tobacco products. Regardless of the actual model being used, one potential criticism of in

vitro cardiovascular disease cell culture models is the nature of their culture under static conditions. In vivo, endothelial cells are exposed to haemodynamic stress imparted on the vessel wall by the flowing blood. This stress is not a phenomenon found equally at all points in the vascular tree and both ex vivo and in vivo studies have provided support for the association of increased haemodynamic stress found STK38 at branch points and curvatures in arteries with increased susceptibility to atherosclerotic lesion formation at these sites ( Cockcroft et al., 2009). Ideally, in vitro models would allow for the exposure of endothelial cells under flow conditions which would mimic those found in vivo. Such models are technically difficult to develop. Although a number of systems allow for the culture of endothelial cells under flow conditions there are drawbacks to such an approach. One drawback is the large volumes of media required for peristaltic flow pumps which precludes the measurement of inflammatory factors. Another is the potential for artefactual mechanical activation of cardiovascular cells in these systems ( Cockcroft et al., 2009).