Pulsatile ex vivo perfusion of human saphenous vein grafts under controlled pressure conditions increases MMP-2 expression

We have developed an ex vivo perfusion system, which uses standardized and strictly controlled hemodynamic parameters for the pulsatile and non-static perfusion of HSVGs to guarantee a reliable analysis of molecular parameters under different pressure conditions. Cell viability of HSVGs (n = 12) was determined by the metabolic conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) into a purple formazan dye.Under physiological flow rates (10 mmHg) HSVGs remained viable for two weeks. Their exposure to arterial conditions (100 mmHg) was possible for one week without important reduction in viability. Baseline expression of matrix metalloproteinase-2 (MMP-2) after venous perfusion (2.2 ± 0.5, n = 5) was strongly up-regulated after exposure to arterial conditions for three days (19.8 ± 4.3) or five days (23.9 ± 6.1, p < 0.05). Zymographic analyses confirmed this increase on the protein level. Our results suggest that expression and activity of MMP-2 are strongly increased after exposure of HSVGs to arterial hemodynamic conditions compared to physiological conditions.Therefore, our system might be helpful to more precisely understand the molecular mechanisms leading to an early failure of HSVGs.Coronary artery bypass grafting (CABG) using venous grafts is a standard procedure in the treatment of advanced coronary artery disease. However, vein graft occlusion implanted in an arterial pressure environment is still a major problem [1]. Approximately 15 to 20% of human saphenous vein grafts (HSVGs) occlude within one month and 25% within the first year. Ten years after CABG about 50% of the HSVGs are occluded and 25% have been severely stenosed [2-6]. Early changes in vein grafts include endothelial disruption leaving the graft vulnerable to thrombotic incidents and smooth muscle cell (SMC) migration and proliferation from the media into the intima within the first week after grafting [1,7]. The vein graft intimal thickening and remodeling occurs a