Production and inhibition of the gelatinolytic matrix metalloproteinases in a human model of vein graft stenosis

Abstract

Objectives: human vein graft stenoses are caused by intimal hyperplasia, a process which is characterised by extensive degradation and accumulation of extracellular matrix. This study investigated the role of other matrix metalloproteinases (MMPs) - the principal physiological mediators of extracellular matrix degradation - in the development of intiminal hyperplasia in cultured long saphenous vein. Design: experimental study. Material and methods : paired venous segments with the endothelium intact or denuded were cultured in standard conditions for 14 days. At the termination of culture, MMPs were extracted from one half of the tissue, whist the remainder of the vein was prepared for histological examination. Results: stereologic analysis revealed that the endothelium intact veins developed a significantly thicker neointima when compared to the denuded venous segments (20 μm v. 0 μm, p = 0.006). Quantification of MMPs by substrate gel enzymography demonstrated that the development of a neointima was associated with increased production of the gelatinolytic MMP-9 (p = 0.03) in intact veins. Inmmunocytochemistry showed what the MMP-9 localised to the internal elastic lumina, which suggested a role in facilitating smooth-muscle-cell migration into the intima. The role of MMPs-2 and -9 in intimal hyperplasia was further investigated by culturing intact venous segments with a therapeutic concentration of doxycycline - a potent MMP inhibitor. These experiments demonstrated that a therapeutic dose of doxycycline significantly reduced neointimal thickness (control 21 μm, doxycycline 10 mg/l-5.5 μm), in conjunction with a significant reduction in the production of MMP-9. Conclusions: these data suggest that elevated levels of MMPs may play a significant role in the development of human intimal hyperplasia and that inhibition of these enzymes may offer a potential therapeutic strategy for the prevention of hyperplastic lesions.