Production and inhibition of the gelatinolytic matrix metalloproteinases in a human model of vein graft stenosis.
OBJECTIVE
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 the matrix metalloproteinases (MMPs) - the principal physiological mediators of extracellular matrix degradation - in the development of intimal hyperplasia in cultured human long saphenous vein.


METHODS
experimental study.


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, whilst 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 micron v. 0 micron, 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. Immunocytochemistry showed that 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 micron, doxycycline 10 mg/l-5.5 micron), 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.
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D017364:Collagenases / Q000032:analysis / Q000096:biosynthesis*; D004318:Doxycycline / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology; D005780:Gelatin / Q000493:pharmacokinetics*; D018093:Gelatinases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D006083:Graft Occlusion, Vascular / Q000201:enzymology*; D006801:Humans; D006965:Hyperplasia / Q000201:enzymology; D007150:Immunohistochemistry; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D009924:Organ Culture Techniques / Q000379:methods; D012501:Saphenous Vein / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D018709:Statistics, Nonparametric; D017539:Tunica Intima / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology