Background-Endothelial dysfunction is known to exaggerate coronary artery disease, sometimes leading to irreversible myocardial damage. In such cases, repetitive coronary revascularization including coronary artery bypass grafting is needed, which may cause a shortage of graft conduits. On the other hand, endothelial nitric oxide synthase (eNOS) is an attractive target of cardiovascular gene therapy. The vascular prostheses, of which the inner surfaces are covered with mesenchymal stem cells (MSCs) overexpressing eNOS, are expected to offer feasible effects of NO and angiogenic effects of MSCs on the native coronary arterial beds, as well as improvement of self-patency. Herein, we attempted to develop small caliber vascular prostheses generating the bioactive proteins. Also, we attempted to transduce eNOS cDNA into MSCs. Methods and Results-The MSCs were isolated from rat bone marrow and transduced with each adenovirus harboring rat eNOS cDNA and -galactosidase (-gal) (eNOS/MSCs and -gal/MSCs). The -gal/MSCs were impregnated into vascular prostheses, then the expressions of -gal on the inner surfaces of them were evaluated by 5-bromo-4-chloro-3-indolyl -D-galactoside staining. The NOS activity of eNOS/MSCs was assayed by monitoring the conversion of 3 H-arginine to 3 H-citrulline. The inner surfaces of the vascular prostheses were covered with MSCs expressing -gal. The amount of the 3 H-citrulline increased, and eNOS/MSCs were determined to generate enzymatic activity of eNOS. This activity was completely inhibited by N G -nitro-L-arginine methyl ester. Key Words: mesenchymal stem cells Ⅲ endothelial nitric oxide synthase Ⅲ gene-transduction Ⅲ adenovirus Ⅲ small caliber expanded polytetrafluoroethylene (ePTFE) vascular prosthesis I mpaired endothelial function induces several cardiovascular diseases, including atherosclerosis, hypertension, heart failure, arteriosclerosis obliterans, and coronary heart disease. Endothelial dysfunction also causes NO insufficiency, resulting in the limitation of NO-mediated signal transduction and excretion of bioactive hormone-like products. 1 The attenuated production of NO and the hormone-like products exaggerates these cardiovascular diseases further. Coronary artery disease is one of the most critical diseases enhanced by endothelial dysfunction, sometimes leading to irreversible myocardial damage. Some patients require repetitive coronary revascularization, including coronary artery bypass grafting, which may lead to a shortage of graft materials.
Conclusions-TheSmall caliber vascular prostheses, which would be suitable for graft conduits for coronary artery bypass grafting or arteriosclerosis obliterans below the knee, have an extremely high failure rate that is attributed to thrombus formation and occlusion. Many attempts have been made to increase the patency of small-caliber vascular prostheses over the years. Recently, vascular grafts releasing NO infer possibilities preventing thrombosis and stenosis. 2,3 However, they are far from practical use, because the ma...