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Updated: May 12, 2026
Generation and Grafting of Tissue-engineered Vessels in a Mouse Model
Published on: March 18, 2015
Synthetic vascular prosthesis impregnated with mesenchymal stem cells overexpressing endothelial nitric oxide
Sachiko Kanki-Horimoto1, Hitoshi Horimoto, Shigetoshi Mieno
1Department of Thoracic and Cardiovascular Surgery, Osaka Medical College, 2-7 Daigakumachi Takatsuki, Osaka, 569-8686, Japan. tho064@poh.osaka-med.ac.jp
Insights
Engineered vascular grafts seeded with mesenchymal stem cells (MSCs) overexpressing endothelial nitric oxide synthase (eNOS) show promise for improving graft patency and protecting blood vessels.
Area of Science:
- Biomedical Engineering
- Cardiovascular Research
- Regenerative Medicine
Background:
- Endothelial dysfunction exacerbates coronary artery disease, often necessitating repeat bypass surgeries and leading to graft conduit shortages.
- Endothelial nitric oxide synthase (eNOS) is a key target for cardiovascular gene therapy.
- Small-caliber vascular prostheses are needed to address graft conduit limitations.
Purpose of the Study:
- To develop small-caliber vascular prostheses seeded with mesenchymal stem cells (MSCs) engineered to overexpress eNOS.
- To evaluate the feasibility of transducing MSCs with eNOS cDNA for enhanced therapeutic effects.
Main Methods:
- Mesenchymal stem cells (MSCs) were isolated and transduced with adenoviruses carrying rat eNOS cDNA or beta-galactosidase (beta-gal).
- Vascular prostheses were seeded with beta-gal/MSCs to confirm inner surface coverage and protein expression.
- Nitric oxide synthase (NOS) activity of eNOS/MSCs was measured by monitoring the conversion of 3H-arginine to 3H-citrulline.
Main Results:
- Vascular prostheses seeded with MSCs successfully expressed beta-gal on their inner surfaces.
- eNOS/MSCs demonstrated significant NOS enzymatic activity, confirmed by increased 3H-citrulline production.
- This eNOS activity was effectively inhibited by N(G)-nitro-L-arginine methyl ester, validating the results.
Conclusions:
- Gene-transduced MSCs seeded on vascular prostheses can generate bioactive proteins.
- The development of eNOS-MSC-seeded vascular prostheses holds promise for improved graft patency.
- These engineered grafts may offer significant vasculoprotective effects for patients with coronary artery disease.
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 beta-galactosidase (beta-gal) (eNOS/MSCs and beta-gal/MSCs). The beta-gal/MSCs were impregnated into vascular prostheses, then the expressions of beta-gal on the inner surfaces of them were evaluated by 5-bromo-4-chloro-3-indolyl beta-D-galactoside staining. The NOS activity of eNOS/MSCs was assayed by monitoring the conversion of 3H-arginine to 3H-citrulline. The inner surfaces of the vascular prostheses were covered with MSCs expressing beta-gal. The amount of the 3H-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.
Conclusions:
The inner surface of expanded polytetrafluoroethylene vascular prostheses seeded with lacZ gene-transduced MSCs exhibited recombinant proteins. Development of eNOS/MSC-seeded vascular prostheses would promise much longer graft patency and vasculoprotective effects.
