Synthetic vascular prosthesis impregnated with mesenchymal stem cells overexpressing endothelial nitric oxide synthase
Synthetic vascular prosthesis impregnated with mesenchymal stem cells overexpressing endothelial nitric oxide synthase
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DOI：
10.1161/circulationaha.105.001586
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发表时间：
2006-07-04
期刊：
影响因子：
37.8
通讯作者：
Katsumata, Takahiro
中科院分区：
文献类型：
--
作者：
Kanki-Horimoto, Sachiko;Horimoto, Hitoshi;Katsumata, Takahiro
背景-内皮功能障碍是众所周知的冠状动脉疾病，有时会导致不可逆的心肌损伤。在这种情况下，需要重复冠状动脉血运重建术，包括冠状动脉旁路移植术，这可能导致移植物管道短缺。另一方面，内皮型一氧化氮合酶（eNOS）是心血管基因治疗的一个有吸引力的靶点。内表面覆盖有过表达eNOS的间充质干细胞（MSC）的血管假体预计将提供可行的NO效应和MSC对天然冠状动脉床的血管生成效应，以及改善自身通畅性。在此，我们尝试开发产生生物活性蛋白的小口径血管假体。方法和结果-从大鼠骨髓中分离出MSC，并用携带大鼠eNOS cDNA和β-半乳糖苷酶（β-gal）的每种腺病毒转导（eNOS/MSC和β-gal/MSC）。将β-gal/MSCs复合于人工血管中，用5-溴-4-氯-3-吲哚-β-D-半乳糖苷染色法检测人工血管内表面β-gal的表达。通过监测H-3-精氨酸向H-3-瓜氨酸的转化来测定eNOS/MSC的NOS活性。血管假体的内表面覆盖有表达β-gal的MSC。H-3-瓜氨酸的量增加，并且确定eNOS/MSC产生eNOS的酶活性。这种活性被完全抑制由N-G-硝基-L-精氨酸methylester. Conclusions的内表面的膨胀聚四氟乙烯血管假体接种lacZ基因转导的MSC表现出重组蛋白。eNOS/MSC种植的人造血管的发展将保证更长的移植物通畅性和血管保护作用。
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 H-3-arginine to H-3-citrulline. The inner surfaces of the vascular prostheses were covered with MSCs expressing beta-gal. The amount of the H-3-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.