Department of Pharmaceutical Sciences, Northeastern University, Boston, MA 02115, USA.
Biomed Eng Online. 2010 Sep 27;9:56. doi: 10.1186/1475-925X-9-56.
In this study, we have examined local non-viral gene delivery, transfection, and therapeutic efficacy of endothelial nitric oxide synthase (eNOS) encoding plasmid DNA administered using coated stents in a rabbit iliac artery restenosis model.
Lipopolyplexes (LPPs) with eNOS expressing plasmid DNA were immobilized on stainless steel stents using poly(D,L-lactide-co-glycolide) (PLGA) and type B gelatin coatings. The gene-eluting stents were implanted bilaterally in the denuded iliac arteries and eNOS transfection and therapeutic efficacy were examined 14 days after implantation.
The results show that non-viral lipopolyplex-coated stents can efficiently tranfect eNOS locally in the arterial lumen assessed by PCR and ELISA. Human eNOS ELISA levels were significantly raised 24 hours after transfection compared to controls (125 pg eNOS compared to <50 pg for all controls including naked DNA). Local eNOS production suppressed smooth muscle cell proliferation and promoted re-endothelialization of the artery showing a significant reduction in restenosis of 1.75 neointima/media ratio for stents with lipoplexes encoding eNOS compared with 2.3 neointima/media ratio for stents with lipoplexes encosing an empty vector.
These results support the hypothesis that a potent non-viral gene vector encoding for eNOS coated onto a stent can inhibit restenosis through inhibition of smooth muscle cell growth and promotion of a healthy endothelium.
在这项研究中，我们研究了局部非病毒基因传递、转染和内皮型一氧化氮合酶（eNOS）编码质粒 DNA 的治疗效果，该质粒 DNA 采用涂覆支架在兔髂动脉再狭窄模型中给药。
使用聚（D，L-丙交酯-共-乙交酯）（PLGA）和 B 型明胶涂层将含有 eNOS 表达质粒 DNA 的脂质体（LPPs）固定在不锈钢支架上。将基因洗脱支架双侧植入裸露的髂动脉中，并在植入后 14 天检查 eNOS 转染和治疗效果。
结果表明，非病毒脂质体涂覆支架可以通过 PCR 和 ELISA 有效地在动脉管腔中局部转染 eNOS。与对照组相比（所有对照组的人 eNOS ELISA 水平均<50pg，包括裸 DNA），转染后 24 小时 eNOS ELISA 水平显著升高。局部 eNOS 产生抑制平滑肌细胞增殖并促进动脉再内皮化，与包载空载体脂质体的支架相比，包载编码 eNOS 的脂质体的支架的新生内膜/中膜比值显著降低（1.75 比 2.3）。
这些结果支持这样一种假设，即涂覆在支架上的强效非病毒基因载体编码的 eNOS 可以通过抑制平滑肌细胞生长和促进健康的内皮来抑制再狭窄。
