目的:评估内皮型一氧化氮合酶(Endothelial nitric oxide synthase,eNOS)基因治疗对大鼠缺血后肢血管新生的影响。方法:局麻下将30只雄性SD大鼠后肢缺血模型制作后,随机分成实验组和对照组,每组15只。模型制作后1周,采用肌肉注射的方法...目的:评估内皮型一氧化氮合酶(Endothelial nitric oxide synthase,eNOS)基因治疗对大鼠缺血后肢血管新生的影响。方法:局麻下将30只雄性SD大鼠后肢缺血模型制作后,随机分成实验组和对照组,每组15只。模型制作后1周,采用肌肉注射的方法,实验组缺血后肢接受载有eNOS基因的5型重组腺病毒治疗,对照组接受生理盐水治疗。eNOS基因治疗后4周,评估SD大鼠踝部动脉压、微循环灌注、数字减影血管造影、组织微血管计数以及eNOS蛋白的表达。结果:eNOS基因治疗后4周,和对照组相比,接受eNOS基因治疗的大鼠缺血肢体,表现了更好的血流恢复(踝部动脉压(mmHg):58.2±4.7 vs 86.8±4.3,P<0.01;微循环灌注:142.0%±21.5%vs 219.6%±26.2%,P<0.01)、侧枝开放和血管新生(血管造影:6.7±1.1 vs 14.4±1.7,P<0.01;微血管/肌纤维比值:0.34±0.03 vs 0.56±0.02,P<0.01)以及eNOS蛋白的高表达(0.46±0.02 vs 0.73±0.02,P<0.01)。结论:eNOS基因治疗促进SD大鼠缺血后肢的血管新生。展开更多
Background People recently realized that it is important for artificial vascular biodegradable graft to bionically mimic the functions of the native vessel. In order to overcome the high risk of thrombosis and keep th...Background People recently realized that it is important for artificial vascular biodegradable graft to bionically mimic the functions of the native vessel. In order to overcome the high risk of thrombosis and keep the patency in the clinical small-diameter vascular graft （SDVG） transplantation, a double-layer bionic scaffold, which can offer anticoagulation and mechanical strength simultaneously, was designed and fabricated via electrospinning technique. Methods Heparin-conjugated polycaprolactone （hPCL） and polyurethane （PU）-collagen type I composite was used as the inner and outer layers, respectively. The porosity and the burst pressure of SDVG were evaluated. Its biocompatibility was demonstrated by the 3-（4,5-dimethyl-2-thiazol）-2,5-diphenyl-2H tetrazolium bromide （MTT） test in vitro and subcutaneous implants in vivo respectively. The grafts of diameter 2.5 mm and length 4.0 cm were implanted to replace the femoral artery in Beagle dog model. Then, angiography was performed in the Beagle dogs to investigate the patency and aneurysm of grafts at 2, 4, and 8 weeks post-transplantation. After angiography, the patent grafts were explanted for histological analysis. Results The double-layer bionic SDVG meet the clinical mechanical demand. Its good biocompatibility was proven by cytotoxicity experiment （the cell＇s relative growth rates （RGR） of PU-collagen outer layer were 102.8%, 109.2% and 103.5%, while the RGR of hPCL inner layer were 99.0%, 100.0% and 98.0%, on days 1, 3, and 5, respectively） and the subdermal implants experiment in the Beagle dog. Arteriography showed that all the implanted SDVGs were patent without any aneurismal dilatation or obvious anastomotic stenosis at the 2nd, 4th, and 8th week after the operation, except one SDVG that failed at the 2nd week. Histological analysis and SEM showed that the inner layer was covered by new endothelial-like cells. Conclusion The double-layer bionic SDVG is a promising candidate as a replacement of native small-diameter vascular graft.展开更多
文摘目的:评估内皮型一氧化氮合酶(Endothelial nitric oxide synthase,eNOS)基因治疗对大鼠缺血后肢血管新生的影响。方法:局麻下将30只雄性SD大鼠后肢缺血模型制作后,随机分成实验组和对照组,每组15只。模型制作后1周,采用肌肉注射的方法,实验组缺血后肢接受载有eNOS基因的5型重组腺病毒治疗,对照组接受生理盐水治疗。eNOS基因治疗后4周,评估SD大鼠踝部动脉压、微循环灌注、数字减影血管造影、组织微血管计数以及eNOS蛋白的表达。结果:eNOS基因治疗后4周,和对照组相比,接受eNOS基因治疗的大鼠缺血肢体,表现了更好的血流恢复(踝部动脉压(mmHg):58.2±4.7 vs 86.8±4.3,P<0.01;微循环灌注:142.0%±21.5%vs 219.6%±26.2%,P<0.01)、侧枝开放和血管新生(血管造影:6.7±1.1 vs 14.4±1.7,P<0.01;微血管/肌纤维比值:0.34±0.03 vs 0.56±0.02,P<0.01)以及eNOS蛋白的高表达(0.46±0.02 vs 0.73±0.02,P<0.01)。结论:eNOS基因治疗促进SD大鼠缺血后肢的血管新生。
文摘Background People recently realized that it is important for artificial vascular biodegradable graft to bionically mimic the functions of the native vessel. In order to overcome the high risk of thrombosis and keep the patency in the clinical small-diameter vascular graft （SDVG） transplantation, a double-layer bionic scaffold, which can offer anticoagulation and mechanical strength simultaneously, was designed and fabricated via electrospinning technique. Methods Heparin-conjugated polycaprolactone （hPCL） and polyurethane （PU）-collagen type I composite was used as the inner and outer layers, respectively. The porosity and the burst pressure of SDVG were evaluated. Its biocompatibility was demonstrated by the 3-（4,5-dimethyl-2-thiazol）-2,5-diphenyl-2H tetrazolium bromide （MTT） test in vitro and subcutaneous implants in vivo respectively. The grafts of diameter 2.5 mm and length 4.0 cm were implanted to replace the femoral artery in Beagle dog model. Then, angiography was performed in the Beagle dogs to investigate the patency and aneurysm of grafts at 2, 4, and 8 weeks post-transplantation. After angiography, the patent grafts were explanted for histological analysis. Results The double-layer bionic SDVG meet the clinical mechanical demand. Its good biocompatibility was proven by cytotoxicity experiment （the cell＇s relative growth rates （RGR） of PU-collagen outer layer were 102.8%, 109.2% and 103.5%, while the RGR of hPCL inner layer were 99.0%, 100.0% and 98.0%, on days 1, 3, and 5, respectively） and the subdermal implants experiment in the Beagle dog. Arteriography showed that all the implanted SDVGs were patent without any aneurismal dilatation or obvious anastomotic stenosis at the 2nd, 4th, and 8th week after the operation, except one SDVG that failed at the 2nd week. Histological analysis and SEM showed that the inner layer was covered by new endothelial-like cells. Conclusion The double-layer bionic SDVG is a promising candidate as a replacement of native small-diameter vascular graft.
