Development of mouse tissue transfer model of VEGF gene therapy

术前或术中将dna -脂质体复合物注射于HR-1品系小鼠（雄性，体重30g）腹股沟皮瓣或腹股沟区复合移植物皮下。6μg VEGFh165-pBLAST质粒及Plus用600μl D-MEM稀释试剂，与24μl Lipofectin （Invivogen）混合制成dna -脂质体复合物。对照组注射D-MEM。[经验1]皮下注射组微血管密度增加至第3天。皮下注射组水平延伸腹股沟皮瓣皮瓣瓣存活率显著高于对照组（分别为60.0±0.002%和48.3±0.19%）。对照组和实验组的皮肤存活率无统计学差异（t=0.67）。复合植片和对照组的皮叶存活率分别为18.4±0.12%和24.7±0.09%。两组间差异无统计学意义（t=0.40）[经验2]皮下注射组炎性细胞和成纤维细胞在硅胶板周围聚集形成囊层。微血管密度的增加主要发生在结构外围的表面。硅胶板中心以上的微血管也被注意到。皮下注射组垂直延伸腹股沟皮瓣皮瓣瓣存活率为60.0±0.001%。VEGF处理复合移植物的存活率高于对照组（10.3%和3.4%），但差异无统计学意义（p < 0.05）。注射VEGF质粒-脂质体复合物后未见任何副作用。

DNA-liposome complex was injected on HR-1 strain mouse (male, body weight 30g) subcutaneously on the groin flap or composite graft on the groin area before or on the operation. 6μg VEGFh165-pBLAST plasmid and Plus. Reagent was diluted with 600μl D-MEM and DNA-liposome complex was made by mixing with 24μl Lipofectin (Invivogen). D-MEM was injected in control group.[experience 1] The increase in microvessel density in the subdermal injected group up to day 3 from was observed.The skin paddle survival on the horizontal extended groin flap in the subdermal injected group was larger than controls(60.0±0.002%% and 48.3±0.19% of total skin paddle respectively). No statistically significant difference was noted between the skin paddle survival in controls and treatments (t=0.67).The skin paddle survival on the composite graft and controls were 18.4±0.12% and 24.7±0.09% of total skin paddle respectively. No statistically significant difference was noted between two groups (t=0.40)[experience 2] Aggregation of inflammatory cell and fibroblast formed capsule layer around the silicone plate in the subdermal injected group. The increase in microvessel density was observed on the surface side of the periphery of the structure. The microvascular findings were also noted above the center of silicone plate.The skin paddle survival on the vertical extended groin flap in the subdermal injected group was 60.0±0.001% of total skin paddle. No statistically significant difference was noted compared with controls (59.6±0.16%)The survival of composite graft treated with VEGF was greater than that of controls(10.3% and 3.4%), but statistical significance was not reached.Any side effect was not observed after VEGF plasmid-liposome complex injection.