Development of mouse tissue transfer model of VEGF gene therapy

将DNA-脂质体复合体注射于HR-1系小鼠(雄性，体重30g)的腹股沟皮瓣下或腹股沟复合移植物上。6μg VEGFh165-pBLAST表达载体和Plus.试剂用600μ的L D-MEM稀释，与24μ的L脂质体(Invivogen)混合制成脂质体。[经验1]皮下注射组至术后3d，微血管密度明显增加，皮下注射组腹股沟水平延伸皮瓣成活明显高于对照组(分别为60.0±0.002%和48.3±0.19%)。对照组和治疗组皮瓣存活率差异无统计学意义(t=0.67)，复合移植物和对照组皮瓣存活率分别为18.4±0.12%和24.7±0.09%。皮下注射组炎性细胞聚集，成纤维细胞在硅胶板周围形成囊层，差异无统计学意义(t=0.40)[经验2]。微血管密度在结构外围的表面侧增加。皮下注射组垂直延伸腹股沟皮瓣上皮瓣存活率为60.00±0.001%。与对照组(59.6±0.16%)比较，差异无统计学意义(P>0.05)。注射血管内皮生长因子的复合移植物存活率(10.3%和3.4%)高于对照组(10.3%和3.4%)，但无统计学意义。

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.