New support system of vascular surgery using novel bio-materials

(1)利用新型生物材料开发新的治疗性血管生成本研究利用酸性明胶水凝胶微球（AGHM）和聚合物纳米胶束颗粒系统（NP）作为新型生物材料。在AGHM中，重组bFGF蛋白与AGHM结合，并通过兔慢性肢体缺血模型的靶动脉施用bFGF浸渍的AGHM。注射后28天，几项评估显示缺血肢体的血液灌注和新血管形成显著改善。同时，在NP中，首先将具有标记基因DNA的复合物注射到肌肉组织中，并评估基因转移效率。但标记基因在细胞中未检测到明显表达。为了提高基因转移的效率，我们向NP（RGD-NP、rRGD-NP和AWBP-NP）中加入配体肽（RGD肽、环状RGD肽或动脉壁结合肽）。然而，这些添加配体的NP没有显示出对肌肉组织的基因转移效率的显著改善。因此，我们采用反向转染法研究了NP基因转移的新途径，并在体外条件下获得了良好的结果。(2)应用新型生物材料进行基因转染治疗血管内膜增生为建立基因转染治疗血管内膜增生的方法，我们将NP与标记基因DNA复合物应用于兔颈动脉新生内膜层。虽然在本研究中检测了非配体NP、RGD-NP、rRGD-NP和AWBP-NP，但与对照相比没有检测到基因转移的显著增加。然后，我们还进行了新的聚离子胶束，PEG-DET的评价，并观察到良好的基因传递到兔颈动脉新生内膜。

(1) Development of new therapeutic angiogenesis using novel bio-materialsThe present study utilized acidic gelatin hydrogel microspheres (AGHM) and polymer nano-micelle particle system (NP) as novel bio-material. In AGHM, recombinant bFGF protein was bound to AGHM, and the bFGF-impregnated AGHM was administrated through targeted artery in rabbit model of chronic limb ischemia. Twenty eight days after the injection, several assessments showed significant improvement of blood perfusion and neovascularization in the ischemic limb. Meanwhile in NP, the complex with marker gene DNA was first injected to muscle tissue, and gene transfer efficiency was evaluated. However, no significant expression of marker gene was detected. To improve the efficacy of gene transfer, we added ligand peptides (RGD peptide, ring-formed RGD peptide or artery wall binding peptide) to the NP (RGD-NP, rRGD-NP and AWBP-NP). Though, these ligand-added NPs showed no significant improvement of gene transfer efficiency to muscle tissue. Therefore, we studied new gene transfer approach of NP by using reverse transfection method, and obtained favorable result under in vitro condition.(2) Gene transfer to intimal hyperplasia using novel bio-materialsTo develop gene transfer method to intimal hyperplasia of vessels, we applied NP complexed with marker gene DNA to neointimal layer of rabbit carotid artery. Although non-ligand NP, RGD-NP, rRGD-NP and AWBP-NP were examined in this study, no significant increase of gene transfer was detected as compared with control. Then, we also carried out the evaluation of new poly-ion micelle, PEG-DET, and observed favorable gene delivery to neointima of rabbit carotid artery.