New support system of vascular surgery using novel bio-materials

(1)利用新型生物材料开发新型血管生成治疗技术。本研究利用酸性明胶水凝胶微球（AGHM）和高分子纳米胶束颗粒系统（NP）作为新型生物材料。在AGHM中，将重组bFGF蛋白与AGHM结合，通过靶向动脉给药制备bFGF浸染的AGHM，建立兔慢性肢体缺血模型。注射后28天，几项评估显示缺血肢体的血液灌注和新生血管明显改善。同时，在NP中，首先将带有标记基因DNA的复合物注射到肌肉组织中，并评估基因转移效率。但未检测到标记基因的显著表达。为了提高基因转移的效果，我们在NP中添加了配体肽（RGD肽、环状RGD肽或动脉壁结合肽）（RGD-NP、rRGD-NP和AWBP-NP）。然而，这些配体添加的NPs没有显示出基因向肌肉组织转移效率的显著提高。因此，我们利用反向转染的方法研究了NP基因转移的新途径，并在体外条件下取得了良好的效果。(2)利用新型生物材料进行血管内膜增生基因转移为了建立血管内膜增生基因转移方法，我们将NP复合标记基因DNA应用于兔颈动脉新生内膜。虽然本研究检测了非配体NP、RGD-NP、rRGD-NP和AWBP-NP，但与对照组相比，未发现基因转移明显增加。然后，我们还对新型多离子胶团PEG-DET进行了评价，并观察到其对兔颈动脉新生内膜的有利基因传递。

(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.