Novel tissue regeneration using polymer nano-micelle gene delivery system

(1)特异性基因非病毒载体系统的建立为制备特异性基因输送的纳米胶束载体，我们在多离子复合纳米胶束体系(RGD胶束、环-RGD胶束和AWBP胶束)中添加了配基多肽(RGD多肽、RGD环状多肽或动脉壁结合多肽)。对几种培养细胞的评价表明，RING-RGD胶束的基因转移效率明显高于无配体胶束和RGD胶束。此外，在培养的平滑肌细胞中，AWBP胶束比其他胶束具有显著的基因转移作用。(2)重建组织血管形成基因传递方法的发展尽管我们用RGD胶束和RING-RGD胶束评估了体内对肌肉组织的基因传递效率，但与非配基胶束相比，RGD胶束和RING-RGD胶束并没有明显的优势。我们还检测了AWBP胶束对损伤动脉壁的基因传递效果，但没有检测到明显的效果。因此，我们利用逆转染法建立了纳米胶束基因载体的基因转移新途径。此外，我们还对新型聚离子胶束进行了评价，观察到了良好的基因输送到兔颈动脉。(3)将含有三种可能促进骨再生的基因的再生骨PEGDET植入合适的支架内，观察到良好的骨再生。

(1)Development of non-viral vector system for specific gene deliveryTo develop nano-micelle gene vector for specific gene delivery, we added ligand peptides (RGD peptide, ring-formed RGD peptide or artery wall binding peptide) to the poly-ion complex nano-micelle system (RGD micelle, ring-RGD micelle and AWBP micelle). The evaluations using several culture cell lines showed that gene transfer efficiency by ring-RGD micelle was significantly higher than those by micelle without ligand and RGD micelle. Further, in culture smooth muscle cells, AWBP micelle showed remarkable gene transfer as compared with other micelles.(2)Developmento of gene delivery method for vascularization of reconstructive tissueAlthough we assessed in vivo gene delivery efficiency to muscle tissue by using RGD micelle and ring-RGD micelle, there was no significant advantage in RGD micelle and ring-RGD micelle as compared with non-ligand micelle. We also tested gene delivery effect by AWBP micelle to the injured arterial wall, but no significant effect was detected. Therefore, we established new gene transfer approach of nano-micelle gene vector by using reverse transfection method. Further, we also carried out the evaluation of new poly-ion micelle, PEG-DET, and observed favorable gene delivery to rabbit carotid artery.(3)Vascularization of regenerative bonePEG-DET containing three kinds of gene, which potentially stimulate bone regeneration, was implanted to mice with appropriate scaffold, and favorable bone regeneration was observed in the scaffold.