Novel tissue regeneration using polymer nano-micelle gene delivery system

（1）特异性基因递送的非病毒载体系统的研制为了研制特异性基因递送的纳米胶束基因载体，我们在聚离子复合物纳米胶束系统（RGD胶束、环-RGD胶束和AWBP胶束）中加入配体肽（RGD肽、环状RGD肽或动脉壁结合肽）。用几种培养细胞系的评价表明，环-RGD胶束的基因转移效率显着高于无配体胶束和RGD胶束。此外，在培养的平滑肌细胞，AWBP胶束显示出显着的基因转移相比，其他胶束。（2）重建组织血管化的基因递送方法的建立虽然我们评估了使用RGD胶束和环-RGD胶束对肌肉组织的体内基因递送效率，但与无配体胶束相比，RGD胶束和环-RGD胶束没有显著优势。我们还测试了AWBP胶束对损伤动脉壁的基因递送效果，但没有检测到显著的效果。因此，我们利用反向转染法建立了纳米胶束基因载体的基因转移新途径。此外，我们还进行了新的聚离子胶束，PEG-DET的评价，并观察到有利的基因传递到兔颈动脉。（3）再生骨的血管化将含有三种可能促进骨再生的基因的PEG-DET与合适的支架一起植入小鼠体内，观察到支架中良好的骨再生。

(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.