Novel tissue regeneration using polymer nano-micelle gene delivery system

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(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.