Chitosan Grafted with Phosphorylcholine and Macrocyclic Polyamine as an Effective Gene Delivery Vector: Preparation, Characterization and In Vitro Transfection

摘要

Herein, an effective gene delivery vector phosphorylcholine and macrocyclic polyamine grafted chitosan (PC-g(6)-Cs-g(2)-Cyclen) was developed. Chemical characterization of product PC-g(6)-Cs-g(2)-Cyclen was performed by NMR, FT-IR, gel permeation chromatography (GPC), and X-ray photoelectron spectroscopy (XPS) analysis. PC-g(6)-Cs-g(2)-Cyclen could more efficiently bind and protect plasmid DNA than macrocyclic polyamine grafted chitosan (Cs-g-Cyclen) and phosphorylcholine grafted chitosan (Cs-g-PC), as evaluated by agarose gel electrophoresis, circular dichroism spectra, and fluorescence quenching assays. PC-g(6)-Cs-g-(2)Cyclen could wrap DNA into uniform nanoparticles in the size of 112.6 +/- 8.5nm and possessed net cationic charge. UV spectroscopy and MTT assays showed excellent water-solubility and cell viability for PC-g(6)-Cs-g(2)-Cyclen. In addition, three polymer/DNA complexes showed 5.1-15.1-fold greater uptake activity and 10-14-fold higher transfection efficiency in 293 T cells as compared to chitosan/DNA complex, in which PC-g(6)-Cs-g(2)-Cyclen demonstrated the highest transfection activity. These date demonstrated that PC-g(6)-Cs-g(2)-Cyclen is a promising vector candidate for gene delivery.