Construction of Polyethyleneimine-β-cyclodextrin/pDNA Multilayer Structure for Improved In Situ Gene Transfection

This study reports in situ gene delivery from gene-functionalized poly(D,L-lactic acid) (PDLLA, M-w of around 2.0 x 10(5) g mol(-1)) films, which were constructed via layer-by-layer (LbL) assembly technique with low molecular weight polyethylenimine-beta-cyclodextrin (PEI-CD) conjugate and plasmid DNA (pDNA). PEI-CD was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR), respectively. The buildup of multilayered PEI-CD/pDNA pairs onto PDLLA films was monitored with contact angle measurements and UV-Vis spectrometer, respectively. A sustained release of pDNA from multilayered films was observed for 28 h. The mechanism of in situ gene delivery on PDLLA film was investigated in this study as well. Spherical PEI-CD/pDNA complexes were formed and released following the deconstruction of multilayered films, which was confirmed by transmission electron microscopy (TEM) and gel electrophoresis, respectively. Surface mediated in situ gene transfection. was achieved when culturing hepatoma G2 (HepG2) and human embryonic kidney 293 (HEK293) onto PEI-CD/pDNA multilayered films. Furthermore, PEI-CD improved the gene transfection efficiency when compared with that of PEI. Such gene-functionalized biomaterial reported here has potential application in tissue engineering and implant technology.