PURPOSE: The objective of this study is to characterize the micelles of novel block copolymer of poly (p-Dioxanone-co-L-Lactide)-block-Poly (ethylene glycol) (PPDO/PLLA-b-PEG-) and evaluate its ability to induce gene transfection. METHODS: The ability of the block copolymer to self-assemble was determined by viscometery, dye solublization, NMR spectra and dynamic light scattering. The Trypan blue assay for in vitro biocompatibility of the block copolymer was carried out with NIH 3T3, CT-26 and MCF-7 cells, and beta-glactosidase assay was applied to measure the transfection efficiency of the block copolymer on MCF-7 breast cancer cell. RESULTS: Depending on the block lengths and molecular weights, solubility of the polymeric samples in water was varied. Diluted aqueous solution properties of the copolymer were studied. 1,6-Diphenyl-1,3,5-hexatriene solubilization and 1H NMR spectra carried out in CDCl3 and D2O, were used to prove the existence of hydrophobic domains as the core of micelle. Average particle size of 60-165 nm with low polydispersity, and lower negative zeta potential of -3 to -14 mV were observed on the aqueous copolymer dispersion. Copolymer was found with almost no cytotoxic effect and was able to promote the transfection efficiency (about 3-fold) in MCF-7 cells. CONCLUSIONS: The PPDO/PLLA-b-PEG copolymer has ability to assemble into nanoscopic structures in aqueous environment, which enable to enhance gene transfection.
PURPOSE: The objective of this study is to characterize the micelles of novel block copolymer of poly (p-Dioxanone-co-L-Lactide)-block-Poly (ethylene glycol) (PPDO/PLLA-b-PEG-) and evaluate its ability to induce gene transfection. METHODS: The ability of the block copolymer to self-assemble was determined by viscometery, dye solublization, NMR spectra and dynamic light scattering. The Trypan blue assay for in vitro biocompatibility of the block copolymer was carried out with NIH 3T3, CT-26 and MCF-7 cells, and beta-glactosidase assay was applied to measure the transfection efficiency of the block copolymer on MCF-7 breast cancer cell. RESULTS: Depending on the block lengths and molecular weights, solubility of the polymeric samples in water was varied. Diluted aqueous solution properties of the copolymer were studied. 1,6-Diphenyl-1,3,5-hexatriene solubilization and 1H NMR spectra carried out in CDCl3 and D2O, were used to prove the existence of hydrophobic domains as the core of micelle. Average particle size of 60-165 nm with low polydispersity, and lower negative zeta potential of -3 to -14 mV were observed on the aqueous copolymer dispersion. Copolymer was found with almost no cytotoxic effect and was able to promote the transfection efficiency (about 3-fold) in MCF-7 cells. CONCLUSIONS: The PPDO/PLLA-b-PEG copolymer has ability to assemble into nanoscopic structures in aqueous environment, which enable to enhance gene transfection.
Authors: N Saito; T Okada; H Horiuchi; N Murakami; J Takahashi; M Nawata; H Ota; K Nozaki; K Takaoka Journal: Nat Biotechnol Date: 2001-04 Impact factor: 54.908