Min-Da Shau

The characteristics and transfection efficiency of PEI modified by biodegradable poly(β-amino ester)

To improve the cytotoxicity of PEI25k and the transfection efficiency of poly(β-amino ester) with DNA, we synthesized a poly(β-amino ester), PEDP, bearing ester linkages in the backbone and tertiary amines in the backbone and side chain and prepared a binary mixture, PEDP–PEI25k, using physical blending meyhod. Both poly(β-amino ester) PEDP and binary mixture PEDP–PEI25k, readily self-assembled with plasmid DNA (pCMV-β gal) in a HEPES buffer, were characterized by dynamic light scattering. The results reveal that PEDP–PEI25k was able to self-assemble plasmid DNA into PEDP–PEI25k/DNA nano-complexes small enough to enter a cell through endocytosis. Titration studies were performed to determine the buffering capacities of PEDP and PEDP–PEI25k. The COS-7 cell viabilities in the presence of PEDP and PEDP–PEI25k were studied. At low mass ratio of PEDP/PEI25k (1/1), it is found that the transfection curve of PEDP–PEI25k/DNA bearing a maximum peak is similar to that of PEI25k/DNA. In addition, the PEDP–PEI25k/DNA complexes were able to transfect COS-7 cells in vitro with a high efficiency comparable to a well-known gene carrier PEI25k/DNA. The results indicate that binary mixture PEDP–PEI25k is an attractive cationic carrier for gene delivery and an interesting candidate for further study.

A thermodynamic study of cationic polymer-plasmid DNA complexes by highly-sensitive differential scanning calorimetry.

The characteristics of polymer-DNA complexes formed by positively-negatively charged interaction have a great influence on their transfection potential. Since the limit changes in thermal transitions which were hardly measured in conventional calorimetry, now in this study they have been successfully carried out by highly-sensitive differential scanning calorimetry for better understanding the pDMAEMA-plasmid DNA complexing process. Thermal behaviors of plasmid DNA, polymer and their formed complexes were recorded to give insights into their conformational changes when temperature was raised. In results, the supercoiled or open-circular plasmid DNA is not thermal reversible indicated by the decrease of denaturation peak and disappearance of DNA conformational transition related to its twist status at 50-70 degrees C. The cationic polymer is thermally stable by showing reversible transition peaks after two heating processes. For the cationic polymer-plasmid DNA complexes, electrostatic forces lead to a higher denaturation temperature of plasmid DNA and transition temperature of polymer. Also, heat can cause a topological change in plasmid DNA and then change their mutual complexation capacity.

Synthesis and characterizations of new glycidyl-based cationic poly(aminoester) and study on gene delivery

New glycidyl-based (epoxide-based) poly(aminoester) (EPAE) containing hydroxyl and amino groups in the backbone and side chain was synthesized. EPAE self-assembled readily with the plasmid DNA(pCMV-betagal) in HEPES buffer and was characterized by dynamic light scattering, Zeta-potential, fluorescence images, and XTT cell viability assays. To evaluate the effect of molecular weight of EPAE system on transfection, EPAE polymers with three different molecular weights (EPAE22k, EPAE18k, and EPAE8k) were also prepared. This study found that all EPAE polymers were able to bind plasmid DNA and yielded positively charged complexes with a nano-sized particles (200 nm). The EPAE22k/DNA and EPAE18k/DNA complexes were able to transfect COS-7 cell in vitro with higher transfection efficiency than other EPAE8k/DNA. These results demonstrated that molecular weight of EPAE system had a significant effect on transferring ability. Examination of the cytotoxicity of PEI25k and EPAEs system revealed that EPAEs system had lower cytotoxicity. In this article, EPAEs seemed to be a novel cationic poly(aminoester) for gene delivery and an interesting candidate for further study.

A new cationic poly(ester-anhydride): Synthesis, structure characterization, and biological properties

To improve the cytotoxicity and biocompatibility of cationic polymers used as gene carriers, new cationic poly(ester-anhydride) copolymers (Pea-PAs) bearing a quaternary ammonium group in the backbone were synthesized through a reaction between an aminoester dicarboxylic acid with a tertiary amine in the backbone and excess acetyl chloride. Their structure was characterized by nuclear magnetic resonance (NMR), Fourier transform infrared spectrometry (FTIR), and gel permeation chromatography (GPC). They degraded hydrolytically in 20 mM HEPES buffer at pH 7.4 at 37°C with a half-life of more than 70 h. Their cytotoxicity was substantially lower than that of PEI. The blood clotting index (BCI) and haemolysis ratio (HR) also showed that these newly synthesized Pea-PA polymer are biopolymers with good blood compatibility.