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Cationic polybutyl cyanoacrylate nanoparticles for DNA delivery.

Duan J, Zhang Y, Chen W, Shen C, Liao M, Pan Y, Wang J, Deng X, Zhao J - J. Biomed. Biotechnol. (2009)

Bottom Line: Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours.Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation.The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Nanobiological Technology, Ministry of Health National Hepatobiliary and Enteric Surgery Research Center, Central South University, Changsha, Hunan 410008, China.

ABSTRACT
To enhance the intracellular delivery potential of plasmid DNA using nonviral vectors, we used polybutyl cyanoacrylate (PBCA) and chitosan to prepare PBCA nanoparticles (NPs) by emulsion polymerization and prepared NP/DNA complexes through the complex coacervation of nanoparticles with the DNA. The object of our work is to evaluate the characterization and transfection efficiency of PBCA-NPs. The NPs have a zeta potential of 25.53 mV at pH 7.4 and size about 200 nm. Electrophoretic analysis suggested that the NPs with positive charges could protect the DNA from nuclease degradation and cell viability assay showed that the NPs exhibit a low cytotoxicity to human hepatocellular carcinoma (HepG2) cells. Qualitative and quantitative analysis of transfection in HepG2 cells by the nanoparticles carrying plasmid DNA encoding for enhanced green fluorescent protein (EGFP-N1) was done by digital fluorescence imaging microscopy system and fluorescence-activated cell sorting (FACS). Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours. Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation. The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery.

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(a) Qualitative evaluation of EGFP-N1 transgene expression in HepG2 cells with chitosan-coated PBCA-NPs, fluorescent image (×200) of the cells was obtained after 96 hours upon incubation. (b) Quantitative evaluation  of EGFP-N1 transgene expression in HepG2 cells as a function of time measured by flow cytometric analysis with chitosan-coated PBCA-NPs. HepG2 cells without any treatment were used as control.
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fig9: (a) Qualitative evaluation of EGFP-N1 transgene expression in HepG2 cells with chitosan-coated PBCA-NPs, fluorescent image (×200) of the cells was obtained after 96 hours upon incubation. (b) Quantitative evaluation of EGFP-N1 transgene expression in HepG2 cells as a function of time measured by flow cytometric analysis with chitosan-coated PBCA-NPs. HepG2 cells without any treatment were used as control.

Mentions: The pictures of fluorescence microscope to detect the green fluorescencesent out by transfected cells indicated relatively higher transfectionefficiencies of NP/DNA complexes (Figure 9(a)). To determine the gene transfer capabilityof NPs in vitro exactly, flow cytometry experiments were conducted to determinethe EGFP transfection levels in HepG2 cells. It was found that the transgeneexpression levels increased with the time, and reached a climax at 72 hours,then decreased. The results of the complexes are fused with cell membrane easilythrough electrostatical interaction can be explained by the fact that complexespossess the positive potential in neutral conditions.


Cationic polybutyl cyanoacrylate nanoparticles for DNA delivery.

Duan J, Zhang Y, Chen W, Shen C, Liao M, Pan Y, Wang J, Deng X, Zhao J - J. Biomed. Biotechnol. (2009)

(a) Qualitative evaluation of EGFP-N1 transgene expression in HepG2 cells with chitosan-coated PBCA-NPs, fluorescent image (×200) of the cells was obtained after 96 hours upon incubation. (b) Quantitative evaluation  of EGFP-N1 transgene expression in HepG2 cells as a function of time measured by flow cytometric analysis with chitosan-coated PBCA-NPs. HepG2 cells without any treatment were used as control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2655361&req=5

fig9: (a) Qualitative evaluation of EGFP-N1 transgene expression in HepG2 cells with chitosan-coated PBCA-NPs, fluorescent image (×200) of the cells was obtained after 96 hours upon incubation. (b) Quantitative evaluation of EGFP-N1 transgene expression in HepG2 cells as a function of time measured by flow cytometric analysis with chitosan-coated PBCA-NPs. HepG2 cells without any treatment were used as control.
Mentions: The pictures of fluorescence microscope to detect the green fluorescencesent out by transfected cells indicated relatively higher transfectionefficiencies of NP/DNA complexes (Figure 9(a)). To determine the gene transfer capabilityof NPs in vitro exactly, flow cytometry experiments were conducted to determinethe EGFP transfection levels in HepG2 cells. It was found that the transgeneexpression levels increased with the time, and reached a climax at 72 hours,then decreased. The results of the complexes are fused with cell membrane easilythrough electrostatical interaction can be explained by the fact that complexespossess the positive potential in neutral conditions.

Bottom Line: Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours.Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation.The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Nanobiological Technology, Ministry of Health National Hepatobiliary and Enteric Surgery Research Center, Central South University, Changsha, Hunan 410008, China.

ABSTRACT
To enhance the intracellular delivery potential of plasmid DNA using nonviral vectors, we used polybutyl cyanoacrylate (PBCA) and chitosan to prepare PBCA nanoparticles (NPs) by emulsion polymerization and prepared NP/DNA complexes through the complex coacervation of nanoparticles with the DNA. The object of our work is to evaluate the characterization and transfection efficiency of PBCA-NPs. The NPs have a zeta potential of 25.53 mV at pH 7.4 and size about 200 nm. Electrophoretic analysis suggested that the NPs with positive charges could protect the DNA from nuclease degradation and cell viability assay showed that the NPs exhibit a low cytotoxicity to human hepatocellular carcinoma (HepG2) cells. Qualitative and quantitative analysis of transfection in HepG2 cells by the nanoparticles carrying plasmid DNA encoding for enhanced green fluorescent protein (EGFP-N1) was done by digital fluorescence imaging microscopy system and fluorescence-activated cell sorting (FACS). Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours. Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation. The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery.

Show MeSH
Related in: MedlinePlus