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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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Related in: MedlinePlus

Agarose gel electrophoresis of NP/DNA complexes. Lane 1: DNA molecular weight marker; lane 2: naked plasmid DNA; lane 3: chitosan-coated PBCA-NPs; lanes 4–7 correspond to DNA with progressively increasing proportions of NPs at the charge ratio of 5:1, 10:1, 15:1, and 30:1 (w/w), respectively.
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fig4: Agarose gel electrophoresis of NP/DNA complexes. Lane 1: DNA molecular weight marker; lane 2: naked plasmid DNA; lane 3: chitosan-coated PBCA-NPs; lanes 4–7 correspond to DNA with progressively increasing proportions of NPs at the charge ratio of 5:1, 10:1, 15:1, and 30:1 (w/w), respectively.

Mentions: pDNA complex withpreviously prepared nanoparticles was achieved by the formation of ion-pairs betweenthe positively charged amino groups located on the particle surface and thenegatively charged phosphodiester backbone of the plasmid DNA [45]. The electrophoretic mobility of DNA, complex with PBCA-NPs, decreasesgradually on the agarose gel with increasing charge ratio (Figure 4). However,in the control experiment, there was no DNA in agarose gel. This method also allowed showing the dependencyof this interaction on pH, reflecting the deprotonation of the modified silicaparticles at alkaline pH (data not shown). The particle-DNA binding was alsoinhibited by the presence of high salt concentrations (data not shown). This suggeststhat this interaction is based on electrostatic forces, resulting ininterpolyelectrolyte complexes.


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)

Agarose gel electrophoresis of NP/DNA complexes. Lane 1: DNA molecular weight marker; lane 2: naked plasmid DNA; lane 3: chitosan-coated PBCA-NPs; lanes 4–7 correspond to DNA with progressively increasing proportions of NPs at the charge ratio of 5:1, 10:1, 15:1, and 30:1 (w/w), respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Agarose gel electrophoresis of NP/DNA complexes. Lane 1: DNA molecular weight marker; lane 2: naked plasmid DNA; lane 3: chitosan-coated PBCA-NPs; lanes 4–7 correspond to DNA with progressively increasing proportions of NPs at the charge ratio of 5:1, 10:1, 15:1, and 30:1 (w/w), respectively.
Mentions: pDNA complex withpreviously prepared nanoparticles was achieved by the formation of ion-pairs betweenthe positively charged amino groups located on the particle surface and thenegatively charged phosphodiester backbone of the plasmid DNA [45]. The electrophoretic mobility of DNA, complex with PBCA-NPs, decreasesgradually on the agarose gel with increasing charge ratio (Figure 4). However,in the control experiment, there was no DNA in agarose gel. This method also allowed showing the dependencyof this interaction on pH, reflecting the deprotonation of the modified silicaparticles at alkaline pH (data not shown). The particle-DNA binding was alsoinhibited by the presence of high salt concentrations (data not shown). This suggeststhat this interaction is based on electrostatic forces, resulting ininterpolyelectrolyte complexes.

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