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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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Cytotoxicity of cationic chitosan coated PBCA-NPs analyzed by MTT assay, 24 hours posttreatment (n = 8). Columns, mean; bars, SD, P < .001 as evaluated by one-way ANOVA using SPSS 17.0 version.
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fig6: Cytotoxicity of cationic chitosan coated PBCA-NPs analyzed by MTT assay, 24 hours posttreatment (n = 8). Columns, mean; bars, SD, P < .001 as evaluated by one-way ANOVA using SPSS 17.0 version.

Mentions: In vitrotoxicity of NP/DNA complexes was evaluated by MTT assay in HepG2 cells usingincreasing doses of NPs. MTT assay shows that the cytotoxicity of the NPsdepended on their concentration ranging from 0.1–8.0 μg/μL (Figure 6). The statistic analysis demonstrates thatthere are significant differences between concentrations (one-way ANOVA, P < .001). These data can indicate that the NPs below 2 μg/μL had little adverse effect on the HepG2 cells viability,suggesting that the doses of in vitro HepG2 cells uptake study lessthan 0.5 μg/μL presented no toxicity. This isimportant because most of the cationic polymers and lipids, which are commonly usedfor gene transfection, have toxic effects on cells at a higher concentrationdue to electrostatic interaction with negatively charged cellular membrane [48, 49]. Thus, the PBCA-NPs is a promising carrier for gene delivery with low cytotoxicity. However, its long-term in vivo toxicity and immunogenicity should be furtherinvestigated.


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)

Cytotoxicity of cationic chitosan coated PBCA-NPs analyzed by MTT assay, 24 hours posttreatment (n = 8). Columns, mean; bars, SD, P < .001 as evaluated by one-way ANOVA using SPSS 17.0 version.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig6: Cytotoxicity of cationic chitosan coated PBCA-NPs analyzed by MTT assay, 24 hours posttreatment (n = 8). Columns, mean; bars, SD, P < .001 as evaluated by one-way ANOVA using SPSS 17.0 version.
Mentions: In vitrotoxicity of NP/DNA complexes was evaluated by MTT assay in HepG2 cells usingincreasing doses of NPs. MTT assay shows that the cytotoxicity of the NPsdepended on their concentration ranging from 0.1–8.0 μg/μL (Figure 6). The statistic analysis demonstrates thatthere are significant differences between concentrations (one-way ANOVA, P < .001). These data can indicate that the NPs below 2 μg/μL had little adverse effect on the HepG2 cells viability,suggesting that the doses of in vitro HepG2 cells uptake study lessthan 0.5 μg/μL presented no toxicity. This isimportant because most of the cationic polymers and lipids, which are commonly usedfor gene transfection, have toxic effects on cells at a higher concentrationdue to electrostatic interaction with negatively charged cellular membrane [48, 49]. Thus, the PBCA-NPs is a promising carrier for gene delivery with low cytotoxicity. However, its long-term in vivo toxicity and immunogenicity should be furtherinvestigated.

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