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PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells.

Liang GF, Zhu YL, Sun B, Hu FH, Tian T, Li SC, Xiao ZD - Nanoscale Res Lett (2011)

Bottom Line: The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes.Moreover, nanocomplex shows better serum compatibility than commercial liposome.Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. zdxiao@seu.edu.

ABSTRACT
The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly(D,L-lactide-co-glycolide) (PLGA)-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI)- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS) was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

No MeSH data available.


Related in: MedlinePlus

Agarose gel electrophoresis assay of PLGA/PEI/pDNA nanocomplexes. Lane 1, pDNA alone; lanes 2 to 7, PLGA/PEI /pDNA complexes N/P ratio 1, 2, 3, 4, 6, and 10.
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Figure 2: Agarose gel electrophoresis assay of PLGA/PEI/pDNA nanocomplexes. Lane 1, pDNA alone; lanes 2 to 7, PLGA/PEI /pDNA complexes N/P ratio 1, 2, 3, 4, 6, and 10.

Mentions: To explore complex formation of pDNA and PLGA/PEI nanoparticles, PLGA/PEI nanoparticles were mixed with pDNA at various ratios. Complex formation was assessed by a gel retardation assay. The pDNA was vortexed with the nanoparticles in nuclease-free water at different N/P ratios. As shown in Figure 2, the nanoparticles are able to fully retard the mobility of DNA in agarose gel when the N/P ratio is 10:1 or higher than 10:1. While N/P is lower than 10:1, the retardation is not full. DNA bands are visible in N/P complexes in lanes 2 to 4 (Figure 2), indicating the presence of free DNA in nanocomplexes of 1 and 4 N/P ratios. This result is comparable with the band observed in lane 1, which contain only free plasmid. Few free DNA bands was observed in subsequent lanes of N/P ratios 6 and 10, indicating complete complexation of all free plasmid. The results indicate that the nanocomplexes can be easily prepared by simply mixing cationic polymer and DNA solution, and the results also showed that the optimal N/P is 6:1.


PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells.

Liang GF, Zhu YL, Sun B, Hu FH, Tian T, Li SC, Xiao ZD - Nanoscale Res Lett (2011)

Agarose gel electrophoresis assay of PLGA/PEI/pDNA nanocomplexes. Lane 1, pDNA alone; lanes 2 to 7, PLGA/PEI /pDNA complexes N/P ratio 1, 2, 3, 4, 6, and 10.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3211866&req=5

Figure 2: Agarose gel electrophoresis assay of PLGA/PEI/pDNA nanocomplexes. Lane 1, pDNA alone; lanes 2 to 7, PLGA/PEI /pDNA complexes N/P ratio 1, 2, 3, 4, 6, and 10.
Mentions: To explore complex formation of pDNA and PLGA/PEI nanoparticles, PLGA/PEI nanoparticles were mixed with pDNA at various ratios. Complex formation was assessed by a gel retardation assay. The pDNA was vortexed with the nanoparticles in nuclease-free water at different N/P ratios. As shown in Figure 2, the nanoparticles are able to fully retard the mobility of DNA in agarose gel when the N/P ratio is 10:1 or higher than 10:1. While N/P is lower than 10:1, the retardation is not full. DNA bands are visible in N/P complexes in lanes 2 to 4 (Figure 2), indicating the presence of free DNA in nanocomplexes of 1 and 4 N/P ratios. This result is comparable with the band observed in lane 1, which contain only free plasmid. Few free DNA bands was observed in subsequent lanes of N/P ratios 6 and 10, indicating complete complexation of all free plasmid. The results indicate that the nanocomplexes can be easily prepared by simply mixing cationic polymer and DNA solution, and the results also showed that the optimal N/P is 6:1.

Bottom Line: The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes.Moreover, nanocomplex shows better serum compatibility than commercial liposome.Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. zdxiao@seu.edu.

ABSTRACT
The biggest challenge in the field of gene therapy is how to effectively deliver target genes to special cells. This study aimed to develop a new type of poly(D,L-lactide-co-glycolide) (PLGA)-based nanoparticles for gene delivery, which are capable of overcoming the disadvantages of polyethylenimine (PEI)- or cationic liposome-based gene carrier, such as the cytotoxicity induced by excess positive charge, as well as the aggregation on the cell surface. The PLGA-based nanoparticles presented in this study were synthesized by emulsion evaporation method and characterized by transmission electron microscopy, dynamic light scattering, and energy dispersive spectroscopy. The size of PLGA/PEI nanoparticles in phosphate-buffered saline (PBS) was about 60 nm at the optimal charge ratio. Without observable aggregation, the nanoparticles showed a better monodispersity. The PLGA-based nanoparticles were used as vector carrier for miRNA transfection in HepG2 cells. It exhibited a higher transfection efficiency and lower cytotoxicity in HepG2 cells compared to the PEI/DNA complex. The N/P ratio (ratio of the polymer nitrogen to the DNA phosphate) 6 of the PLGA/PEI/DNA nanocomplex displays the best property among various N/P proportions, yielding similar transfection efficiency when compared to Lipofectamine/DNA lipoplexes. Moreover, nanocomplex shows better serum compatibility than commercial liposome. PLGA nanocomplexes obviously accumulate in tumor cells after transfection, which indicate that the complexes contribute to cellular uptake of pDNA and pronouncedly enhance the treatment effect of miR-26a by inducing cell cycle arrest. Therefore, these results demonstrate that PLGA/PEI nanoparticles are promising non-viral vectors for gene delivery.

No MeSH data available.


Related in: MedlinePlus