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

Nanoscale Research Letters, Dec 2011

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.

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

Nanoscale Research Letters PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells Gao Feng Liang 0 Yan Liang Zhu 0 Bo Sun 0 Fei Hu Hu 0 Tian Tian 0 Shu Chun Li 0 Zhong Dang Xiao 0 0 State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University , Nanjing, 210096 , China 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 liposomebased 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. - Introduction MicroRNAs (miRNAs) are small, highly conserved, noncoding RNAs that regulate gene expression at the posttranscriptional level. They involve in various cellular mechanisms including development, differentiation, proliferation, and apoptosis. The pivotal roles of these miRNAs in human cancers have been discovered [1,2], and the therapeutic applications of miRNA have been developed using various viral vectors [3,4]. However, the disadvantages of viral vectors limited their application in gene delivery, such as immunogenic/ inflammatory responses, low loading capacity, large scale manufacturing, and quality control [5]. Consequently, more attention have been paid on non-viral gene delivery vectors in recent years, such as liposomes (lipoplexes), polycationic polymers (polyplexes), and organic or inorganic nanoparticles (nanoplexes) [6]. To enhance gene delivery effect, various cationic complexes have been developed for delivering plasmid DNA, antisense, or siRNA into cells [7-9]. Poly(D,L-lactide-co-glycolide) (PLGA) were extensively assessed for their ability of delivering variety of therapeutic agents [10-12]. PLGA nanoparticles were shown to escape from the endo-lysosomal compartment to the cytoplasmic compartment and release their contents over extended periods of time [13]. These features rendered PLGA nanoparticles as potential tool for gene delivery efficiently. Polyethylenimine (PEI) is water-soluble, linear, or branched polymers with a protonable amino group [14,15]. Due to their high cationic charge density at physiological pH, PEIs are able to form non-covalent complexes with DNA, siRNA, and antisense oligodeoxynucleotide. Therefore, PEIs hold a prominent position among the polycationic polymers used for gene delivery [16-18]. The intracellular release of PEI/nucleic acids complexes from endosomes is considered as relying on the protonation of amines in the PEI molecule, which leading to osmotic swelling and subsequent burst of the endosomes. Moreover, PEIs also facilitate nucleic acid entry into the nucleus [19,20]. However, it has been reported that long PEI chains are highly effective in gene transfection, but more cytotoxic [14,21,22]. In order to overcome these hurdles in gene therapy and improve gene delivery efficiency, we developed novel non-liposome-based cationic polymers which are composed of PLGA as the core and cationic PEI as the shell. The biodegradable PLGA nanoparticles, modified with a polyplexed PEI coating, were tested by loading the expression vector (pDNA) of miR-26a, which is capable of inducing cell cycle arrest in HepG2 cells. In this study, nanoparticles of controlled size and persistent shape have been obtained by an emulsion evaporation method and characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), and energy dispersive spectroscopy (...truncated)


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Gao Feng Liang, Yan Liang Zhu, Bo Sun, Fei Hu Hu, Tian Tian, Shu Chun Li, Zhong Dang Xiao. PLGA-based gene delivering nanoparticle enhance suppression effect of miRNA in HePG2 cells, Nanoscale Research Letters, 2011, pp. 447, Volume 6, Issue 1, DOI: 10.1186/1556-276X-6-447