Optimization of stearylamine-based nanoemulsions obtained by spontaneous emulsification process as nucleic acids delivery systems

Fraga, Michelle; Laux, Manoela; Zandoná, Bernardo Remuzzi; Santos, Gabriella Rejane dos; Giuberti, Cristiane dos Santos; Oliveira, Mônica Cristina; Matte, Úrsula; Teixeira, Hélder Ferreira

doi:10.1016/s1773-2247(08)50078-5

Cited by 10 publications

(15 citation statements)

References 30 publications

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“…6,15 Following up on these results, in this study we investigated the effect of different isolated phospholipids on the physicochemical properties, cytotoxicity, and transfection of nanoemulsions/DNA plasmid complexes on Hep G2 cells.…”

Section: Discussionmentioning

confidence: 99%

“…displacement methods seems to be mainly influenced by the diffusion speed of the organic solvent in water. 6,18 In the final pH of the formulations, the zeta potential, which was stabilized by zwitterionic PC or phosphatidylethanolamine in a binary mixture with SA, was highly positive (.50 mV). Formulations containing isolated phospholipids showed significantly higher zeta potentials than that detected for the PC/SA formulation.…”

Section: Discussionmentioning

confidence: 99%

“…2,4,5 Nonviral vectors have important safety advantages over viral approaches, including reduced pathogenicity and capacity for insertional mutagenesis, as well as low cost and ease of production. 6,7 However, application of non viral vectors has been held back by their poor efficiency in delivery to cells and transient expression of transgenes. Therefore, new strategies are needed for the application of these nonviral vectors as nucleic acid delivery systems.…”

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confidence: 99%

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Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells

Fraga¹,

Bruxel²,

Lagranha³

et al. 2011

IJN

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Background Cationic nanoemulsions have been recently considered as potential delivery systems for nucleic acids. This study reports the influence of phospholipids on the properties of cationic nanoemulsions/DNA plasmid complexes. Methods Nanoemulsions composed of medium-chain triglycerides, stearylamine, egg lecithin or isolated phospholipids, ie, DSPC, DOPC, DSPE, or DOPE, glycerol, and water were prepared by spontaneous emulsification. Gene transfer to Hep G2 cells was analyzed using real-time polymerase chain reaction. Results The procedure resulted in monodispersed nanoemulsions with a droplet size and zeta potential of approximately 250 nm and +50 mV, respectively. The complexation of cationic nanoemulsions with DNA plasmid, analyzed by agarose gel retardation assay, was complete when the complex was obtained at a charge ratio of ≥1.0. In these conditions, the complexes were protected from enzymatic degradation by DNase I. The cytotoxicity of the complexes in Hep G2 cells, evaluated by MTT assay, showed that an increasing number of complexes led to progressive toxicity. Higher amounts of reporter DNA were detected for the formulation obtained with the DSPC phospholipid. Complexes containing DSPC and DSPE phospholipids, which have high phase transition temperatures, were less toxic in comparison with the formulations obtained with lecithin, DOPC, and DOPE. Conclusion The results show the effect of the DNA/nanoemulsion complexes composition on the toxicity and transfection results.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells

Fraga¹,

Bruxel²,

Lagranha³

et al. 2011

IJN

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“…The ratio of the anti-cancer drug to conjugate with the QDs is 1,500(epirubicin):1(QDs). A series of cytotoxicity testes are conducted and verified for the QDs, pure epirubicin and the nano drugs using a dimythylthiazol diphenyl tetrazolium bromide (MTT) assay (Fraga et al 2008) at different treatment dosages (0M, 10 −11 M, 10 −10 M, 10 −9 M, 10 −8 M for the QDs and the nano drugs and 0M, 1.5 × 10 −8 M, 1.5 × 10 −7 M, 1.5 × 10 −6 M, 1.5×10 −5 M for the epirubicin). The MTT assay is conducted as described previously.…”

Section: Sample Preparationmentioning

confidence: 99%

A microfluidic cell culture platform for real-time cellular imaging

Hsieh

Huang

et al. 2009

Biomed Microdevices

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This study reports a new microfluidic cell culture platform for real-time, in vitro microscopic observation and evaluation of cellular functions. Microheaters, a micro temperature sensor, and micropumps are integrated into the system to achieve a self-contained, perfusion-based, cell culture microenvironment. The key feature of the platform includes a unique, ultra-thin, culture chamber with a depth of 180 mum, allowing for real-time, high-resolution cellular imaging by combining bright field and fluorescent optics to visualize nanoparticle-cell/organelle interactions. The cell plating, culturing, harvesting and replenishing processes are performed automatically. The developed platform also enables drug screening and real-time, in situ investigation of the cellular and sub-cellular delivery process of nano vectors. The mitotic activity and the interaction between cells and the nano drug carriers (conjugated quantum dots-epirubicin) are successfully monitored in this device. This developed system could be a promising platform for a wide variety of applications such as high-throughput, cell-based studies and as a diagnostic cellular imaging system.

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“…Non-viral vectors have important safety advantages over viral approaches, including their reduced pathogenicity and capacity for insertional mutagenesis, as well as their low cost and ease of production (Fraga et al, 2008). The application of non-viral vectors to humans has, however, been held back by the poor efficiency of their delivery to cells and the transient expression of the transgenes.…”

Section: Non-viral Approachesmentioning

confidence: 99%