Understanding the mechanism of protamine in solid lipid nanoparticle-based lipofection: The importance of the entry pathway

Delgado, Diego; Pozo-Rodríguez, Ana del; Solinís, Maria Ángelés; Rodríguez-Gascón, Alicia

doi:10.1016/j.ejpb.2011.06.005

Cited by 90 publications

(83 citation statements)

References 36 publications

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“…In preceding studies we have developed nonviral vectors composed of cationic lipids, more precisely solid lipid nanoparticles (SLNs) (del Pozo-Rodríguez et al, 2009;Delgado et al, 2011;Gascó n et al, 2011). These SLNs have shown the capacity to transfect in vitro retinal pigment epithelium (RPE) cells, used as a model for inherited retinal diseases (del PozoRodríguez et al, 2008).…”

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confidence: 99%

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

et al. 2012

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The goal of the present study was to analyze the potential application of nonviral vectors based on solid lipid nanoparticles (SLN) for the treatment of ocular diseases by gene therapy, specifically X-linked juvenile retinoschisis (XLRS). Vectors were prepared with SLN, dextran, protamine, and a plasmid (pCMS-EGFP or pCEP4-RS1). Formulations were characterized and the in vitro transfection capacity as well as the cellular uptake and the intracellular trafficking were studied in ARPE-19 cells. Formulations were also tested in vivo in Wistar rat eyes, and the efficacy was studied by monitoring the expression of enhanced green fluorescent protein (EGFP) after intravitreal, subretinal, and topical administration. The presence of dextran and protamine in the SLN improved greatly the expression of retinoschisin and EGFP in ARPE-19 cells. The nuclear localization signals of protamine, its ability to protect the DNA, and a shift in the entry mechanism from caveola-mediated to clathrinmediated endocytosis promoted by the dextran, justify the increase in transfection. After ocular administration of the dextran-protamine-DNA-SLN complex to rat eyes, we detected the expression of EGFP in various types of cells depending on the administration route. Our vectors were also able to transfect corneal cells after topical application. We have demonstrated the potential usefulness of our nonviral vectors loaded with XLRS1 plasmid and provided evidence for their potential application for the management or treatment of degenerative retinal disorders as well as ocular surface diseases.

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Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

et al. 2012

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“…As shown in Figure 7, green color of NBD-PC NLCs was localized in cytoplasm, whereas endosomes were stained and visualized as red fluorescence from LysoTracker Red. Overlap of red and green fluorescence was observed as yellow color in cells incubated with NBD-PC-NLCs for 30 min, which showed that the nanoparticles had penetrated rapidly and were localized in the endosomes after internalization following postulated endocytosis mechanisms (Delgado et al, 2011).…”

Section: Cellular Uptake and In Vitro Cytotoxicitymentioning

confidence: 99%

Development of lipid nanoparticles for a histone deacetylases inhibitor as a promising anticancer therapeutic

et al. 2014

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Background: Vorinostat (VRS), a histone deacetylases inhibitor, has significant cytotoxic potential in a large number of human cancer cell lines. Objective: To clarify its promising anticancer potential and to improve its drawback related to physical properties and in vivo performance of VRS. Methods: VRS was successfully incorporated into nanostructured lipid carriers (NLCs) by the hot microemulsion method using sonication following a homogenization technique. Results: After the optimization process, VRS-loaded NLCs (VRS-NLCs) were obtained as ideal quality nanoparticles with a spherical shape, small size ($150 nm), negative charge ($À22 mV), and narrow size distribution. In addition, the high entrapment efficiency ($99%) and sustained drug release profile were recorded. Cytotoxicity study in three different cell lines (A549, MCF-7, and SCC-7) demonstrated higher cytotoxicity of VRS-NLCs than free drug. Finally, the AUC of VRS (118.16 ± 17.35 mgh/mL) was enhanced $4.4 times compared with that of free drug (27.03 ± 3.25 mgh/mL). Conclusion: These results suggest the potential of NLCs as an oral delivery system for enhancement of cellular uptake, in vitro cytotoxicity in cancer cell lines and the oral bioavailability of VRS.

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“…[19][20][21][22] The stable nanoparticles were formed by addition of protamine to pDNA at various theoretical charge ratios ( Table 1). As the charge ratio of protamine was increased, the zeta potential of pDNA/protamine complexes also increased.…”

Section: Discussionmentioning

confidence: 99%

Ternary Complex of Plasmid DNA with Protamine and γ-Polyglutamic Acid for Biocompatible Gene Delivery System

Kanda

Kodama

Kurosaki

et al. 2013

Biological & Pharmaceutical Bulletin

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The purpose of the present study was to investigate the usefulness of the ternary complex with protamine and γ-polyglutamic acid (γ-PGA), which are biodegradable materials for foods and medical products, as a safe gene delivery vector. We formed cationic binary complexes (plasmid DNA (pDNA)/protamine complexes) with high transfection efficiency. The binary complex showed slight toxicity probably related to its total cationic charge. Then, we formed ternary complexes (pDNA/protamine/γ-PGA complexes) by addition of anionic polymer, γ-PGA, and they showed no cytotoxicity. The transfection efficiency of the pDNA/protamine/γ-PGA complexes was as high as that of the pDNA/protamine complexes, although their zeta potentials were different. Inhibition study of the gene expressions in B16-F10 cells suggested that pDNA/protamine complexes were taken up by caveolae-mediated endocytosis and macropinocytosis. On the other hand, pDNA/protamine/ γ-PGA complexes were taken up by clathrin-mediated endocytosis and macropinocytosis. Thus, we succeeded in developing the ternary complex as a safe gene delivery vector with biocompatible materials.

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Understanding the mechanism of protamine in solid lipid nanoparticle-based lipofection: The importance of the entry pathway

Cited by 90 publications

References 36 publications

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

Development of lipid nanoparticles for a histone deacetylases inhibitor as a promising anticancer therapeutic

Ternary Complex of Plasmid DNA with Protamine and γ-Polyglutamic Acid for Biocompatible Gene Delivery System

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