Employment of cationic solid-lipid nanoparticles as RNA carriers.
Gene transfer represents an important advance in the treatment of both genetic and acquired diseases. In this article, the suitability of cationically modified solid-lipid nanoparticles (SLN) as a nonviral vector for gene delivery was investigated, in order to obtain stable materials able to condense RNA. Cationic SLN were produced by microemulsion using Compritol ATO 888 as matrix lipid, Pluronic F68 as tenside, and dimethyldioctadecylammonium bromide (DDAB) as cationic lipid. The resulting particles were approximately 100 nm in size and showed a highly positive surface charge (+41 mV) in water. Size and shape were further characterized by scanning electron microscopy (SEM) measurements. Moreover, we utilized the sea urchin as a model system to test their applicability on a living organism. To evaluate cationic SLN ability to complex the in vitro transcribed Paracentrotus lividus bep3 RNA, we utilized both light scattering and gel mobility experiments, and protection by nuclease degradation was also investigated. By microinjection experiment, we demonstrated that the nanoparticles do not inference with the viability of the P. lividus embryo and the complex nanoparticles-bep3 permits movement of the RNA during its localization in the egg, suggesting that it could be a suitable system for gene delivery. Taken together, all these results indicate that the cationic SNL are a good RNA carrier for gene transfer system and the sea urchin a simple and versatile candidate to test biological properties of nanotechnology devices.
Montana
Giovanna
G
Istituto di Biomedicina e Immunologia Molecolare, Istituto per lo Studio dei Materiali Nanostrutturati, sezione di Palermo, Palermo, Italy.
Bondì
Maria L
ML
Carrotta
Rita
R
Picone
Pasquale
P
Craparo
Emanuela F
EF
San Biagio
Pier L
PL
Giammona
Gaetano
G
Di Carlo
Marta
M
D000818:Animals; D002412:Cations / Q000737:chemistry*; D002470:Cell Survival; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D004655:Emulsions; D005227:Fatty Acids / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008845:Microinjections; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles; D036103:Nanotechnology; D010063:Ovum / Q000378:metabolism; D020442:Poloxamer / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012313:RNA / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology; D012617:Sea Urchins / Q000196:embryology; D014162:Transfection
D002412:Cations; D004337:Drug Carriers; D004655:Emulsions; D005227:Fatty Acids; D008055:Lipids; D008565:Membrane Proteins; D000644:Quaternary Ammonium Compounds; C105368:bep3 protein, Paracentrotus lividus; D020442:Poloxamer; C086409:glyceryl behenate; C015831:dimethyldioctadecylammonium; D012313:RNA