Abstract
In recent years, non-viral delivery systems for plasmid DNA have become particularly important. They can overcome the disadvantages of viral systems such as insertional mutagenesis and unpredicted immunogenicity. Some additional advantages of non-viral gene delivery systems are; good stability, low cost, targetability, delivery of a high amount of genetic materials. The aim of the study was to develop novel non-viral nanosystems suitable for gene delivery. Two formulations were developed for this purpose: water-in-oil microemulsion (ME) and solid lipid nanoparticles (SLN). The microemulsion was composed of Peceol, Tween 80, Plurol oleique, ethanol and water. The SLN was consisting of Precirol, Esterquat-1 (EQ1), Tween 80, Lecithin, ethanol and water. Characterization studies were carried out by measuring particle size, zeta potential, viscosity and pH. TEM imaging was performed on SLN formulations. Protection against DNase I degradation was examined. Cytotoxicity and transfection efficacy of selected formulations were tested on L929 mouse fibroblast cells. Particle sizes of complexes were below 100 nm and with high positive zeta potential. TEM images revealed that SLNs are spherical. The SLN:DNA complexes have low toxicity and good transfection ability. All results showed that the developed SLN formulations can be considered as suitable non-viral gene delivery systems.
Highlights
Gene therapy using plasmid DNA molecules for specific diseases gained great importance in last decades
Ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), Didecyldimethylammonium bromide (DDAB), Cetyl trimethylammonium bromide (CTAB), Tween 80, Chitosan, Phenol, 1-propanol and Span 80 were purchased from Sigma Aldrich GmbH (Steinheim, Germany)
Ethidium bromide was purchased from Fluka Chemie GmbH (Steinheim, Germany). pEGFP-C1 plasmid DNA and maxiprep plasmid DNA purification kit were purchased from Invitrogen (Carlsbad, CA, USA)
Summary
Gene therapy using plasmid DNA molecules for specific diseases gained great importance in last decades. The main purpose of gene therapy is replacing diseased genes with new healthy ones. Selection of a proper gene delivery system is a very important issue when considering gene therapy because naked plasmid DNA (pDNA) is rapidly degraded following administration to the human body (Park et al, 2015). There are several methods developed for gene delivery. These methods can be divided into two main groups; 1) methods applying viral delivery systems, 2) methods applying non-viral delivery systems. Viral gene delivery systems have been the main choice during early gene therapy trials (Jomary et al, 1994; Edelstein et al, 2004).
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