Preparation and characterization of non-viral gene delivery systems with pEGFP-C1 Plasmid DNA

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.

Solid Lipid Nanoparticle Carrier Platform Containing Synthetic TLR4 Agonist Mediates Non-Viral DNA Vaccine Delivery

“…Lipoplex-A particle-DNA complexes were found to be of similar size but were notably more heterogenous in shape, with undefined edges. Previous studies have found that nanoparticle shape and size change upon DNA loading, which may be due to complete coating of nanoparticles thus obscuring the edges and giving the lipoplexes a quasi-spherical appearance [14,26]. Lipoplex-A samples also contained small aggregates of DNA visible as smaller particle contaminants in the sample, assumed to be a product of the TEM preparation process, owing to the wetting effect of the hydrophobic grid surface on unbound hydrophilic DNA as previously observed [27].…”

“…SLN-A particles also demonstrated their ability to bind and protect DNA from DNase degradation. The ability of lipid nanoparticles to protect DNA from nuclease degradation has been documented and is attributed to strong electrostatic interaction with DNA at the particle surface, preventing access to and degradation of bound DNA by nucleases [12,14,15].…”

“…Cationic solid lipid nanoparticles have been widely applied for gene delivery by condensing and delivering negatively charged DNA or RNA to form nanostructured complexes called ‘lipoplexes’. Solid lipid nanoparticle lipoplexes have been demonstrated to effectively deliver siRNA and plasmid DNA in vitro [ 9 , 12 , 13 , 14 , 15 , 16 ] but there are no reports of adjuvant-modified solid lipid nanoparticles (SLNs) as a DNA vaccine delivery system. Monophosphoryl lipid A (MPL-A) is a non-toxic chemically modified lipopolysaccharide (LPS) analogue which acts as a potent Toll-like receptor 4 (TLR4) agonist, and has been licensed for use in commercial vaccines Ceravix (Human Papillomavirus) and Shingrix (Herpes zoster virus) [ 17 , 18 ].…”

Nanocarriers-Mediated Drug Delivery Systems for Anticancer Agents: An Overview and Perspectives

“…The SLNs-based delivery system also has the potential for cellular-based drug directing by altering cellular signaling and gene expression. 58 Besides, SLNs are also deliberated as appropriate substitute carriers for parenteral delivery, 59 gene delivery, 60 pulmonary delivery, 61 and topical and transdermal delivery. 62 , 63 In this review, we will only discuss chemotherapeutic applications.…”

Antibiotikodun Lipido Solidozko Nanopartikulak bakterioen aurkako eraginkortasuna hobetzeko

“…SLNak ekoizteko nagusiki erabiltzen diren lipido solidoak[15,16,18]. SLNak ekoizteko nagusiki erabiltzen diren emultsionatzaile eta ko-emultsionatzaileak[15,16,18,[37][38][39][40].…”