Abstract
In gene delivery, non-viral vectors have become the preferred carrier system for DNA delivery. They can overcome major viral issues such as immunogenicity and mutagenicity. Cationic lipid-mediated gene transfer is one of the most commonly used non-viral vectors, which have been shown to be a safe and effective carrier. However, their use in gene delivery often exhibits low transfection efficiency and stability. The aim of this study was to examine the effectiveness of novel non-viral gene delivery systems. This study has investigated the encapsulation and transfection efficiency of cationic liposomes prepared from DOTAP and carboxymethyl-β-cyclodextrin (CD). The encapsulation efficiency of the CD-lipoplex complexes were also studied with and without the addition of Pluronic-F127, using both microfluidic and thin film hydration methods. In vitro transfection efficiencies of these complexes were determined in COS7 and SH-SY5Y cell lines. Formulation stability was evaluated using liposomes size, zeta potential and polydispersity index. In addition, the external morphology was studied using transmission electron microcopy (TEM). Results revealed that formulations produced by microfluidic method had smaller, more uniform and homogenious size and zeta-potential as well as higher encapsulation efficiency when compared with liposomes manufactured by thin film hydration method. Overall, the results of this study show that carboxymethyl-β-cyclodextrin increased lipoplexes’ encapsulation efficiency using both NanoAssemblr and rotary evaporator manufacturing processes. However, this increase was reduced slightly following the addition of Pluronic-F127. The addition of carboxymethyl-β-cyclodextrin to cationic liposomes resulted in an increase in transfection efficiency in mammalian cell lines. However, this increase appeared to be cell line specific, COS7 showed higher transfection efficiency compared to SH-SY5Y.
Highlights
IntroductionNon-viral vectors have become the preferred carrier system for DNA delivery
In gene delivery, non-viral vectors have become the preferred carrier system for DNA delivery
To investigate this further carboxymethyl-β-cyclodextrin (CD) was incorporated with cationic lipid (DOTAP), netural lipid (DOPE) and cholesterol to form liposomes to study the effect on gene cationic liposomes transfection and to evaluate the degree of gene encapsulation efficiency
Summary
Non-viral vectors have become the preferred carrier system for DNA delivery. Cationic lipid-mediated gene transfer is one of the most commonly used non-viral vectors, which have been shown to be a safe and effective carrier. Their use in gene delivery often exhibits low transfection efficiency and stability. The results of this study show that carboxymethyl-β-cyclodextrin increased lipoplexes’ encapsulation efficiency using both NanoAssemblr and rotary evaporator manufacturing processes This increase was reduced slightly following the addition of Pluronic-F127. Cycloextrin was included in this study, as cyclodextrins have prospective endosomal disrupting effects These effects might be through the release of membrane components from endosomal membranes after endocytosis Incorporation of CD with an excipient such as folic acid or Pluronic-F127 can positively affect the stability and effectiveness of gene formulations
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