Abstract
Cationic lipid-mediated gene transfer is one of the most commonly used non-viralvectors. It has been shown to be a safe and effective carrier. However, its use ingene delivery was hampered by its low transfection efficiency and stability.DOTAP, DOPE, cholesterol (CHO) and carboxymethyl-β-cyclodextrin (CD) wereused to prepare cationic liposomes. Cationic liposomes were prepared using both,thin film hydration and a microfluidic method. Formulation stability wasevaluated using liposome size, zeta potential and polydispersity index (PDI).Promega QuantiFluor® ONE dsDNA System was used to investigate theencapsulation efficiency. COS7 and SH-SY5Y cell lines were used to determinetransfection efficiency. Results show that carboxymethyl-β-cyclodextrin increasedencapsulation efficiency by 15.5% and 8% using NanoAssemblr® and rotaryevaporator, respectively compared to liposomes without CD. The addition ofcarboxymethyl-β-cyclodextrin to cationic liposomes resulted in an increase intransfection efficiency in both cell lines.B
Highlights
Liposomes are a form of spherical vesicles that consist either of one, few or many phospholipid bilayers interacting with one another in an energetically favourable manner
Cationic liposomes transfection efficiency can be affected by the composition of the transfection reagent, liposome size and zeta potential, as well as liposome to DNA ratio
Results of lipoplexes were close to the value obtained from liposomes alone; there was a noticeable reduction in the zeta potential following the addition of pDNA
Summary
Cationic lipid-mediated gene transfer is one of the most commonly used non-viral vectors. Its use in gene delivery was hampered by its low transfection efficiency and stability. DOTAP, DOPE, cholesterol (CHO) and carboxymethyl-β-cyclodextrin (CD) were used to prepare cationic liposomes. Cationic liposomes were prepared using both, thin film hydration and a microfluidic method. Formulation stability was evaluated using liposome size, zeta potential and polydispersity index (PDI). Promega QuantiFluor® ONE dsDNA System was used to investigate the encapsulation efficiency. COS7 and SH-SY5Y cell lines were used to determine transfection efficiency. Results show that carboxymethyl-β-cyclodextrin increased encapsulation efficiency by 15.5% and 8% using NanoAssemblrTM and rotary evaporator, respectively compared to liposomes without CD. The addition of carboxymethyl-β-cyclodextrin to cationic liposomes resulted in an increase in transfection efficiency in both cell lines
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