Evaluation of novel cationic gene based liposomes with cyclodextrin prepared by thin film hydration and microfluidic systems

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.

Effects of Liposome and Cardiolipin on Folding and Function of Mitochondrial Erv1

“…The average diameter of the liposomes in each formulation was measured using dynamic light scattering ( Figure 2 B) and was found to be the same, with or without the presence of cardiolipin. The mean diameter was 160 ± 1 nm, which was as expected, based on previous results [ 35 ].…”

All-in-one microfluidic assembly of insulin-loaded pH-responsive nano-in-microparticles for oral insulin delivery

“…[22][23][24] However, even with high potential to deliver proteins, conventional liposomes are still likely to disintegrate under gastric conditions 25,26 and can show poor permeation across the intestinal epithelial cell membrane, 27 thus resorting to coatings with other molecules, such as polymers 28,29 or protein corona. 30,31 Yet, liposomes are usually prepared by bulk methods such as thin-film hydration, 32,33 ethanol injection, 34 and reverse phase evaporation, 35,36 in which some of them require a post-processing step to obtain a better control of size and polydispersity index (PDI), such as high pressure extrusion. 37 Microfluidics, a sophisticated technique for particle production, has gained substantial attention to prepare liposomes.…”

Diflunisal‐loaded poly(propylene sulfide) nanoparticles decrease S. aureus‐mediated bone destruction during osteomyelitis

“…After determining optimal parameters for drug loading, nanoparticles were fabricated in large batches by microfluidics processes as described previously for animal studies 33 . Briefly, PPS 135 ‐ b ‐p(Cy7 1 ‐ ran ‐DMA 149 ) (60.0 mg) was codissolved with pHPMA‐Bz (6.0 mg) and/or diflunisal (6.0 mg) in methanol (0.6 ml) and mixed with sterile PBS using a benchtop NanoAssemblr (Precision Nanosystems, Inc.).…”

“…18,21,32 After determining optimal parameters for drug loading, nanoparticles were fabricated in large batches by microfluidics processes as described previously for animal studies. 33 sessed as conducted previously. 34 In total, 86 animals were used to complete these studies.…”