Effects of cationic lipids in cationic liposomes and disaccharides in the freeze-drying of siRNA lipoplexes on gene silencing in cells by reverse transfection

2019 ◽  
Vol 30 (3) ◽  
pp. 235-245
Author(s):  
Yoshiyuki Hattori ◽  
Subin Hu ◽  
Hiraku Onishi
Keyword(s):  
Gene Silencing ◽  
Freeze Drying ◽  
Cationic Liposomes ◽  
Cationic Lipids ◽  
Reverse Transfection ◽  
In Cells

scholarly journals Сationic liposomes as delivery systems for nucleic acids

2020 ◽  
Vol 15 (1) ◽  
pp. 7-27
Author(s):  
A. A. Mikheev ◽  
E. V. Shmendel ◽  
E. S. Zhestovskaya ◽  
G. V. Nazarov ◽  
M. A. Maslov
Keyword(s):  
Gene Therapy ◽  
Nucleic Acids ◽  
Serum Proteins ◽  
Transfection Efficiency ◽  
Biological Fluids ◽  
Genetic Material ◽  
Cationic Liposomes ◽  
Delivery Systems ◽  
Cationic Lipids

Objectives. Gene therapy is based on the introduction of genetic material into cells, tissues, or organs for the treatment of hereditary or acquired diseases. A key factor in the success of gene therapy is the development of delivery systems that can efficiently transfer genetic material to the place of their therapeutic action without causing any associated side effects. Over the past 10 years, significant effort has been directed toward creating more efficient and biocompatible vectors capable of transferring nucleic acids (NAs) into cells without inducing an immune response. Cationic liposomes are among the most versatile tools for delivering NAs into cells; however, the use of liposomes for gene therapy is limited by their low specificity. This is due to the presence of various biological barriers to the complex of liposomes with NA, including instability in biological fluids, interaction with serum proteins, plasma and nuclear membranes, and endosomal degradation. This review summarizes the results of research in recent years on the development of cationic liposomes that are effective in vitro and in vivo. Particular attention is paid to the individual structural elements of cationic liposomes that determine the transfection efficiency and cytotoxicity. The purpose of this review was to provide a theoretical justification of the most promising choice of cationic liposomes for the delivery of NAs into eukaryotic cells and study the effect of the composition of cationic lipids (CLs) on the transfection efficiency in vitro.Results. As a result of the analysis of the related literature, it can be argued that one of the most promising delivery systems of NAs is CL based on cholesterol and spermine with the addition of a helper lipid DOPE. In addition, it was found that varying the composition of cationic liposomes, the ratio of CL to NA, or the size and zeta potential of liposomes has a significant effect on the transfection efficiency.Conclusions. Further studies in this direction should include optimization of the conditions for obtaining cationic liposomes, taking into account the physicochemical properties and established laws. It is necessary to identify mechanisms that increase the efficiency of NA delivery in vitro by searching for optimal structures of cationic liposomes, determining the ratio of lipoplex components, and studying the delivery efficiency and properties of multicomponent liposomes.

Ultracryotomy of biological tissues to preserve membrane structure

1976 ◽  
Vol 20 (3) ◽  
pp. 687-698
Author(s):  
S. Hodson ◽  
L. Williams
Keyword(s):  
Plasma Membrane ◽  
Biological Tissue ◽  
Membrane Structure ◽  
Freeze Drying ◽  
Biological Tissues ◽  
Ice Crystal ◽  
Transfer Stage ◽  
In Cells

A vacuum transfer stage is described which permits visualization of ultracryotome sections without the considerable distortions found in sections which have either thawed or rehydrated after freeze drying. Membrane structure-nucleus, nucleolus, mitochondria with their cristae and plasma membrane-was observed only in cells at the surface of the tissue which had undergone the fastest freezing rates. Inspection of knife damage in sections through these superficial cells showed that glass or diamond knives which are perfectly adequate for sectioning resin-embedded tissues are less suited to sectioning frozen biological tissue. Deeper in the tissue, where the freezing rates were slower, ice crystal cavities destroyed all membranous structures.

The development of tertiary amine cationic lipids for safe and efficient siRNA delivery

2019 ◽  
Vol 7 (7) ◽  
pp. 2777-2792 ◽  
Author(s):  
Ziming Lin ◽  
Moxyel Bao ◽  
Zexuan Yu ◽  
Lingjing Xue ◽  
Caoyun Ju ◽  
...  
Keyword(s):  
Tertiary Amine ◽  
Sirna Delivery ◽  
Cationic Lipid ◽  
Cationic Liposomes ◽  
Cationic Lipids

Tertiary amine-derived cationic lipid serves as the primary lipid of cationic liposomes, which can balance the effectiveness and safety of siRNA vectors.

Niosomes Containing Spermine-Based Cationic Lipid with Different Linkers for siRNA Delivery

2019 ◽  
Vol 819 ◽  
pp. 169-174
Author(s):  
Supusson Pengnam ◽  
Praneet Opanasopit ◽  
Theerasak Rojanarata ◽  
Nattisa Ni-yomtham ◽  
Boon Ek Yingyongnarongkul ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Cellular Uptake ◽  
Sirna Delivery ◽  
Weight Ratio ◽  
Cationic Lipid ◽  
Cationic Lipids ◽  
Molar Ratio ◽  
Ionic Surfactant ◽  
Low Toxicity ◽  
Cationic Niosomes

Niosomes are a lipid nanoparticle which have been widely used as non-viral carrier for therapeutic DNA or siRNA. They are formulated from non-ionic surfactant and other helper lipids. The aim of this study were to formulate niosome containing spermine-based cationic lipid with different linkers and to evaluate the efficiency of siRNA delivery in cervical cancer cell (HeLa cell). The niosomes were formulated from cholesterol (Chol), Span 20 and different cationic lipid (Ay, By, Cy and Dy) at various molar ratios. The properties of niosomes and ability of niosome to complex with siRNA were characterized. The cellular uptake, gene silencing efficiency and cytotoxicity were also determined. From the results, niosomes formulated at Chol:Span20:lipid molar ratio of 2.5:2.5:2 showed positive zeta potential and they were in nanosize (<200 nm). The binding ability of cationic niosomes to siRNA depended on types of cationic lipid. Among niosome/siRNA complexes, the niosome By/siRNA complex provided the highest gene silencing efficiency at weight ratio of 20. The highest cellular uptake also obtained by using niosome By as a carrier. The cytotoxicity revealed that cationic niosomes had low toxicity (cell viability > 80%). In conclusion, the cationic niosomes prepared from Chol, Span 20 and spermine-based cationic lipids are able to complex with siRNA and suitable for siRNA delivery with low toxicity.

scholarly journals Intracellular Distribution of Oligonucleotides Delivered by Cationic Liposomes: Light and Electron Microscopic Study1

1997 ◽  
Vol 45 (2) ◽  
pp. 265-274 ◽  
Author(s):  
Katriina Lappalainen ◽  
Riitta Miettinen ◽  
Jari Kellokoski ◽  
Ilpo Jääskeläinen ◽  
Stina Syrjänen
Keyword(s):  
Cellular Uptake ◽  
Ultrastructural Level ◽  
Cationic Liposomes ◽  
Intracellular Distribution ◽  
Cationic Lipids ◽  
Electron Microscopic ◽  
Net Charge ◽  
Transport Efficiency ◽  
Perinuclear Area

Synthesized oligonucleotides are used in anti-sense and anti-gene technology to control gene expression. Because cells do not easily take up oligonucleotides, cationic liposomes have been employed to facilitate their transport into cells. Athough cationic liposomes have been used in this way for several years, the precise mechanisms of the delivery of oligonucleotides into cells are not known. Because no earlier reports have been published on the liposomal delivery of oligonucleotides at the ultrastructural level, we performed a study, using electron microscopy, on the cellular uptake and intracellular distribution of liposomal digoxigenin-labeled oligodeoxynucleotides (ODNs) at several concentrations (0.1, 0.2, and 1.0 μM) in CaSki cells. Two cationic lipids (10 μM) were compared for transport efficiency: polycationic 2,3-dioleoyloxy- N-[2(sperminecarboxamido)ethyl]- N,N-dimethyl-1-propanaminium trifluoroacetate (DOSPA) and monocationic dimethyl-dioctadecylammonium bromide (DDAB). Both liposomes contained dioleoyl-phosphatidylethanolamine (DOPE) as a helper lipid. Endocytosis was found to be the main pathway of cellular uptake of liposomal ODNs. After release from intracellular vesicles, ODNs were carried into the perinuclear area. The nuclear membrane was found to be a barrier against the penetration of ODNs delivered by liposomes into the nucleus. Release from vesicles and transport into the nuclear area was faster when the oligo-DDAB/DOPE complex had a positive net charge (0.1 and 0.2 μM ODN concentrations), and only under this condition were some ODNs found in nucleoplasm. Although DOSPA/DOPE could also efficiently deliver ODNs into the cytosol, no ODNs were found in nucleoplasm. These findings suggest that both the type of liposome and the charge of the oligo-liposome complex are important for determination of the intracellular distribution of ODNs.

Effect of Acyl Chain Length of Spermine Derivatives on Transfection Efficiency

2012 ◽  
Vol 506 ◽  
pp. 445-448 ◽  
Author(s):  
Orapan Paecharoenchai ◽  
Nattisa Niyomtham ◽  
A. Apirakaramwong ◽  
B. Yingyongnarongkul ◽  
Praneet Opanasopit
Keyword(s):  
Transfection Efficiency ◽  
Gene Transfection ◽  
Acyl Chain ◽  
Weight Ratio ◽  
Cationic Liposomes ◽  
Cationic Lipids ◽  
Head Group ◽  
Chemical Structures ◽  
Cytotoxicity Studies ◽  
Cell Line Hela

In this study, cationic liposomes prepared from egg phosphatidylcholine (PC) and novel spermine-based cationic lipids at a molar ratio of 5:1 were formulated. The chemical structures of these cationic lipids consisted of spermine head group and four hydrocarbon tails with differences in acyl chain (C14, C16 and C18). The effects of acyl chain and weight ratio of liposomes to DNA on transfection efficiency and cytotoxicity were investigated on a human cervical carcinoma cell line (HeLa cells) using the pDNA encoding green fluorescent protein (pEGFP-C2). The results from agarose gel electrophoresis illustrated that all cationic liposomes were able to condense with pDNA. The transfection efficiency of these cationic liposomes was in the following order: C18 (3,497±120 cells/cm2) > C14 (809±52 cells/cm2) > C16 (91±5 cells/cm2). The highest transfection efficiency was observed in the formulation of cationic liposomes with C18 tail at weight ratio of 15. In cytotoxicity studies, all formulations showed low cytotoxicity. In conclusion, these cationic liposomes containing novel cationic lipids (C18), showed promising potential as a gene carrier by efficient DNA condensation and mediated higher level of gene transfection.

Sign in / Sign up

Export Citation Format

Share Document