Mechanisms of protection of cationic lipid-DNA complexes during lyophilization

Allison, S. Dean; Anchordoquy, Thomas J.

doi:10.1002/(sici)1520-6017(200005)89:5<682::aid-jps14>3.3.co;2-r

Cited by 16 publications

(48 citation statements)

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“…This lyoprotective effect may be explained by several mechanisms. These include prevention of mechanical damage and rupture of the lipid bilayer, maintenance of membrane flexibility (13,17), water replacement, prevention of aggregation due to diluting of complexes within the freezeconcentrated solutions or dried cake (17) and prevention of aggregation by formation of a steric barrier between particles by polymer such as polyethylene glycol (29,30).…”

Section: Gel Electrophoresismentioning

confidence: 99%

“…Most plasmid delivery systems including PEI, polysine, adenovirus and lipoplexes require lyo/cryoprotection to maintain transfection efficiency (7). Cryo/lyoprotectants limit mechanical damage and rupture of the lipid bilayer, caused by ice crystals, during the freeze-drying and the rehydration process by maintaining the membrane in a flexible state (13,17).…”

Section: Introductionmentioning

confidence: 99%

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Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

et al. 2007

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Abstract. The purpose of this research was to describe the application of lyophilization in the delivery of siRNA using cationic lipids by addressing the long-term formulation/stability issues associated with cationic lipids and to understand the mechanism of lyoprotection. siRNA liposomes complexes were formed in different potential cyro/lyoprotectants and subjected to either lyophilization or freeze thaw cycles. siRNA, liposomes and/or lipoplexes were tested for activity, SYBR Green I binding, cellular uptake and particle size. The lipoplexes when lyophilized in the presence of sugars as lyoprotectants could be lyophilized and reconstituted without loss of transfection efficacy but in ionic solutions they lost 65-75% of their functionality. The mechanism of this loss of activity was further investigated. The lyophilization process did not alter siRNA's intrinsic biological activity as was evident by the ability of lyophilized siRNA to retain functionality and SYBR green I binding ability. While the lipoplex size dramatically increased (∼50-70 times) after lyophilization in the absence of non-ionic lyoprotectants. This increase in size correlated to the decrease in cellular accumulation of siRNA and a decrease in activity. In conclusion, siRNAs can be applied in cationic lipid lyophilized formulations and these complexes represent a potential method of increasing the stability of pre-formed complex.

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Section: Gel Electrophoresismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

et al. 2007

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“…31 Lyophilization is a useful method for preserving nonviral gene vectors. 32,33 However, significantly elevated target gene expression levels were observed in freshly prepared pDNA/chitosan complexes compared to those in lyophilized pDNA/chitosan complexes. 10 Accordingly, lyophilized pDNA complexes were coated with CS to enhance their transfection efficiencies; the pDNA/chitosan/CS complexes exhibited high transfection abilities even after lyophilization.…”

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo gene delivery using chitosan/hyaluronic acid nanoparticles: Influences of molecular mass of hyaluronic acid and lyophilization on transfection efficiency

Sato

Nakata

Yang

et al. 2017

The Journal of Gene Medicine

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“…It would be desirable to manufacture ready‐made solutions for use in a clinical setting. However, solutions of pDNA complexed with non‐viral gene delivery vehicles including polycations lose their ability to transfect cells over time 5–8. An alternative strategy of combining non‐viral gene delivery carriers with plasmid DNA immediately prior to administration would be challenging to implement in a clinical setting.…”

Section: Introductionmentioning

confidence: 99%

“…An alternative strategy of combining non‐viral gene delivery carriers with plasmid DNA immediately prior to administration would be challenging to implement in a clinical setting. Lyophilization of pDNA complexes offers a potential response to these challenges by allowing for stable storage of dry complexes and highly reproducible in situ sample preparation 5–8…”

Section: Introductionmentioning

confidence: 99%

Polycation Structure Mediates Expression of Lyophilized Polycation/pDNA Complexes

Hahn

Kong

Mooney

2010

Macromolecular Bioscience

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Lyophilization of polycation/pDNA complexes provides stable, long-term storage of complexes prior to clinical use but also reduces gene delivery efficiency. We examined whether polycation structure mediates effects of lyophilization on gene expression. Linear and branched PEI of the same molecular weight were used as a model system. Interestingly, pDNA/linear PEI complexes led to much smaller effects on gene expression following lyophilization compared with branched PEI complexes. The effect of polycation structure correlated with changes in dissociation ability of pDNA/PEI complexes. These results will be useful for developing new gene delivery vehicles.

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Mechanisms of protection of cationic lipid-DNA complexes during lyophilization

Cited by 16 publications

References 0 publications

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

Effect of Lyophilization and Freeze-thawing on the Stability of siRNA-liposome Complexes

In vitro and in vivo gene delivery using chitosan/hyaluronic acid nanoparticles: Influences of molecular mass of hyaluronic acid and lyophilization on transfection efficiency

Polycation Structure Mediates Expression of Lyophilized Polycation/pDNA Complexes

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