Effects of a Delocalizable Cation on the Headgroup of Gemini Lipids on the Lipoplex-Type Nanoaggregates Directly Formed from Plasmid DNA
| Autor: | Mónica Muñoz-Úbeda, Emilio Aicart, Sougata Datta, Clara Aicart-Ramos, Santanu Bhattacharya, Pablo Castro-Hartmann, Paturu Kondaiah, Elena Junquera, Ana L. Barrán-Berdón, Santosh K. Misra |
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| Rok vydání: | 2013 |
| Předmět: |
Polymers and Plastics
Surface Properties Stereochemistry Biocompatible Materials Bioengineering Effective nuclear charge Cell Line Biomaterials Cations Materials Chemistry Zeta potential Humans Particle Size Gel electrophoresis Liposome Molecular Structure Chemistry Small-angle X-ray scattering DNA Lipids Fluorescence Nanostructures Crystallography HEK293 Cells Lipofectamine Liposomes lipids (amino acids peptides and proteins) Fluorescence anisotropy HeLa Cells Plasmids |
| Zdroj: | Biomacromolecules. 14:3951-3963 |
| ISSN: | 1526-4602 1525-7797 |
| Popis: | Lipoplex-type nanoaggregates prepared from pEGFP-C3 plasmid DNA (pDNA) and mixed liposomes, with a gemini cationic lipid (CL) 1,2-bis(hexadecyl imidazolium) alkanes], referred as (C(16)Im)(2)C-n (where C-n is the alkane spacer length, n = 2, 3, 5, or 12, between the imidazolium heads) and DOPE zwitterionic lipid, have been analyzed by zeta potential, gel electrophoresis, SAXS, cryo-TEM, fluorescence anisotropy, transfection efficiency, fluorescence confocal microscopy, and cell viability/cytotoxicity experiments to establish a structure-biological activity relationship. The study, carried out at several mixed liposome compositions, alpha, and effective charge ratios, rho(eff), of the lipoplex, demonstrates that the transfection of pDNA using CLs initially requires the determination of the effective charge of both. The electrochemical study confirms that CLs with a delocalizable positive charge in their headgroups yield an effective positive charge that is 90% of their expected nominal one, while pDNA is compacted yielding an effective negative charge which is only 10-25% than that of the linear DNA. SAXS diffractograms show that lipoplexes formed by CLs with shorter spacer (n = 2, 3, or 5) present three lamellar structures, two of them in coexistence, while those formed by CL with longest spacer (n = 12) present two additional inverted hexagonal structures. Cryo-TEM micrographs show nanoaggregates with two multilamellar structures, a cluster-type (at low alpha value) and a fingerprint-type, that coexist with the cluster-type at moderate alpha composition. The optimized transfection efficiency (TE) of pDNA, in HEK293T, HeLa, and H1299 cells was higher using lipoplexes containing gemini CLs with shorter spacers at low a value. Each lipid formulation did not show any significant levels of toxicity, the reported lipoplexes being adequate DNA vectors for gene therapy and considerably better than both Lipofectamine 2000 and CLs of the 1,2-bis(hexadecyl ammnoniun) alkane series, recently reported. |
| Databáze: | OpenAIRE |
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