Characterization of monoolein-based lipoplexes using fluorescence spectroscopy

Bibliographic Details
Main Author: Silva, J. P. Neves
Publication Date: 2008
Other Authors: Coutinho, Paulo J. G., Oliveira, M. Elisabete
Format: Article
Language: eng
Source: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Download full: http://hdl.handle.net/1822/9409
Summary: Lipoplexes are commonly used as delivery systems in vitro and in vivo, the role of a neutral lipid as helper being of extreme importance in these systems. Cationic liposomes composed of dioctadecyldimethylammonium bromide (DODAB) with monoolein (MO) as a helper, at different molar ratios (1:2; 1:1 and 1:0.5) were prepared, and subsequently titrated to DNA. The structural and physicochemical properties of the lipid/DNA complexes were assessed by Ethidium Bromide (EtBr) exclusion, 90º Static Light Scattering (90º SLS) assays and Fluorescence Resonance Energy Transfer (FRET). In EtBr exclusion assays, the steady-state fluorescence spectra of EtBr were decomposed into the sum of two lognormal emissions, emanating from two different environments – H2O and DNA, and the effect of charge ratio (+/-) was observed. 90º SLS assays gave an important contribution, detecting size variations in systems with different MO fractions on the lipoplexes. In FRET assays, 2-(3-(diphenylhexatrienyl)propanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (DPH-HPC) was used as donor and EtBr as acceptor. The DNA component previously calculated by EtBr exclusion, was used to determine the energy transfer efficiency, as an indirect measurement of the lipoplexes structural and physicochemical properties. Our results demonstrate that the inclusion of monoolein in the cationic liposomes formulation significantly modifies the rate of DNA complexation, being DODAB:MO (1:1) the system with higher DNA condensation efficiency.
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spelling Characterization of monoolein-based lipoplexes using fluorescence spectroscopyMonoolein-based lipoplexesEtBr exclusionFRET90º SLS90 degrees SLSScience & TechnologyLipoplexes are commonly used as delivery systems in vitro and in vivo, the role of a neutral lipid as helper being of extreme importance in these systems. Cationic liposomes composed of dioctadecyldimethylammonium bromide (DODAB) with monoolein (MO) as a helper, at different molar ratios (1:2; 1:1 and 1:0.5) were prepared, and subsequently titrated to DNA. The structural and physicochemical properties of the lipid/DNA complexes were assessed by Ethidium Bromide (EtBr) exclusion, 90º Static Light Scattering (90º SLS) assays and Fluorescence Resonance Energy Transfer (FRET). In EtBr exclusion assays, the steady-state fluorescence spectra of EtBr were decomposed into the sum of two lognormal emissions, emanating from two different environments – H2O and DNA, and the effect of charge ratio (+/-) was observed. 90º SLS assays gave an important contribution, detecting size variations in systems with different MO fractions on the lipoplexes. In FRET assays, 2-(3-(diphenylhexatrienyl)propanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (DPH-HPC) was used as donor and EtBr as acceptor. The DNA component previously calculated by EtBr exclusion, was used to determine the energy transfer efficiency, as an indirect measurement of the lipoplexes structural and physicochemical properties. Our results demonstrate that the inclusion of monoolein in the cationic liposomes formulation significantly modifies the rate of DNA complexation, being DODAB:MO (1:1) the system with higher DNA condensation efficiency.Fundação para a Ciência e a Tecnologia (FCT)SpringerUniversidade do MinhoSilva, J. P. NevesCoutinho, Paulo J. G.Oliveira, M. Elisabete2008-032008-03-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/9409eng“Journal of Fluorescence”. ISSN 1053-0509. 18:2 (2008) 555-562.1053-050910.1007/s10895-007-0299-518157688www.springerlink.cominfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:27:29Zoai:repositorium.sdum.uminho.pt:1822/9409Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:22:04.792476Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
title Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
spellingShingle Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
Silva, J. P. Neves
Monoolein-based lipoplexes
EtBr exclusion
FRET
90º SLS
90 degrees SLS
Science & Technology
title_short Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
title_full Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
title_fullStr Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
title_full_unstemmed Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
title_sort Characterization of monoolein-based lipoplexes using fluorescence spectroscopy
author Silva, J. P. Neves
author_facet Silva, J. P. Neves
Coutinho, Paulo J. G.
Oliveira, M. Elisabete
author_role author
author2 Coutinho, Paulo J. G.
Oliveira, M. Elisabete
author2_role author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Silva, J. P. Neves
Coutinho, Paulo J. G.
Oliveira, M. Elisabete
dc.subject.por.fl_str_mv Monoolein-based lipoplexes
EtBr exclusion
FRET
90º SLS
90 degrees SLS
Science & Technology
topic Monoolein-based lipoplexes
EtBr exclusion
FRET
90º SLS
90 degrees SLS
Science & Technology
description Lipoplexes are commonly used as delivery systems in vitro and in vivo, the role of a neutral lipid as helper being of extreme importance in these systems. Cationic liposomes composed of dioctadecyldimethylammonium bromide (DODAB) with monoolein (MO) as a helper, at different molar ratios (1:2; 1:1 and 1:0.5) were prepared, and subsequently titrated to DNA. The structural and physicochemical properties of the lipid/DNA complexes were assessed by Ethidium Bromide (EtBr) exclusion, 90º Static Light Scattering (90º SLS) assays and Fluorescence Resonance Energy Transfer (FRET). In EtBr exclusion assays, the steady-state fluorescence spectra of EtBr were decomposed into the sum of two lognormal emissions, emanating from two different environments – H2O and DNA, and the effect of charge ratio (+/-) was observed. 90º SLS assays gave an important contribution, detecting size variations in systems with different MO fractions on the lipoplexes. In FRET assays, 2-(3-(diphenylhexatrienyl)propanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine (DPH-HPC) was used as donor and EtBr as acceptor. The DNA component previously calculated by EtBr exclusion, was used to determine the energy transfer efficiency, as an indirect measurement of the lipoplexes structural and physicochemical properties. Our results demonstrate that the inclusion of monoolein in the cationic liposomes formulation significantly modifies the rate of DNA complexation, being DODAB:MO (1:1) the system with higher DNA condensation efficiency.
publishDate 2008
dc.date.none.fl_str_mv 2008-03
2008-03-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1822/9409
url http://hdl.handle.net/1822/9409
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv “Journal of Fluorescence”. ISSN 1053-0509. 18:2 (2008) 555-562.
1053-0509
10.1007/s10895-007-0299-5
18157688
www.springerlink.com
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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