Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions

Detalhes bibliográficos
Autor(a) principal: Martinez Junior, André Miguel [UNESP]
Data de Publicação: 2022
Outros Autores: Lima, Aline Margarete Furuyama [UNESP], Martins, Grazieli Olinda [UNESP], Tiera, Vera Aparecida de Oliveira [UNESP], Benderdour, Mohamed, Fernandes, Julio Cesar, Tiera, Marcio José [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.3390/md20080476
http://hdl.handle.net/11449/241629
Resumo: Nowadays, the therapeutic efficiency of small interfering RNAs (siRNA) is still limited by the efficiency of gene therapy vectors capable of carrying them inside the target cells. In this study, siRNA nanocarriers based on low molecular weight chitosan grafted with increasing proportions (5 to 55%) of diisopropylethylamine (DIPEA) groups were developed, which allowed precise control of the degree of ionization of the polycations at pH 7.4. This approach made obtaining siRNA nanocarriers with small sizes (100–200 nm), positive surface charge and enhanced colloidal stability (up to 24 h) at physiological conditions of pH (7.4) and ionic strength (150 mmol L−1) possible. Moreover, the PEGylation improved the stability of the nanoparticles, which maintained their colloidal stability and nanometric sizes even in an albumin-containing medium. The chitosan-derivatives displayed non-cytotoxic effects in both fibroblasts (NIH/3T3) and macrophages (RAW 264.7) at high N/P ratios and polymer concentrations (up to 0.5 g L−1). Confocal microscopy showed a successful uptake of nanocarriers by RAW 264.7 macrophages and a promising ability to silence green fluorescent protein (GFP) in HeLa cells. These results were confirmed by a high level of tumor necrosis factor-α (TNFα) knockdown (higher than 60%) in LPS-stimulated macrophages treated with the siRNA-loaded nanoparticles even in the FBS-containing medium, findings that reveal a good correlation between the degree of ionization of the polycations and the physicochemical properties of nanocarriers. Overall, this study provides an approach to enhance siRNA condensation by chitosan-based carriers and highlights the potential of these nanocarriers for in vivo studies.
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spelling Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological ConditionsDIPEAgene therapynanoparticlesphysiological pHsiRNANowadays, the therapeutic efficiency of small interfering RNAs (siRNA) is still limited by the efficiency of gene therapy vectors capable of carrying them inside the target cells. In this study, siRNA nanocarriers based on low molecular weight chitosan grafted with increasing proportions (5 to 55%) of diisopropylethylamine (DIPEA) groups were developed, which allowed precise control of the degree of ionization of the polycations at pH 7.4. This approach made obtaining siRNA nanocarriers with small sizes (100–200 nm), positive surface charge and enhanced colloidal stability (up to 24 h) at physiological conditions of pH (7.4) and ionic strength (150 mmol L−1) possible. Moreover, the PEGylation improved the stability of the nanoparticles, which maintained their colloidal stability and nanometric sizes even in an albumin-containing medium. The chitosan-derivatives displayed non-cytotoxic effects in both fibroblasts (NIH/3T3) and macrophages (RAW 264.7) at high N/P ratios and polymer concentrations (up to 0.5 g L−1). Confocal microscopy showed a successful uptake of nanocarriers by RAW 264.7 macrophages and a promising ability to silence green fluorescent protein (GFP) in HeLa cells. These results were confirmed by a high level of tumor necrosis factor-α (TNFα) knockdown (higher than 60%) in LPS-stimulated macrophages treated with the siRNA-loaded nanoparticles even in the FBS-containing medium, findings that reveal a good correlation between the degree of ionization of the polycations and the physicochemical properties of nanocarriers. Overall, this study provides an approach to enhance siRNA condensation by chitosan-based carriers and highlights the potential of these nanocarriers for in vivo studies.Department of Chemistry and Environmental Sciences IBILCE São Paulo State University—UNESP, SPOrthopedic Research Laboratory Hôpital du Sacré-Coeur de Montréal Université de Montréal-CanadaDepartment of Chemistry and Environmental Sciences IBILCE São Paulo State University—UNESP, SPUniversidade Estadual Paulista (UNESP)Université de Montréal-CanadaMartinez Junior, André Miguel [UNESP]Lima, Aline Margarete Furuyama [UNESP]Martins, Grazieli Olinda [UNESP]Tiera, Vera Aparecida de Oliveira [UNESP]Benderdour, MohamedFernandes, Julio CesarTiera, Marcio José [UNESP]2023-03-01T21:13:54Z2023-03-01T21:13:54Z2022-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.3390/md20080476Marine Drugs, v. 20, n. 8, 2022.1660-3397http://hdl.handle.net/11449/24162910.3390/md200804762-s2.0-85137561081Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMarine Drugsinfo:eu-repo/semantics/openAccess2023-03-01T21:13:54Zoai:repositorio.unesp.br:11449/241629Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T19:06:29.683245Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
title Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
spellingShingle Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
Martinez Junior, André Miguel [UNESP]
DIPEA
gene therapy
nanoparticles
physiological pH
siRNA
title_short Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
title_full Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
title_fullStr Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
title_full_unstemmed Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
title_sort Impact of Degree of Ionization and PEGylation on the Stability of Nanoparticles of Chitosan Derivatives at Physiological Conditions
author Martinez Junior, André Miguel [UNESP]
author_facet Martinez Junior, André Miguel [UNESP]
Lima, Aline Margarete Furuyama [UNESP]
Martins, Grazieli Olinda [UNESP]
Tiera, Vera Aparecida de Oliveira [UNESP]
Benderdour, Mohamed
Fernandes, Julio Cesar
Tiera, Marcio José [UNESP]
author_role author
author2 Lima, Aline Margarete Furuyama [UNESP]
Martins, Grazieli Olinda [UNESP]
Tiera, Vera Aparecida de Oliveira [UNESP]
Benderdour, Mohamed
Fernandes, Julio Cesar
Tiera, Marcio José [UNESP]
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (UNESP)
Université de Montréal-Canada
dc.contributor.author.fl_str_mv Martinez Junior, André Miguel [UNESP]
Lima, Aline Margarete Furuyama [UNESP]
Martins, Grazieli Olinda [UNESP]
Tiera, Vera Aparecida de Oliveira [UNESP]
Benderdour, Mohamed
Fernandes, Julio Cesar
Tiera, Marcio José [UNESP]
dc.subject.por.fl_str_mv DIPEA
gene therapy
nanoparticles
physiological pH
siRNA
topic DIPEA
gene therapy
nanoparticles
physiological pH
siRNA
description Nowadays, the therapeutic efficiency of small interfering RNAs (siRNA) is still limited by the efficiency of gene therapy vectors capable of carrying them inside the target cells. In this study, siRNA nanocarriers based on low molecular weight chitosan grafted with increasing proportions (5 to 55%) of diisopropylethylamine (DIPEA) groups were developed, which allowed precise control of the degree of ionization of the polycations at pH 7.4. This approach made obtaining siRNA nanocarriers with small sizes (100–200 nm), positive surface charge and enhanced colloidal stability (up to 24 h) at physiological conditions of pH (7.4) and ionic strength (150 mmol L−1) possible. Moreover, the PEGylation improved the stability of the nanoparticles, which maintained their colloidal stability and nanometric sizes even in an albumin-containing medium. The chitosan-derivatives displayed non-cytotoxic effects in both fibroblasts (NIH/3T3) and macrophages (RAW 264.7) at high N/P ratios and polymer concentrations (up to 0.5 g L−1). Confocal microscopy showed a successful uptake of nanocarriers by RAW 264.7 macrophages and a promising ability to silence green fluorescent protein (GFP) in HeLa cells. These results were confirmed by a high level of tumor necrosis factor-α (TNFα) knockdown (higher than 60%) in LPS-stimulated macrophages treated with the siRNA-loaded nanoparticles even in the FBS-containing medium, findings that reveal a good correlation between the degree of ionization of the polycations and the physicochemical properties of nanocarriers. Overall, this study provides an approach to enhance siRNA condensation by chitosan-based carriers and highlights the potential of these nanocarriers for in vivo studies.
publishDate 2022
dc.date.none.fl_str_mv 2022-08-01
2023-03-01T21:13:54Z
2023-03-01T21:13:54Z
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://dx.doi.org/10.3390/md20080476
Marine Drugs, v. 20, n. 8, 2022.
1660-3397
http://hdl.handle.net/11449/241629
10.3390/md20080476
2-s2.0-85137561081
url http://dx.doi.org/10.3390/md20080476
http://hdl.handle.net/11449/241629
identifier_str_mv Marine Drugs, v. 20, n. 8, 2022.
1660-3397
10.3390/md20080476
2-s2.0-85137561081
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Marine Drugs
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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