Ionically crosslinked chitosan/tripolyphosphate nanoparticles for oligonucleotide and plasmid DNA delivery.
Identifieur interne : 002786 ( Main/Exploration ); précédent : 002785; suivant : 002787Ionically crosslinked chitosan/tripolyphosphate nanoparticles for oligonucleotide and plasmid DNA delivery.
Auteurs : Noemi Csaba [Espagne] ; Magnus Köping-Högg Rd ; Maria Jose AlonsoSource :
- International journal of pharmaceutics [ 1873-3476 ] ; 2009.
Descripteurs français
- KwdFr :
- Animaux, Chitosane (), Chitosane (administration et posologie), Chitosane (métabolisme), Chitosane (toxicité), Endocytose, Facteurs temps, Gènes rapporteurs, Humains, Lignée cellulaire, Masse moléculaire, Microscopie confocale, Nanoparticules, Oligonucléotides (), Oligonucléotides (administration et posologie), Oligonucléotides (métabolisme), Plasmides (), Plasmides (administration et posologie), Plasmides (métabolisme), Polyphosphates (), Polyphosphates (administration et posologie), Polyphosphates (métabolisme), Polyphosphates (toxicité), Réactifs réticulants (), Réactifs réticulants (administration et posologie), Réactifs réticulants (métabolisme), Réactifs réticulants (toxicité), Régulation de l'expression des gènes, Souris, Souris de lignée BALB C, Sérumalbumine bovine (), Transfection (), Transport biologique, beta-Galactosidase (biosynthèse), beta-Galactosidase (génétique).
- MESH :
- administration et posologie : Chitosane, Oligonucléotides, Plasmides, Polyphosphates, Réactifs réticulants.
- biosynthèse : beta-Galactosidase.
- génétique : beta-Galactosidase.
- métabolisme : Chitosane, Oligonucléotides, Plasmides, Polyphosphates, Réactifs réticulants.
- toxicité : Chitosane, Polyphosphates, Réactifs réticulants.
- Animaux, Chitosane, Endocytose, Facteurs temps, Gènes rapporteurs, Humains, Lignée cellulaire, Masse moléculaire, Microscopie confocale, Nanoparticules, Oligonucléotides, Plasmides, Polyphosphates, Réactifs réticulants, Régulation de l'expression des gènes, Souris, Souris de lignée BALB C, Sérumalbumine bovine, Transfection, Transport biologique.
English descriptors
- KwdEn :
- Animals, Biological Transport, Cell Line, Chitosan (administration & dosage), Chitosan (chemistry), Chitosan (metabolism), Chitosan (toxicity), Cross-Linking Reagents (administration & dosage), Cross-Linking Reagents (chemistry), Cross-Linking Reagents (metabolism), Cross-Linking Reagents (toxicity), Endocytosis, Gene Expression Regulation, Genes, Reporter, Humans, Mice, Mice, Inbred BALB C, Microscopy, Confocal, Molecular Weight, Nanoparticles, Oligonucleotides (administration & dosage), Oligonucleotides (chemistry), Oligonucleotides (metabolism), Plasmids (administration & dosage), Plasmids (chemistry), Plasmids (metabolism), Polyphosphates (administration & dosage), Polyphosphates (chemistry), Polyphosphates (metabolism), Polyphosphates (toxicity), Serum Albumin, Bovine (chemistry), Time Factors, Transfection (methods), beta-Galactosidase (biosynthesis), beta-Galactosidase (genetics).
- MESH :
- chemical , administration & dosage : Chitosan, Cross-Linking Reagents, Oligonucleotides, Polyphosphates.
- chemical , biosynthesis : beta-Galactosidase.
- chemical , chemistry : Chitosan, Cross-Linking Reagents, Oligonucleotides, Polyphosphates, Serum Albumin, Bovine.
- chemical , genetics : beta-Galactosidase.
- chemical , metabolism : Chitosan, Cross-Linking Reagents, Oligonucleotides, Polyphosphates.
- chemical , toxicity : Chitosan, Cross-Linking Reagents, Polyphosphates.
- administration & dosage : Plasmids.
- chemistry : Plasmids.
- metabolism : Plasmids.
- methods : Transfection.
- Animals, Biological Transport, Cell Line, Endocytosis, Gene Expression Regulation, Genes, Reporter, Humans, Mice, Mice, Inbred BALB C, Microscopy, Confocal, Molecular Weight, Nanoparticles, Time Factors.
Abstract
Ionically crosslinked nanoparticles based on high and low molecular weight chitosans (CS) were formulated with plasmid DNA or dsDNA oligomers using the ionic gelation technique with pentasodium tripolyphospate (TPP) as crosslinking agent. The resulting CS/TPP nanoparticles were investigated with regard to their physical-chemical properties, in vitro transfection efficiency, toxicity, cellular uptake, and in vivo gene expression following intratracheal administration to mice. The effects of co-formulating the nanoparticles with a model protein, BSA, were also studied. CS/TPP nanoparticles showed high encapsulation efficiencies both for plasmid DNA and dsDNA oligomers (20-mers), independent of CS molecular weight. TEM images revealed a spherical shape of the CS/TPP nanoparticles in contrast to the heterogeneous and irregular morphology displayed by conventional chitosan polyplexes. The nanoparticles showed high physical stability and no DNA release could be detected in diverse release media, nor even after incubation with heparin. Low molecular weight (LMW) CS/TPP nanoparticles gave high gene expression levels in HEK 293 cells already 2 days after transfection, reaching a plateau of sustained and high gene expression between 4 and 10 days. The inclusion of BSA into the nanostructures did not alter the inherent transfection efficiency of the nanoparticles. Confocal studies suggest endocytotic cellular uptake of the nanoparticles and a subsequent release into the cytoplasm within 14 h. LMW CS/TPP nanoparticles mediated a strong beta-galactosidase expression in vivo after intratracheal administration. The results of this study forward ionically crosslinked CS/TPP nanoparticles as a biocompatible non-viral gene delivery system and generate a solid ground for further optimization studies, for example with regard to steric stabilization and targeting.
DOI: 10.1016/j.ijpharm.2009.07.028
PubMed: 19660537
Affiliations:
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Rd, Magnus" sort="Koping Hogg Rd, Magnus" uniqKey="Koping Hogg Rd M" first="Magnus" last="Köping-Högg Rd">Magnus Köping-Högg Rd</name></author><author><name sortKey="Alonso, Maria Jose" sort="Alonso, Maria Jose" uniqKey="Alonso M" first="Maria Jose" last="Alonso">Maria Jose Alonso</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19660537</idno><idno type="pmid">19660537</idno><idno type="doi">10.1016/j.ijpharm.2009.07.028</idno><idno type="wicri:Area/PubMed/Corpus">001F98</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001F98</idno><idno type="wicri:Area/PubMed/Curation">001F98</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001F98</idno><idno type="wicri:Area/PubMed/Checkpoint">001F03</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001F03</idno><idno 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Santiago de Compostela</wicri:regionArea><placeName><region nuts="2" type="communauté">Galice</region><settlement type="city">Santiago de Compostela</settlement></placeName><orgName type="university">Université de Saint-Jacques-de-Compostelle</orgName></affiliation></author><author><name sortKey="Koping Hogg Rd, Magnus" sort="Koping Hogg Rd, Magnus" uniqKey="Koping Hogg Rd M" first="Magnus" last="Köping-Högg Rd">Magnus Köping-Högg Rd</name></author><author><name sortKey="Alonso, Maria Jose" sort="Alonso, Maria Jose" uniqKey="Alonso M" first="Maria Jose" last="Alonso">Maria Jose Alonso</name></author></analytic><series><title level="j">International journal of pharmaceutics</title><idno type="eISSN">1873-3476</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Biological Transport</term><term>Cell Line</term><term>Chitosan (administration & dosage)</term><term>Chitosan (chemistry)</term><term>Chitosan (metabolism)</term><term>Chitosan (toxicity)</term><term>Cross-Linking Reagents (administration & dosage)</term><term>Cross-Linking Reagents (chemistry)</term><term>Cross-Linking Reagents (metabolism)</term><term>Cross-Linking Reagents (toxicity)</term><term>Endocytosis</term><term>Gene Expression Regulation</term><term>Genes, Reporter</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Microscopy, Confocal</term><term>Molecular Weight</term><term>Nanoparticles</term><term>Oligonucleotides (administration & dosage)</term><term>Oligonucleotides (chemistry)</term><term>Oligonucleotides (metabolism)</term><term>Plasmids (administration & dosage)</term><term>Plasmids (chemistry)</term><term>Plasmids (metabolism)</term><term>Polyphosphates (administration & dosage)</term><term>Polyphosphates (chemistry)</term><term>Polyphosphates (metabolism)</term><term>Polyphosphates (toxicity)</term><term>Serum Albumin, Bovine (chemistry)</term><term>Time Factors</term><term>Transfection (methods)</term><term>beta-Galactosidase (biosynthesis)</term><term>beta-Galactosidase (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Chitosane ()</term><term>Chitosane (administration et posologie)</term><term>Chitosane (métabolisme)</term><term>Chitosane (toxicité)</term><term>Endocytose</term><term>Facteurs temps</term><term>Gènes rapporteurs</term><term>Humains</term><term>Lignée cellulaire</term><term>Masse moléculaire</term><term>Microscopie confocale</term><term>Nanoparticules</term><term>Oligonucléotides ()</term><term>Oligonucléotides (administration et posologie)</term><term>Oligonucléotides (métabolisme)</term><term>Plasmides ()</term><term>Plasmides (administration et posologie)</term><term>Plasmides (métabolisme)</term><term>Polyphosphates ()</term><term>Polyphosphates (administration et posologie)</term><term>Polyphosphates (métabolisme)</term><term>Polyphosphates (toxicité)</term><term>Réactifs réticulants ()</term><term>Réactifs réticulants (administration et posologie)</term><term>Réactifs réticulants (métabolisme)</term><term>Réactifs réticulants (toxicité)</term><term>Régulation de l'expression des gènes</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Sérumalbumine bovine ()</term><term>Transfection ()</term><term>Transport biologique</term><term>beta-Galactosidase (biosynthèse)</term><term>beta-Galactosidase (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="administration & dosage" xml:lang="en"><term>Chitosan</term><term>Cross-Linking Reagents</term><term>Oligonucleotides</term><term>Polyphosphates</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>beta-Galactosidase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Chitosan</term><term>Cross-Linking Reagents</term><term>Oligonucleotides</term><term>Polyphosphates</term><term>Serum Albumin, Bovine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>beta-Galactosidase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Chitosan</term><term>Cross-Linking Reagents</term><term>Oligonucleotides</term><term>Polyphosphates</term></keywords><keywords scheme="MESH" type="chemical" qualifier="toxicity" xml:lang="en"><term>Chitosan</term><term>Cross-Linking Reagents</term><term>Polyphosphates</term></keywords><keywords scheme="MESH" qualifier="administration & dosage" xml:lang="en"><term>Plasmids</term></keywords><keywords scheme="MESH" qualifier="administration et posologie" xml:lang="fr"><term>Chitosane</term><term>Oligonucléotides</term><term>Plasmides</term><term>Polyphosphates</term><term>Réactifs réticulants</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>beta-Galactosidase</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Plasmids</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>beta-Galactosidase</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Plasmids</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Transfection</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Chitosane</term><term>Oligonucléotides</term><term>Plasmides</term><term>Polyphosphates</term><term>Réactifs réticulants</term></keywords><keywords scheme="MESH" qualifier="toxicité" xml:lang="fr"><term>Chitosane</term><term>Polyphosphates</term><term>Réactifs réticulants</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Biological Transport</term><term>Cell Line</term><term>Endocytosis</term><term>Gene Expression Regulation</term><term>Genes, Reporter</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Microscopy, Confocal</term><term>Molecular Weight</term><term>Nanoparticles</term><term>Time Factors</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Chitosane</term><term>Endocytose</term><term>Facteurs temps</term><term>Gènes rapporteurs</term><term>Humains</term><term>Lignée cellulaire</term><term>Masse moléculaire</term><term>Microscopie confocale</term><term>Nanoparticules</term><term>Oligonucléotides</term><term>Plasmides</term><term>Polyphosphates</term><term>Réactifs réticulants</term><term>Régulation de l'expression des gènes</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Sérumalbumine bovine</term><term>Transfection</term><term>Transport biologique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Ionically crosslinked nanoparticles based on high and low molecular weight chitosans (CS) were formulated with plasmid DNA or dsDNA oligomers using the ionic gelation technique with pentasodium tripolyphospate (TPP) as crosslinking agent. The resulting CS/TPP nanoparticles were investigated with regard to their physical-chemical properties, in vitro transfection efficiency, toxicity, cellular uptake, and in vivo gene expression following intratracheal administration to mice. The effects of co-formulating the nanoparticles with a model protein, BSA, were also studied. CS/TPP nanoparticles showed high encapsulation efficiencies both for plasmid DNA and dsDNA oligomers (20-mers), independent of CS molecular weight. TEM images revealed a spherical shape of the CS/TPP nanoparticles in contrast to the heterogeneous and irregular morphology displayed by conventional chitosan polyplexes. The nanoparticles showed high physical stability and no DNA release could be detected in diverse release media, nor even after incubation with heparin. Low molecular weight (LMW) CS/TPP nanoparticles gave high gene expression levels in HEK 293 cells already 2 days after transfection, reaching a plateau of sustained and high gene expression between 4 and 10 days. The inclusion of BSA into the nanostructures did not alter the inherent transfection efficiency of the nanoparticles. Confocal studies suggest endocytotic cellular uptake of the nanoparticles and a subsequent release into the cytoplasm within 14 h. LMW CS/TPP nanoparticles mediated a strong beta-galactosidase expression in vivo after intratracheal administration. The results of this study forward ionically crosslinked CS/TPP nanoparticles as a biocompatible non-viral gene delivery system and generate a solid ground for further optimization studies, for example with regard to steric stabilization and targeting.</div></front></TEI><affiliations><list><country><li>Espagne</li></country><region><li>Galice</li></region><settlement><li>Santiago de Compostela</li></settlement><orgName><li>Université de Saint-Jacques-de-Compostelle</li></orgName></list><tree><noCountry><name sortKey="Alonso, Maria Jose" sort="Alonso, Maria Jose" uniqKey="Alonso M" first="Maria Jose" last="Alonso">Maria Jose Alonso</name><name sortKey="Koping Hogg Rd, Magnus" sort="Koping Hogg Rd, Magnus" uniqKey="Koping Hogg Rd M" first="Magnus" last="Köping-Högg Rd">Magnus Köping-Högg Rd</name></noCountry><country name="Espagne"><region name="Galice"><name sortKey="Csaba, Noemi" sort="Csaba, Noemi" uniqKey="Csaba N" first="Noemi" last="Csaba">Noemi Csaba</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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