An improved 3D tetraculture system mimicking the cellular organisation at the alveolar barrier to study the potential toxic effects of particles on the lung
Identifieur interne : 000533 ( Main/Curation ); précédent : 000532; suivant : 000534An improved 3D tetraculture system mimicking the cellular organisation at the alveolar barrier to study the potential toxic effects of particles on the lung
Auteurs : Sebastian G. Klein [Luxembourg (pays), Allemagne] ; Tommaso Serchi [Luxembourg (pays)] ; Lucien Hoffmann [Luxembourg (pays)] ; Brunhilde Blömeke [Allemagne] ; Arno C. Gutleb [Luxembourg (pays)]Source :
- Particle and Fibre Toxicology [ 1743-8977 ] ; 2013.
Abstract
Exposure to fine and ultra-fine ambient particles is still a problem of concern in many industrialised parts of the world and the intensified use of nanotechnology may further increase exposure to small particles. Complex
The spatial distribution of the cells in the tetraculture was analysed by confocal laser scanning microscopy (CLSM), showing a confluent layer of endothelial and epithelial cells on both sides of the transwell. Macrophage-like cells and mast cells can be found on top of the epithelial cells. The cells formed colonies under submerged conditions, which disappeared at the ALI. To evaluate the response to oxidative stress, the dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used together with 2,2’-azobis-2-methyl-propanimidamide-dihydrochloride (AAPH) as inducer of oxidative stress. The tetraculture showed less induction of reactive oxygen species (ROS) production after being treated with a positive control compared to the monocultures of EA.hy 926, THP-1 and HMC-1. Submerged cultures showed elevated ROS and IL-8 levels compared to ALI cultures. The Vitrocell™ aerosol exposure system was not significantly influencing the viability. Using this system, cells were exposed to an aerosol of 50 nm SiO2-Rhodamine NPs in PBS. The distribution of the NPs in the tetraculture after exposure was evaluated by CLSM. Fluorescence from internalized particles was detected in CD11b-positive THP-1 cells only.
The system can be used in conjunction with a native aerosol exposure system and may finally lead to a more realistic judgement regarding the hazard of new compounds and/or new nano-scaled materials in the future. The results for the ROS production and IL-8 secretion suggest that submerged exposure may lead to an overestimation of observed effects.
Url:
DOI: 10.1186/1743-8977-10-31
PubMed: 23890538
PubMed Central: 3733942
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Klein</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="I2">Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier</wicri:regionArea><wicri:noRegion>Trier</wicri:noRegion><orgName type="university">Université de Trèves</orgName><placeName><settlement type="city">Trèves (Allemagne)</settlement><region type="land" nuts="1">Rhénanie-Palatinat</region></placeName></affiliation></author><author><name sortKey="Serchi, Tommaso" sort="Serchi, Tommaso" uniqKey="Serchi T" first="Tommaso" last="Serchi">Tommaso Serchi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author><author><name sortKey="Hoffmann, Lucien" sort="Hoffmann, Lucien" uniqKey="Hoffmann L" first="Lucien" last="Hoffmann">Lucien Hoffmann</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author><author><name sortKey="Blomeke, Brunhilde" sort="Blomeke, Brunhilde" uniqKey="Blomeke B" first="Brunhilde" last="Blömeke">Brunhilde Blömeke</name><affiliation wicri:level="4"><nlm:aff id="I2">Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier</wicri:regionArea><wicri:noRegion>Trier</wicri:noRegion><orgName type="university">Université de Trèves</orgName><placeName><settlement type="city">Trèves (Allemagne)</settlement><region type="land" nuts="1">Rhénanie-Palatinat</region></placeName></affiliation></author><author><name sortKey="Gutleb, Arno C" sort="Gutleb, Arno C" uniqKey="Gutleb A" first="Arno C" last="Gutleb">Arno C. Gutleb</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23890538</idno><idno type="pmc">3733942</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3733942</idno><idno type="RBID">PMC:3733942</idno><idno type="doi">10.1186/1743-8977-10-31</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">000104</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000104</idno><idno type="wicri:Area/Pmc/Curation">000104</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000104</idno><idno type="wicri:Area/Pmc/Checkpoint">000093</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000093</idno><idno type="wicri:Area/Ncbi/Merge">000612</idno><idno type="wicri:Area/Ncbi/Curation">000612</idno><idno type="wicri:Area/Ncbi/Checkpoint">000612</idno><idno type="wicri:Area/Main/Merge">000535</idno><idno type="wicri:Area/Main/Curation">000533</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">An improved 3D tetraculture system mimicking the cellular organisation at the alveolar barrier to study the potential toxic effects of particles on the lung</title><author><name sortKey="Klein, Sebastian G" sort="Klein, Sebastian G" uniqKey="Klein S" first="Sebastian G" last="Klein">Sebastian G. Klein</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation><affiliation wicri:level="4"><nlm:aff id="I2">Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier</wicri:regionArea><wicri:noRegion>Trier</wicri:noRegion><orgName type="university">Université de Trèves</orgName><placeName><settlement type="city">Trèves (Allemagne)</settlement><region type="land" nuts="1">Rhénanie-Palatinat</region></placeName></affiliation></author><author><name sortKey="Serchi, Tommaso" sort="Serchi, Tommaso" uniqKey="Serchi T" first="Tommaso" last="Serchi">Tommaso Serchi</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author><author><name sortKey="Hoffmann, Lucien" sort="Hoffmann, Lucien" uniqKey="Hoffmann L" first="Lucien" last="Hoffmann">Lucien Hoffmann</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author><author><name sortKey="Blomeke, Brunhilde" sort="Blomeke, Brunhilde" uniqKey="Blomeke B" first="Brunhilde" last="Blömeke">Brunhilde Blömeke</name><affiliation wicri:level="4"><nlm:aff id="I2">Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Department of Environmental Toxicology, University of Trier, Universitätsring 15, Trier</wicri:regionArea><wicri:noRegion>Trier</wicri:noRegion><orgName type="university">Université de Trèves</orgName><placeName><settlement type="city">Trèves (Allemagne)</settlement><region type="land" nuts="1">Rhénanie-Palatinat</region></placeName></affiliation></author><author><name sortKey="Gutleb, Arno C" sort="Gutleb, Arno C" uniqKey="Gutleb A" first="Arno C" last="Gutleb">Arno C. Gutleb</name><affiliation wicri:level="1"><nlm:aff id="I1">Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg</nlm:aff><country xml:lang="fr">Luxembourg (pays)</country><wicri:regionArea>Department Environment and Agro-biotechnologies (EVA), Centre de Recherche Public - Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux</wicri:regionArea><wicri:noRegion>L-4422 Belvaux</wicri:noRegion></affiliation></author></analytic><series><title level="j">Particle and Fibre Toxicology</title><idno type="eISSN">1743-8977</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Exposure to fine and ultra-fine ambient particles is still a problem of concern in many industrialised parts of the world and the intensified use of nanotechnology may further increase exposure to small particles. Complex <italic>in vitro</italic> coculture systems may be valuable tools to study particle-induced processes and to extrapolate effects of particles on the lung. A system consisting of four different human cell lines which mimics the cell response of the alveolar surface <italic>in vitro</italic> was developed to study native aerosol exposure (Vitrocell™ chamber). The system is composed of an alveolar type-II cell line (A549), differentiated macrophage-like cells (THP-1), mast cells (HMC-1) and endothelial cells (EA.hy 926), seeded in a 3D-orientation on a microporous membrane.</p></sec><sec><title>Results</title><p>The spatial distribution of the cells in the tetraculture was analysed by confocal laser scanning microscopy (CLSM), showing a confluent layer of endothelial and epithelial cells on both sides of the transwell. Macrophage-like cells and mast cells can be found on top of the epithelial cells. The cells formed colonies under submerged conditions, which disappeared at the ALI. To evaluate the response to oxidative stress, the dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used together with 2,2’-azobis-2-methyl-propanimidamide-dihydrochloride (AAPH) as inducer of oxidative stress. The tetraculture showed less induction of reactive oxygen species (ROS) production after being treated with a positive control compared to the monocultures of EA.hy 926, THP-1 and HMC-1. Submerged cultures showed elevated ROS and IL-8 levels compared to ALI cultures. The Vitrocell™ aerosol exposure system was not significantly influencing the viability. Using this system, cells were exposed to an aerosol of 50 nm SiO<sub>2</sub>-Rhodamine NPs in PBS. The distribution of the NPs in the tetraculture after exposure was evaluated by CLSM. Fluorescence from internalized particles was detected in CD11b-positive THP-1 cells only.</p></sec><sec><title>Conclusion</title><p>The system can be used in conjunction with a native aerosol exposure system and may finally lead to a more realistic judgement regarding the hazard of new compounds and/or new nano-scaled materials in the future. The results for the ROS production and IL-8 secretion suggest that submerged exposure may lead to an overestimation of observed effects.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Gutleb, Ac" uniqKey="Gutleb A">AC Gutleb</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Logan, Wp" uniqKey="Logan W">WP Logan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Krein, A" uniqKey="Krein A">A Krein</name></author><author><name sortKey="Udelhoven, T" uniqKey="Udelhoven T">T Udelhoven</name></author><author><name sortKey="Audinot, Jn" uniqKey="Audinot J">JN Audinot</name></author><author><name sortKey="Hissler, C" uniqKey="Hissler C">C Hissler</name></author><author><name sortKey="Guignard, C" uniqKey="Guignard C">C Guignard</name></author><author><name sortKey="Pfister, L" uniqKey="Pfister L">L Pfister</name></author><author><name sortKey="Migeon, Hn" 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uniqKey="Narayanan P">PK Narayanan</name></author><author><name sortKey="Robinson, Jp" uniqKey="Robinson J">JP Robinson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Berube, K" uniqKey="Berube K">K BéruBé</name></author><author><name sortKey="Aufderheide, M" uniqKey="Aufderheide M">M Aufderheide</name></author><author><name sortKey="Breheny, D" uniqKey="Breheny D">D Breheny</name></author><author><name sortKey="Clothier, R" uniqKey="Clothier R">R Clothier</name></author><author><name sortKey="Combes, R" uniqKey="Combes R">R Combes</name></author><author><name sortKey="Duffin, R" uniqKey="Duffin R">R Duffin</name></author><author><name sortKey="Forbes, B" uniqKey="Forbes B">B Forbes</name></author><author><name sortKey="Gaca, M" uniqKey="Gaca M">M Gaca</name></author><author><name sortKey="Gray, A" uniqKey="Gray A">A Gray</name></author><author><name sortKey="Hall, I" uniqKey="Hall I">I Hall</name></author><author><name sortKey="Kelly, M" uniqKey="Kelly M">M Kelly</name></author><author><name sortKey="Lethem, M" uniqKey="Lethem M">M Lethem</name></author><author><name sortKey="Liebsch, M" uniqKey="Liebsch M">M Liebsch</name></author><author><name sortKey="Merolla, L" uniqKey="Merolla L">L Merolla</name></author><author><name sortKey="Morin, Jp" uniqKey="Morin J">JP Morin</name></author><author><name sortKey="Seagrave, J" uniqKey="Seagrave J">J Seagrave</name></author><author><name sortKey="Swartz, Ma" uniqKey="Swartz M">MA Swartz</name></author><author><name sortKey="Tetley, Td" uniqKey="Tetley T">TD Tetley</name></author><author><name sortKey="Umachandran, M" uniqKey="Umachandran M">M Umachandran</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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