Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles

As the lung is one of the main routes of exposure to manufactured nanoparticles, we developed an in vitro model resembling the alveolo-capillary barrier for the study of nanoparticle translocation. In order to provide a relevant and ethical in vitro model, cost effective and easy-to-implement human...

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Main Authors: Samir Dekali, Christelle Gamez, Thierry Kortulewski, Kelly Blazy, Patrice Rat, Ghislaine Lacroix
Format: Article
Language:English
Published: Elsevier 2014-01-01
Series:Toxicology Reports
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2214750014000092
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author Samir Dekali
Christelle Gamez
Thierry Kortulewski
Kelly Blazy
Patrice Rat
Ghislaine Lacroix
author_facet Samir Dekali
Christelle Gamez
Thierry Kortulewski
Kelly Blazy
Patrice Rat
Ghislaine Lacroix
author_sort Samir Dekali
collection DOAJ
description As the lung is one of the main routes of exposure to manufactured nanoparticles, we developed an in vitro model resembling the alveolo-capillary barrier for the study of nanoparticle translocation. In order to provide a relevant and ethical in vitro model, cost effective and easy-to-implement human cell lines were used. Pulmonary epithelial cells (Calu-3 cell line) and macrophages (THP-1 differentiated cells) were cultivated on the apical side and pulmonary endothelial cells (HPMEC-ST1.6R cell line) on the basal side of a microporous polyester membrane (Transwell®). Translocation of non-functionalized (51 and 110 nm) and aminated (52 nm) fluorescent polystyrene (PS) nanobeads was studied in this system. The use of Calu-3 cells allowed high transepithelial electrical resistance (TEER) values (>1000 Ω cm2) in co-cultures with or without macrophages. After 24 h of exposure to non-cytotoxic concentrations of non-functionalized PS nanobeads, the relative TEER values (%/t0) were significantly decreased in co-cultures. Epithelial cells and macrophages were able to internalize PS nanobeads. Regarding translocation, Transwell® membranes per se limit the passage of nanoparticles between apical and basal side. However, small non-functionalized PS nanobeads (51 nm) were able to translocate as they were detected in the basal side of co-cultures. Altogether, these results show that this co-culture model present good barrier properties allowing the study of nanoparticle translocation but research effort need to be done to improve the neutrality of the porous membrane delimitating apical and basal sides of the model.
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spelling doaj.art-2f2f9bee8e124836aa8858ea34fcf9ce2022-12-22T03:36:49ZengElsevierToxicology Reports2214-75002014-01-011C15717110.1016/j.toxrep.2014.03.003Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticlesSamir Dekali0Christelle Gamez1Thierry Kortulewski2Kelly Blazy3Patrice Rat4Ghislaine Lacroix5INERIS (Institut National de l’Environnement industriel et des RISques), Unité de Toxicologie expérimentale, 60550 Verneuil-en-Halatte, FranceINERIS (Institut National de l’Environnement industriel et des RISques), Unité de Toxicologie expérimentale, 60550 Verneuil-en-Halatte, FranceCEA, DSV, iRCM, Plateforme imagerie photonique, 92260 Fontenay-aux-Roses, FranceINERIS (Institut National de l’Environnement industriel et des RISques), Unité de Toxicologie expérimentale, 60550 Verneuil-en-Halatte, FranceLaboratoire de chimie et toxicologie analytique et cellulaire (C-TAC), Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris Descartes (PRES Sorbonne Paris Cité), 75270 Paris Cedex 06, FranceINERIS (Institut National de l’Environnement industriel et des RISques), Unité de Toxicologie expérimentale, 60550 Verneuil-en-Halatte, FranceAs the lung is one of the main routes of exposure to manufactured nanoparticles, we developed an in vitro model resembling the alveolo-capillary barrier for the study of nanoparticle translocation. In order to provide a relevant and ethical in vitro model, cost effective and easy-to-implement human cell lines were used. Pulmonary epithelial cells (Calu-3 cell line) and macrophages (THP-1 differentiated cells) were cultivated on the apical side and pulmonary endothelial cells (HPMEC-ST1.6R cell line) on the basal side of a microporous polyester membrane (Transwell®). Translocation of non-functionalized (51 and 110 nm) and aminated (52 nm) fluorescent polystyrene (PS) nanobeads was studied in this system. The use of Calu-3 cells allowed high transepithelial electrical resistance (TEER) values (>1000 Ω cm2) in co-cultures with or without macrophages. After 24 h of exposure to non-cytotoxic concentrations of non-functionalized PS nanobeads, the relative TEER values (%/t0) were significantly decreased in co-cultures. Epithelial cells and macrophages were able to internalize PS nanobeads. Regarding translocation, Transwell® membranes per se limit the passage of nanoparticles between apical and basal side. However, small non-functionalized PS nanobeads (51 nm) were able to translocate as they were detected in the basal side of co-cultures. Altogether, these results show that this co-culture model present good barrier properties allowing the study of nanoparticle translocation but research effort need to be done to improve the neutrality of the porous membrane delimitating apical and basal sides of the model.http://www.sciencedirect.com/science/article/pii/S2214750014000092Alveolo-capillary barrierNanoparticlesPolystyreneCalu-3THP-1HPMEC-ST1.6R
spellingShingle Samir Dekali
Christelle Gamez
Thierry Kortulewski
Kelly Blazy
Patrice Rat
Ghislaine Lacroix
Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
Toxicology Reports
Alveolo-capillary barrier
Nanoparticles
Polystyrene
Calu-3
THP-1
HPMEC-ST1.6R
title Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
title_full Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
title_fullStr Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
title_full_unstemmed Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
title_short Assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
title_sort assessment of an in vitro model of pulmonary barrier to study the translocation of nanoparticles
topic Alveolo-capillary barrier
Nanoparticles
Polystyrene
Calu-3
THP-1
HPMEC-ST1.6R
url http://www.sciencedirect.com/science/article/pii/S2214750014000092
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AT kellyblazy assessmentofaninvitromodelofpulmonarybarriertostudythetranslocationofnanoparticles
AT patricerat assessmentofaninvitromodelofpulmonarybarriertostudythetranslocationofnanoparticles
AT ghislainelacroix assessmentofaninvitromodelofpulmonarybarriertostudythetranslocationofnanoparticles