Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.

Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic...

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Main Authors: Maria Mrakovcic, Markus Absenger, Regina Riedl, Claudia Smole, Eva Roblegg, Leopold F Fröhlich, Eleonore Fröhlich
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2013-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0056791&type=printable
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author Maria Mrakovcic
Markus Absenger
Regina Riedl
Claudia Smole
Eva Roblegg
Leopold F Fröhlich
Eleonore Fröhlich
author_facet Maria Mrakovcic
Markus Absenger
Regina Riedl
Claudia Smole
Eva Roblegg
Leopold F Fröhlich
Eleonore Fröhlich
author_sort Maria Mrakovcic
collection DOAJ
description Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitator(TM)). Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing.
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spelling doaj-art-fb291024ec2a41619ddf8b33433f1b862025-08-20T02:05:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5679110.1371/journal.pone.0056791Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.Maria MrakovcicMarkus AbsengerRegina RiedlClaudia SmoleEva RobleggLeopold F FröhlichEleonore FröhlichNano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitator(TM)). Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0056791&type=printable
spellingShingle Maria Mrakovcic
Markus Absenger
Regina Riedl
Claudia Smole
Eva Roblegg
Leopold F Fröhlich
Eleonore Fröhlich
Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
PLoS ONE
title Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
title_full Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
title_fullStr Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
title_full_unstemmed Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
title_short Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
title_sort assessment of long term effects of nanoparticles in a microcarrier cell culture system
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0056791&type=printable
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