A new exposure model for in vitro testing of effects of gaseous pollutants on mammalian cells by means of gas diffusion through plastic films

Alink, G.M.; Hoeven, J.C.M. van der; Debets, F.M.H.; Ven, W.S.M. van de; Koeman, J.H.; Boleij, J.S.M.

doi:10.1016/0045-6535(79)90051-1

Cited by 23 publications

(4 citation statements)

References 17 publications

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“…Our experimental design resembled that of Alink et al (17,18), in that both used FEP-Teflon membranes in chambers situated in a flowing gas environment. Significant technical differences do exist, however.…”

Section: Discussionmentioning

confidence: 99%

Gaseous Oxide Toxicity Evaluated with Cell Monolayers on Collagen-Coated, Gas-Permeable Teflon Membranes

Gabridge¹,

Gladd²

1984

Environmental Health Perspectives

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JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact support@jstor.org.. The National Institute of Environmental Health Sciences (NIEHS) and Brogan & Partners are collaborating with JSTOR to digitize, preserve and extend access to Environmental Health Perspectives.A system was developed to evaluate the cytotoxic potential of gaseous oxides in vitro. Target cells were MRC-5 human lung fibroblasts cultivated as monolayers on gaspermeable, FEP-Teflon membranes. Membranes were secured in Chamber/Dishes with a 25 mm diameter well. To promote attachment of fibroblasts to the membranes, the latter were incubated in collagen (Vitrogen) solutions for 10 min prior to plating the cells. The collagen pretreatment was significantly more effective than poly-Llysine, fetal calf serum, polybrene and bovine serum albumin. Several types (mouse and calf) of acidsoluble and alcohol-soluble collagen fractions were evaluated, and all of them promoted cell attachment with equivalent efficiency. Cells on membranes were exposed to gases in a Plexiglass chamber with a gas flow of 2 L/min. Sulfur dioxide caused a marked loss in cell viability (as indicated by ATP content of the monolayer) after 30 min exposure to 0.01% and 0.005%. A level of 0.001% did not affect viability, and none of the levels tested caused a sloughing of the monolayer after 90 min. Nitrogen dioxide induced a more modest drop in cell viability after 30 min exposure to 0.1%, while 0.005% and 0.05% were nontoxic. No cell sloughing occurred with NO2 exposures, and exposures to CO2 at levels of 20% for 90 min were nontoxic. This system, with cell culture monolayers on gas-permeable Teflon membranes, is simple and convenient. As such, it has potential application to cytotoxicity evaluations with numerous gases.

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Section: Discussionmentioning

confidence: 99%

Gaseous Oxide Toxicity Evaluated with Cell Monolayers on Collagen-Coated, Gas-Permeable Teflon Membranes

Gabridge¹,

Gladd²

1984

Environmental Health Perspectives

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“…Such membranes have been used with gas-sensing electrodes (14) and for delivering oxygen to cells in batch culture (15). Most recently they have been used to evaluate the in vitro cytotoxic potential of ozone, a common gaseous pollutant (16,17). Ozone was shown to decrease both viability and plating efficiency after a 2.5 hr exposure to 0.1 to 0.2 jg ozone/12 cm2 of membrane covered with alveolar Type II cells.…”

Section: Discussionmentioning

confidence: 99%

Gaseous oxide toxicity evaluated with cell monolayers on collagen-coated, gas-permeable teflon membranes.

Gabridge¹,

Gladd²

1984

Environ Health Perspect

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A system was developed to evaluate the cytotoxic potential of gaseous oxides in vitro. Target cells were MRC-5 human lung fibroblasts cultivated as monolayers on gas-permeable , FEP-Teflon membranes. Membranes were secured in Chamber/Dishes with a 25 mm diameter well. To promote attachment of fibroblasts to the membranes, the latter were incubated in collagen ( Vitrogen ) solutions for 10 min prior to plating the cells. The collagen pretreatment was significantly more effective than poly-L-lysine, fetal calf serum, polybrene and bovine serum albumin. Several types (mouse and calf) of acid-soluble and alcohol-soluble collagen fractions were evaluated, and all of them promoted cell attachment with equivalent efficiency. Cells on membranes were exposed to gases in a Plexiglass chamber with a gas flow of 2L/min. Sulfur dioxide caused a marked loss in cell viability (as indicated by ATP content of the monolayer) after 30 min exposure to 0.01% and 0.005%. A level of 0.001% did not affect viability, and none of the levels tested caused a sloughing of the monolayer after 90 min. Nitrogen dioxide induced a more modest drop in cell viability after 30 min exposure to 0.1%, while 0.005% and 0.05% were nontoxic. No cell sloughing occurred with NO2 exposures, and exposures to CO2 at levels of 20% for 90 min were nontoxic. This system, with cell culture monolayers on gas-permeable Teflon membranes, is simple and convenient. As such, it has potential application to cytotoxicity evaluations with numerous gases.

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“…This was achieved by placing the cultures dishes in a 36-1 perspex fumigation box at 3 7GC. 21 The inlet gas mixture contained 85% N2, 5% CO2, 10% 02 and an experimental amount of NO2. NO2 (0.9% NO2 in N2) was purchased from Matheson (Oevel, Belgium).…”

Section: Biochemical Parametersmentioning

confidence: 99%

Biochemical and morphological changes in lung tissue and isolated lung cells of rats induced by short-term nitrogen dioxide exposure

Bree

Rietjens²,

Alink

et al. 2000

Hum Exp Toxicol

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To investigate the effects of repeated exposure to nitrogen dioxide (NO2) on antioxidant enzymes in lung tissue and isolated lung cells, rats were continuously exposed to 20 mg/m3 NO2 (10.6 ppm) for 4 days. The activities of glucose-6-phosphate dehydrogenase (G6PDH), glutathione reductase (GR), and glutathione peroxidase (GSHPx) were measured in the cytosolic fraction of lung tissue of both control and NO2-exposed rats as well as in isolated alveolar macrophages (AMs) and type II cells. Qualitative and quantitative changes in AM and type II cells were studied by electron microscopy and by morphometric analyses using enzyme and immunohistochemistry. NO2 exposure resulted in significantly increased pulmonary activities of G6PDH, GR, and GSHPx, both expressed per lung and per gram of lung weight. Morphometric data show that NO2 exposure significantly increased the number of type II cells, predominantly in the centriacinar region, indicating proliferation of epithelium following cellular injury. Type II cells in lungs of NO2-exposed rats had a squamous, less cuboidal appearance with more lamellar bodies compared to type II cells in lungs of control rats. Compared to control lungs, a higher number of macrophages could be isolated from NO2-exposed lungs, while numbers of type II cells isolated from lungs of control and NO2-exposed rats were the same. Isolated type II cells from control and NO2-exposed rats were polymorphic, with a small number of lamellar bodies and without polarity. Isolated macro-phages were rounded and contained many filopodia. NO2 exposure caused increases in the activities of G6PDH and GSHPx in isolated type II cells and of GSHPx in isolated macrophages, when expressed per number of cells. Macrophages and type II cells isolated from control and NO2-exposed rats and re-exposed in vitro to NO2, showed no differences in phagocytosis and viability features. Our results indicate that NO2-induced increases in pulmonary antioxidant enzymes are also reflected in isolated AM and type II cells. Since these lung cells do not display a decreased sensitivities toward an in vitro NO2 exposure, overall increase in antioxidant enzyme activities do not seem to play the most pivotal role in controlling cellular NO2 sensitivity and oxidant defence. Combined data from biochemical, morphological, and morphometric analyses of lungs and lung cells suggest that lung cell and tissue oxidant sensitivity and defence largely depends on the cell and tissue organisation, i.e., cell numbers and morphology as well as the ratio of surface area to cytoplasmic volume.

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A new exposure model for in vitro testing of effects of gaseous pollutants on mammalian cells by means of gas diffusion through plastic films

Cited by 23 publications

References 17 publications

Gaseous Oxide Toxicity Evaluated with Cell Monolayers on Collagen-Coated, Gas-Permeable Teflon Membranes

Gaseous Oxide Toxicity Evaluated with Cell Monolayers on Collagen-Coated, Gas-Permeable Teflon Membranes

Gaseous oxide toxicity evaluated with cell monolayers on collagen-coated, gas-permeable teflon membranes.

Biochemical and morphological changes in lung tissue and isolated lung cells of rats induced by short-term nitrogen dioxide exposure

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