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Effects of flame made zinc oxide particles in human lung cells - a comparison of aerosol and suspension exposures.

Raemy DO, Grass RN, Stark WJ, Schumacher CM, Clift MJ, Gehr P, Rothen-Rutishauser B - Part Fibre Toxicol (2012)

Bottom Line: Such an approach however, does not reflect particle inhalation.No direct effects could be attributed to ZnO particles.Exposure to suspensions represents a valuable complementary method and allows investigations on particle-associated toxicity by excluding all gas-derived effects.

View Article: PubMed Central - HTML - PubMed

Affiliation: Adolphe Merkle Institute, Bionanomaterials, University of Fribourg, Rte de l'Ancienne Papeterie, P.O. Box 209, CH-1732, Marly, Switzerland.

ABSTRACT

Background: Predominantly, studies of nanoparticle (NPs) toxicology in vitro are based upon the exposure of submerged cell cultures to particle suspensions. Such an approach however, does not reflect particle inhalation. As a more realistic simulation of such a scenario, efforts were made towards direct delivery of aerosols to air-liquid-interface cultivated cell cultures by the use of aerosol exposure systems.This study aims to provide a direct comparison of the effects of zinc oxide (ZnO) NPs when delivered as either an aerosol, or in suspension to a triple cell co-culture model of the epithelial airway barrier. To ensure dose-equivalence, ZnO-deposition was determined in each exposure scenario by atomic absorption spectroscopy. Biological endpoints being investigated after 4 or 24h incubation include cytotoxicity, total reduced glutathione, induction of antioxidative genes such as heme-oxygenase 1 (HO-1) as well as the release of the (pro)-inflammatory cytokine TNFα.

Results: Off-gases released as by-product of flame ZnO synthesis caused a significant decrease of total reduced GSH and induced further the release of the cytokine TNFα, demonstrating the influence of the gas phase on aerosol toxicology. No direct effects could be attributed to ZnO particles. By performing suspension exposure to avoid the factor "flame-gases", particle specific effects become apparent. Other parameters such as LDH and HO-1 were not influenced by gaseous compounds: Following aerosol exposure, LDH levels appeared elevated at both timepoints and the HO-1 transcript correlated positively with deposited ZnO-dose. Under submerged conditions, the HO-1 induction scheme deviated for 4 and 24h and increased extracellular LDH was found following 24h exposure.

Conclusion: In the current study, aerosol and suspension-exposure has been compared by exposing cell cultures to equivalent amounts of ZnO. Both exposure strategies differ fundamentally in their dose-response pattern. Additional differences can be found for the factor time: In the aerosol scenario, parameters tend to their maximum already after 4h of exposure, whereas under submerged conditions, effects appear most pronounced mainly after 24h. Aerosol exposure provides information about the synergistic interplay of gaseous and particulate phase of an aerosol in the context of inhalation toxicology. Exposure to suspensions represents a valuable complementary method and allows investigations on particle-associated toxicity by excluding all gas-derived effects.

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Related in: MedlinePlus

Aerosol Deposition measurement. ZnO was quantified by atomic absorption spectroscopy, taking advantage of the solubility of zinc in acetic acid. A ZnO mass deposition of 1.3 – 6.1 μg/cm2 was measured in the range of 22 – 90 sec reactor runtime, used for subsequent biological experiments. Points indicate individual measurements, bars and whiskers show mean values and standard deviation. Please note the interrupted axis.
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Figure 2: Aerosol Deposition measurement. ZnO was quantified by atomic absorption spectroscopy, taking advantage of the solubility of zinc in acetic acid. A ZnO mass deposition of 1.3 – 6.1 μg/cm2 was measured in the range of 22 – 90 sec reactor runtime, used for subsequent biological experiments. Points indicate individual measurements, bars and whiskers show mean values and standard deviation. Please note the interrupted axis.

Mentions: To correlate reactor runtime with a defined dose, the ZnO deposition per area over a 30 min exposure period was determined by element analysis (Figure2). The average zinc mass per cm2 was measured as 1.3 (SD 0.7), 2.9 (SD 0.6), 6.1 (SD 0.2, n = 2) and 31.1 (SD 4.8) μg, in scenarios with 22, 45, 90 and 270 sec reactor runtime. The reactor off – gas control indicated with 0.1 μg/cm2 (SD 0.2, n = 6) a clean chamber environment.


Effects of flame made zinc oxide particles in human lung cells - a comparison of aerosol and suspension exposures.

Raemy DO, Grass RN, Stark WJ, Schumacher CM, Clift MJ, Gehr P, Rothen-Rutishauser B - Part Fibre Toxicol (2012)

Aerosol Deposition measurement. ZnO was quantified by atomic absorption spectroscopy, taking advantage of the solubility of zinc in acetic acid. A ZnO mass deposition of 1.3 – 6.1 μg/cm2 was measured in the range of 22 – 90 sec reactor runtime, used for subsequent biological experiments. Points indicate individual measurements, bars and whiskers show mean values and standard deviation. Please note the interrupted axis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3585858&req=5

Figure 2: Aerosol Deposition measurement. ZnO was quantified by atomic absorption spectroscopy, taking advantage of the solubility of zinc in acetic acid. A ZnO mass deposition of 1.3 – 6.1 μg/cm2 was measured in the range of 22 – 90 sec reactor runtime, used for subsequent biological experiments. Points indicate individual measurements, bars and whiskers show mean values and standard deviation. Please note the interrupted axis.
Mentions: To correlate reactor runtime with a defined dose, the ZnO deposition per area over a 30 min exposure period was determined by element analysis (Figure2). The average zinc mass per cm2 was measured as 1.3 (SD 0.7), 2.9 (SD 0.6), 6.1 (SD 0.2, n = 2) and 31.1 (SD 4.8) μg, in scenarios with 22, 45, 90 and 270 sec reactor runtime. The reactor off – gas control indicated with 0.1 μg/cm2 (SD 0.2, n = 6) a clean chamber environment.

Bottom Line: Such an approach however, does not reflect particle inhalation.No direct effects could be attributed to ZnO particles.Exposure to suspensions represents a valuable complementary method and allows investigations on particle-associated toxicity by excluding all gas-derived effects.

View Article: PubMed Central - HTML - PubMed

Affiliation: Adolphe Merkle Institute, Bionanomaterials, University of Fribourg, Rte de l'Ancienne Papeterie, P.O. Box 209, CH-1732, Marly, Switzerland.

ABSTRACT

Background: Predominantly, studies of nanoparticle (NPs) toxicology in vitro are based upon the exposure of submerged cell cultures to particle suspensions. Such an approach however, does not reflect particle inhalation. As a more realistic simulation of such a scenario, efforts were made towards direct delivery of aerosols to air-liquid-interface cultivated cell cultures by the use of aerosol exposure systems.This study aims to provide a direct comparison of the effects of zinc oxide (ZnO) NPs when delivered as either an aerosol, or in suspension to a triple cell co-culture model of the epithelial airway barrier. To ensure dose-equivalence, ZnO-deposition was determined in each exposure scenario by atomic absorption spectroscopy. Biological endpoints being investigated after 4 or 24h incubation include cytotoxicity, total reduced glutathione, induction of antioxidative genes such as heme-oxygenase 1 (HO-1) as well as the release of the (pro)-inflammatory cytokine TNFα.

Results: Off-gases released as by-product of flame ZnO synthesis caused a significant decrease of total reduced GSH and induced further the release of the cytokine TNFα, demonstrating the influence of the gas phase on aerosol toxicology. No direct effects could be attributed to ZnO particles. By performing suspension exposure to avoid the factor "flame-gases", particle specific effects become apparent. Other parameters such as LDH and HO-1 were not influenced by gaseous compounds: Following aerosol exposure, LDH levels appeared elevated at both timepoints and the HO-1 transcript correlated positively with deposited ZnO-dose. Under submerged conditions, the HO-1 induction scheme deviated for 4 and 24h and increased extracellular LDH was found following 24h exposure.

Conclusion: In the current study, aerosol and suspension-exposure has been compared by exposing cell cultures to equivalent amounts of ZnO. Both exposure strategies differ fundamentally in their dose-response pattern. Additional differences can be found for the factor time: In the aerosol scenario, parameters tend to their maximum already after 4h of exposure, whereas under submerged conditions, effects appear most pronounced mainly after 24h. Aerosol exposure provides information about the synergistic interplay of gaseous and particulate phase of an aerosol in the context of inhalation toxicology. Exposure to suspensions represents a valuable complementary method and allows investigations on particle-associated toxicity by excluding all gas-derived effects.

Show MeSH
Related in: MedlinePlus