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Toxicity evaluation of manufactured CeO2 nanoparticles before and after alteration: combined physicochemical and whole-genome expression analysis in Caco-2 cells.

Fisichella M, Berenguer F, Steinmetz G, Auffan M, Rose J, Prat O - BMC Genomics (2014)

Bottom Line: Gene expression profiles obtained from cells exposed to NPs before and after their alteration were compared, to highlight differences in cellular functions.No change in the cerium redox state was observed for altered NPs.Conversely, Nanobyk 3810™ coated with ammonium citrate did not display any adverse effect at the same concentration.It can be assumed that the safe design of engineered nanoparticles could include robust protective layers conferring on them greater resistance to alteration during their life cycle.

View Article: PubMed Central - PubMed

Affiliation: CEA, IBEB, SBTN, Laboratoire d'Etude des Protéines Cibles, F-30207 Bagnols-sur-Cèze, France. odette.prat@cea.fr.

ABSTRACT

Background: Engineered nanomaterials may release nanosized residues, by degradation, throughout their life cycle. These residues may be a threat for living organisms. They may be ingested by humans through food and water. Although the toxicity of pristine CeO2 nanoparticles (NPs) has been documented, there is a lack of studies on manufactured nanoparticles, which are often surface modified. Here, we investigated the potential adverse effects of CeO2 Nanobyk 3810™ NPs, used in wood care, and their residues, altered by light or acid.

Results: Human intestinal Caco-2 cells were exposed to residues degraded by daylight or in a medium simulating gastric acidity. Size and zeta potential were determined by dynamic light scattering. The surface structure and redox state of cerium were analyzed by transmission electronic microscopy (TEM) and X-ray absorption spectroscopy, respectively. Viability tests were performed in Caco-2 cells exposed to NPs. Cell morphology was imaged with scanning electronic microscopy. Gene expression profiles obtained from cells exposed to NPs before and after their alteration were compared, to highlight differences in cellular functions.No change in the cerium redox state was observed for altered NPs. All CeO2 NPs suspended in the culture medium became microsized. Cytotoxicity tests showed no toxicity after Caco-2 cell exposure to these various NPs up to 170 μg/mL (24 h and 72 h). Nevertheless, a more-sensitive whole-gene-expression study, based on a pathway-driven analysis, highlighted a modification of metabolic activity, especially mitochondrial function, by altered Nanobyk 3810™. The down-regulation of key genes of this pathway was validated by qRT-PCR. Conversely, Nanobyk 3810™ coated with ammonium citrate did not display any adverse effect at the same concentration.

Conclusion: The degraded nanoparticles were more toxic than their coated counterparts. Desorption of the outside layer was the most likely cause of this discrepancy in toxicity. It can be assumed that the safe design of engineered nanoparticles could include robust protective layers conferring on them greater resistance to alteration during their life cycle.

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High-Resolution Transmission Electron Microscope images of CeO2NPs and Nanobyk™ NPs in deionized water. A) uncoated CeO2 NPs, B) NB NPs before treatment, C) NB NPs after 4 months aging in daylight (NB-DL), and D) NB NPs after acidic degradation (NB-DA). The inter-reticular distance (dhkl) measured at ~3.2 Å is attributed to the (111) crystalline plane of CeO2. No changes in shape, crystal structure, or CeO2 cluster size were observed by HRTEM.
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Fig1: High-Resolution Transmission Electron Microscope images of CeO2NPs and Nanobyk™ NPs in deionized water. A) uncoated CeO2 NPs, B) NB NPs before treatment, C) NB NPs after 4 months aging in daylight (NB-DL), and D) NB NPs after acidic degradation (NB-DA). The inter-reticular distance (dhkl) measured at ~3.2 Å is attributed to the (111) crystalline plane of CeO2. No changes in shape, crystal structure, or CeO2 cluster size were observed by HRTEM.

Mentions: Unaltered Nanobyk™ NPs and pristine CeO2 NPs were characterized in pure water. In both cases, TEM showed well-crystallized clusters of cerianite with an inter-reticular distance (dhkl) measured at around ~3.2 Å (close to the d111 of CeO2). These clusters were pseudospherical with diameters of 3 ± 1 nm (average of 50 measurements) (Figure 1). In the stock suspension, the pristine CeO2 NPs and unaltered Nanobyk™ NPs were colloidally stable, with average hydrodynamic diameters (Dh) of ~7 nm. At pH ~ 7 ± 0.1, the zeta potential of the Nanobyk™ NPs was negative (−45 ± 5 mV) compared with the positive zeta potential of the pristine CeO2 NPs (28 ± 2 mV). After both environmental (light) and gastric (acidic) degradation, the zeta potential of the Nanobyk™ NPs increased (−28 ± 2 mV and −19 ± 2 mV, respectively, at pH 7 ± 0.2). No change in the Ce redox state was observed between the unaltered Nanobyk™, Nanobyk DL and Nanobyk DA (Additional file 1: Figure S1).Figure 1


Toxicity evaluation of manufactured CeO2 nanoparticles before and after alteration: combined physicochemical and whole-genome expression analysis in Caco-2 cells.

Fisichella M, Berenguer F, Steinmetz G, Auffan M, Rose J, Prat O - BMC Genomics (2014)

High-Resolution Transmission Electron Microscope images of CeO2NPs and Nanobyk™ NPs in deionized water. A) uncoated CeO2 NPs, B) NB NPs before treatment, C) NB NPs after 4 months aging in daylight (NB-DL), and D) NB NPs after acidic degradation (NB-DA). The inter-reticular distance (dhkl) measured at ~3.2 Å is attributed to the (111) crystalline plane of CeO2. No changes in shape, crystal structure, or CeO2 cluster size were observed by HRTEM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150968&req=5

Fig1: High-Resolution Transmission Electron Microscope images of CeO2NPs and Nanobyk™ NPs in deionized water. A) uncoated CeO2 NPs, B) NB NPs before treatment, C) NB NPs after 4 months aging in daylight (NB-DL), and D) NB NPs after acidic degradation (NB-DA). The inter-reticular distance (dhkl) measured at ~3.2 Å is attributed to the (111) crystalline plane of CeO2. No changes in shape, crystal structure, or CeO2 cluster size were observed by HRTEM.
Mentions: Unaltered Nanobyk™ NPs and pristine CeO2 NPs were characterized in pure water. In both cases, TEM showed well-crystallized clusters of cerianite with an inter-reticular distance (dhkl) measured at around ~3.2 Å (close to the d111 of CeO2). These clusters were pseudospherical with diameters of 3 ± 1 nm (average of 50 measurements) (Figure 1). In the stock suspension, the pristine CeO2 NPs and unaltered Nanobyk™ NPs were colloidally stable, with average hydrodynamic diameters (Dh) of ~7 nm. At pH ~ 7 ± 0.1, the zeta potential of the Nanobyk™ NPs was negative (−45 ± 5 mV) compared with the positive zeta potential of the pristine CeO2 NPs (28 ± 2 mV). After both environmental (light) and gastric (acidic) degradation, the zeta potential of the Nanobyk™ NPs increased (−28 ± 2 mV and −19 ± 2 mV, respectively, at pH 7 ± 0.2). No change in the Ce redox state was observed between the unaltered Nanobyk™, Nanobyk DL and Nanobyk DA (Additional file 1: Figure S1).Figure 1

Bottom Line: Gene expression profiles obtained from cells exposed to NPs before and after their alteration were compared, to highlight differences in cellular functions.No change in the cerium redox state was observed for altered NPs.Conversely, Nanobyk 3810™ coated with ammonium citrate did not display any adverse effect at the same concentration.It can be assumed that the safe design of engineered nanoparticles could include robust protective layers conferring on them greater resistance to alteration during their life cycle.

View Article: PubMed Central - PubMed

Affiliation: CEA, IBEB, SBTN, Laboratoire d'Etude des Protéines Cibles, F-30207 Bagnols-sur-Cèze, France. odette.prat@cea.fr.

ABSTRACT

Background: Engineered nanomaterials may release nanosized residues, by degradation, throughout their life cycle. These residues may be a threat for living organisms. They may be ingested by humans through food and water. Although the toxicity of pristine CeO2 nanoparticles (NPs) has been documented, there is a lack of studies on manufactured nanoparticles, which are often surface modified. Here, we investigated the potential adverse effects of CeO2 Nanobyk 3810™ NPs, used in wood care, and their residues, altered by light or acid.

Results: Human intestinal Caco-2 cells were exposed to residues degraded by daylight or in a medium simulating gastric acidity. Size and zeta potential were determined by dynamic light scattering. The surface structure and redox state of cerium were analyzed by transmission electronic microscopy (TEM) and X-ray absorption spectroscopy, respectively. Viability tests were performed in Caco-2 cells exposed to NPs. Cell morphology was imaged with scanning electronic microscopy. Gene expression profiles obtained from cells exposed to NPs before and after their alteration were compared, to highlight differences in cellular functions.No change in the cerium redox state was observed for altered NPs. All CeO2 NPs suspended in the culture medium became microsized. Cytotoxicity tests showed no toxicity after Caco-2 cell exposure to these various NPs up to 170 μg/mL (24 h and 72 h). Nevertheless, a more-sensitive whole-gene-expression study, based on a pathway-driven analysis, highlighted a modification of metabolic activity, especially mitochondrial function, by altered Nanobyk 3810™. The down-regulation of key genes of this pathway was validated by qRT-PCR. Conversely, Nanobyk 3810™ coated with ammonium citrate did not display any adverse effect at the same concentration.

Conclusion: The degraded nanoparticles were more toxic than their coated counterparts. Desorption of the outside layer was the most likely cause of this discrepancy in toxicity. It can be assumed that the safe design of engineered nanoparticles could include robust protective layers conferring on them greater resistance to alteration during their life cycle.

Show MeSH
Related in: MedlinePlus