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The impact of species and cell type on the nanosafety profile of iron oxide nanoparticles in neural cells

View Article: PubMed Central - PubMed

ABSTRACT

Background: While nanotechnology is advancing rapidly, nanosafety tends to lag behind since general mechanistic insights into cell-nanoparticle (NP) interactions remain rare. To tackle this issue, standardization of nanosafety assessment is imperative. In this regard, we believe that the cell type selection should not be overlooked since the applicability of cell lines could be questioned given their altered phenotype. Hence, we evaluated the impact of the cell type on in vitro nanosafety evaluations in a human and murine neuroblastoma cell line, neural progenitor cell line and in neural stem cells. Acute toxicity was evaluated for gold, silver and iron oxide (IO)NPs, and the latter were additionally subjected to a multiparametric analysis to assess sublethal effects.

Results: The stem cells and murine neuroblastoma cell line respectively showed most and least acute cytotoxicity. Using high content imaging, we observed cell type- and species-specific responses to the IONPs on the level of reactive oxygen species production, calcium homeostasis, mitochondrial integrity and cell morphology, indicating that cellular homeostasis is impaired in distinct ways.

Conclusions: Our data reveal cell type-specific toxicity profiles and demonstrate that a single cell line or toxicity end point will not provide sufficient information on in vitro nanosafety. We propose to identify a set of standard cell lines for screening purposes and to select cell types for detailed nanosafety studies based on the intended application and/or expected exposure.

Electronic supplementary material: The online version of this article (doi:10.1186/s12951-016-0220-y) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus

IONP-induced alterations in cell area (grey bars) and cell circularity (orange lines) visualized after labelling of the cytoplasm with the CellMask™ Blue probe for the NSC, progenitor cell lines and murine neuroblastoma cell line. Cell circularity is a measure of cell spreading and is a value between zero and one, where one represents a perfect sphere. LA-N-2 cell morphology was analysed in terms of cluster area (grey bars) and number of cells per cluster (orange bars). A decreased cell area and increased cell circularity were detected in the NSC, ReNcell and Neuro-2a cell line. For the C17.2 cells only a diminution in cell area was detected. In the LA-N-2 cell line a reduction in cells per cluster and cluster size were observed. Statistical significance is indicated when appropriate (*p < 0.05), in black for the cell area and orange in case of the cell circularity (respectively cluster area and cells per cluster in case of the LA-N-2 cell line). NTC not treated control
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Fig5: IONP-induced alterations in cell area (grey bars) and cell circularity (orange lines) visualized after labelling of the cytoplasm with the CellMask™ Blue probe for the NSC, progenitor cell lines and murine neuroblastoma cell line. Cell circularity is a measure of cell spreading and is a value between zero and one, where one represents a perfect sphere. LA-N-2 cell morphology was analysed in terms of cluster area (grey bars) and number of cells per cluster (orange bars). A decreased cell area and increased cell circularity were detected in the NSC, ReNcell and Neuro-2a cell line. For the C17.2 cells only a diminution in cell area was detected. In the LA-N-2 cell line a reduction in cells per cluster and cluster size were observed. Statistical significance is indicated when appropriate (*p < 0.05), in black for the cell area and orange in case of the cell circularity (respectively cluster area and cells per cluster in case of the LA-N-2 cell line). NTC not treated control

Mentions: While only a significantly decreased cell area was noted for the C17.2 cell line, both a reduced cell area and an increase in circularity were observed in the NSC, ReNcells and Neuro-2a cells (Fig. 5; Additional file 1: Figure S8). Thus, the cells became both smaller and more spherical in a concentration-dependent fashion (Fig. 6), which was most outspoken in the ReNcells. Such loss of specific morphological features and cell shrinking has already been described in numerous studies for multiple NPs and cell types [3, 8, 22, 42]. Since it is known that cell transformation or immortalization affects cell morphology, it is not surprising that morphology was also differentially affected in the various cell types. For instance the mNSCs were more strongly affected in terms of morphology whereas only minor effects were observed in the C17.2 or Neuro-2a cell line. Since stem cells have a more intricate architecture in comparison to most cell lines, it was not surprising that the morphology of the former was impaired more extensively. Finally, as the LA-N-2 cells tend to grow in clusters we evaluated effects on cell morphology in terms of the total cluster area and number of cells per cluster, which both showed a similar concentration-dependent decrease starting from 3.5 nM IONPs. Since the decrease in cluster area was slightly more severe than the number of cells per cluster, we concluded that the cell area also decreased with every dose tested.Fig. 5


The impact of species and cell type on the nanosafety profile of iron oxide nanoparticles in neural cells
IONP-induced alterations in cell area (grey bars) and cell circularity (orange lines) visualized after labelling of the cytoplasm with the CellMask™ Blue probe for the NSC, progenitor cell lines and murine neuroblastoma cell line. Cell circularity is a measure of cell spreading and is a value between zero and one, where one represents a perfect sphere. LA-N-2 cell morphology was analysed in terms of cluster area (grey bars) and number of cells per cluster (orange bars). A decreased cell area and increased cell circularity were detected in the NSC, ReNcell and Neuro-2a cell line. For the C17.2 cells only a diminution in cell area was detected. In the LA-N-2 cell line a reduction in cells per cluster and cluster size were observed. Statistical significance is indicated when appropriate (*p < 0.05), in black for the cell area and orange in case of the cell circularity (respectively cluster area and cells per cluster in case of the LA-N-2 cell line). NTC not treated control
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5017038&req=5

Fig5: IONP-induced alterations in cell area (grey bars) and cell circularity (orange lines) visualized after labelling of the cytoplasm with the CellMask™ Blue probe for the NSC, progenitor cell lines and murine neuroblastoma cell line. Cell circularity is a measure of cell spreading and is a value between zero and one, where one represents a perfect sphere. LA-N-2 cell morphology was analysed in terms of cluster area (grey bars) and number of cells per cluster (orange bars). A decreased cell area and increased cell circularity were detected in the NSC, ReNcell and Neuro-2a cell line. For the C17.2 cells only a diminution in cell area was detected. In the LA-N-2 cell line a reduction in cells per cluster and cluster size were observed. Statistical significance is indicated when appropriate (*p < 0.05), in black for the cell area and orange in case of the cell circularity (respectively cluster area and cells per cluster in case of the LA-N-2 cell line). NTC not treated control
Mentions: While only a significantly decreased cell area was noted for the C17.2 cell line, both a reduced cell area and an increase in circularity were observed in the NSC, ReNcells and Neuro-2a cells (Fig. 5; Additional file 1: Figure S8). Thus, the cells became both smaller and more spherical in a concentration-dependent fashion (Fig. 6), which was most outspoken in the ReNcells. Such loss of specific morphological features and cell shrinking has already been described in numerous studies for multiple NPs and cell types [3, 8, 22, 42]. Since it is known that cell transformation or immortalization affects cell morphology, it is not surprising that morphology was also differentially affected in the various cell types. For instance the mNSCs were more strongly affected in terms of morphology whereas only minor effects were observed in the C17.2 or Neuro-2a cell line. Since stem cells have a more intricate architecture in comparison to most cell lines, it was not surprising that the morphology of the former was impaired more extensively. Finally, as the LA-N-2 cells tend to grow in clusters we evaluated effects on cell morphology in terms of the total cluster area and number of cells per cluster, which both showed a similar concentration-dependent decrease starting from 3.5 nM IONPs. Since the decrease in cluster area was slightly more severe than the number of cells per cluster, we concluded that the cell area also decreased with every dose tested.Fig. 5

View Article: PubMed Central - PubMed

ABSTRACT

Background: While nanotechnology is advancing rapidly, nanosafety tends to lag behind since general mechanistic insights into cell-nanoparticle (NP) interactions remain rare. To tackle this issue, standardization of nanosafety assessment is imperative. In this regard, we believe that the cell type selection should not be overlooked since the applicability of cell lines could be questioned given their altered phenotype. Hence, we evaluated the impact of the cell type on in vitro nanosafety evaluations in a human and murine neuroblastoma cell line, neural progenitor cell line and in neural stem cells. Acute toxicity was evaluated for gold, silver and iron oxide (IO)NPs, and the latter were additionally subjected to a multiparametric analysis to assess sublethal effects.

Results: The stem cells and murine neuroblastoma cell line respectively showed most and least acute cytotoxicity. Using high content imaging, we observed cell type- and species-specific responses to the IONPs on the level of reactive oxygen species production, calcium homeostasis, mitochondrial integrity and cell morphology, indicating that cellular homeostasis is impaired in distinct ways.

Conclusions: Our data reveal cell type-specific toxicity profiles and demonstrate that a single cell line or toxicity end point will not provide sufficient information on in vitro nanosafety. We propose to identify a set of standard cell lines for screening purposes and to select cell types for detailed nanosafety studies based on the intended application and/or expected exposure.

Electronic supplementary material: The online version of this article (doi:10.1186/s12951-016-0220-y) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus