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1.
Figure 6

Figure 6. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

Components of a computational Framework for Predictive nano-Toxicology.

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
2.
Figure 2

Figure 2. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

Assay Development Workflow. Adapted from: Molecular Screening R.Damoiseaux Handbook of Drug Discovery Wiley and Sons, forthcoming

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
3.
Figure 1

Figure 1. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

Assay Development Workflow for high throughput toxicity screening of Nanomaterials.

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
4.
Figure 7

Figure 7. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

(a) Setup information for a HTS cytotoxicity assay showing plate arrangement. (b) Clustered heat-map corresponding to this data, blue colors indicate no activity, while yellow indicates toxic activity. Visual inspection identifies clusters of biologically active nanoparticles

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
5.
Figure 4

Figure 4. High content screening for assaying cytotoxic events triggered by nanoparticle interaction. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

(i) BEAS-2B cells subjected to nanoparticles, stained with nucleic acid staining Hoechst 33342 and mitochondrial dye JC1. Healthy cells shows blue nuclei and red mitochondria (Negative control, TiO2 and CeO2) while the mitochondrial depolarization causes the cytoplasm to fluoresce green (green cells in ZnO). (ii) BEAS-2B cells treated with nanopaticles and stained with a dye cocktail of Hoechst 33342, fluo-4 (for intracellular Ca2+) and propidium iodide (for accessing membrane damage). Healthy cells shows blue nuclei, while damaged cells shows green cytoplasm and red nuclei (cells in ZnO group). Depending upon the color profile, the percentage of cells affected can be assayed and nanoparticles can be ranked for their cytotoxic potential.

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
6.
Figure 3

Figure 3. Hierarchical oxidative stress model explains cellular events occurring sequentially as the level of oxidative stress increases. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

A mild oxidative stress activates the protective antioxidant machinery in an attempt to restore the redox equilibrium (Tier 1). Higher levels of oxidative stress activate MAPK and Nf-kB cascade of molecular events leading to pro-inflammatory responses (Tier 2). Oxidative stress beyond the tolerable limit of cells, triggers cell death pathways often manifested as increased cytosolic Ca2+ level, lowering of mitochondrial membrane potential, opening of mitochondrial permeability transition and cell membrane damage (Tier 3).

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.
7.
Figure 5

Figure 5. Hazardous nanomaterial properties leading to ROS generation and induction of integrated events in cytotoxicity pathway. From: No Time To Lose - High Throughput Screening To Assess Nanomaterial Safety.

Nanomaterials induce ROS production as a direct consequence of specific material properties or as a consequence of triggering cellular injury responses leading to oxidant radical generation. ROS production could trigger a range of oxidative stress effects as outlined in the hierarchical oxidative stress model. The induction of cellular toxicity at the highest level of oxidative stress involves a number of interrelated cellular responses that include intracellular Ca2+ release and mitochondrial perturbation leading to cell death with accompanying changes in cell membrane integrity and nuclear PI uptake.

R Damoiseaux, et al. Nanoscale. 2011 April;3(4):1345-1360.

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