Evaluation of the in vitro and in vivo proinflammatory activities of gold (+) and gold (-) nanoparticles.

OBJECTIVE AND DESIGN: The aim of this study was to determine potential effects of gold (+) and gold (-) nanoparticles, AuNP(+) and AuNP(-), on neutrophil biology. MATERIAL OR SUBJECTS: Freshly isolated human neutrophils were used for the in vitro aspects and CD-1 mice were used in the in vivo murine air pouch model of acute neutrophilic inflammation. TREATMENT: Human neutrophils were treated with the indicated concentrations of AuNP(+) or AuNP(-) in vitro and mice received 100 or 500 µg/ml AuNP(+) or AuNP(-) into air pouches. METHODS: Cellular uptake of AuNP by neutrophils was confirmed by transmission electron microscopy and the ability of the NP to modulate apoptosis, gelatinase activity, and chemokine production and chemotaxis was determined by cytology, zymography, ELISArray, antibody array, and ELISA and by a micro-chemotaxis chamber, respectively. In vivo, exudates were harvested after 6 h to determine the leukocyte infiltration to detect the production of several cytokines by an antibody array approach and ELISA. One-way analysis of variance was used for statistical analysis. RESULTS: AuNP possess proinflammatory activities in vitro and induce mainly a neutrophil influx in vivo, albeit at different degrees. CONCLUSIONS: AuNP(+) and AuNP(-) should be added as new candidates into a growing list of NP having proinflammatory activities by themselves.

Activation of human AML14.3D10 eosinophils by nanoparticles: Modulatory activity on apoptosis and cytokine production.

Eosinophilic inflammation is frequently observed in response to nanoparticle (NP) exposure in airway rodent models of allergies where the number of eosinophils is increased in lungs. Despite this, it is surprising that the potential cytotoxic effect of NP, as well as their direct role on eosinophils is poorly documented. The present study investigated how different NP can alter the biology of the human eosinophilic cell line AML14.3D10. It was found that among NP forms of CeO2, ZnO, TiO2, and nanosilver of 20 nm (AgNP20) or 70 nm (AgNP70) diameters, only ZnO and AgNP20 induced apoptosis. Caspases-7 and -9 were not activated by the tested NP while caspase-3 was activated by AgNP20 only. However, both ZnO and AgNP20 induced cytoskeletal breakdown as evidenced by the cleavage of lamin B1. Using an ELISArray approach for the simultaneous detection of several analytes (cytokines/chemokines), it was found that only ZnO and AgNP20 increased the production of different analytes including the potent pro-inflammatory CXCL8 (IL-8) chemokine. From the data here, we conclude that toxic effects of some NP could be observed in human eosinophil-like cells and that this could be related, at least partially, by induction of apoptosis and production of cytokines and chemokines involved in inflammation. The results of this study also indicate that distinct NP do not activate similarly human eosinophils, since ZnO and AgNP20 induce apoptosis and cytokine production while others such as TiO2, CeO2, and AgNP70 do not.

Gold nanoparticles induce apoptosis, endoplasmic reticulum stress events and cleavage of cytoskeletal proteins in human neutrophils.

Gold nanoparticles (AuNPs) are promising candidates for developing nanomedicines, for the treatment of different disorders, including inflammatory diseases. However, how AuNPs could alter the biology of human neutrophils, key player cells in inflammation, is a poorly documented area of research. Here we found that, although AuNP of 20 nm (AuNP20) could be internalized in cytosolic vacuoles but that AuNP70 were localized at the cell membrane, both induced apoptosis similarly by a caspase-dependent mechanism. AuNPs induced degradation of the cytoskeletal proteins vimentin, lamin B1 and gelsolin, but, unexpectedly, did not increase their cell surface expression. Consequent with caspase-4 processing, AuNPs were found to activate endoplasmic reticulum (ER)-stress, as evidenced by activation of the three ER sensors, IRE1 (inositol-requiring protein-1), ATF-6 (activating transcription factor-6) and PERK (protein kinase RNA (PKR)-like ER kinase). AuNPs are novel human neutrophil proapoptotic agents indicating that they are toxic to these cells. However, the fact that they do not induce cell surface expression of cytoskeletal proteins could decrease potential adverse effects and toxicity of AuNPs by limiting, for example, the production of autoantibody against cytoskeleton components.

Human S100A9 potentiates IL-8 production in response to GM-CSF or fMLP via activation of a different set of transcription factors in neutrophils.

Inflammation is highly regulated by various agents. Unexpectedly, we report here that the damage-associated molecular pattern S100A9 protein, a potent neutrophil activator and inducer of cytokine production in monocytes, is not a direct activator of cytokine production in human neutrophils. However, S100A9 primed IL-8 production in fMLP- and GM-CSF-stimulated neutrophiles via NF-κB and CREB-1, and NF-κB, STAT3 and STAT5, respectively. Pharmacological inhibition confirmed the importance of these transcription factors by significantly decreasing IL-8 production. This is the first time that a different set of transcription factors are shown to be involved in S100A9-primed neutrophils in response to proinflammatory agonist.