Titanium dioxide (TiO2) nanoparticles induce neutrophil influx and local production of several pro-inflammatory mediators in vivo

doi:10.1016/j.intimp.2011.03.007

International Immunopharmacology

Volume 11, Issue 8, August 2011, Pages 1109-1115

https://doi.org/10.1016/j.intimp.2011.03.007 Get rights and content

Abstract

We have recently described the in vitro effects of titanium dioxide (TiO₂) nanoparticles (NPs) in human neutrophils. The objective of the present study was to determine if TiO₂ NPs induced leukocyte infiltration in the in vivo murine air pouch model of acute inflammation and, if so, to identify which of the main cell populations were attracted. TiO₂ NPs induced leukocyte influx after 3–9 h of treatment, since the number of leukocytes attracted significantly increased when compared to controls. Neutrophils are the most abundant leukocytes attracted by TiO₂ representing more than ~ 80% of cells, with the rest being mononuclear cells. Using an antibody array technique to detect 40 different analytes, we observed that TiO₂ increased the local production of several analytes in the exudates, including diverse chemokines. Using an ELISA assay, we found that the concentration of TiO₂-induced MIP-1β was significantly increased vs controls, corroborating the antibody array results.

Introduction

The field of nanoscience has witnessed a rapid growth in the last decade. Applications are numerous and include aerospace, defense, national security, electronics, biology and biomedicine [1]. For example, in biomedicine, “nanoparticle (NP)-oligonucleotide” hybrids can perform many chemical and biological tasks, including molecule and protein targeting or drug and gene delivery vehicles. These new nanodevices have many novel properties that are valuable for the medical biotechnology sector [2]. Consequently, most researchers working in nanoscience focus mainly on technological applications, rather than on concerns regarding the safety of nanomaterial exposure. In light of this, there has been a recent surge of interest in the expanding field of nanotoxicology [3] investigating the safety evaluation of engineered nanostructures and nanodevices [4].

Titanium dioxide (TiO₂), an odorless white powder that naturally occurs in minerals such as anatase, rutile, and brookite, is widely used as white pigment for paints, paper, plastic, ceramics, etc. However, TiO₂ NPs are transparent at the nanoscale and are able to absorb and reflect UV light. This explains their usefulness in sunscreens and in solar cells used to make solar panels [3]. At the macrometer scale, TiO₂ has always been considered nontoxic and biologically inert [5]. However, at the nanoscale, it has different physical properties. TiO₂ has been shown to be able to penetrate skin [6] and catalyze oxidative damage to DNA in vitro and in vivo [7]. A distribution experiment also revealed that after intraperitoneal injection in mice, TiO₂ NPs were retained in multiple organs and tissues, and induced various degrees of organ lesions. [8]. From an inflammatory perspective, it appears that TiO₂ NPs possess pro-inflammatory properties similar to those exhibited by other nanomaterials. Some pro-inflammatory effects of TiO₂ NPs were observed in vitro in pulmonary cells and in animals following lung instillation. In fact, the vast majority of studies investigating the inflammatory properties of nanomaterials target pulmonary cells, airways and lungs [5], [9], [10], [11]. However, NPs can also gain entry into human systems through ingestion and via dermal routes [6], [12]. Curiously, there are few in vivo and in vitro studies interested in cytokine production to assess potential inflammatory responses following cell stimulation by TiO₂. In one report, TiO₂ NPs administered intravenously in rats caused no significant chemokine, cytokine or any other parameter of immune modulation variation [12]. In another study, a 24 h treatment of A549 human lung epithelial cells with TiO₂ NPs caused a significant increase in interleukin (IL)-8 [11].

It is a well-known fact that polymorphonuclear neutrophil (PMN) cell numbers increase in TiO₂-lung inflammation in vivo [8], [10], [13]. However, research on this phenomenon in other tissues is scarce. There is also a lack of literature concerning the role of TiO₂ on neutrophil cell physiology, particularly in PMNs of human origin. We have addressed this problem and found that TiO₂ exerts important human neutrophil agonistic properties in vitro, including the capacity to induce morphological cell change and phosphorylation events, to delay apoptosis and to increase the production of cytokines and chemokines including the potent IL-8 chemokine [14]. The objective of the present study was to determine if TiO₂ NPs induced leukocyte infiltration in the murine air pouch model of acute inflammation.

Section snippets

Nanoparticles

Titanium dioxide (−) (TiO₂) nanoparticles (NPs) were purchased from Vivenano (Toronto, ONT) as an aqueous suspension of TiO₂ NPs stabilized by polyacrylate sodium that are stable over a wide range of pH. The particles sizes are 1–10 nm (90%) as determined by transmission electron microscopy. Structure of the NPs is anatase crystal as confirmed by X-ray crystallography. They were used as received from the manufacturer without modification.

Murine air pouch model

CD1 female mice, 6 to 8 weeks of age, were obtained from

TiO₂ induces leukocyte infiltration in vivo: predominance of neutrophils

We have routinely used the murine air pouch model to detect leukocyte infiltration after 6–9 h of treatment with a variety of compounds, including cytokines [17], [18]. Because of this, we initiated a series of experiments in order to determine whether or not TiO₂ NPs attract leukocytes into air pouches. As illustrated in Fig. 1A. TiO₂ NPs attract an increased number of leukocytes into the air pouch after 9 h in a concentration-dependent fashion. A statistically significant increased number of

Discussion

Although it is important to fully characterize NPs for some applications, it is clear that when such NPs are administered in vivo, they will be completely different (e.g. formation of aggregates, complex with macromolecules and proteins, etc.). Therefore, we did not characterize further the NPs than the manufacturer. However, we clearly establish that these TiO₂ NPs possess in vivo pro-inflammatory activity since they attract PMNs and, to a lesser extent, mononuclear cells in the murine air

Acknowledgments

This study was partly supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) and Environmental Health Research Network (RRSE) from Fonds de la Recherche en Santé du Québec (FRSQ). DMG holds a Fondation Armand-Frappier M.Sc award. We thank Mary Gregory for reading this manuscript.

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Nanotechnology is a growing science that may provide several new applications for medicine, food preservation, diagnostic technologies, and sanitation. Despite its beneficial applications, there are several questions related to the safety of nanomaterials for human use. The development of nanotechnology is associated with some concerns because of the increased risk of carcinogenesis following exposure to nanomaterials. The increased levels of reactive oxygen species (ROS) that are due to exposure to nanoparticles (NPs) are primarily responsible for the genotoxicity of metal NPs. Not all, but most metal NPs are able to directly produce free radicals through the release of metal ions and through interactions with water molecules. Furthermore, the increased production of free radicals and the cell death caused by metal NPs can stimulate reduction/oxidation (redox) reactions, leading to the continuous endogenous production of ROS in a positive feedback loop. The overexpression of inflammatory mediators, such as NF-kB and STATs, the mitochondrial malfunction and the increased intracellular calcium levels mediate the chronic oxidative stress that occurs after exposure to metal NPs. In this paper, we review the genotoxicity of different types of metal NPs and the redox mechanisms that amplify the toxicity of these NPs.
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Titanium dioxide (TiO2) nanoparticles induce neutrophil influx and local production of several pro-inflammatory mediators in vivo

Abstract

Introduction

Section snippets

Nanoparticles

Murine air pouch model

TiO2 induces leukocyte infiltration in vivo: predominance of neutrophils

Discussion

Acknowledgments

Toxicol Lett

FEBS Lett

Sci Total Environ

Toxicol Appl Pharmacol

Toxicol In Vitro

Transpl Immunol

Nanoparticles and nanodevices in biological applications

Biology of TiO2-oligonucleotide nanocomposites

Nat Mater

Safety of nanoparticles

Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles

Environ Health Perspect