Toxicité des nanoparticules : cas des nanotubes de carbone

International audienceConstituted only by carbon atoms, carbon nanotubes (CNT) are hydrophobic and hardly detectable in biological tissues. These properties make biokinetics and toxicology studies more complex. In this work, we propose to study the effect of bovine serum albumin as a dispersing agent for CNT. The toxicological effect of a multi walled CNT (MWCNT) and a method to investigate the biopersistance of CNT in organism, based on detection of nickel, a metallic impurity present in the MWCNT we investigated were then analyses. Our results, in rats that received 100 micro g of MWCNT by a unique intratracheal instillation, reveal that MWCNT do not induce histopathological lesions, modifications of physiological pulmonary parameters and inflammation. Moreover, MWCNT do not significantly cross the pulmonary barrier and can be eliminated by a slow mechanism. Using electonic microscopy and infrared spectroscopy, we showed that MWCNT may be cleaved and chemically modified in the lung. Analyses of MWCNT lung clearance mechanism lead us to propose a hypothesis based on the phagocytosis, apoptosis of alveolar macrophages, possible degradation of MWCNT by alveolar macrophages, and elimination of apoptotic cells.Les nanotubes de carbone (NTC) sont difficilement détectables dans les matrices biologiques. Ceci rend l'étude de leur toxicité et de leur biodistribution plus difficile. Lors de ce travail, nous avons étudié, dans un premier temps, l'effet de l'albumine sérique de veau (BSA) sur la dispersion des NTC puis dans un second temps, la toxicité, la biodistribution ainsi que la clairance d'un NTC multi-feuillet (MWCNT) chez le rat en utilisant le nickel, une impureté métallique présente dans le nanotube étudié. Après une unique instillation intratrachéale de 100 micro g de MWCNT, nos résultats ne montrent ni inflammation, ni lésions pulmonaires, ni modifications des paramètres physiologiques pulmonaires. De plus, l'absence de passage de la barrière alvéolo-capillaire et la mise en place d'un long mécanisme de clairance ont été observées dans le poumon. Afin de mieux comprendre ce mécanisme et à l'aide de la microscopie electronique et de la spectroscopie infrarouge, nous avons montré que les MWCNT sont chimiquement modifiés et sont clivés dans le poumon. Ces résultats, ainsi que l'étude de la phagocytose des MWCNT et de l'apoptose des macrophages alvéolaires, ont permis d'émettre l'hypothèse d'un mécanisme de clairance selon laquelle l'élimination des MWCNT dans le poumon serait liée à la phagocytose, l'apoptose, la dégradation de MWCNT par les macrophages alvéolaires puis la phagocytose de cellules apoptotiques

Ozone et système immunitaire

National audienceThis review focuses on rite effect on health of changes in the immune system secondary to ozone exposure and on various mechanistic hypotheses pur forward. Beyond the problems related to the variability of study criteria (e.g. age, sex, concentration and duration of different types of exposure, the slightly volatile nature of ozone and the complexity of the immune system), ozone may induce immunostimulation as shown by intensified allergic phenomena of immunosuppression expressed by increased sensitivity to bacterial infections. Different functions of the immune response (Sor example macrophage and polynuclear phagocytic and bactericidal activity, NK activity, cytokine and antibody production...) are affected. In terms of risk, the consequences of these changes depend on their intensity, their perennial nature and their association with particular genetic characteristics or other forms of external aggression, for example infection. The effect of exposure to a mixture of pollutants with unknown interactions should also be taken into consideration. Finally, the problem of normal bur possibly exaggerated immune response to a compound whose allergenicity may have been modified by ozone must also be taken into accountCette revue fait le point sur les altérations de la santé attribuables à des perturbations du Système Immunitaire consécutives à une exposition à l'ozone ainsi que sur les diverses hypothèses mécanistiques étudiées. Au-delà des problèmes posés par des critères de variabilité (âge, sexe, concentration et durée d'exposition différentes, caractère peu rémanent de l'ozone, complexité du système immunitaire...) l'ozone peut induire une immunostimulation se manifestant par une intensification des phénomènes allergiques ou une immunosuppression caractérisée par une augmentation de la sensibilité aux infections bactériennes. Des fonctions de différents acteurs de la réponse immunitaire sont atteintes: capacité de phagocytose et de bactéricidie par les macrophages et polynucléaires, activité NK production de cytokines, d'anticorps... En terme de risque, les conséquences de ces altérations dépendront de leur intensité, de leur pérennité et de leur association avec des caractères génétiques particuliers ou avec d'autres agressions, par exemple d'origines infectieuses. De même, il faut considérer que nous sommes souvent exposés à un mélange de polluants dont les interactions nous restent, à ce jour, inconnues. Il faut enfin évoquer le problème de la réponse, normale mais éventuellement exacerbée, du système immunitaire à un composé dont l'allergénicité peut avoir été modifiée par l'ozon

Effet of combined nitrogen dioxide and carbon nanoparticle exposure on lung function during ovalbumin sensitization in brown norway rat

International audienceThe interaction of particulate and gaseous pollutants in their effects on the severity of allergic inflammation and airway responsiveness are not well understood. We assessed the effect of exposure to NO2 in the presence or absence of repetitive treatment with carbon nanoparticle (CNP) during allergen sensitization and challenges in Borwn-Norway (BN) rat, in order to assess their interactions on lung function and airway responses (AR) to allergen and methacholine (MCH), end-expiratory lung volume (EELV), bronchoalveolar lavage fluid (BALF) cellular content, serum and BALF cytokine levels and histological changes. Animals were divided into the following groups (n = 6): Control; CNP (Degussa-FW2): 13 nm, 0.5 mg/kg instilled intratracheally ×3 at 7-day intervals; OVA: ovalbumin-sensitised; OVA+CNP: both sensitized and exposed to CNP. Rats were divided into equal groups exposed either to air or to NO2, 10 ppm, 6 h/d, 5d/wk for 4 weeks. Exposure to NO2, significantly enhanced lung inflammation and airway reactivity, with a significantly larger effect in animals sensitized to allergen, which was related to a higher expression of TH1 and TH2-type cytokines. Conversely, exposure to NO2 in animals undergoing repeated tracheal instillation of CNP alone, increased BALF neutrophilia and enhanced the expression of TH1 cytokines: TNF-a and IFN-?, but did not show an additive effect on airway reactivity in comparison to NO2 alone. The exposure to NO2 combined with CNP treatment and allergen sensitization however, unexpectedly resulted in a significant decrease in both airway reactivity to allergen and to methacholine, and a reduction in TH2-type cytokines compared to allergen sensitization alone. EELV was significantly reduced with sensitization, CNP treatment or both. These data suggest an immunomodulatory effect of repeated tracheal instillation of CNP on the proinflammatory effects of NO2 exposure in sensitized BN rat. Furthermore, our findings suggest that NO2, CNP and OVA sensitization may significantly slow overall lung growth in parenchymally mature animals

Retentissement d'une acidification du milieu sur la glycosylation membranaire de cellules épithéliales de trachée en culture

International audienceLes pluies acides representent une des formes les plus graves de la pollution atmospherique. Elles sont generees essendellement par solubilisation dans l'eau de deux gaz polluants: le S02 qui peut se transformer en acide sulfurique et le dioxide d'azote qui peut generer de l'acide nitrique. Ces acides conferent alors aux precipitations un pH moyen de 3,6 pouvant aller jusqu'a 2 et, de ce fait, contribuent a abaisser le pH de nombreux milieux biologiques. Ceux ci comprennent des ecosystemes entiers (lacs) mais egalement certains tissus humains comme les voies respiratoires. Des etudes ont montre que I'abaissement du pH pouvait jouer un role significatif dans l'alteration de certaines fonctions essentielles pour de nombreuses cellules, en particulier, les cellules epitheliales. Certaines de ces fonctions sont associees ä l'expression membranaire de structures complexes dans lesquelles la partie glucidique est primordiale et, dans ce cas, les acides sialiques sont incrimines

Interference of a short-term exposure to nitrogen dioxide with allergic airways responses to allergenic challenges in BALB/c mice.

Nitrogen dioxide (NO(2)) is a common indoor and outdoor air pollutant whose role in the induction of asthma is unclear. We investigated the effects of NO(2) on the development of asthma-like responses to allergenic challenge in BALB/c mice. Ovalbumin (OVA)-immunized mice were intranasally challenged with OVA or saline solution just before starting a 3 h exposure to 5 or 20 ppm NO(2) or air. Twenty parts per million of NO(2) induced a significant increase of bronchopulmonary hyperreactivity in OVA-challenged mice and of permeability according to the fibronectin content of the bronchoalveolar lavage fluid (BALF) 24 h after exposure, as compared with air or 5 ppm NO(2). Eosinophilia (cell counts in the BALF and eosinophil peroxidase of lung tissue) was detected at 24 and 72 h with similar levels for air and 20 ppm NO(2), whereas a marked reduction was unexpectedly observed for 5 ppm NO(2). At 24 h, interleukin-5 in the BALF was markedly reduced at 5 ppm compared with 20 ppm NO(2) and was also more intense for 20 ppm NO(2) than for the air group. In contrast to specific IgG1 titers, anti-OVA IgE titers and interleukin-4 in the BALF were not affected by NO(2) exposure. Irrespective of the concentration of NO(2), OVA-challenged mice did not develop late mucosal metaplasia compared with those exposed to OVA-air. These results indicate that a short exposure to NO(2) can exacerbate or inhibit some features of the development of allergic disease in mice and may depend on the concentration of pollutant

In vivo biodistribution and biological impact of injected carbon nanotubes using magnetic resonance techniques

International audienceSingle-walled carbon nanotubes (SWCNT) hold promise for applications as contrast agents and target delivery carriers in the field of nanomedicine. When administered in vivo, their biodistribution and pharmacological profile needs to be fully characterized. The tissue distribution of carbon nanotubes and their potential impact on metabolism depend on their shape, coating, and metallic impurities. Because standard radiolabeled or fluorescentlylabeled pharmaceuticals are not well suited for long-term in vivo follow-up of carbon nanotubes, alternative methods are required

Comparative toxicity of 24 manufactured nanoparticles in human alveolar epithelial and macrophage cell lines

<p>Abstract</p> <p>Background</p> <p>A critical issue with nanomaterials is the clear understanding of their potential toxicity. We evaluated the toxic effect of 24 nanoparticles of similar equivalent spherical diameter and various elemental compositions on 2 human pulmonary cell lines: A549 and THP-1. A secondary aim was to elaborate a generic experimental set-up that would allow the rapid screening of cytotoxic effect of nanoparticles. We therefore compared 2 cytotoxicity assays (MTT and Neutral Red) and analyzed 2 time points (3 and 24 hours) for each cell type and nanoparticle. When possible, TC50 (Toxic Concentration 50 i.e. nanoparticle concentration inducing 50% cell mortality) was calculated.</p> <p>Results</p> <p>The use of MTT assay on THP-1 cells exposed for 24 hours appears to be the most sensitive experimental design to assess the cytotoxic effect of one nanoparticle. With this experimental set-up, Copper- and Zinc-based nanoparticles appear to be the most toxic. Titania, Alumina, Ceria and Zirconia-based nanoparticles show moderate toxicity, and no toxicity was observed for Tungsten Carbide. No correlation between cytotoxicity and equivalent spherical diameter or specific surface area was found.</p> <p>Conclusion</p> <p>Our study clearly highlights the difference of sensitivity between cell types and cytotoxicity assays that has to be carefully taken into account when assessing nanoparticles toxicity.</p

Biodistribution and clearance of instilled carbon nanotubes in rat lung

<p>Abstract</p> <p>Background</p> <p>Constituted only by carbon atoms, CNT are hydrophobic and hardly detectable in biological tissues. These properties make biokinetics and toxicology studies more complex.</p> <p>Methods</p> <p>We propose here a method to investigate the biopersistence of CNT in organism, based on detection of nickel, a metal present in the MWCNT we investigated.</p> <p>Results and conclusion</p> <p>Our results in rats that received MWCNT by intratracheal instillation, reveal that MWCNT can be eliminated and do not significantly cross the pulmonary barrier but are still present in lungs 6 months after a unique instillation. MWCNT structure was also showed to be chemically modified and cleaved in the lung. These results provide the first data of CNT biopersistence and clearance at 6 months after respiratory administration.</p

Decrease in ovalbumin-induced pulmonary allergic response by benzaldehyde but not acetaldehyde exposure in a guinea pig model

International audienceThe pulmonary effects of two environmentally relevant aldehydes were investigated in non-sensitized or ovalbumin (OA)-sensitized guinea pigs (GPs). Four-week-old male Hartley GPs, weighing about 400 g, were intraperitoneally injected with 1 ml of an NaCl solution containing 100 mug OA and 100 mg Al/(OH)(3). They were then exposed to either acetaldehyde (200 ppb) or benzaldehyde (500 ppb) (or 4 wk (6 h/d, 5 d/wk). At the end of exposure, GPs were challenged with an OA aerosol (0.1% in NaCl) and pulmonary functions were measured. The day after, guinea pigs were anesthetized and several endpoints related to inflammatory anti allergic responses were assessed in blood, whole-lung histology, and bronchoalveolar lavage (BAL). Sensitized nonexposed GPs showed bronchial hyperresponsiveness to OA and an increased number of eosinophils in blood and BAL, together with a rise in total protein and leukotrienes (LTB4 and LTC4/D-4/E-4) in BAL. In nonsensitized GPs, exposure to acetaldehyde or benzaldehyde did not induce any change in the tested parameters;, with the exception of irritation of the respiratory tract as detected by histology and an increased number of alveolar macrophages in animals exposed to acetaldehyde. In sensitized GPs, exposure to acetaldehyde induced a moderate irritation of the respiratory tract but no change in biological parameters linked to the inflammatory and allergic responses, In contrast, exposure to benzaldehyde induced a decrease both in OA-induced bronchoconstriction and in eosinophil and neutrophil numbers in BAL, an increase in the bronchodilatator mediator prostaglandin E-2 (PGE(2)) and a decrease in the bronchoconstrictor mediators LTC4/D-4/E-4. Further investigations are needed to determine if the attenuated response observed in sensitized GPs exposed to benzaldehyde is due to an alteration of the mechanism of sensitization or to a more direct effect on various mechanisms of the allergic response

Intratracheally administered titanium dioxide or carbon black nanoparticles do not aggravate elastase-induced pulmonary emphysema in rats.

International audienceABSTRACT: BACKGROUND: Titanium dioxide (TiO2) and carbon black (CB) nanoparticles (NPs) have biological effects that could aggravate pulmonary emphysema. The aim of this study was to evaluate whether pulmonary administration of TiO2 or CB NPs in rats could induce and/or aggravate elastase-induced emphysema, and to investigate the underlying molecular mechanisms. METHODS: On day 1, Sprague-Dawley rats were intratracheally instilled with 25 U kg1 pancreatic porcine elastase or saline. On day 7, they received an intratracheal instillation of TiO2 or CB (at 100 and 500 mug) dispersed in bovine serum albumin or bovine serum albumin alone. Animals were sacrificed at days 8 or 21, and bronchoalveolar lavage (BAL) cellularity, histological analysis of inflammation and emphysema, and lung mRNA expression of heme oxygenase-1 (HO-1), interleukin-1beta (IL-1beta), macrophage inflammatory protein-2, monocyte chemotactic protein-1, and matrix metalloprotease (MMP)-1, and -12 were measured. In addition, pulmonary MMP-12 expression was also analyzed at the protein level by immunohistochemistry. RESULTS: TiO2 NPs per se did not modify the parameters investigated, but CB NPs increased perivascular/peribronchial infiltration, and macrophage MMP-12 expression, without inducing emphysema. Elastase administration increased BAL cellularity, histological inflammation, HO-1, IL-1beta and macrophage MMP-12 expression and induced emphysema. Exposure to TiO2 NPs did not modify pulmonary responses to elastase, but exposure to CB NPs aggravated elastase-induced histological inflammation without aggravating emphysema. CONCLUSIONS: TiO2 and CB NPs did not aggravate elastase-induced emphysema. However, CB NPs induced histological inflammation and MMP-12 mRNA and protein expression in macrophages