Detection and analysis of nanoparticles in patients: A critical review of the status quo of clinical nanotoxicology

International audienceOn the cusp of massive commercialization of nanotechnology-enhanced products and services, the physical and chemical analysis of nanoparticles in human specimens merits immediate attention from the research community as a prerequisite for a confident clinical interpretationof their occurrence in the human organism. In this review, we describe the caveats in current practices of extracting and isolating nanoparticles from clinical samples and show that they do not help truly define the clinical significance of any detected exogenous nano-sized objects. Finally, we suggest a systematic way of tackling these demanding scientific tasks. More specifically, a precise and true qualitative evaluation of nanoparticles in human biological samples still remains difficult to achieve because of various technical reasons. Such a procedure is more refined when the nature of the pollutants is known, like in the case of nano-sized wear debris originating from biomedical prostheses. Nevertheless, nearly all available analytical methods provide unknown quantitative accuracy and qualitative precision due to the challenging physical and chemical nature of nanoparticles. Without trustworthy information to detect and describe the nanoparticulate load of clinical samples, it is impossible to accurately assess its pathological impact on isolated cases or allow for relevant epidemiological surveys on large populations. Therefore, we suggest that the many and various specimens stored in hospitals be used for the refinement of methods of exhaustive quantitative and qualitative characterization of prominent nanoparticles in complex human milieu

Etude de l'activité biologique de poudres industrielles fines et ultrafines de carbure de silicium : caractérisation physico-chimique.

National audienceL'industrie élabore et utilise des poudres de SiC de plus en plus fines. Depuis leur synthèse jusqu'à leur utilisation finale, les poudres subissent diverses opérations et traitements au cours desquels elles sont susceptibles de se disséminer dans l'atmosphère des locaux industriels, provoquant ainsi un risque d'inhalation. Les particules fines peuvent provoquer une réaction inflammatoire au niveau des poumons. L'objectif de cette étude est d'évaluer la toxicité et d'expliquer la réactivité biologique de différentes variétés de poudres industrielles de SiC

Molecular Imaging of Microglial Activation in Amyotrophic Lateral Sclerosis

There is growing evidence of activated microglia and inflammatory processes in the cerebral cortex in amyotrophic lateral sclerosis (ALS). Activated microglia is characterized by increased expression of the 18 kDa translocator protein (TSPO) in the brain and may be a useful biomarker of inflammation. In this study, we evaluated neuroinflammation in ALS patients using a radioligand of TSPO, 18F-DPA-714. Ten patients with probable or definite ALS (all right-handed, without dementia, and untreated by riluzole or other medication that might bias the binding on the TSPO), were enrolled prospectively and eight healthy controls matched for age underwent a PET study. Comparison of the distribution volume ratios between both groups were performed using a Mann-Whitney’s test. Significant increase of distribution of volume ratios values corresponding to microglial activation was found in the ALS sample in primary motor, supplementary motor and temporal cortex (p = 0.009, p = 0.001 and p = 0.004, respectively). These results suggested that the cortical uptake of 18F-DPA-714 was increased in ALS patients during the ‘‘time of diagnosis’’ phase of the disease. This finding might improve our understanding of the pathophysiology of ALS and might be a surrogate marker of efficacy of treatment on microglial activation

Biological response to purification and acid functionalization of carbon nanotubes

The final publication is available at Springer via: http://link.springer.com/article/10.1007/s11051-014-2507-yInternational audienceAcid functionalization has been considered as an easy way to enhance the dispersion and biodegradation of carbon nanotubes (CNT). However, inconsistencies between toxicity studies of acid functionalized CNT remain unexplained. This could be due to a joint effect of the main physicochemical modifications resulting from an acid functionalization: addition of surface acid groups and purification from catalytic metallic impurities. In this study, the impact on CNT biotoxicity of these two physiochemical features was assessed separately. The in vitro biological response of RAW 264.7 macrophages was evaluated after exposure to 15-240 µg x mL−1 of two types of multi-walled CNT. For each type of CNT (small: 20 nm diameter, and big: 90 nm diameter), three different surface chemical properties were studied (total of six CNT samples): pristine, acid functionalized and desorbed. Desorbed CNT were purified by the acid functionalization but presented a very low amount of surface acid groups due to a thermal treatment under vacuum. A Janus effect of acid functionalization with two opposite impacts is highlighted. The CNT purification decreased the overall toxicity, while the surface acid groups intensified it when present at a specific threshold. These acid groups especially amplified the pro-inflammatory response. The threshold mechanism which seemed to regulate the impact of acid groups should be further studied to determine its value and potential link to the other physicochemical state of the CNT. The results suggest that, for a safer-design approach, the benefit-risk balance of an acid functionalization has to be considered, depending on the CNT primary state of purification. Further research should be conducted in this direction

Testicular biodistribution of silica-gold nanoparticles after intramuscular injection in mice

International audienceWith the continuing development of nanomaterials, the assessment of their potential impact on human health, and especially human reproductive toxicity, is a major issue. The testicular biodistribution of nanoparticles remains poorly studied. This study investigated whether gold-silica nanoparticlescould be detected in mouse testes after intramuscular injection, with a particular focus on their ability to cross the blood– testis barrier. To that purpose, well-characterized 70-nm gold core–silica shell nanoparticles were used to ensure sensitive detection using high-resolution techniques. Testes were collected at different time points corresponding to spermatogenesis stages in mice. Transmission electronmicroscopy and confocal microscopy were used for nanoparticle detection, and nanoparticle quantification was performed by atomic emission spectroscopy. All these techniques showed that no particles were able to reach the testes. Results accorded with the normal histological appearance of testes even at 45 days post sacrifice.High-resolution techniques did not detect 70-nm silica-gold nanoparticles in mouse testes after intramuscular injection.These results are reassuring about the safety of nanoparticles with regard to male human reproduction, especially in the context of nanomedicine

Thermal annealing of carbon nanotubes reveals a strong toxicological impact of the structural defects

Le document inséré dans cette page comprend des données supplémentaire à l'article " Thermal annealing of carbon nanotubes reveals a toxicological impact of the structural defects" (DOI: 10.1007/s11051-015-2999-0) paru dans Journal of Nanoparticles ResearchInternational audienceThe biological response to pristine and annealed multi-walled carbon nanotubes (MWCNT) was assessed on murine macrophages (RAW 264.7). First, the physicochemical features of the as-produced MWCNT and annealed at 2125 °C for 1 h were fully characterized. A decrease in structural defects, hydrophobicity and catalytic impurities was detected after annealing. Thereafter, their impact on cytotoxicity, oxidative stress, and pro-inflammatory response was investigated at concentrations ranging from 15 to 120 µg mL−1. No effect of the 2125 °C treatment was detected on the cytotoxicity. In contrast, the annealed carbon nanotubes showed a significant increase of the pro-inflammatory response. We assumed that this behavior was due to the reduction in structural defects that may modify the layer of adsorbed biomolecules. Surprisingly, the purification of metallic catalysts did not have any significant impact on the oxidative stress. We suggested that the structural improvements from the 2125 °C treatment can decrease the carbon nanotube scavenging capacity and thus allow a higher free radical release which may counterbalance the decrease of oxidative stress due to a lower content of metallic impurities

Quantification of nanoparticle endocytosis based on double fluorescent pH-sensitive nanoparticles

International audienceAmorphous silica is a particularly interesting material because of its inertness and chemical stability. Silica nanoparticles have been recently developed for biomedical purposes but their innocuousness must be carefully investigated before clinical use. The relationship between nanoparticles physicochemical features, their uptake by cells and their biological activity represents a crucial issue, especially for the development of nanomedicine. This work aimed at adapting a method for the quantification of nanoparticle endocytosis based on pH-sensitive and double fluorescent particles. For that purpose, silica nanoparticles containing two fluorophores: FITC and pHrodoTM were developed, their respective fluorescence emission depends on the external pH. Indeed, FITC emits a green fluorescence at physiological pH and pHrodoTM emits a red fluorescence which intensity increased with acidification. Therefore, nanoparticles remained outside the cells could be clearly distinguished from nanoparticles uptaken by cells as these latter could be spotted inside cellular acidic compartments (such as phagolysosomes, micropinosomes...). Using this model, the endocytosis of 60 nm nanoparticles incubated with the RAW 264.7 macrophages was quantified using time-lapse microscopy and compared to that of 130 nm submicronic particles. The amount of internalized particles was also evaluated by fluorimetry. The biological impact of the particles was also investigated in terms of cytotoxicity, pro-inflammatory response and oxidative stress. Results clearly demonstrated that nanoparticles were more uptaken and more reactive than submicronic particles. Moreover, we validated a method of endocytosis quantification

Toxicity of boehmite nanoparticles: impact of the ultrafine fraction and of the agglomerates size on cytotoxicity and pro-inflammatory response

International audienceBoehmite (γ-AlOOH) nanoparticles (NPs) are used in a wide range of industrial applications. However, little is known about their potential toxicity. This study aimed at a better understanding of the relationship between the physico-chemical properties of these NPs and their in vitro biological activity. After an extensive physico-chemical characterization, the cytotoxicity, pro-inflammatory response and oxidative stress induced by a bulk industrial powder and its ultrafine fraction were assessed using RAW264.7 macrophages. Although the bulk powder did not trigger a significant biological activity, pro-inflammatory response was highly enhanced with the ultrafine fraction. This observation was confirmed with boehmite NPs synthesized at the laboratory scale, with well-defined and tightly controlled physico-chemical features: toxicity was increased when NPs were dispersed. In conclusion, the agglomerates size of boehmite NPs has a major impact on their toxicity, highlighting the need to study not only raw industrial powders containing NPs but also the ultrafine fractions representative of respirable particles. Read More: http://informahealthcare.com/doi/abs/10.3109/08958378.2014.92599

Testicular biodistribution of 450 nm fluorescent latex particles after intramuscular injection in mice.

International audienceThe significant expansion in the use of nanoparticles and submicron particles during the last 20 years has led to increasing concern about their potential toxicity to humans and particularly their impact on male fertility. Currently, an insufficient number of studies have focused on the testicular biodistribution of particles. The aim of our study was to assess the distribution of 450 nm fluorescent particles in mouse testes after intramuscular injection. To this end, testes were removed from 5 groups of 3 mice each at 1 h (H1), 4 days (D4), 21 days (D21), 45 days (D45) and 90 days (D90) after the injection of 7.28 × 10(9) particles in the tibialis anterior muscles of each mouse. We examined histological sections from these samples by epifluorescence microscopy and confocal microscopy and identified testicular biodistribution of a small number of particles in groups H1, D4, D21, D45 and D90. Using CD11b immunostaining, we showed that particles were not carried into the testis by macrophages. The intratesticular repartition of particles mainly followed testicular vascularization. Finally, we found some particles in seminiferous tubules but could not determine if the blood-testis barrier was crossed

Metals distribution in colorectal biopsies: New insight on the elemental fingerprint of tumour tissue

International audienceBackground: Colorectal cancer is considered to be an environmental disease. In this context, the study of environmental risk factors associated with the presence of chemical elements is important, as well as improving our knowledge of the elemental fingerprint of tumor tissuecompared to non-cancer tissue.Aims: The objective was to evaluate the element distribution in colorectal adenocarcinoma biopsies, adjacent non-tumor tissues, and healthy controls (non-cancer colorectal biopsies including occlusion or ischemic colons).Methods: The study is a case-control study which compared the element distribution in colon biopsies from two groups of patients: with colorectal cancer and without colorectal cancer. Patients with colorectal cancer provided 2 different groups of samples: colorectal cancer biopsies and adjacent non-tumor tissues. 15 metal concentrations (Al, B, Cd, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Se, Si, Ti, V, and Zn) in colorectal biopsies were quantified by using acid digestion procedures and then inductively coupled plasma (ICP) atomic emission spectrometry.Results: A total of 104 patients were included. 76 patients in the colorectal cancer group (i.e. tumor and adjacent non-tumor tissues) and 28 patients in the healthy control group (i.e. noncancer colorectal biopsies). Among the 15 elements analyzed by ICP spectrometry, only boron, chromium, zinc, silicon, and magnesium were found in colorectal tissue at clearly detectable concentrations. Our data indicated that colorectal tumor biopsies have significantly elevated concentrations of magnesium as compared to adjacent non-tumor or healthy tissues. Zinc concentration followed the same trend but differences were not statistically significant. In addition, silicon appears to be more accumulated in colorectal cancer tissue than in healthy non-cancer tissue, while chromium was mostly found in adjacent non-tumor tissue. Conclusion: Magnesium, chromium, zinc and silicon were found in noteworthy concentrations in colorectal tumor. Their potential role in colorectal carcinogenesis should be explored