The role of HMGB1 on SiC NPs-induced inflammation response in lung epithelial-macrophage co-culture system.

In recent years, carbonized silicon nanoparticles (SiC NPs) have found widespread scientific and engineering applications, raising concerns about potential human health risks. SiC NPs may induce pulmonary damage through sustained inflammatory responses and oxidative stress, with unclear toxicity mechanisms. This study uses an in vitro co-culture model of alveolar macrophages (NR8383) and alveolar epithelial cells (RLE-6TN) to simulate the interaction between airway epithelial cells and immune cells, providing initial insights into SiC NP-triggered inflammatory responses. The research reveals that increasing SiC NP exposure prompts NR8383 cells to release high mobility group box 1 protein (HMGB1), which migrates into RLE-6TN cells and activates the receptor for advanced glycation end-products (RAGE) and Toll-like receptor 4 (TLR4). RAGE and TLR4 synergistically activate the MyD88/NF-κB inflammatory pathway, ultimately inducing inflammatory responses and oxidative stress in RLE-6TN cells, characterized by excessive ROS generation and altered cytokine levels. Pretreatment with RAGE and TLR4 inhibitors attenuates SiC-induced HMGB1 expression and downstream pathway proteins, reducing inflammatory responses and oxidative damage. This highlights the pivotal role of RAGE-TLR4 crosstalk in SiC NP-induced pulmonary inflammation, providing insights into SiC NP cytotoxicity and nanomaterial safety guidelines.

Chronic toxicity of core-shell SiC/TiO2 (nano)-particles to Daphnia magna under environmentally relevant food rations in the presence of humic acid.

To date, research on the toxicity and potential environmental impacts of nanomaterials has predominantly focused on relatively simple and single-component materials, whilst more complex nanomaterials are currently entering commercial stages. The current study aimed to assess the long-term and size-dependent (60 and 500 nm) toxicity of a novel core-shell nanostructure consisting of a SiC core and TiO2 shell (SiC/TiO2, 5, 25, and 50 mg L-1) to the common model organism Daphnia magna. These novel core-shell nanostructures can be categorized as advanced materials. Experiments were conducted under environmentally realistic feeding rations and in the presence of a range of concentrations of humic acid (0.5, 2, 5, and 10 mg L-1 TOC). The findings show that although effect concentrations of SiC/TiO2 were several orders of magnitude lower than the current reported environmental concentrations of more abundantly used nanomaterials, humic acid can exacerbate the toxicity of SiC/TiO2 by reducing aggregation and sedimentation rates. The EC50 values (mean ± standard error) based on nominal SiC/TiO2 concentrations for the 60 nm particles were 28.0 ± 11.5 mg L-1 (TOC 0.5 mg L-1), 21.1 ± 3.7 mg L-1 (TOC 2 mg L-1), 18.3 ± 5.4 mg L-1 (TOC 5 mg L-1), and 17.8 ± 2.4 mg L-1 (TOC 10 mg L-1). For the 500 nm particles, the EC50 values were 34.9 ± 16.5 mg L-1 (TOC 0.5 mg L-1), 24.8 ± 5.6 mg L-1 (TOC 2 mg L-1), 28.0 ± 10.0 mg L-1 (TOC 5 mg L-1), and 23.2 ± 4.1 mg L-1 (TOC 10 mg L-1). We argue that fate-driven phenomena are often neglected in effect assessments, whilst environmental factors such as the presence of humic acid may significantly influence the toxicity of nanomaterials.

Exposure to silicon carbide and cancer risk: a systematic review.

PURPOSE: To conduct a systematic review of epidemiologic studies on risk of cancer from exposure to silicon carbide (SiC). METHODS: We followed established guidelines to search electronic databases for studies on populations exposed to SiC. We conducted meta-analyses when the data justified it. RESULTS: We identified two studies of SiC production workers and several studies of users. The studies of production workers indicated an increased risk of lung cancer. The increased risk was restricted to workers with elevated dust exposure and, in the most informative study from Norway, was linked to estimated cristobalite exposure, a form of crystalline silica. Increased risk was not linked to SiC particles, once cristobalite exposure was controlled for. Studies of SiC users in various industries did not reveal an increased risk of lung cancer. CONCLUSIONS: The increased risk of lung cancer detected in the SiC production industry appears to be associated with high exposure levels to total dust, including crystalline silica and cristobalite which occurred in this industry in the past decades. It may not persist under current exposure circumstances, characterized by lower levels and use of personal protection equipment. Commercial users of SiC-based products were not affected.

Evaluations of the Carcinogenicity of Carbon Nanotubes, Fluoro-Edinite, and Silicon Carbide by the International Agency for Research on Cancer (IARC).

We reported the evaluations of the carcinogenicity of fluoro-edinite, silicon carbide, and carbon nanotubes performed by IARC working group in October 2014. For carbon nanotubes (CNTs), multi-walled carbon nanotube (MWCNT)-7 was classified as Group 2B, and MWCNTs without MWCNT-7 and single-walled carbon nanotubes (SWCNTs) were classified as not classifiable in terms of their carcinogenicity to humans. There is sufficient evidence of carcinogenicity for MWCNT-7 in experimental animals, limited evidence for other MWCNTs, and inadequate evidence for SWCNTs. The mechanic evidence for CNTs was not strong. Fluoro-edinite was classified as carcinogenic to humans (Group 1) on the basis of sufficient evidence of carcinogenicity to humans and experimental animals. Silicon carbide was classified into silicon carbide fibers and whiskers. Silicon carbide fibers were evaluated as possibly carcinogenic to humans (Group 2B) on the basis of limited evidence of carcinogenicity to humans. Silicon carbide whiskers were evaluated as probably carcinogenic to humans (Group 2A) on the basis of sufficient evidence of carcinogenicity to experimental animals and the similarity of their physicochemical properties to those of asbestos in terms of the mechanism of carcinogenicity. We report the process of progression in meeting and discuss how to determine the evidence and the evaluation of the carcinogenicity of the three materials.

Oxidative stress in bacteria (Pseudomonas putida) exposed to nanostructures of silicon carbide.

Silicon carbide (SiC) nanostructures produced by combustion synthesis can cause oxidative stress in the bacterium Pseudomonas putida. The results of this study showed that SiC nanostructures damaged the cell membrane, which can lead to oxidative stress in living cells and to the loss of cell viability. As a reference, micrometric SiC was also used, which did not exhibit toxicity toward cells. Oxidative stress was studied by analyzing the activity of peroxidases, and the expression of the glucose-6-phosphate dehydrogenase gene (zwf1) using real-time PCR and northern blot techniques. Damage to nucleic acid was studied by isolating and hydrolyzing plasmids with the formamidopyrimidine [fapy]-DNA glycosylase (also known as 8-oxoguanine DNA glycosylase) (Fpg), which is able to detect damaged DNA. The level of viable microbial cells was investigated by propidium iodide and acridine orange staining.

Effect of SiC interlayer between Ti6Al4V alloy and hydroxyapatite films.

Bioactive coatings are frequently used to improve the osseointegration of the metallic implants used in dentistry or orthopaedics. Among different types of bioactive coatings, hydroxyapatite (Ca10(PO4)6(OH)2) is one of the most extensively used due to its chemical similarities to the components of bones and teeth. In this article, production and characterization of hydroxyapatite films deposited on Ti6Al4V alloy prepared by magnetron sputtering were reported. Besides, SiC was deposited on substrate surface to study the interlayer effect. Obtained coatings were annealed at 600 °C for 30 and 120 min in a mixed atmosphere of N2 + H2O vapours with the heating rate of 12 °C min(-1). The effects of SiC interlayer and heat treatment parameters on the structural, mechanical and corrosion properties were investigated. After heat treatment process, the crystalline hydroxyapatite was obtained. Additionally, cell viability tests were performed. The results show that the presence of the SiC interlayer contributes a decrease in surface roughness and improves the mechanical properties and corrosion performance of the hydroxyapatite coatings. Biological properties were not affected by the presence of the SiC interlayer.

In vitro cellular responses to silicon carbide particles manufactured through the Acheson process: impact of physico-chemical features on pro-inflammatory and pro-oxidative effects.

Silicon carbide (SiC) an industrial-scale product manufactured through the Acheson process, is largely employed in various applications. Its toxicity has been poorly investigated. Our study aims at characterizing the physico-chemical features and the in vitro impact on biological activity of five manufactured SiC powders: two coarse powders (SiC C1/C2), two fine powders (SiC F1/F2) and a powder rich in iron impurities (SiC I). RAW 264.7 macrophages were exposed to the different SiC particles and the cellular responses were evaluated. Contrary to what happens with silica, no SiC cytotoxicity was observed but pro-oxidative and pro-inflammatory responses of variable intensity were evidenced. Oxidative stress (H2O2 production) appeared related to SiC particle size, while iron level regulated pro-inflammatory response (TNFα production). To investigate the impact of surface reactivity on the biological responses, coarse SiC C1 and fine SiC F1 powders were submitted to different thermal treatments (650-1400 °C) in order to alter the oxidation state of the particle surface. At 1400 °C a decrease in TNFα production and an increase in HO·, COO(·-) radicals production were observed in correlation with the formation of a surface layer of crystalline silica. Finally, a strong correlation was observed between surface oxidation state and in vitro toxicity.

Toxicological investigations on the respirable fraction of silicon carbide grain products by the in vitro vector model.

Increased lung cancer incidence with workers at the production site of crude silicon carbide (SiC) using the Acheson process has been reported. Several agents derived from the process were discussed as causative factors. Recently concern had been expressed about the presence of cleavage fragments (CFs) in commercial products fulfilling the WHO criteria for fibers. This study has focused on the toxicological significance of such CFs. The test samples were respirable fractions of five different commercial samples of SiC grains. The CF content (scanning electron microscopy) was in the range 17-493 fibers/µg. Crystalline silica and whiskers could not be detected. Quartz DQ12, cristobalite, SiC whisker, UICC crocidolite and electrocorundum were used as control reference samples. Biological activity was assessed with the in vitro vector model (VM) on ex vivo rat and guinea pig alveolar macrophages (AMs). The dose range of the VM is derived from calculated AM loads from intratracheal instillation experiments and confirmed by measured AM loads from inhalation studies on alumina monohydrate particles with low biological activity: ≤120 pg/AM. The response of the references was clearly different from that of the SiC grains which yielded low toxicity overall. However, the parameter reactive oxygen species secreted by AMs was elevated at the higher SiC doses, but not related to the CF content of these samples. Our data showed that CFs seem to have no biological relevance. This is in agreement with results from recent studies in which no carcinogenic activity had been demonstrated for CFs.

Toxicity assessment of SiC nanofibers and nanorods against bacteria.

In the present study, evidence of the antibacterial effects of silicon carbide (SiC) nanofibers (NFSiC) and nanorods (NRSiC) obtained by combustion synthesis has been presented. It has been shown that the examined bacteria, Pseudomonas putida, could bind to the surface of the investigated SiC nanostructures. The results of respiration measurements, dehydrogenase activity measurements, and evaluation of viable bacteria after incubation with NFSiC and NRSiC demonstrated that the nanostructures of SiC affect the growth and activity of the bacteria examined. The direct count of bacteria stained with propidium iodide after incubation with SiC nanostructures revealed that the loss of cell membrane integrity could be one of the main effects leading to the death of the bacteria.

Cutaneous responses to environmental stressors.

Living organisms are continuously exposed to environmental pollutants. Because of its critical location, the skin is a major interface between the body and the environment and provides a biological barrier against an array of chemical and physical environmental pollutants. The skin can be defined as our first defense against the environment because of its constant exposure to oxidants, including ultraviolet (UV) radiation and other environmental pollutants such as diesel fuel exhaust, cigarette smoke (CS), halogenated hydrocarbons, heavy metals, and ozone (O3). The exposure to environmental pro-oxidant agents leads to the formation of reactive oxygen species (ROS) and the generation of bioactive molecules that can damage skin cells. This short review provides an overview of the effects and mechanisms of action of CS, O3, and UV on cutanous tissues.

Effects of SiC nanoparticles orally administered in a rat model: biodistribution, toxicity and elemental composition changes in feces and organs.

BACKGROUND: Silicon carbide (SiC) presents noteworthy properties as a material such as high hardness, thermal stability, and photoluminescent properties as a nanocrystal. However, there are very few studies in regard to the toxicological potential of SiC NPs. OBJECTIVES: To study the toxicity and biodistribution of silicon carbide (SiC) nanoparticles in an in vivo rat model after acute (24h) and subacute (28days) oral administrations. The acute doses were 0.5, 5, 50, 300 and 600mg·kg(-1), while the subacute doses were 0.5 and 50mg·kg(-1). RESULTS: SiC biodistribution and elemental composition of feces and organs (liver, kidneys, and spleen) have been studied by Particle-Induced X-ray Emission (PIXE). SiC and other elements in feces excretion increased by the end of the subacute assessment. SiC did not accumulate in organs but some elemental composition modifications were observed after the acute assessment. Histopathological sections from organs (stomach, intestines, liver, and kidneys) indicate the absence of damage at all applied doses, in both assessments. A decrease in the concentration of urea in blood was found in the 50mg·kg(-1) group from the subacute assessment. No alterations in the urine parameters (sodium, potassium, osmolarity) were found. CONCLUSION: This is the first study that assesses the toxicity, biodistribution, and composition changes in feces and organs of SiC nanoparticles in an in vivo rat model. SiC was excreted mostly in feces and low traces were retrieved in urine, indicating that SiC can cross the intestinal barrier. No sign of toxicity was however found after oral administration.

Mortality from non-malignant respiratory diseases among workers in the Norwegian silicon carbide industry: associations with dust exposure.

OBJECTIVES: Increased mortality from asthma, chronic bronchitis and emphysema has previously been reported among workers in the silicon carbide (SiC) industry. The objective of the present study was to evaluate the influence of specific exposure factors on mortality from obstructive lung diseases (OLD), using a newly revised job-exposure matrix. MATERIALS AND METHODS: 1687 long-term workers employed in 1913-2003 in the Norwegian SiC industry were characterised with respect to cumulative exposure to quartz, cristobalite, SiC particles and SiC fibres. Standardised mortality ratios (SMRs) for underlying causes of death, 1951-2007, were calculated stratified by category of cumulative exposure, and Poisson regression analyses of OLD were performed using cumulative exposure variables. RESULTS: An increased total mortality (SMR 1.1, 95% CI 1.0 to 1.2) and increased mortality from cancer, non-malignant respiratory diseases and external factors, were observed. The SMR of OLD was increased at the highest level of cumulative exposure to all investigated exposure factors. In the internal analyses, a twofold increased risk of OLD was observed with increasing levels of cumulative exposure to SiC particles. In a multivariate model, SiC particles showed the most stable increased risk estimate when controlled for other exposure factors, among workers with less than 15 years of employment. Among workers with more than 15 years of employment, crystalline silica, primarily cristobalite, seemed to be the most important exposure factor. CONCLUSION: Exposure to SiC and crystalline silica may contribute to OLD development among SiC industry workers in different time windows, and possibly through different mechanisms.

Toxicity of silicon carbide nanowires to sediment-dwelling invertebrates in water or sediment exposures.

Silicon carbide nanowires (SiCNW) are insoluble in water. When released into an aquatic environment, SiCNW would likely accumulate in sediment. The objective of this study was to assess the toxicity of SiCNW to four freshwater sediment-dwelling organisms: amphipods (Hyalella azteca), midges (Chironomus dilutus), oligochaetes (Lumbriculus variegatus), and mussels (Lampsilis siliquoidea). Amphipods were exposed to either sonicated or nonsonicated SiCNW in water (1.0 g/L) for 48 h. Midges, mussels, and oligochaetes were exposed only to sonicated SiCNW in water for 96 h. In addition, amphipods were exposed to sonicated SiCNW in whole sediment for 10 d (44% SiCNW on dry wt basis). Mean 48-h survival of amphipods exposed to nonsonicated SiCNW in water was not significantly different from the control, whereas mean survival of amphipods exposed to sonicated SiCNW in two 48-h exposures (0 or 15% survival) was significantly different from the control (90 or 98% survival). In contrast, no effect of sonicated SiCNW was observed on survival of midges, mussels, or oligochaetes. Survival of amphipods was not significantly reduced in 10-d exposures to sonicated SiCNW either mixed in the sediment or layered on the sediment surface. However, significant reduction in amphipod biomass was observed with the SiCNW either mixed in sediment or layered on the sediment surface, and the reduction was more pronounced for SiCNW layered on the sediment. These results indicated that, under the experimental conditions, nonsonicated SiCNW in water were not acutely toxic to amphipods, sonicated SiCNW in water were acutely toxic to the amphipods, but not to other organisms tested, and sonicated SiCNW in sediment affected the growth but not the survival of amphipods.

In vitro evaluation of SiC nanoparticles impact on A549 pulmonary cells: cyto-, genotoxicity and oxidative stress.

Silicon carbide (SiC) is considered a highly biocompatible material, consequently SiC nanoparticles (NPs) have been proposed for potential applications in diverse areas of technology. Since no toxicological data are available for these NPs, the aim of this study was to draw their global toxicological profile on A549 lung epithelial cells, using a battery of classical in vitro assays. Five SiC-NPs, with varying diameters and Si/C ratios were used, and we show that these SiC-NPs are internalized in cells where they cause a significant, though limited, cytotoxic effect. Cell redox status is deeply disturbed: SiC-NP exposure cause reactive oxygen species production, glutathione depletion and inactivation of some antioxidant enzymes: glutathione reductase, superoxide dismutase, but not catalase. Finally, the alkaline comet assay shows that SiC-NPs are genotoxic. Taken together, these data prove that SiC-NPs biocompatibility should be revisited.

Reproductive and developmental toxicity of degradation products of refrigerants in experimental animals.

The present paper summarizes the results of animal studies on the reproductive and developmental toxicity of the degradation products of refrigerants, including trifluoroacetic acid (TFA), carbon dioxide (CO(2)), carbon monoxide (CO), carbonyl fluoride (CF), hydrogen fluoride (HF) and formic acid (FA). Excessive CO(2) in the atmosphere is testicular and reproductive toxic, embryolethal, developmentally neurotoxic and teratogenic in experimental animals. As for CO, maternal exposure causes prenatal and postnatal lethality and growth retardation, skeletal variations, cardiomegaly, blood biochemical, immunological and postnatal behavioral changes, and neurological impairment in offspring of several species. Very early studies of CO in rats and guinea pigs reported fetal malformations in exposed dams. The results of toxicological studies on sodium fluoride (NaF) were used to obtain insight into the toxicity of CF and HF, because CF is rapidly hydrolyzed in contact with water yielding CO(2) and HF, and NaF is similar in kinetics and dynamics to HF. Increased fetal skeletal variation, but not malformation, was noted after the maternal administration of NaF. Rat multiple-generation studies revealed that NaF caused retarded ossification and degenerative changes in the lung and kidney in offspring. There is a lack of information about the toxicity of TFA and FA.

Histopathological changes in rat lung following intratracheal instillation of silicon carbide whiskers and potassium octatitanate whiskers.

We evaluated the pattern of pulmonary inflammation for the assessment of the biological hazards of two man-made mineral fibers. Rats were exposed by intratracheal instillation to a 2 mg dose of each of two kinds of man-made mineral fibers (PT1, potassium octatitanate whisker; and SiCW, silicon carbide whisker), or three kinds of comparable respirable particles (crystalline silica, crocidolite asbestos, and titanium dioxide, TiO(2)). The lung tissue was evaluated at 3 day, 1 wk, and 1, 3 and 6 mo after exposure. Digital images taken of the lung sections were examined by morphometric point counting method (PCM). PT1 and SiCW showed a similar inflammatory pattern, which contains temporal inflammation such as moderate alveolitis within 1 wk after the exposure, and in later phase aggregation foci of instilled fibers. Differences in repair patterns of these two man-made mineral fibers showed that the toxicity of these two fibers is less toxic than for crocidolite or crystalline silica. Although SiCW showed a higher inflammation score than TiO(2) within 1 mo after instillation, the inflammation scores and fibrotic changes of PT1 and SiCW were not significant as TiO(2) at 3 mo and 6 mo in this study. Careful use should be recommended when these materials are used in the workplace.

Calcitonin gene-related peptide (CGRP) as hazard marker for lung injury induced by dusts.

Calcitonin gene-related peptide (CGRP), which has a function as a growth factor of epithelial cells, is thought to play a role in pulmonary epithelium repair. In order to establish whether or not CGRP is associated with repair in lung damaged by dust, we examined gene expression of CGRP in the lungs of animal models exposed to different dusts. Male Wistar rats were administered 2 mg of crystalline silica, crocidolite, potassium octatitanate whisker (PT-1), and silicon carbide whisker (SiCW) suspended in saline by a single intratracheal instillation and were sacrificed at 3 d, 1 wk, 1 mo, 3 mo, and 6 mo of recovery time. Pathological findings of advanced pulmonary fibrosis were present in the rats exposed to crystalline silica and crocidolite through the experiment, whereas findings of mild or reversible pulmonary fibrosis were present in those exposed to SiCW and PT-1. The expression of CGRP in rat lung was observed by reverse-transcription polymerase chain reaction (RT-PCR) and enzyme immunometric assay (EIA). In RT-PCR, CGRP gene expression was decreased at the interval of 3 d and 1 wk in the case of crystalline silica and crocidolite; on the other hand, it was increased at 3 d and 1 wk in SiCW and at 3 d, 1 wk, and 3 mo in PT-1-exposed rats. CGRP protein level in lungs exposed to PT-1 and SiCW was also higher than that to silica and crocidolite at 3 d of recovery time. These data suggest that CGRP is associated with repair in lung damaged by different dusts, and that CGRP could be used as a sensitive biomarker to indicate the pathogenicity of dusts.

Pulmonary effects and biopersistence of deposited silicon carbide whisker after 1-year inhalation in rats.

Silicon carbide whisker (SiCW) is a durable asbestos substitute. To clarify the biopersistence, the changes of geometrical configurations of deposited whiskers, and potential carcinogenicity and fibrogenicity, 42 Wistar male rats were exposed to SiCW for 6 h/day, 5 days/wk for 1 yr by inhalation. The mass median aerodynamic diameter (geometric standard deviation, GSD), the geometric mean fiber diameter (GSD), and the geometric mean fiber length (GSD) were 2.4 microm (2.4), 0.5 microm (1.5) and 2.8 microm (2.3), respectively. The daily average exposure concentrations were 2.6 +/- 0.4 mg/m3 (98 +/- 19 fibers/ml) and the rats were sacrificed at 6 days and 3, 6, and 12 mo after the exposure. The SiCW amount deposited in each rat lung at 6 days after the exposure, determined by an x-ray diffraction method, was 5.3 +/- 1.4 mg. The biological half-time (BHT) was 16 mo calculated from the deposited SiCW at 6 days and 3, 6, and 12 mo. The BHT was more prolonged than normal physiological clearance. The diameter of SiCW in the lung at each sacrificed time during 12 mo of clearance after the inhalation did not change. However, as for the length of SiCW in the lung, longer SiCW tended to be retained in the lung as the clearance time increased, especially after 6 mo. The histopathological examination revealed bronchoalveolar hyperplasia (BAH) in 2 rats at 1 yr after the exposure and severe fibrotic changes around aggregated SiCW.