RESUMEN
To elucidate the effect of size on the pulmonary toxicity of single-wall carbon nanotubes (SWCNTs), we prepared two types of dispersed SWCNTs, namely relatively thin bundles with short linear shapes (CNT-1) and thick bundles with long linear shapes (CNT-2), and conducted rat intratracheal instillation tests and in vitro cell-based assays using NR8383 rat alveolar macrophages. Total protein levels, MIP-1α expression, cell counts in BALF, and histopathological examinations revealed that CNT-1 caused pulmonary inflammation and slower recovery and that CNT-2 elicited acute lung inflammation shortly after their instillation. Comprehensive gene expression analysis confirmed that CNT-1-induced genes were strongly associated with inflammatory responses, cell proliferation, and immune system processes at 7 or 30 d post-instillation. Numerous genes were significantly upregulated or downregulated by CNT-2 at 1 d post-instillation. In vitro assays demonstrated that CNT-1 and CNT-2 SWCNTs were phagocytized by NR8383 cells. CNT-2 treatment induced cell growth inhibition, reactive oxygen species production, MIP-1α expression, and several genes involved in response to stimulus, whereas CNT-1 treatment did not exert a significant impact in these regards. These results suggest that SWCNTs formed as relatively thin bundles with short linear shapes elicited delayed pulmonary inflammation with slower recovery. In contrast, SWCNTs with a relatively thick bundle and long linear shapes sensitively induced cellular responses in alveolar macrophages and elicited acute lung inflammation shortly after inhalation. We conclude that the pulmonary toxicity of SWCNTs is closely associated with the size of the bundles. These physical parameters are useful for risk assessment and management of SWCNTs.
Asunto(s)
Nanotubos de Carbono/toxicidad , Administración por Inhalación , Animales , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Recuento de Células , Línea Celular , Quimiocina CCL3/inmunología , Perfilación de la Expresión Génica , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/patología , Masculino , Ratas Wistar , Especies Reactivas de Oxígeno/inmunologíaRESUMEN
Concern over the influence of carbon nanotubes (CNTs) on human health has arisen due to advances; however, little is known about the potential toxicity of CNTs. In this study, impurity-free single-wall carbon nanotubes (SWCNTs), with different physical properties in cell culture medium, were prepared by a novel dispersion procedure. SWCNTs with small bundles (short linear shape) and SWCNTs with large bundles (long linear shape) did not cause a significant inhibition of cell proliferation, induction of apoptosis or arrest of cell cycle progression in A549 alveolar epithelial cells. Expression of many genes involved in the inflammatory response, apoptosis, response to oxidative stress and degradation of the extracellular matrix were not markedly upregulated or downregulated. However, SWCNTs with relatively large bundles significantly increased the level of intracellular reactive oxygen species (ROS) in a dose-dependent manner, and the levels of these ROS were higher than those of SWCNTs with relatively small bundles or commercial SWCNTs with residual metals. Transmission electron microscopy (TEM) revealed that impurity-free SWCNTs were observed in the cytoplasm and vacuoles of cells after 24 h. These results suggested that the physical properties, especially the size and length of the bundles of the SWCNTs dispersed in cell culture medium, contributed to a change in intracellular ROS generation, even for the same bulk SWCNTs. Additionally, the residual metals associated with the manufacturing of SWCNTs may not be a definitive parameter for intracellular ROS generation in A549 cells.
Asunto(s)
Nanotubos de Carbono , Alveolos Pulmonares/citología , Células Cultivadas , Medios de Cultivo , Células Epiteliales/citología , Citometría de Flujo , Microscopía Electrónica de TransmisiónRESUMEN
Diethylhexyl phthalate (DEHP) emission to air and transfer to house dust from a polyvinyl chloride (PVC) sheet were quantified for periods of 1, 3, 7, and 14 days using a passive flux sampler (PFS). Japanese Industrial Standards (JIS) test powders class 15 was used as the test house dust in settled weights of 0.3, 1, 3, and 12 mg/cm2. DEHP concentrations in the surface air on the PVC sheet were estimated as 2.6-3.3 µg/m3 according to an emission test without dust. Although DEHP transfer rates from the PVC sheet to the house dust decreased over time, the adsorption did not reach an equilibrium state within 14 days. The transfer rates per dust weight increased with decreasing dust weight on the PVC sheet. The transfer rates per PVC sheet area increased nonlinearly with increasing dust weight on the PVC sheet. DEHP emission from a PVC sheet to air was one to three orders of magnitude lower than DEHP transfer from a PVC sheet to dust. In the case of 0.3 mg/cm2 of settled house dust for 7 days, the emission rates to air were 35, 15, 9.1, 6.4, and 3.8 µg/m2/h for a diffusion distance of 0.90, 1.85, 2.75, 3.80, and 5.75 mm, respectively, and the transfer rate to dust was 5.3 × 102 µg/m2/h (no difference among the five diffusion distances). Compared to residents who clean the floor every day, exposure to DEHP in house dust could be 10 times higher for residents who clean the floor once every two weeks based on the time-weighted average concentrations in house dust.
RESUMEN
Relationships between the physical properties of carbon nanotubes (CNTs) and their toxicities have been studied. However, little research has been conducted to investigate the pulmonary and pleural inflammation caused by short-fiber single-walled CNTs (SWCNTs) and multi-walled CNTs (MWCNTs). This study was performed to characterize differences in rat pulmonary and pleural inflammation caused by intratracheal instillation with doses of 0.15 or 1.5mg/kg of either short-sized SWCNTs or MWCNTs. Data from bronchoalveolar lavage fluid analysis, histopathological findings, and transcriptional profiling of rat lungs obtained over a 90-day period indicated that short SWCNTs caused persistent pulmonary inflammation. In addition, the short MWCNTs markedly impacted alveoli immediately after instillation, with the levels of pulmonary inflammation following MWCNT instillation being reduced in a time-dependent manner. MWCNT instillation induced greater levels of pleural inflammation than did short SWCNTs. SWCNTs and MWCNTs translocated in mediastinal lymph nodes were observed, suggesting that SWCNTs and MWCNTs underwent lymphatic drainage to the mediastinal lymph nodes after pleural penetration. Our results suggest that short SWCNTs and MWCNTs induced pulmonary and pleural inflammation and that they might be transported throughout the body after intratracheal instillation. The extent of changes in inflammation differed following SWCNT and MWCNT instillation in a time-dependent manner.
Asunto(s)
Pulmón/efectos de los fármacos , Nanotubos de Carbono/toxicidad , Pleura/efectos de los fármacos , Pleuresia/inducido químicamente , Neumonía/inducido químicamente , Animales , Citocinas/genética , Citocinas/metabolismo , Perfilación de la Expresión Génica , Mediadores de Inflamación/metabolismo , Exposición por Inhalación , Pulmón/metabolismo , Pulmón/patología , Sistema Linfático/efectos de los fármacos , Sistema Linfático/metabolismo , Masculino , Pleura/metabolismo , Pleura/patología , Pleuresia/genética , Pleuresia/metabolismo , Pleuresia/patología , Neumonía/genética , Neumonía/metabolismo , Neumonía/patología , Ratas Wistar , Factores de TiempoRESUMEN
A cDNA containing the gene for Japanese flounder IgD consisted of 3240 bp encoding 998 amino acid residues. The amino acid sequence of the constant region of Japanese flounder IgD shares 38-80% identity with the sequences of previously reported teleost IgDs. The structure of the constant region of Japanese flounder IgD, which contains the micro1, delta1, delta2, delta3, delta4, delta5, delta6, delta7, and TM regions, is similar to the structures of the constant regions of the IgDs of channel catfish and Atlantic salmon. Southern blot hybridisation showed that the Japanese flounder IgD gene exists as a single locus. The Japanese flounder IgD gene was mainly detected in peripheral blood leucocytes (PBLs) and small amounts were detected in the spleen, head and trunk kidney, although IgM mRNA was detected in similar amounts in PBLs, the head kidney, and spleen. The copy number of IgM mRNA in Japanese flounder PBL was 56-fold higher than that of IgD.