Raw single-walled carbon nanotube-induced cytotoxic effects in human bronchial epithelial cells: comparison to asbestos

Single-walled carbon nanotubes (SWCNT) are being developed to be used in many industrial and biomedical applications. However, SWCNT's durability and likely fibrous morphology have raised health concerns. The present investigations were focused on understanding the cellular and molecular mechanisms induced by raw SWCNT (SWCNT) in human bronchial-epithelial cells (BEAS-2B). Asbestos (crocidolite) was used as a positive control. Exposure of BEAS-2B cells to SWCNT induced apoptosis, DNA damage, and oxidative stress. The generation of hydroxyl radical (^?OH) and increase of superoxide dismutase (SOD) activity were concentration-dependent. The increase in apoptosis was associated with activation of caspase-3, caspase-7, and poly (ADP-ribose) polymerase-1 (PARP-1). A short recovery period of 6?h of cells from SWCNT exposure resulted in reversal of caspase-3 and caspase-7, and a partial reversal of PARP-1 activation. The activation of PARP-1, caspase-3, and caspase-7 was only partially diminished after a recovery of 6?h from the exposure to crocidolite. Exposure of BEAS-2B cells to SWCNT resulted in the phosphorylation of protein p42/44 (p42/44) and protein p38 (p38). SWCNT did not induce protein serine-threonine kinase (AKT) phosphorylation. For all the above end points, crocidolite induced a greater response compared to SWCNT. SWCNT induced a significant activation of activator protein-1 (AP-1) and nuclear factor kappa B (NF-κB), and the effect was inhibited by mitogen-activated protein kinase (MAPK) inhibitors. SWCNT also induced significant increase in the expression levels of c-Jun, βIGH3, and CD44 genes. The results of this study show that the molecular mechanism for raw SWCNT-mediated toxicity in BEAS-2B cells is through the activation of caspase-3, caspase-7, and PARP-1. Furthermore, the mechanism of AP-1 and NF-κB activation is through MAPK. This bioactivity of raw SWCNT is associated with the generation of oxidative stress and DNA damage. Considering the role of airway epithelium as a critical barrier for normal pulmonary function and focal point for tumor development, this study demonstrates that raw SWCNT activate molecular events which may be linked to adverse biological responses implicated in pulmonary diseases.     

Surface characterization of acid-oxidized multi-walled carbon nanotubes

Multi-walled carbon nanotubes (MWNTs) oxidized with sulfuric acid, nitric acid, and the mixture of sulfuric and nitric acids were characterized by thermogravimetric analysis (TGA), Raman spectroscopy, elemental analysis, X-ray diffraction, and X-ray photoelectron spectroscopy. The TGA data showed that the MWNTs were more resistant to oxidation than C₆₀ or activated carbon fibers. Catalyst particles could be removed by the oxidants containing sulfuric acid, and thus indicative of the tip opening of MWNTs. The sulfuric acid had a propensity to create defect sites and introduce the surface oxides at those defects that already exist or be newly generated on MWNTs. However, the acid mixture could open the caps of MWNTs but preserve the structure homogeneity. The treatment with nitric acid gave rise to the highest bulk oxygen content in MWNTs, while the most abundant surface oxides were provided by sulfuric acid oxidation. In addition, nitric acid exhibited the best ability to transform the phenolic groups into carboxyl groups.     

Single walled carbon nanotubes induce cytotoxicity and oxidative stress in HEK293 cells

The aim of the present study was to evaluate the potential toxicity and general mechanisms involved in single walled carbon nanotubes (SWCNTs)-induced cytotoxicity using human embryonic kidney cell line (HEK293) cells. Carbon nanotubes (coded as CNT) used in this study were synthesized by the chemical vapor deposition method. To elucidate the possible mechanisms underlying SWCNT-induced cytotoxicity, cell viability, cell membrane damage (lactate dehydrogenase activity (LDH) assay), reduced glutathione (GSH), interleukin-8 (IL-8) and lipid peroxidation products levels were quantitatively assessed following SWCNT exposure for 48 hr using HEK293 cells. Exposure of cells to SWCNT at 3–300 μg/ml produced significant reduction in cell viability in a concentration-dependent manner. The IC₅₀ value of SWCNT was found to be 87.58 μg/ml. Exposure of HEK cells to SWCNT at 10–100 μg/ml resulted in concentration-dependent cell membrane damage, increased production of IL-8, elevated levels of thiobarbituric acid reactive substances like malondialdehyde and decreased intracellular GSH levels. In summary, exposure to SWCNT resulted in a concentration-dependent cytotoxicity in cultured HEK293 cells that was associated with increased oxidative stress.     

Cytotoxicity and cell membrane depolarization induced by aluminum oxide nanoparticles in human lung epithelial cells A549

The cytotoxicity of 13 and 22 nm aluminum oxide (Al₂O₃) nanoparticles was investigated in cultured human bronchoalveolar carcinoma-derived cells (A549) and compared with 20 nm CeO₂ and 40 nm TiO₂ nanoparticles as positive and negative control, respectively. Exposure to both Al₂O₃ nanoparticles for 24 h at 10 and 25 µg mL^?1 doses significantly decreased cell viability compared with control. However, the cytotoxicity of 13 and 22 nm Al₂O₃ nanoparticles had no difference at 5–25 µg mL^?1 dose range. The cytotoxicity of both Al₂O₃ nanoparticles were higher than negative control TiO₂ nanoparticles but lower than positive control CeO₂ nanoparticles (TiO₂ < Al₂O₃ < CeO₂). A real-time single cell imaging system was employed to study the cell membrane potential change caused by Al₂O₃ and CeO₂ nanoparticles using a membrane potential sensitive fluorescent probe DiBAC₄(3). Exposure to the 13 nm Al₂O₃ nanoparticles resulted in more significant depolarization than the 30 nm Al₂O₃ particles. On the other hand, the 20 nm CeO₂ particles, the most toxic, caused less significant depolarization than both the 13 and 22 nm Al₂O₃. Factors such as exposure duration, surface chemistry, and other mechanisms may contribute differently between cytotoxicity and membrane depolarization.     

纳米SiO2与常规SiO2颗粒对Hela细胞的细胞毒性作用

为了探讨纳米SiO2和常规SiO2颗粒对Hela细胞的细胞毒性作用,采用不同浓度的纳米SiO2和常规SiO2颗粒(0.05、0.1、0.2、0.4、0.8、1.6μg·μL-1)对Hela细胞进行12h染毒,应用MTT法检测细胞毒性效应.研究发现,较低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2对Hela细胞无明显细胞毒性(p>0.05);较高浓度时,纳米SiO2(≥0.4μg·μL-1)和常规SiO2(≥0.8μg·μL-1)对Hela细胞具有明显细胞毒性作用(p<0.01),并且随浓度增大细胞毒性增强;当浓度≥0.4μg·μL-1时,纳米SiO2的细胞毒性明显高于相同浓度的常规SiO2(p<0.05).以上结果表明,纳米SiO2和常规SiO2颗粒均能对Hela细胞产生细胞毒性,且纳米SiO2的细胞毒性强于常规SiO2;低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2具有很好的生物相容性.     

乙腈溶剂中单壁碳纳米管与常见有机化合物的相互作用

以单壁碳纳米管(SWCNTs)为固定相,采用高压匀浆法,制备HPLC色谱柱.测定乙腈相中18种有机化合物在SWCNTs色谱柱上的保留因子,研究SWCNTs与化合物的相互作用.结果表明,有机化合物取代基的种类和个数、杂原子的个数影响乙腈相中有机化合物与SWCNTs的相互作用.采用线性溶解能关系(LSER)定量表征SWCNTs与有机化合物的相互作用,所构建的LSER模型具有较强的拟合能力(R2=0.844)和稳健性(Q2LOO=0.768),发现π-/n-电子对作用和空穴/弥散作用是控制SWCNTs与有机化合物相互作用的主要因素.     

Cadmium sulfide nanoparticle induces oxidative stress and pro-inflammatory effects in human lung adenocarcinoma epithelial cells

Cadmium sulfide nanoparticles (CdSNP) are increasingly used in biological applications. This study was undertaken to understand the mechanisms underlying adverse effects of CdSNP using human lung adenocarcinoma epithelial (A549) cells. Cellular toxicity was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide and neutral red assays. Results showed that CdSNP reduced mitochondrial function and induced lysosomal activity in concentration and time-dependent manner. CdSNP produced oxidative stress as evidenced by reduction of glutathione (GSH) levels and increase in reactive oxygen species and lipid peroxidation levels. Induction of caspase-3 enzymes and condensed, fragmented nuclei was observed in CdSNP-treated cells. Furthermore, the levels of interleukin-8, tumor growth factor and DNA fragmentation were significantly higher in CdSNP exposed cells. Data indicated that toxicity of CdSNP noted in A549 cells may be mediated through oxidative stress. This study warrants more comprehensive assessment of CdSNP prior to industrial applications.     

Catalytic wet air oxidation of phenol,nitrobenzene and aniline over the multi-walled carbon nanotubes (MWCNTs) as catalysts

Wet air oxidation (WAO) is one of effective technologies to eliminate hazardous, toxic and highly concentrated organic compounds in the wastewater. In the paper, multi-walled carbon nanotubes (MWCNTs), functionalized by O₃, were used as catalysts in the absence of any metals to investigate the catalytic activity in the catalytic wet air oxidation (CWAO) of phenol, nitrobenzene (NB) and aniline at the mild operating conditions (reaction temperature of 155°C and total pressure of 2.5 MPa) in a batch reactor. The MWCNTs were characterized with scanning electron microscopy (SEM), transmission electron microscopy (TEM), gas adsorption measurements (BET), fourier transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS). The functionalized MWCNTs showed good catalytic performance. In the CWAO of phenol over the functionalized MWCNTs, total phenol removal was obtained after 90 min run, and the reaction apparent activation energy was ca. 40 kJ·mol^-1. The NB was not removed in the CWAO of single NB, while ca. 97% NB removal was obtained and 40% NB removal was attributed to the catalytic activity after 180 min run in the presence of phenol. Ca. 49% aniline conversion was achieved after 120 min run in the CWAO of aniline.     

基于密度泛函理论模拟单壁碳纳米管对五种核酸碱基的吸附

基于密度泛函理论,模拟了单壁碳纳米管(SWNTs)对5种碱基的吸附作用.考察了SWNTs直径、电荷转移量、碱基最高占据分子轨道能(EHOMO)和最低未占据分子轨道能(ELUMO)与SWNTs吸附碱基的吸附能之间的关系.结果表明,随着SWNTs直径的增大,SWNTs吸附碱基的吸附能降低.SWNT(6,6)吸附5种碱基的最低吸附能Emin与由碱基转移到SWNTs的电荷转移量(Q)及碱基的EHOMO线性负相关,相关系数分别为-0.966和-0.804(P<0.05).吸附后SWNTs与碱基的前线轨道无重叠,且SWNTs电子结构未受影响,表明吸附行为属于物理吸附.     

活性氧参与多壁碳纳米管诱导的RAW264.7细胞毒性

碳纳米管以其独特的结构和性能,在生物医药和电子等领域广泛应用,而其生态安全性也成为科学界关注的焦点。为探究多壁碳纳米管(MWCNTs)诱导的细胞毒性机制,将小鼠肺泡巨噬细胞(RAW264.7)暴露于6个浓度梯度(0、25、50、100、150和200μg.mL-1)的MWCNTs中,应用噻唑蓝(MTT)法测定细胞存活率,用2’,7’-二氯荧光素二乙酸(DCFH-DA)荧光染色法测定细胞内活性氧的生产量,用流式细胞方法测定MWCNTs对细胞周期的影响。同时使用抗氧化剂氮乙酰半胱氨酸(NAC)验证MWCNTs诱导的细胞氧化损伤的作用机理。结果显示,MWCNTs对RAW264.7的细胞毒性呈剂量依赖性。暴露于不同浓度的MWCNTs(25、50、100、150和200μg.mL-1)下24h后,细胞活力分别为对照的74%、62%、59%、51%和45%。MWCNTs对RAW264.7的周期阻滞作用主要发生在G0/G1期。200μg.mL-1的MWCNTs处理3h后活性氧较对照组上升6.6倍。NAC对MWCNTs细胞毒作用有明显的抑制作用,且NAC能减弱MWCNTs对RAW264.7的细胞周期阻滞作用。研究表明,活性氧能够介导MWCNTs对小鼠巨噬细胞RAW264.7的损伤,并且MWCNTS通过细胞周期G0/G1期的阻滞,诱导细胞凋亡。     

Cytotoxicity of benalaxyl,metalaxyl, and triadimefon on Chinese hamster ovary cells

The cytotoxicity of the fungicides benalaxyl, metalaxyl, and triadimefon was evaluated in vitro using the Chinese hamster ovary (CHO-K1) cell line. The midpoint cytotoxicity values of neutral red (NR) incorporation (NRI₅₀), total cellular protein content (TCP₅₀), and the methyl tetrazolium assay (MTT₅₀) were estimated. Benalaxyl was the most cytotoxic fungicide, followed by metalaxyl and triadimefon. Fetal calf serum (10%) caused a reduction in benalaxyl, metalaxyl, and triadimefon cytotoxicity by factors of 1.8, 1.3, and 1.3. The effects of sublethal concentrations (NRI₂₅) of the three fungicides on the glutathione redox cycle components glutathione S-transferase, glutathione reductase, glutathione peroxidase, and total glutathione content were studied. The ameliorative effects of extracellular glutathione (1 mmol L^?1), vitamin C (70 µmol L^?1), and vitamin E (30 µmol L^?1) were also investigated. The three antioxidants led to significant effects on the glutathione redox cycle components.     

In vitro cytotoxicity of multi-wall carbon nanotubes on human cell lines

The aim of this study was to evaluate the in vitro toxicity of two multi-wall carbon nanotubes on four different cell lines: human alveolar epithelial (A549) cells, hepatocytes (Hep 3B cells), human embryonic kidney cells, and intestinal (P407 cells) cells. The adverse effects of carbon nanoparticles were analyzed after 24 h incubation with different cell lines using the trypan blue dye exclusion method. Incubation of carbon nanotubes with different cells produced a concentration-dependent inhibition of growth of the cells. The TC₅₀ or IC₅₀ values (toxic concentration 50, i.e., concentration of particles inducing 50% cell mortality) of two nanoparticles were (1) found to be in the range 23.5–30.5 µg mL^?1, and (2) less than that of quartz (known toxic agent, 28.8–66.9 µg mL^?1), indicating the greater cytotoxic effect of carbon nanoparticles than quartz particles.     

Adsorption of toluene, ethylbenzene and xylene isomers on multi-walled carbon nanotubes oxidized by different concentration of NaOCl

Multi-walled carbon nanotubes (MWCNTs) were fabricated and oxidized by different concentrations of sodium hypochlorite (NaOCl) solutions. The untreated MWCNTs and modified MWCNTs were employed as adsorbents to study their characterizations and adsorption performance of toluene, ethylbenzene and xylene isomers (TEX) in an aqueous solution. The physicochemical properties of MWCNTs were greatly affected after oxidation, which influences TEX adsorption capacity. The 3% NaOCl-oxidized MWCNTs shows the greatest enhancement in TEX adsorption, followed by the 30% NaOCl. More interestingly, the 15% NaOCl-oxidized MWCNTs has lower adsorption capacities than untreated MWCNTs. The adsorption mechanism of TEX on treated MWCNTs is attributed to the combined action of hydrophobic interaction, π-π bonding interaction between the aromatic ring of TEX and the oxygen-containing functional groups of MWCNTs and electrostatic interaction. 3% NaOCl solution could not only introduce much oxygen-containing functional groups on MWCNTs, but also lead to less damage for the pore structure. This suggests that the CNTs-3% NaOCl is efficient adsorbent for TEX and that they may possess good potential for TEX removal in wastewater treatment.     

Assessment of DNA damage and cytotoxicity of palmatine on human skin epithelial carcinoma cells

The present investigation was carried out to examine the cytotoxic and genotoxic effects of a Tinospora cordifolia crude methanolic extract (palmatine) on human skin epithelial carcinoma cells (A431). T. cordifolia is one of the indispensable medicinal plants used in Ayurvedic medicine for treatment of various diseases and recommended for improving the immune system. Cytotoxicity and genotoxicity evaluation was carried out using A431 cells treated with different concentrations of palmatine. The duration of the treatment was 24 and 48 hr. A cellular proliferative capacity test showed that palmatine produced cytotoxicity in concentration- and time-dependent manner. Further, palmatine induced significant intracellular reactive oxygen species generation and elevated lipid peroxidation, as well as activities of catalase and superoxide dismutase. DNA fragmentation analysis using the comet assay showed that palmatine induced genotoxicity in a concentration- and time-dependent manner. Evidence indicates palmatine is capable of induction of oxidative stress resulting in cell death and genomic instability.     

碳纳米管负载的镍及其氧化物对高毒气体PH3的催化分解性能

采用沉积-沉淀法制备了均一负载于碳纳米管(CNTs)载体的金属Ni和NiO,将其应用于PH3催化分解反应.通过XRD、TEM、XPS、BET等一系列检测手段,对样品的相结构、形貌、组分和比表面积进行了表征.研究结果表明,反应过程中,Ni和NiO很快被磷化为高活性的金属磷化物NiP2,作为反应的活性相.在420℃温度下,两样品对PH3的催化分解率均可达到99.5%以上.将反应中原位生成的NiP2进行催化反应,较钝化-还原的处理方式更有利于样品催化性能的保持.     

DNA damage in mouse organs and in human sperm cells by bisphenol A

Abstract

The in vivo genotoxic potential of bisphenol A using the comet assay in mice and in human sperm cells in vitro without metabolizing enzymes was studied. Male mice were exposed by oral gavage to the following doses of bisphenol A (0 125, 250 and 500?mg/kg body weight). DNA damage was investigated in liver, kidney, testes, urinary bladder, colon and lungs cells. In testicular cells, a significant increase in DNA strand breaks was observed in the lowest, but not in the medium or highest dose groups. Histopathological investigation of the testicular samples did not show any treatment dose-related effects. No DNA strand breaks were observed in any of the other investigated tissues. In human sperm cells in vitro, bisphenol A did not induce DNA strand breaks.     

Effect of TiO2 nanoparticles and multiwall carbon nanotubes on the freshwater diatom Nitzschia frustulum: evaluation of growth,cellular components and morphology

Great attention should be paid to the possible impacts of various nanoparticles on aquatic organism. Freshwater diatoms are essential components of phytoplankton and play a critical role in bioassessment of nanoparticle exposure in the environment. In this study, cell growth was inhibited by TiO₂-NPs and MWCNTs, the 24?h EC₅₀ values of TiO₂-NPs and MWCNTs to Nitzschia frustulum were 20.75 and 24.64?mg?L^?1, respectively. Significant decreases of Chl a content after TiO₂-NPs exposures were detected and the Chl a content of N. frustulum was obviously increased by MWCNTs treatment at lower concentration. The ROS was detected in N. frustulum after TiO₂-NPs and MWCNTs exposures. The MDA content was significantly induced by TiO₂-NPs at lower concentrations of 24 and 48?h exposure; meanwhile, it increased at all tested concentrations at 24?h MWCNTs exposure. The SOD enzyme was induced by 72 and 96?h TiO₂-NPs exposure, and increased by MWCNTs treatment at 96?h in N. frustulum. Scanning electron microscopy results revealed that N. frustulum had obvious cell deformation after TiO₂-NPs treatment. The result showed that the physiological and biochemical response mechanisms after NPs exposure of diatom were species-specific, and in relation to the exposure concentration and time.     

Indium tin oxide nanoparticles-mediated DNA fragmentation and cell death by apoptosis in human lung epithelial cells

Indium tin oxide (ITO) nanoparticles (NP) have extensive applications in industrial fields, and concerns regarding their potential toxicity in humans and environmental impact have increased. Since exposure to ITO NP is mainly via skin and inhalation, this study was conducted utilizing human lung epithelial (A549) cell line. Cells were exposed to different concentrations of the ITO NP for 24 and 48 hr. A severe cytotoxic response of ITO NP was observed as evident by the (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and neutral red uptake assays after 48 hr exposure. ITO NP significantly reduced glutathione levels with a concomitant increase in lipid hydroperoxide levels, superoxide activity, and reactive oxygen species (ROS) generation after exposure. A significant induction in caspase activity and formation of condensed chromosomal bodies was also observed after ITO NP (10 or 25 µg/ml) exposure. Furthermore, a significant induction in DNA damage was observed by the Comet assay in cells exposed to ITO NP. Our data demonstrate that ITO NP display cytotoxic and genotoxic potential. However, increase in ROS levels and oxidative stress leading to oxidative DNA damage and condensed chromosomal bodies formation, suggests involvement of apotosis. Thus, ITO NP-mediated effects on cell viability indicate cytotoxicity, and therefore, exposures need to be carefully monitored in the industrial sector.     

Involvement of mitochondrial dysfunction in nanosized lead oxide induced cellular damage in human lung alveolar epithelial cells

High levels of industrial lead (Pb) exposure have decreased in the last 10 years as an outcome of removal of the metal from gasoline and paints. However, environmental Pb exposures remain extensive and may be correlated with adverse human health outcomes. The present study was designed to examine molecular mechanisms underlying cytotoxicity of lead oxide nanoparticles (PbONPs) on human lung alveolar epithelial (A549) cells. When A549 cells were incubated with PbONPs, the production of reactive oxygen species was enhanced as observed by 2',7'-dichlorodihydrofluorescein diacetate. PbONPs significantly reduced proliferation of A549 cells and increased caspase3 activity. In addition, exposure of PbONPs decreased levels of glutathione, and increased lipid peroxide levels and activities of superoxide dismutase and catalase. Exposure of PbONPs enhanced DNA damage as evidenced by tail DNA (%) and olive tail moment. Taken together, these finding indicated that PbONPs diminished cell proliferation and increased apoptotic cell death of A549 cells.