Eunhye Jo

Combined repeated-dose toxicity study of silver nanoparticles with the reproduction/developmental toxicity screening test

Abstract Combined repeated-dose toxicity study of citrate-capped silver nanoparticles (7.9 ± 0.95 nm) with reproduction/developmental toxicity was investigated in rats orally treated with 62.5, 125 and 250 mg/kg, once a day for 42 days for males and up to 52 days for females. The test was performed based on the Organization for Economic Cooperation and Development test guideline 422 and Good Laboratory Practice principles. No death was observed in any of the groups. Alopecia, salivation and yellow discolouration of the lung were observed in a few rats but the symptoms were not dose-dependent. Haematology, serum biochemical investigation and histopathological analysis revealed no statistically significant differences between control group and the treated groups. Toxicity endpoints of reproduction/developmental screening test including mating, fertility, implantation, delivery and foetus were measured. There was no evidence of toxicity.

Effects of sulfathiazole, oxytetracycline and chlortetracycline on steroidogenesis in the human adrenocarcinoma (H295R) cell line and freshwater fish Oryzias latipes

Pharmaceuticals in the environment are of growing concern for their potential consequences on human and ecosystem health. Alterations in the endocrine system in humans or wildlife are of special interest because these alterations could eventually lead to changes in reproductive fitness. Using the H295R cell line, the potential endocrine disrupting effects of six pharmaceuticals including diclofenac, erythromycin, sulfamethazine, sulfathiazole, oxytetracycline, and chlortetracycline were investigated. After exposure to each target pharmaceutical for 48 h, production of 17beta-estradiol (E2) and testosterone (T), aromatase (CYP19) enzyme activity, or expression of steroidogenic genes were measured. Concentrations of E2 in blood plasma were determined in male Japanese medaka fish after 14 d exposure to sulfathiazole, oxytetracycline, or chlortetracycline. Among the pharmaceuticals studied, sulfathiazole, oxytetracycline and chlortetracycline all significantly affected E2 production by H295R cells. This mechanism of the effect was enhanced aromatase activity and up-regulation of mRNAs for CYP17, CYP19, and 3betaHSD, all of which are important components of steroidogenic pathways. Sulfathiazole was the most potent compound affecting steroidogenesis in H295R cells, followed by chlortetracycline and oxytetracycline. Sulfathiazole significantly increased aromatase activity at 0.2 mg/l. In medaka fish, concentrations of E2 in plasma increased significantly during 14-d exposure to 50 or 500 mg/l sulfathiazole, or 40 mg/l chlortetracycline. Based on the results of this study, certain pharmaceuticals could affect steroidogenic pathway and alter sex hormone balance. Concentrations of the pharmaceuticals studied that have been reported to occur in rivers of Korea are much less than the thresholds for effects on the endpoints studied here. Thus, it is unlikely that these pharmaceuticals are causing adverse effects on fish in those rivers.

Potential Toxicity of Differential Functionalized Multiwalled Carbon Nanotubes (MWCNT) in Human Cell Line (BEAS2B) and Caenorhabditis elegans

The aim of this study was to evaluate in vitro (human bronchial epithelial cells, BEAS2B cells) and in vivo (the nematode Caenorhabditis elegans, C. elegans) toxicity outcomes following exposure to pristine as well as surface-functionalized multiwalled carbon nanotubes (MWCNT) following hydroxylation-oxygenation (O+), amination (NH2), or carboxylation (COOH) of the carbon nanotubes (CNT). Cell viability and proliferation were measured by Ez-Cytox, trypan blue exclusion, and colony formation assays. The genotoxic potential of the MWCNT was determined by using the alkaline comet assay. In addition, survival and reproduction were used as endpoints for detection of toxicity of MWCNT in C. elegans. The carboxylated (COOH)-MWCNT was found most toxic as evidenced by cytotoxic and genotoxic among all tested compounds. The order of sensitivity was COOH > O+ > NH2 > pristine. There were almost no marked changes in survival following exposure of C. elegans to MWCNT. It is of interest that only pristine MWCNT exerted significant reduction in reproductive capacity of C. elegans. Surface functionalization significantly influenced the bioactivity of MWCNT, which displayed species as well as target-organ specificity. The mechanisms underlying these specific modes of nano-biological interactions need to be elucidated.

Toxicity of Zinc Oxide Nanoparticles in Rats Treated by Two Different Routes: Single Intravenous Injection and Single Oral Administration

Toxicokinetics of zinc oxide nanoparticles (ZnONP) was studied in rats via a single intravenous (iv) injection and a single oral administration (3 mg/kg or 30 mg/kg), respectively. Blood concentrations of zinc (Zn) were monitored for 7 d and tissue distribution were determined in liver, kidneys, lung, spleen, thymus, brain, and testes. To ascertain the excretion of ZnONP, Zn levels in urine and feces were measured for 7 d. ZnONP were not readily absorbed from the gastrointestinal tract (GIT) after oral administration and were excreted mostly in feces. When the nanoparticles were injected iv to rats at a dose of 30 mg/kg, peak concentration appeared at 5 min but returned to normal range by d 2 (48 h after injection). ZnONP were distributed mainly to liver, kidneys, lung, and spleen, but not to thymus, brain, and testes. The distribution level was significantly decreased to normal by d 7. Feces excretion levels after iv injection supported biliary excretion of ZnONP. In rats injected iv with 30 mg/kg, mitotic figures in hepatocytes were significantly increased and multifocal acute injuries with dark brown pigment were noted in lungs, while no significant damage was observed in rats treated orally with the same dosage. Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/uteh

Stepwise Embryonic Toxicity of Silver Nanoparticles on Oryzias latipes

The developmental toxicity of silver nanoparticles (AgNPs) was investigated following exposure of Oryzias latipes (medaka) embryos to 0.1−1 mg/L of homogeneously dispersed AgNPs for 14 days. During this period, developmental endpoints, including lethality, heart rate, and hatching rate, were evaluated by microscopy for different stages of medaka embryonic development. To compare toxic sensitivity, acute adult toxicity was assessed. There was no difference in acute lethal toxicity between embryo and adult medaka. Interestingly, we found that the increase in stepwise toxicity was dependent on the developmental stage of the embryo. Lethal embryonic toxicity increased from exposure days 1 to 3 and exposure days 5 to 8, whereas there was no change from exposure days 3 to 5. In addition, 7 d exposure to 0.8 mg/L AgNPs resulted in significant heart beat retardation in medaka embryos. AgNPs also caused a dose-dependent decrease in the hatching rate and body length of larvae. These results indicate that AgNP exposure causes severe developmental toxicity to medaka embryos and that toxicity levels are enhanced at certain developmental stages, which should be taken into consideration in assessments of metallic NPs toxicity to embryos.

Aluminum Nanoparticles Induce ERK and p38MAPK Activation in Rat Brain

Aluminum nanoparticles (Al-NPs) are one of the most widely used nanomaterial in cosmetics and medical materials. For this reason, Al-NP exposure is very likely to occur via inhalation in the environment and the workplace. Nevertheless, little is known about the mechanism of Al-NP neurotoxicity via inhalation exposure. In this study, we investigated the effect AL-NPs on the brain. Rats were exposed to Al-NPs by nasal instillation at 1 mg/kg body weight (low exposure group), 20 mg/kg body weight (moderate exposure group), and 40 mg/kg body weight (high exposure group), for a total of 3 times, with a 24-hr interval after each exposure. Inductively coupled plasma mass spectrometry (ICP-MS) analysis indicated that the presence of aluminum was increased in a dose-dependent manner in the olfactory bulb (OFB) and the brain. In microarray analysis, the regulation of mitogen-activated protein kinases (MAPK) activity (GO: 0043405), including Ptprc, P2rx7, Map2k4, Trib3, Trib1, and Fgd4 was significantly over-expressed in the treated mice than in the controls (p = 0.0027). Moreover, Al-NPs induced the activation of ERK1 and p38 MAPK protein expression in the brain, but did not alter the protein expression of JNK, when compared to the control. These data demonstrate that the nasal exposure of Al-NPs can permeate the brain via the olfactory bulb and modulate the gene and protein expression of MAPK and its activity.

Autophagy negatively regulates tumor cell proliferation through phosphorylation dependent degradation of the Notch1 intracellular domain

Autophagy is a highly conserved mechanism that degrades long-lived proteins and dysfunctional organelles, and contributes to cell fate. In this study, autophagy attenuates Notch1 signaling by degrading the Notch1 intracellular domain (Notch1-IC). Nutrient-deprivation promotes Notch1-IC phosphorylation by MEKK1 and phosphorylated Notch1-IC is recognized by Fbw7 E3 ligase. The ubiquitination of Notch1-IC by Fbw7 is essential for the interaction between Notch1-IC and p62 and for the formation of aggregates. Inhibition of Notch1 signaling prevents the transformation of breast cancer cells, tumor progression, and metastasis. The expression of Notch1 and p62 is inversely correlated with Beclin1 expression in human breast cancer patients. These results show that autophagy inhibits Notch1 signaling by promoting Notch1-IC degradation and therefore plays a role in tumor suppression.

Stability of carboxyl-functionalized carbon black nanoparticles: the role of solution chemistry and humic acid

Carboxyl-functionalized carbon black nanoparticles (CB-NPs) are widely used in various industries. Studies on the dispersion and aggregation of nanomaterials in the aquatic environment are being actively conducted these days. In this study, the aggregation and sedimentation of carboxyl-functionalized CB-NPs were investigated according to the changes in the solution chemistry (0.1–10 mM NaCl and 0.01–1 mM CaCl2) and in the presence/absence of natural organic matter (1 and 5 mg L−1 humic acid) in the aquatic environment. Overall, humic acid was found to have the greatest effect on the stability of CB-NPs under the aquatic conditions investigated. Specifically, the sedimentation caused by CB-NP aggregation was more actively observed in NaCl than in CaCl2. With the increase in the ionic strength of the NaCl solution, the aggregation rate of CB-NPs also increased, whereas in the CaCl2 solution, the CB-NPs suspension stability was almost insensitive to the ionic strength. The reason was that the divalent cation Ca2+ was specifically adsorbed onto CB-NPs in the CaCl2 solution to reverse the negative CB-NP zeta potential, and increase the electrostatic repulsive force between CB-NPs. In the presence of humic acid in the NaCl and CaCl2 solutions, stability improved in the whole ionic strength range. A comparison of the classical Derjaguin–Landau–Verwey–Overbeek (DLVO) theory with the modified DLVO theory considering the steric force revealed that the enhanced stability of a CB-NP suspension in the presence of humic acid is attributed to a steric repulsive force as well as a stronger electrostatic repulsive force caused by humic acid adsorption.

Transport of carboxyl-functionalized carbon black nanoparticles in saturated porous media: Column experiments and model analyses

The aim of this study was to investigate the transport behavior of carboxyl-functionalized carbon black nanoparticles (CBNPs) in porous media including quartz sand, iron oxide-coated sand (IOCS), and aluminum oxide-coated sand (AOCS). Two sets of column experiments were performed under saturated flow conditions for potassium chloride (KCl), a conservative tracer, and CBNPs. Breakthrough curves were analyzed to obtain mass recovery and one-dimensional transport model parameters. The first set of experiments was conducted to examine the effects of metal (Fe, Al) oxides and flow rate (0.25 and 0.5 mL min(-1)) on the transport of CBNPs suspended in deionized water. The results showed that the mass recovery of CBNPs in quartz sand (flow rate=0.5 mL min(-1)) was 83.1%, whereas no breakthrough of CBNPs (mass recovery=0%) was observed in IOCS and AOCS at the same flow rate, indicating that metal (Fe, Al) oxides can play a significant role in the attachment of CBNPs to porous media. In addition, the mass recovery of CBNPs in quartz sand decreased to 76.1% as the flow rate decreased to 0.25 mL min(-1). Interaction energy profiles for CBNP-porous media were calculated using DLVO theory for sphere-plate geometry, demonstrating that the interaction energy for CBNP-quartz sand was repulsive, whereas the interaction energies for CBNP-IOCS and CBNP-AOCS were attractive with no energy barriers. The second set of experiments was conducted in quartz sand to observe the effect of ionic strength (NaCl=0.1 and 1.0mM; CaCl2=0.01 and 0.1mM) and pH (pH=4.5 and 5.4) on the transport of CBNPs suspended in electrolyte. The results showed that the mass recoveries of CBNPs in NaCl=0.1 and 1.0mM were 65.3 and 6.4%, respectively. The mass recoveries of CBNPs in CaCl2=0.01 and 0.1mM were 81.6 and 6.3%, respectively. These results demonstrated that CBNP attachment to quartz sand can be enhanced by increasing the electrolyte concentration. Interaction energy profiles demonstrated that the interaction energy profile for CBNP-quartz sand was compressed and that the energy barrier decreased as the electrolyte concentration increased. Furthermore, the mass recovery of CBNPs in the presence of divalent ions (CaCl2=0.1 mM) was far lower than that in the presence of monovalent ions (NaCl=0.1 mM), demonstrating a much stronger effect of Ca(2+) than Na(+) on CBNP transport. Mass recovery of CBNPs at pH 4.5 was 55.6%, which was lower than that (83.1%) at pH 5.4, indicating that CBNP attachment to quartz sand can be enhanced by decreasing the pH. The sticking efficiencies (α) calculated from the mass recovery by colloid filtration theory were in the range from 2.1×10(-2) to 4.5×10(-1), which were far greater than the values (2.56×10(-6)-3.33×10(-2)) of theoretical sticking efficiencies (αtheory) calculated from the DLVO energy by the Maxwell model.

Safety evaluation of zinc oxide nanoparticles in terms of acute dermal toxicity, dermal irritation and corrosion, and skin sensitization

To clarify the health risks related to the use of zinc oxide (ZnO) nanoparticles, acute dermal toxicity, dermal irritation and corrosion, and skin sensitization were evaluated according to the guideline for Organization for Economic Cooperation and Development (OECD) and Good Laboratory Practice (GLP). In acute dermal toxicity test using rats, there were no treatment-related mortality, clinical signs of toxicity, body weight changes, and gross findings at a dose level of 2000 mg/kg. Similarly, dermal irritation and corrosion tests using rabbits revealed no mortality, clinical signs of toxicity, and acute irritation or corrosion reaction on the skin. In a skin sensitization test using guinea pigs, there were no significant mortality, clinical signs of toxicity, body weight changes, and skin reactions 24 and 48 h after the removal of challenge patch, which was scored “0” based on the Magnusson and Kligman grading scale. Our findings suggest that the nano-sized ZnO used in this study is relatively safe since it did not induce acute dermal toxicity, dermal irritation and corrosion, and skin sensitization.