Yiping Li

Translocation, transfer, and in vivo safety evaluation of engineered nanomaterials in the non-mammalian alternative toxicity assay model of nematode Caenorhabditis elegans

Translocation, transfer, and safety evaluation of engineered nanomaterials in non-mammalian alternative assay model of Caenorhabditis elegans are summarized and discussed. Due to the extensive applications of engineered nanomaterials (ENMs) in industry, agriculture, medicine and public health, environmental exposure of ENMs to human and environmental organisms is inevitable, therefore studies on translocation, transfer, and toxicology of ENMs receive wide interest. Modelling the nematode C. elegans, a widely recognized non-mammalian alternative toxicity assay system has been proven to be valuable in environmental safety evaluation, toxicological study, and examination of translocation and transfer of toxicants. In this review, we summarize and discuss the values and contributions of C. elegans in studies on environmental safety evaluation, toxicology, and translocation and transfer of ENMs. We introduced acute and chronic toxicity assay systems and discussed environmental safety assessment of specific ENMs at predicted environmental relevant concentrations, and influences of chemical properties of ENMs, exposure routes, developmental, genetic or physiological state, and environmental factors on nanotoxicity formation in nematodes. We then discussed the toxicological mechanisms of ENMs mainly with respect to roles of oxidative stress in normal and stress conditions and signaling pathways, reproductive toxicology, neurotoxicology, and translocation, distribution, transfer and metabolism of ENMs in C. elegans. Ways to eliminate or reduce the toxicities of specific ENMs by chemical modification with surface designs were further discussed. Finally, four important challenges have been raised and discussed by surrounding the aspects of environmental safety assessment, toxicological mechanisms, and designs of new safe ENMs.

Comparison of toxicities from three metal oxide nanoparticles at environmental relevant concentrations in nematode Caenorhabditis elegans

Nematode Caenorhabditis elegans has been developed in a variety of environmental studies to address adverse effects of a wide range of toxicants. In the present study, we compared the toxicities of three metal oxide nanoparticles (NPs) including TiO(2)-NPs, ZnO-NPs, and SiO(2)-NPs with the same nanosize (30 nm) after prolonged exposure from L1-larvae to adult at environmental relevant concentrations. Our data indicated that the adverse effects were detected in nematodes exposed to TiO(2)-NPs and ZnO-NPs at concentrations more than 0.05 μg/L and SiO(2)-NPs at concentrations more than 5 μg/L with locomotion behavior and ROS production as endpoints. With growth, locomotion behavior, reproduction, and ROS production as endpoints, toxicity order for the examined metal oxide NPs was: ZnO-NPs>TiO(2)-NPs>SiO(2)-NPs. In nematodes exposed to the examined metal oxide NPs, ROS production was significantly correlated with lethality, growth, reproduction, and locomotion behavior. Moreover, treatment with antioxidants of ascorbate or NAC effectively inhibited the formation of oxidative stress and retrieved the adverse effects of TiO(2)-NPs, ZnO-NPs, and SiO(2)-NPs on survival, growth, reproduction and locomotion behaviors in nematodes. Our data demonstrated the subtle toxicity differences of different NPs exposure at environmental relevant concentrations in C. elegans.

Evaluation of Environmental Safety Concentrations of DMSA Coated Fe2O3-NPs Using Different Assay Systems in Nematode Caenorhabditis elegans

Dimercaptosuccinic acid (DMSA) coating improves the uptake efficiency presumably by engendering the Fe2O3-NPs. In the present study, we investigated the possible environmental safety concentrations of Fe2O3-NPs using different assay systems in nematode Caenorhabditis elegans with lethality, development, reproduction, locomotion behavior, pharyngeal pumping, defecation, intestinal autofluorescence and reactive oxygen species (ROS) production as the endpoints. After exposure from L4-larvae for 24-hr, DMSA coated Fe2O3-NPs at concentrations more than 50 mg/L exhibited adverse effects on nematodes. After exposure from L1-larvae to adult, DMSA coated Fe2O3-NPs at concentrations more than 500 μg/L had adverse effects on nematodes. After exposure from L1-larvae to day-8 adult, DMSA coated Fe2O3-NPs at concentrations more than 100 μg/L resulted in the adverse effects on nematodes. Accompanied with the alterations of locomotion behaviors, ROS production was pronouncedly induced by exposure to DMSA coated Fe2O3-NPs in the examined three assay systems, and the close associations of ROS production with lethality, growth, reproduction, locomotion behavior, pharyngeal pumping, defecation, or intestinal autofluorescence in nematodes exposed to DMSA coated Fe2O3-NPs were confirmed by the linear regression analysis. Moreover, mutations of sod-2 and sod-3 genes, encoding Mn-SODs, showed more susceptible properties than wild-type when they were used for assessing the DMSA coated Fe2O3-NPs-induced toxicity, and the safety concentrations for DMSA coated Fe2O3-NPs should be defined as concentrations lower than 10 μg/L in sod-2 and sod-3 mutant nematodes.

Molecular Control of TiO2-NPs Toxicity Formation at Predicted Environmental Relevant Concentrations by Mn-SODs Proteins

With growing concerns of the safety of nanotechnology, the in vivo toxicity of nanoparticles (NPs) at environmental relevant concentrations has drawn increasing attentions. We investigated the possible molecular mechanisms of titanium nanoparticles (Ti-NPs) in the induction of toxicity at predicted environmental relevant concentrations. In nematodes, small sizes (4 nm and 10 nm) of TiO2-NPs induced more severe toxicities than large sizes (60 nm and 90 nm) of TiO2-NPs on animals using lethality, growth, reproduction, locomotion behavior, intestinal autofluorescence, and reactive oxygen species (ROS) production as endpoints. Locomotion behaviors could be significantly decreased by exposure to 4-nm and 10-nm TiO2-NPs at concentration of 1 ng/L in nematodes. Among genes required for the control of oxidative stress, only the expression patterns of sod-2 and sod-3 genes encoding Mn-SODs in animals exposed to small sizes of TiO2-NPs were significantly different from those in animals exposed to large sizes of TiO2-NPs. sod-2 and sod-3 gene expressions were closely correlated with lethality, growth, reproduction, locomotion behavior, intestinal autofluorescence, and ROS production in TiO2-NPs-exposed animals. Ectopically expression of human and nematode Mn-SODs genes effectively prevented the induction of ROS production and the development of toxicity of TiO2-NPs. Therefore, the altered expression patterns of Mn-SODs may explain the toxicity formation for different sizes of TiO2-NPs at predicted environmental relevant concentrations. In addition, we demonstrated here a strategy to investigate the toxicological effects of exposure to NPs upon humans by generating transgenic strains in nematodes for specific human genes.

Response of MicroRNAs to In Vitro Treatment with Graphene Oxide

Graphene oxide (GO) can be potentially used in biomedical and nonbiomedical products. The in vivo studies have demonstrated that GO is predominantly deposited in the lung. In the present study, we employed SOLiD sequencing technique to investigate the molecular control of in vitro GO toxicity in GLC-82 pulmonary adenocarcinoma cells by microRNAs (miRNAs), a large class of short noncoding RNAs acting to post-transcriptionally inhibit gene expression. In GLC-82 cells, GO exposure at concentrations more than 50 mg/L resulted in severe reduction in cell viability, induction of lactate dehydrogenase leakage, reactive oxygen species production and apoptosis, and dysregulation of cell cycle. GO was localized in cytosol, mitochondria, endoplasmic reticulum, and nucleus of cells. Based on SOLiD sequencing, we identified 628 up-regulated and 25 down-regulated miRNAs in GO-exposed GLC-82 cells. Expression of some selected dysregulated miRNAs was concentration-dependent in GO-exposed GLC-82 cells. The dysregulated miRNAs and their predicted targeted genes were involved in many biological processes. By combining both information on targeted genes for dysregulated miRNAs and known signaling pathways for apoptosis control, we hypothesize that the dysregulated miRNAs could activate both a death receptor pathway by influencing functions of tumor necrosis factor α receptor and caspase-3 and a mitochondrial pathway by affecting functions of p53 and Bcl-2 in GO-exposed GLC-82 cells. Our results provide an important molecular basis at the miRNA level for explaining in vitro GO toxicity. Our data will be also useful for developing new strategies to reduce GO toxicity such as surface chemical modification.

Small sizes of TiO2-NPs exhibit adverse effects at predicted environmental relevant concentrations on nematodes in a modified chronic toxicity assay system.

In Caenorhabditis elegans, although acute toxicity of TiO(2) nanoparticles (TiO(2)-NPs) at high concentrations has been investigated, we still know little about chronic toxicity of TiO(2)-NPs. Our data here showed that acute TiO(2)-NPs exposure in the range of μg/L had no obviously adverse effects on nematodes, but the chronic toxicities of large sizes (60 nm and 90 nm) of TiO(2)-NPs in the range of μg/L were detected in nematodes in a modified chronic toxicity assay system. Moreover, chronic toxicities of small sizes (4 nm and 10nm) of TiO(2)-NPs in the range of ng/L were observed in nematodes with locomotion behavior and ROS production as endpoints. In nematodes chronically exposed to small sizes of TiO(2)-NPs at predicted environmental relevant concentrations, locomotion behavior was significantly (P<0.01) correlated with ROS production. Furthermore, treatment with antioxidants (ascorbate and N-acetyl-l-cysteine) inhibited both the induction of ROS production and the decrease of locomotion behaviors observed in nematodes chronically exposed to small sizes of TiO(2)-NPs at predicted environmental relevant concentrations. Therefore, chronic exposure to small sizes of TiO(2)-NPs at predicted environmental relevant concentrations can cause adverse effects on nematodes, and formation of such adverse effects may be largely due to the induction of oxidative stress.

Susceptible genes regulate the adverse effects of TiO2-NPs at predicted environmental relevant concentrations on nematode Caenorhabditis elegans

UNLABELLED Contributions from mutations of susceptible genes to TiO2-NPs toxicity at environmental relevant concentrations (ERCs) and the underlying mechanism are largely unclear. After prolonged exposure, among the examined 19 mutants associated with oxidative stress or stress response, we show that sod-2, sod-3, mtl-2, and hsp-16.48 were susceptible genes for TiO2-NPs toxicity on reproduction and locomotion behavior, sod-2, sod-3, and mtl-2 were susceptible genes for TiO2-NPs toxicity on survival and intestinal development, and mtl-2 was susceptible gene for TiO2-NPs toxicity on development. Mutations of these susceptible genes, together with sensitive endpoints, could be used to evaluate TiO2-NPs toxicity at the concentration of 0.0001μg/L. Our results imply the usefulness of identified susceptible genes in assessing the potential nanotoxicity of engineered nanomaterial (ENM) at ERCs. One important mechanism to explain property of identified susceptible genes for TiO2-NPs toxicity was that mutations of these susceptible genes enhanced the uptake of TiO2-NPs into body of nematodes. FROM THE CLINICAL EDITOR This team of authors identified susceptibility genes influencing the uptake and consequential toxicity of TiO2 nanoparticles in a nematode, highlighting the general importance of investigating genetic influence on nanoparticle delivery.

Translocation and neurotoxicity of CdTe quantum dots in RMEs motor neurons in nematode Caenorhabditis elegans

We employed Caenorhabditis elegans assay system to investigate in vivo neurotoxicity of CdTe quantum dots (QDs) on RMEs motor neurons, which are involved in controlling foraging behavior, and the underlying mechanism of such neurotoxicity. After prolonged exposure to 0.1-1 μg/L of CdTe QDs, abnormal foraging behavior and deficits in development of RMEs motor neurons were observed. The observed neurotoxicity from CdTe QDs on RMEs motor neurons might be not due to released Cd(2+). Overexpression of genes encoding Mn-SODs or unc-30 gene controlling cell identity of RMEs neurons prevented neurotoxic effects of CdTe QDs on RMEs motor neurons, suggesting the crucial roles of oxidative stress and cell identity in regulating CdTe QDs neurotoxicity. In nematodes, CdTe QDs could be translocated through intestinal barrier and be deposited in RMEs motor neurons. In contrast, CdTe@ZnS QDs could not be translocated into RMEs motor neurons and therefore, could only moderately accumulated in intestinal cells, suggesting that ZnS coating might reduce neurotoxicity of CdTe QDs on RMEs motor neurons. Therefore, the combinational effects of oxidative stress, cell identity, and bioavailability may contribute greatly to the mechanism of CdTe QDs neurotoxicity on RMEs motor neurons. Our results provide insights into understanding the potential risks of CdTe QDs on the development and function of nervous systems in animals.

Quantum dots exposure alters both development and function of D-type GABAergic motor neurons in nematode Caenorhabditis elegans

We examined the in vivo quantum dots (QDs) neurotoxicity and the underlying mechanism using Caenorhabditis elegans D-type GABAergic motor neurons as the assay system. Prolonged exposure to low concentrations of CdTe QDs caused damage on both the development and function of D-type motor neurons, and resulted in translocation of CdTe QDs into D-type motor neurons. In addition to oxidative stress, cell identity was also involved in the induction of the toxicity of QDs on D-type motor neurons. ZnS surface coating blocked CdTe QDs translocation and maintained cell identity, thereby suppressing CdTe QDs neurotoxicity. For the underlying mechanism, we hypothesized that both translocation into the targeted neurons and alterations in the development and function of those targeted neurons contribute to the induction of CdTe QDs neurotoxicity. Considering the conserved property of GABAergic neurons during evolution, our data will shed light on our understanding of the potential risks of QDs to the nervous systems of animals.

High Concentration of Vitamin E Decreases Thermosensation and Thermotaxis Learning and the Underlying Mechanisms in the Nematode Caenorhabditis elegans

α-tocopherol is a powerful liposoluble antioxidant and the most abundant isoform of vitamin E in the body. Under normal physiological conditions, adverse effects of relatively high concentration of vitamin E on organisms and the underlying mechanisms are still largely unclear. In the present study, we used the nematode Caenorhabditis elegans as an in vivo assay system to investigate the possible adverse effects of high concentration of vitamin E on thermosensation and thermotaxis learning and the underlying mechanisms. Our data show that treatment with 100–200 µg/mL of vitamin E did not noticeably influence both thermosensation and thermotaxis learning; however, treatment with 400 µg/mL of vitamin E altered both thermosensation and thermotaxis learning. The observed decrease in thermotaxis learning in 400 µg/mL of vitamin E treated nematodes might be partially due to the moderate but significant deficits in thermosensation, but not due to deficits in locomotion behavior or perception to food and starvation. Treatment with 400 µg/mL of vitamin E did not noticeably influence the morphology of GABAergic neurons, but significantly decreased fluorescent intensities of the cell bodies in AFD sensory neurons and AIY interneurons, required for thermosensation and thermotaxis learning control. Treatment with 400 µg/mL of vitamin E affected presynaptic function of neurons, but had no remarkable effects on postsynaptic function. Moreover, promotion of synaptic transmission by activating PKC-1 effectively retrieved deficits in both thermosensation and thermotaxis learning induced by 400 µg/mL of vitamin E. Therefore, relatively high concentrations of vitamin E administration may cause adverse effects on thermosensation and thermotaxis learning by inducing damage on the development of specific neurons and presynaptic function under normal physiological conditions in C. elegans.