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      Biological response and developmental toxicity of zebrafish embryo and larvae exposed to multi-walled carbon nanotubes with different dimension

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      Heliyon
      Elsevier
      Toxicology

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          Abstract

          The development and use of nanomaterials are increasing significantly. Among nanomaterials, carbon nanotubes are of particular interest due to its distinctive physicochemical properties. This material composed of sheets of graphite has very high thermal conductivity, metallic-type electrical conductivity, stiffness, toughness and unique ability to bond to itself in an extended network with extraordinary strength. Its application in the industry is continuously growing, which could lead to the accumulation in the environment and a consequent impact on both humans and ecosystems. Considering that environmental systems are dynamic, it is difficult to predict the risks associated with the release of nanomaterials to the environment. Bioindicators are useful tools as primary signals of environmental risk, and their responses reveal the organism and ecosystem health.

          In the present study, we evaluated the impact of multi-walled carbon nanotubes with different dimensions and agglomeration pattern on zebrafish embryo and larvae; mainly, studies were focused on physiological and behavioral responses. In embryos, measurements were hatching rate, morphology changes, and viability. In larvae, locomotor activity, heart rate, innate inflammatory response, general and tissue-specific morphology were measured. MWCNT-S (short, wide and mostly dispersed) caused depression of the locomotor activity of larvae, indicating an alteration of the central nervous system, and depression of neutrophil migration activity. MWCNT-L (long, thin and agglomerated) caused malformations during larval development, a decrease of neutrophil migration and alteration of cardiac rhythm. Results obtained for both carbon nanotubes were different, highlighting the importance of dimensions of the same nanomaterial, and also the kind of agglomeration and shape adopted, for the toxic effects on organisms.

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          Most cited references27

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          From ecotoxicology to nanoecotoxicology.

          For hazard assessment of NPs quantitative nanoecotoxicological data are required. The objective of this review was to evaluate the currently existing literature data on toxicity (L(E)C50 values) of synthetic NPs in environmentally relevant species in order to: (i) identify tentatively most harmful NPs and most sensitive organism groups, and (ii) to provide relevant ecotoxicological information for further risk assessment. The focus was set on selected synthetic NPs (nano TiO(2), nano ZnO, nano CuO, nano Ag, SWCNTs, MWCNs and C60-fullerenes) and organism groups representing main food-chain levels (bacteria, algae, crustaceans, ciliates, fish, yeasts and nematodes). Altogether 77 effect values were found, mostly for nano TiO(2) (31%), C60 (18%), nano ZnO (17%), nano Ag (13%), SWCNTs and nano CuO (both 9%). Only 3% of the available quantitative ecotoxicological information concerned MWCNTs. Organism-wise, 33% of the data concerned crustaceans, 27% bacteria, 14% algae and 13% fish. For all organism groups studied, solubility of CuO- and ZnO-NPs was a key factor in their aquatic toxicity. On the basis of the 34 median L(E)C50 values derived from 77 individual values, NPs were ranked according to their lowest median L(E)C50 value for the above described organism groups: the most harmful were nano Ag and nano ZnO that were classified "extremely toxic", (L(E)C50<0.1mg/l), followed by C60 fullerenes and nano CuO that were classified "very toxic", (L(E)C50 0.1-1mg/l). SWCNTs and MWCNTs were classified "toxic" (L(E)C50 1-10mg/l). Nano TiO(2) was classified as "harmful", (L(E)C50 10-100mg/l). Throughout, algae and crustaceans (daphnids) were most sensitive and thus probably most vulnerable organism groups in aquatic exposure to NPs. Very low L(E)C50 values should deserve thorough attention of environmental risk assessors for evaluation of the potential adverse effects of synthetic NPs on ecosystems. As the quantitative nanoecotoxicological data are still rare, further studies are needed.
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            Morphologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish.

            The zebrafish is a useful model organism for developmental and genetic studies. The morphology and function of zebrafish myeloid cells were characterized. Adult zebrafish contain 2 distinct granulocytes, a heterophil and a rarer eosinophil, both of which circulate and are generated in the kidney, the adult hematopoietic organ. Heterophils show strong histochemical myeloperoxidasic activity, although weaker peroxidase activity was observed under some conditions in eosinophils and erythrocytes. Embryonic zebrafish have circulating immature heterophils by 48 hours after fertilization (hpf). A zebrafish myeloperoxidase homologue (myeloid-specific peroxidase; mpx) was isolated. Phylogenetic analysis suggested it represented a gene ancestral to the mammalian myeloperoxidase gene family. It was expressed in adult granulocytes and in embryos from 18 hpf, first diffusely in the axial intermediate cell mass and then discretely in a dispersed cell population. Comparison of hemoglobinized cell distribution, mpx gene expression, and myeloperoxidase histochemistry in wild-type and mutant embryos confirmed that the latter reliably identified a population of myeloid cells. Studies in embryos after tail transection demonstrated that mpx- and peroxidase-expressing cells were mobile and localized to a site of inflammation, indicating functional capability of these embryonic granulocytes. Embryonic macrophages removed carbon particles from the circulation by phagocytosis. Collectively, these observations have demonstrated the early onset of zebrafish granulopoiesis, have proved that granulocytes circulate by 48 hpf, and have demonstrated the functional activity of embryonic granulocytes and macrophages. These observations will facilitate the application of this genetically tractable organism to the study of myelopoiesis.
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              Carbon Nanotubes: Applications in Pharmacy and Medicine

              Carbon nanotubes (CNTs) are allotropes of carbon, made of graphite and constructed in cylindrical tubes with nanometer in diameter and several millimeters in length. Their impressive structural, mechanical, and electronic properties are due to their small size and mass, their strong mechanical potency, and their high electrical and thermal conductivity. CNTs have been successfully applied in pharmacy and medicine due to their high surface area that is capable of adsorbing or conjugating with a wide variety of therapeutic and diagnostic agents (drugs, genes, vaccines, antibodies, biosensors, etc.). They have been first proven to be an excellent vehicle for drug delivery directly into cells without metabolism by the body. Then other applications of CNTs have been extensively performed not only for drug and gene therapies but also for tissue regeneration, biosensor diagnosis, enantiomer separation of chiral drugs, extraction and analysis of drugs and pollutants. Moreover, CNTs have been recently revealed as a promising antioxidant. This minireview focuses the applications of CNTs in all fields of pharmacy and medicine from therapeutics to analysis and diagnosis as cited above. It also examines the pharmacokinetics, metabolism and toxicity of different forms of CNTs and discusses the perspectives, the advantages and the obstacles of this promising bionanotechnology in the future.
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                Author and article information

                Contributors
                Journal
                Heliyon
                Heliyon
                Heliyon
                Elsevier
                2405-8440
                21 August 2019
                August 2019
                21 August 2019
                : 5
                : 8
                : e02308
                Affiliations
                [1]Laboratorio de Bio-nanotecnología, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Grupo vinculado GBEyB, IMBICE-CONICET-CICPBA, Bernal, Argentina
                Author notes
                []Corresponding author. carolina.martinez@ 123456unq.edu.ar
                Article
                S2405-8440(19)35968-7 e02308
                10.1016/j.heliyon.2019.e02308
                6716136
                31485519
                3b5dd8c3-0262-4813-b3d0-435ef0fb2d91
                © 2019 Published by Elsevier Ltd.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 9 January 2019
                : 22 February 2019
                : 12 August 2019
                Categories
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                toxicology
                toxicology

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