Critical Review on the Toxicity of Some Widely Used Engineered

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Review

A critical review on the toxicity of some widely used engineered nanoparticles V Srivastava, Deepak Gusain, and Yogesh Chandra Sharma Ind. Eng. Chem. Res., Just Accepted Manuscript • DOI: 10.1021/acs.iecr.5b01610 • Publication Date (Web): 12 May 2015 Downloaded from http://pubs.acs.org on May 14, 2015

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Industrial & Engineering Chemistry Research

A critical review on the toxicity of some widely used engineered nanoparticles Varsha Srivastava, Deepak Gusain, Yogesh Chandra Sharma* Green Chemistry and Renewable Energy Laboratories Department of Chemistry Indian Institute of Technology (Banaras Hindu University) Varanasi Varanasi 221 005, India. * Corresponding Author Tel No +91 5426701865, Fax No +91 5422368428, E Mail [email protected]

ABSTRACT With tremendous increase in development of nanotechnology, there is a developing enthusiasm towards the application of nanoparticles in diverse areas. Carbon nanotubes, fullerenes, quantum dots, dendrimers, iron oxide, silica, gold and silver nanoparticles are frequently used in different applications such as drug delivery, as ceramic materials, semiconductors, electronics, in medicine, cosmetics, etc. Some of these nanoparticles have shown major toxic effects on fauna, flora and human beings like inflammation, cytotoxicity, tissue ulceration and reduction of cell viability. SWCNT and MWCNT can induce oxidative stress and fibrosis in the lungs of rat and mice. SWCNTs can also induce oxidative stress to the nervous system in human beings. Inflammatory injury and respiratory distress can be observed due to TiO2 nanoparticles with small diameter. Nanoparticles can also pose detrimental effects on plants such as decreased growth rate, genomic and proteomic changes, etc. Toxicity of nanoparticles arises because of their specific characteristics such as greater ‘surface area to volume ratio’ compared with bulk particles of the same chemistry. The objective of this review is to critically evaluate the current literature on the toxicity of nanoparticles. Key words: Carbon nanotubes; Nanotechnology; Nanomaterial; Nanotoxicity; Silver nanoparticles, TiO2 nanoparticles; ZnO nanoparticles. 1 ACS Paragon Plus Environment


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1. INTRODUCTION In today’s world, nanoparticles (NPs) and nanotechnology have attracted attention of the scientific community globally and have emerged as a fast developing and fascinating area of research. In December 1959, Nobel Laureate Richard Feynman gave an infamous lecture entitled “There’s plenty of room at the bottom” at the California Institute of Technology during a meeting with the American Physical Society.1 At this meeting, he first mentioned the process which could potentially manipulate individual atoms and molecules and hence advance the field of synthetic chemistry. He was one of the first to recognize the potential of nano-scale materials for our industrial society. In order to describe the process of moving or manipulating atoms at the nano-scale (1-100 nm), this type of technology would later be officially termed “nanotechnology” in 1974.2 Nanotechnology can be defined as follows3: “Nanotechnology is the design, characterization, production and application of structures, devices and systems by controlling shape and size at nanometer scale”. Nanoparticles, the building blocks for nanotechnology, are engineered materials with at least one dimension less than 100 nm. Nanoparticles may be having one (e.g., nanolayers), two (nanowires and nanotubes) or three dimensions on the nanoscale (nanoparticles, quantum dots, metal nanoparticles and fullerenes).4 In general, these Engineered nanoparticles (ENPs) can be categorized into carbon-based materials such as fullerenes and carbon nanotubes: single walled carbon nanotube (SWCNT) and multiwalled carbon nanotube (MWCNT) and inorganic nanoparticles including the ones based on metal oxides (TiO2, ZnO and Al2O3, Fe3O4, Fe2O3, CeO2, etc.), metals (gold, silver, aluminium, and iron), quantum dots (cadmium sulfide and cadmium selenide), dendrimers (which are nano-sized polymers built from branched units capable of being tailored to

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perform specific chemical functions) and composites, which combine nanoparticles with other nanoparticles or with larger, bulk-type materials.5,6 At nanoscale, materials show novel attributes and, because of their small size, they possess ‘substantial surface zone to volume’ ratio which renders engineered nanoparticles (ENPs) more biologically reactive.7,8 Because of their high surface area, nanoparticles have a more prominent number of active sites for interaction with diverse chemical species.9,10 In addition to a large surface area, these particles show unique characteristics, such as catalytic potential and high reactivity, which make them better materials than usual bulk materials. ENPs are actually not new and have existed in the environment since the beginning of Earth’s history. Introduction of nanoparticles into our environment may be natural or anthropogenic. The ENPs enter the environment through natural sources such as volcanoes, forest fires, gas to particle conversion and also through anthropogenic sources like power plants, airplane jet metal fumes, combustion, ENPs (CNTs, quantum dots, metal nanoparticles etc.), etc.7,8 The review intends to emphasize vividly the adverse health effects of the frequently applied ENPs. There are many review articles on synthesis, characterization and application of nano particles, but there are very limited number of reviews in this important area which relates to human life. The review will certainly compel the scientists to ponder over the indiscriminate applications and consequences of application of ENPs. Further work is needed for recovery safe disposal of nanoparticles. Until 2000, the research work on ENPs was mostly focused on their synthesis, characterization and applications. However, few articles related to toxicity of ENPs were also reported.4,7 It was after the year 2000 when studies on the toxicity of ENPs started appearing in literature. In present review, the authors have concentrated on the articles on the topic for 1990 to 2012 duration.



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It was reported that the same nanoparticles obtained/synthesized from different sources may have different physicochemical properties and this may further affect ENPs’ interactions with organisms.7 Recently nanoparticles have received significant concern due to their rapidly expanding applications in different areas. Sol-gel method, inert gas condensation, spark discharge generation, ion sputtering, spray pyrolysis, laser pyrolysis, photothermal synthesis, thermal plasma synthesis, flame synthesis, low-temperature reactive synthesis, flame spray pyrolysis, sol-gel process, mechanical alloying/milling, pulsed laser ablation,

mechano-

chemical synthesis and electro-deposition are the various methods which have been used for the synthesis of nanoparticles.9 Nanoparticles of alumina can be prepared by sol-gel method, hydrolysis, supersonic thermal plasma expansion process, mechanical milling, combustion synthesis, and hydrothermal method.10–15 Carbon nanotubes (CNTs) are grand development of nanotechnology. CNTs are prepared by several methods such as thermal decomposition, precipitation route, sol–gel route, modified sol–gel route, microwave assisted combustion route, spray-drying, sonochemical, wet chemical method, colloidal synthesis of nanoparticles method etc.16–27 Various other nanoparticles such as magnesium aluminate (MgAl2O4), nanosilica, zirconia, tin oxide (SnO2), zinc oxide (ZnO), gold, titanium dioxide (TiO2), iron oxide, ceria nanoparticles, copper nanoparticles, cadmium sulphide (CdS) etc. have also been prepared for different application by using different methods.28-66 3. APPLICATION OF NANOPARTICLES Due to the attractive and unique properties of nanoparticles, they have been used in a variety of applications, including cosmetics, suntan lotions, paints, self-cleaning windows, stainresistant clothing, fillers, opacifiers, catalysts

and semiconductors.8,67 Metal oxide

nanoparticles are extensively used in different applications like food, material, chemical and biological sciences.68 Nanoparticles are also used in goods such as tennis, golf, and bowling


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balls; in the fabrication of high-performance tires; pharmaceutical products, and new therapeutic treatments. Recently, nanoparticles are also being used in filters and membranes for water purification and other environmental solutions.Variety of nanoparticles are used in chemical-biological arms detectors or for the fabrication of lighter but more powerful weapons.69 The most common material according to “The Nanotechnology Consumer Products Inventory” are the carbon nanoparticles.70 Silver is the second most common NP which is used in variety of applications. Nanoparticles have also been proved to be a prominent alternate of conventional adsorbent for the treatment of wastewater.71, 72 Titanium dioxide and zinc oxide NPs are frequently used in personal-care products such as sunscreens , beauty products, toothpaste etc.73,74 In addition, silver NPs are increasingly used as antimicrobial additives in detergents, food packaging and textiles.70 Potential global market value for nanotechnology-related products in 2011–2015 was estimated to be up to100 billion US dollars per annum.75 Various applications of different nanoparticles have been summarized in Table 1 . Among various nanoparticles, CNTs have distinct position and have various applications such as in biology and medicine, as a composite and as an adsorbent material for the removal of pollutants from water.76-81 Tin oxide (SnO2) nanoparticles are used in transparent conducting coatings of glass, gas sensors and solar cells.82,83 Various nanoparticles such as Al2O3, zirconia, CeO2, SiO2, and TiO2, have attracted more attention of researchers for varied applications.84-94 Carbon nanohorns can be used for drug delivery.95-97 ZnO, Fe3O4, metallic 0

copper nanoparticles, Ag, magnesium–aluminum oxide, MgAl2O4,CdS,zero-valent iron (Fe ), gold nanoparticles (II), ZnS, fullerene (C60) are some of the other nanoparticles which have various applications in different fields.98-112 5 ACS Paragon Plus Environment


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Table 1 Due to their unique characteristics, ENPs have been reported to offer high capacity for the removal of various pollutants from aqueous solutions and effluents and have attracted scientific community. The ENPs have been used for the removal of pollutant materials covering a broad spectrum. They have also been used for the removal of inorganic as well as organic pollutants from water/waste water. Cupric oxide nanoparticles were found to be very efficient for the removal of both pentavalent and trivalent forms of arsenic from aqueous solutions.113 Nanoparticles of alumina in the size range of 2-30 nm were used for the removal of DEClP (diethylchlorophosphate).114 It is clear from the Table 2 that there are wide variety of nanoparticles which have been used for the removal of Cr(VI), congo red and Cu(II).115121

The Spirulina platensis nanoparticles were obtained by a mechanical method for the

biosorption of food dyes.122 Magnetic nanoparticles were found to be very efficient for the treatment of Floride, Au(III) and uranium ions.123-125 Superparamagnetic nanoparticles with average size distribution of 9 ± 2.5 nm have been utilized for acridine orange dye removal.126 Nanoparticles of Fe3O4@PAA with an average diameter of 50 nm had been successfully synthesized by hydrothermal method were used as adsorbent for the removal of rhodamine 6G (R6G).127 Another nanoparticle viz. hexadecyl functionalized magnetic silica nanoparticles was also proved to be a good adsorbent for the removal of rhodamine 6G (R6G) from aqueous solutions.128 Further, adsorption capacity of Fe3O4@C nanoparticles with average size ∼250 nm for methylene blue and cresol red was determined to be 44.38 mg/g and 11.22 mg/g respectively.129 The removal efficiency of different nanoparticles such as cadmium sulfide nanoparticles, akaganeite nanocrystals, ZrO2 , CdSe ,BiFeO3 ,chitosan nanoparticles, NiO nanoparticles for


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different metallic species, organic pollutant, and different kind of dyes were also investigated by different researchers (Table 2).113-153 Table 2. Recently application of coated nanoparticles has attracted several researchers due to their enhanced efficiency for different applications. It was observed that surface coating of nanoparticle gives better results in comparison to their bare form. By coating with suitable material, specific surface characteristic can be developed which can make them a suitable candidate for a particular application. However, in some cases coated nanoparticle have been proved more toxic than bare nanoparticles. Some of the coated nanoparticles with their coating materials and applications are summarized in Table 3.154-178 Table 3 Recently, application of nanoparticles for biomedical applications is increasing day by day due to their better efficiency in comparison to their bulk materials. Few years ago, only some nanopartcsles were frequently used for different applications like CNTs, fullerene, quantum dots, iron oxide, silica, silver nanoparticle zero valent nanoiron etc. But presently, researchers are developing various types of nanoparticles with enhanced properties which make them better for biomedical applications. Bagre et al., 2013, developed alginate coated chitosan core shell nanoparticles for oral delivery of enoxaparin.154 In another study, β-lactoglobulincoated gold nanoparticles have been used for investigation of the fate of ingested inorganic nanoparticles in gastrointestine.156 Polymerized-glucose coated Fe3O4 magnetic nanoparticles were synthesized for drug delivery.160 Salicylic acid-coated magnetic nanoparticles have been synthesized by Zhongwu Zhou et al. in 2013 for Genomic DNA extraction170. Dibetics is very common and for oral insulin delivery, PEG coated silica nanoparticle was synthesised by Ana Luiza R et al., 2014175. In other studies, oleic acid-coated Fe3O4 nanoparticles and 7 ACS Paragon Plus Environment


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Polyethylene glycol (PEG) coated

Fe3O4 nanoparticles

were prepared for biomedical

applications.176 4. ENVIRONMENTAL TOXICITY OF NANOPARTICLES During 1990-2000, the researchers concentrated on the development of new processes for the synthesis of various nanoparticles and their applications in various fields, but at the same time they have inadvertently increasing the amount of nanoparticles in the atmosphere. Later on, around the year 2000, scientists started thinking about the safe disposal of nanoparticles. Due to the active development and application of nanotechnology, nanoparticles have emerged as a new class of environmental pollutants that may significantly impact the environment and human health. Increasing application of ENPs in commercial products and industrial applications has eventually resulted in their release into atmospheric, terrestrial, and aquatic environments. There are various routes of exposure of nanoparticles such as occupational exposure in which any person comes in contact with nanoparticles during manufacturing and research, and consumer exposure in which person comes in contact with engineered nanoparticles during use of different personal care products having engineered nanoparticles.179-181 In the third type, the exposure may be due to the entire ecosystem to engineered nanoparticles through the water and soil.8 Nanotoxicology refers to the investigation of interactions of nanostructures with biological systems, explaining the relationship between the physical and chemical properties viz. surface chemistry, composition, size, shape and aggregation with induction of toxic biological responses.70 According to Kirchner et al., there are three possible ways by which nanoparticles can affect any organism. Sometime due to release of toxic metal used in nanosynthesis; nanoparticles can attach to the surface of cell membranes and finally their 8 ACS Paragon Plus Environment

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size or shape can also pose different effect on any organism such as CNT has more toxic effect then than carbon-black and graphite.182 Sometimes, toxicity can be due to the toxicity of precursors which have been used for their preparation. Thus nanoparticles may differ in their toxicological effects which depend on the variety and size of the particles, test organism species, and test methods. Though the nanoparticles may primarily target the respiratory organs, they

also could get into the

gastrointestinal tract by many ways, such as indirectly via mucociliary movement or directly via oral intake of drugs, water and food.183 Presence of nanoparticles in ecosystem can pose threatening results. CNTs, fullerenes, silver nanoparticles, TiO2, ZnO and nano sized iron particles are frequently used in various applications and their presence in the environment directly affects human beings, animals, plants and aquatic species. The increased use of metal oxide nanoparticles in various fields such as catalysis, sensors, environmental remediation and commercial products leads the generation of higher amount of nanoparticles into the environment.184 TiO2, Fe2O3, ZnO and CuO nanoparticles are utilized in cosmetics and antimicrobial products, so there is a strong possibility that these nanoparticles will ultimately enter aquatic ecosystems through waste water discharges and wash offs during recreational activities such as swimming and water skiing. Nanoparticles have many applications in different fields and their extensive use may pose toxic effects on flora, fauna and humans. Several in vivo and in vitro studies have been carried out to observe the toxicity of nanoparticles. It was reported that the results of in vitro study show good correlation with in vivo study but some times results of in vitro study do not implement on in vivo study. To observe the correlation between in vitro and in vivo study, Sayes et al. (2007) compared in vitro and in vivo pulmonary toxicity profile of some carbonyl iron, crystalline silica, amorphous silica nano



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zinc oxide and fine-sized on rats.185 They observed that there was little correlation between the measurement of in vivo and in vitro studies. In 2013, in vitro and in vivo studies were demonstrated to investigate the effect of selenium nanoparticle on Leishmania major.186 In another study, in vitro and in vivo studies for determination of toxicity of metallic nanoparticles were performed and it was observed that the nature of nanoparticle like size, shape and stability can affect toxicity level.187 But results for in vivo and in vitro study was not in good correlation. It may have been due to tested cell line. In vitro and in vivo genotoxicity and cytotoxcicity of silver nanoparticles was demonstrated by Ghosh et al., in 2012 where they reported good correlation between the in vitro and in vivo experiments.188 Kwon et al. studied the genotoxic potential of ZnO nanoparticles for this purpose they selected four kinds of ZnO nanoparticles (20 nm and 70 nm size, +ve and –ve) and almost similar result were obtained for in vitro and in vivo studies.189 Occupational Safety and Health Administration (OSHA) standard has decided

some standard for

nanoparticles

human exposure.190 According to OSHA, maximum limit for nanodust to human exposure is 3x10-5 x10-3 µg/cm3/h and 2-300 particles /cell/h, respectively. The main focus of this review paper is to debate the toxicity of widely used

ENPs

like CNTs, fullerenes, quantum dots, TiO2, Ag, ZnO, Iron/ iron compound and some other other nanoparticles on flora and fauna. 4.1. Toxicity of carbon nanotubes (CNTs) Among the various types of nanoparticles, carbon nanotubes (CNTs) are the most promising nanoparticles due to their specific mechanical, electrical and magnetic characteristics. Carbon nanotubes are allotropic modifications of carbon that can be represented as a sheet of graphene (single layer of graphite) rolled into a cylinder. They are chemically and thermally


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very stable.191-193 It was first discovered by Ijima in 1991.194 CNTs are cylindrical molecules composed solely of carbon atoms. CNTs are distinct from carbon fibers, which are strands of layered graphite sheets. They can be imagined as a seamless cylinder formed from a graphite sheet with a hexagonal lattice structure.195 There are two main forms of manufactured CNTs, the single-walled or SWCNT, and multiwalled or MWCNT.196-198 In terms of structure, the SWCNT is a single-layered graphene sheet which is rolled-up as cylindrical shapes, with a diameter of approximately 1 nm and a length of several micrometers, whereas the MWCNT contains two or more concentric layers with various lengths and diameters.199 Both SWCNTs and MWCNTs have attracted widespread interest for commercial and industrial applications due to their novel properties and unique electronic properties. CNTs exhibit several unique physical and chemical properties which allow their application in numerous technological applications. The unique properties of CNTs in addition to the wide range of functionality afforded by chemical modification, allow them for many applications.70-75 The CNTs have unique absorption in the near-infrared region, which enables its utility for biological sensing.200,201 Due to nanosize, CNTs

have the potential to interact with

macromolecules such as proteins and DNA.202 The near infrared optical absorption of carbon nanotubes being used for laser heating cancer therapy and the unusual one dimension hollow nanostructure particularly makes CNTs useful as novel drug and gene delivery tools.203,204 Because of the rich electronic properties of CNTs, they have been explored for the development of highly sensitive and specific nanoscale biosensors.205 CNTs have also been used for manufacturing various electro analytical nanotube devices and as electro mechanical actuators for artificial muscles.206,207 The more extensive scope of expanding nanotechnology applications for Cnts will probably bring about the expanded



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potential for both human and environmental exposures to this nanomaterial. Therefore, it's necessary to examine the toxicity and also the biocompatibility of CNTs. Several researchers have investigated their direct or indirect deleterious effects on the lungs, skin and other parts of the body due to the entry of CNTs.208,209 It was demonstrated that the toxicity of CNTs depends upon size, dosage and medium.210-217 In another study, Shen et al.(2009), reported that neutral and negative charged MWCNTs were nontoxic to cell lines at a concentration of up to 100 mg/L, but the positive charged ones were found to be toxic to cells at 10 mg/L.218 Some studies on rats and mice have shown that SWCNT and MWCNT induce oxidative stress, inflammation, granulomas and fibrosis in the lungs.219-221 Smith et al.,(2007) obtained evidence of oxidative injury in rainbow trout (Oncorhynchus mykiss) exposed to SWCNTs.222 They concluded that CNTs acted as respiratory toxicants in rainbow trouts. Multi-walled carbon nanotubes and carbon nanofibers have been found to be significantly toxic to human lung tumor cells as early as 24h after exposure.223 Single-walled nanotubes have been found to be toxic in some systems. 224 Tiana etal.,(2006) studied the toxicological effects of five carbon nanoparticles SWCNTs, active carbon,carbon black,MWCNTsand carbon graphiteon human fibroblast cells in vitro.225

It was observed that refined nanoparticles showed more toxic effect than its

unrefined forms. Toxic effects of both SWCNTs and MWCNTsare displayed in Table 4.

226-

266

Table 4. CNTs were found to induce a dose and time dependent increase of intracellular reactive oxygen species (ROS) which is ultimately responsible for cell damage.266 It was reported that CNTs can induce growth inhibition in the case of protozoan.267 However, growth stimulation


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of a unicellular protozoan by CNT was also observed in a growth medium containing yeast extract. This was explained by the uptake of CNT-peptone conjugates where the additional peptone was responsible for the stimulation.268 In an alternate study, it was exhibited that coated CNTs were promptly taken up by Daphnia magna.269 However, impact of nanoparticles were seen at the highest concentration. Acute toxicity was only observed at the highest concentration. Japanese investigators have hypothesized that single MWCNT together with the larger agglomerates, or by themselves, could be responsible for inducing mesotheliomas.270 Increasing applications of CNTs in biomedical applications may lead to potential toxic effects to human health. 271 They revealed that SWCNTs may induce oxidative stress to the nervous system. The effect of MWCNTs on Sprague–Dawley rats were demonstrated by Muller et al (2005).272 It was observed that both MWCNT and ground CNT were responsible for the inflammatory and fibrotic reactions. TNF-α was found in the lung of treated animals which confirms that both CNT and ground CNT can stimulate the production of TNF-α in the animals. Hirano et al. (2008) studied the effect of MWCNT on the plasma membrane of macrophages. The size of MWCNTs was 67 nm.273 It was observed that in the MWCNT-exposed macrophages, several proteins were adsorbed onto MWCNTs. It was reported that the plasma membrane of macrophages was disrupted and infiltrated when exposed to MWCNT fibers. Hirano et al.(2010), studied the cytotoxicity of MWCNTs in human bronchial epithelial cells.274 It was found that CNTs can induce pulmonary disease just like the asbestos. Cheng et al.(2009) reported the effect of MWCNTs on zebrafish (Danio rerio).275 Bio-distribution and long-term effects of functionalized MWCNTs in developing zebrafish was demonstrated using an in vivo study. It was observed that the offsprings of zebrafish loaded with BSAMWCNTs at 1- cell stage had lower survival rates. 13 ACS Paragon Plus Environment


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Cui et al.,(2005) found that SWCNTs inhibited the proliferation of human embryo kidney cells (HEK293) cells by inducing cell apoptosis and decreasing cellular adhesive ability.259 Ultrafine carbon particles can have impact on central nervous system because they are capable to cross the blood–brain barrier and can have impact on the central nervous system.276 Warheit et al.,(2004,2005) studied the pulmonary toxicity in male rats. 277,278 They reported a high mortality rate was observed due to mechanical blockage of the upper airway. Another effect of SWCNTs was an increase in pulmonary cell proliferation and multifocal pulmonary granulomas. Significant increase in lung weight and transient increase in bronchoalveolar lavage were also found. Pulmonary toxicity following acute exposure to three SWCNT preparations in male mice was explored by Lam et al.279 Inflammation and pulmonary granulomas were observed for unrefined nanotubes and purified nanotubes (PNT). Shvedova et al.(2006) investigated the cytotoxicity of raw-CNTs on human keratinocyte cell. This study showed that dermal exposure to raw CNTs may lead to dermal toxicity from accelerated oxidative stress, loss of cell viability, and morphological changes.280 A comparative study of the cytotoxicity of their nanomaterials viz. SWCNT, MWCNT and the C60 fullerene

was carried out on guinea pigs.281. It was observed that

fullerene did not show any cytotoxicity on alveolar macrophages in guinea pigs. While SWCNT and MWCNT showed cytotoxicity on guinea pigs. It was noted that SWCNT caused higher toxicity in comparison to MWCNT. It was observed that by increasing the dose of SWCNTs, the cell nucleus experiences degeneration, enlargement, and rarefaction of nuclear matrix. Muller et al., investigated that a single intratracheal instillation of MWCNT increased the frequency of micronucleated type II pneumocytes in rat lungs in vivo associated with a marked pulmonary inflammation.282 Singh et al., (2009) reviewed the genotoxicity of some


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engineering nanoparticles283 and found that ENPs can cause chromosomal fragmentation, DNA strand breakages, point mutations and alterations in gene expression profiles. Takagi et al. (2008) reported intraperitoneal application of MWCNTs in p53+/- mouse.270 MWCNTs was found to be able to induce mesothelioma and showed carcinogenic effects on mice. Poland et al. (2008) reported that when carbon nanotubes were introduced into the abdominal cavity of mice, they can induce asbestos like pathogenic changes in the mesothelial lining of the abdominal cavity.284. Radomski et al. (2005) reported that SWCNT and MWCNT induce platelet aggregation and vascular thrombosis.285 4.2. Toxicity of fullerenes Fullerenes are molecules with 60 atoms of carbon, commonly denoted as C60.

It

(Buckminster fullerene) was first discovered by Kroto et al. (1985).286. Fullerene consist of closed spherical shells comprised only of carbon atoms. There are also higher mass fullerenes with different geometric structures, such as, C70, C76, C78 and C80.287 However, the most widely explored is the C60 molecule. Fullerenes are a class of materials which shows unique physical properties. Even after applying extreme pressures they never lose their shape and regain their original shape on releasing the pressure. Due to the specific characteristics, C60 has many industrial and medical applications. For example, the use of C60 is being investigated for use in optics and superconductors, and for drug delivery.112,288 Fullerenes are known to be an empty structure with dimensions like several biologically active molecules.289 Recently a number of cosmetic products such as face creams that contain C60 nanoparticles have been introduced in the market.290 The main exposure routes of fullerene nanoparticles are inhalation,dermal contact and ingestion.



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An in vitro and in vivo investigation of the impacts of fullerene on development of mice was explored by Tsuchiya et al. (1996).291 Fullerene(C60) solubilized with polyvinyl pyrrolidone inhibited cellular differentiation. Proliferation of mesencephalic cells was also observed due to C60 fullerenes solubilized with polyvinyl pyrrolidone. C60 was distributed throughout the embryo at 50 mg/kg and the yolk sac was reported to be impaired. In vitro mutagenic activity in in 3 Salmonella strains exposed to C60 was reported by Sera et al. (1996).292 Iwata et al.(1998)studied the harmfulness of C60 on the hepatic catalyst and reported that the C60 fullerene can decrease the hepatic enzyme action of glutathione in humans, mice and rats.293In vitro exposure to the C60 fullerene induced oxidative damage in rat hepatic microsomes.294 Mutagenic activity of three C60 fullerene derivatives on Salmonella typhimurium were studied by Babynin et al.(2002). For this study, three fullerene derivative viz. (i) dimethoxyphosphoryl-carbethoxy-methanofullerene, (ii) dimethoxyphosphoryl-carbmethoxymethanofullerene

and

(iii)

1-methyl-2-(3,5-di-tertbutyl-4-hydroxy-phenyl)-3,4-

fulleropyrrolidine were selected.295 It was observed that, (i) and (iii) of these derivatives showed negative results while the second derivatives were found to be anti-mutagenic. In an alternate study mutagenic activity of C60 fullerene containing malonic acid molecules was demonstrated.296 Cytotoxicity (Cl50) of four different kinds of water-soluble fullerenes was explored by Sayes et al. (2004).297 Effect of fullerene was tested on human cells and they reported that toxicity depends on the nature of functional groups. Oberdorster (2004) showed that C60 fullerenes induced changes in brain of the fish even at very low aquatic exposure level.298 Significant lipid peroxidation was found in the brains of large mouth bass after 48 h of exposure to 0.5 mg/L of uncoated C60 fullerenes. Lovern and


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Klaper(2006) demonstrated that diameter of fullerenes is one of the important parameters which is responsible for the toxicity. They investigated the differential toxicity for 10–20 nm fullerenes and 20–100 nm fullerenes and demonstrated that smaller fullerenes proved to be more toxic with a LC50 at 0.46 mg/L while the larger fullerenes had a LC50 of 7.90 mg/ L.299 Toxic effects of fullerene C60 and modified fullerene C60 in an in vivo/in vitro study are presented in Table 5. 300-317 A recent research on toxicity of fullerene C60 with benzo[a] pyrene in Danio rerio(zebrafish) hepatocytes suggested that C60 can decrease cell viability.318 Table 5. 4.3. Toxicity of quantum dots Quantum dots are spherical nanocrystals from 1 to 10 nm in diameter(Aitken).319,320 Quantum dots have been developed within the variety of insulators, semiconductors, metals, magnetic materials or metallic oxides. Typical structure of quantum dots has been reported by scientific workers. 319,320 They possess unique electronic, optical, magnetic and catalytic properties.321 Semiconductor quantum dots show specific quantal effect which is dimension-dependent. These ENPs have attracted a special interest for their promising applications in molecular biology, medicine and information technology.322,323 In spite of having promising applications in different fields, these ENPs show ecotoxicity.324 The ecotoxicological effects of CdTe quantum dots to freshwater mussel Elliptio complanata have been reported showing that these ENPs are immune toxic to freshwater mussels and can cause oxidative stress in gills causing DNA damage.325



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Wang et al., (2008) observed toxicity of two commercially used nanoparticles viz. titanium dioxide(TiO2) and quantum dots (QDs) using the unicellular green alga Chlamydomona sreinhardtii.326They concluded that QDs were found to be more toxic to Chlamydomonascells in comparison to TiO2. The cytotoxicity of Cadmium selenide (CdSe) nanocrystal solutions and of CdSe/ ZnS for tumour cells and human fibroblasts was studied by Kirchner et al. (2005).182 It was reported that surface chemistry also influenced cytotoxicity of any nanoparticles. Feng et al.(2012) reported that cadmium tellurium(CdTe) QDs exhibited a dose-dependent inhibitory effect on cell growth of Escherichia coli (E. coli).327 It was also observed that toxicity also depend on the particle diameter. Smaller sized quantum dots were found to be more toxic for E.coli. Derfus et al.(2004) reported the cytotoxicity of CdSe quantum dots in an in vitro study.328 The viability of hepatocytes incubated in a solution containing quantum dots decreased according to its concentration (0.0625 < 0.25 < 1 mg/mL). They also suggested that the quantum dots that had been exposed to UV radiations for 8 hours reduced the cellular viability significantly. It was demonstrated that quantum dots can induce cell death by lipid peroxidation of human neuroblastoma cells.329 CdSe quantum dots can be cytotoxic due to the release of Cd2+ ions. 330-332 It can also be toxic on direct interaction with cells.182, 333,334 Green and Howman (2005) carried out an in vitro experiment in which they incubated coiled double-stranded DNA in a cadmium selenide solution encapsulated in zinc sulphite functionalized with surface biotin.335 They found that the quantum dots altered the DNA by producing SO2 free radicals, resulting from ZnS oxidation. In vitro cytotoxicity of CdSe/ZnS quantum dots coated with mercaptoundecanoic acid and sheep serum albumin was demonstrated by Shiohara et al. (2004).336 In this study, they produced three forms of quantum dots, which showed different photoluminescence. Effect


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of different concentration of CdSe/ZnS nanoparticles on human hepatocytes, primate kidney cells and cervical cancer was demonstrated and it was reported that here was a decrease in the viability of the 3 cell lines at concentrations of 0.1 and 0.2 mg/mL, which further increased with the increasing concentration. In one study, Cadmium selenide (CdSe) nanoparticles were found to be responsible for the enhancement of vacuolar membrane permeabilization (VMP). It was observed that exposure to nanoparticles increased in ROS accumulated cells.337 Immunocytotoxic, cytogenotoxic and genotoxic effects of cadmium telluride QDs (CdTe QDs) on the marine mussel Mytilusgalloprovincialis were explored by Rocha et al.,(2014). It was reported that cadmium accumulated in mussel soft tissues and hemolymph which is the reason of immunocytotoxic, and genotoxic effects in M. galloprovincialis .338 4.4. Toxicity of TiO2 nanoparticles Titanium dioxide nanoparticles (TiO2 ENPs) are among the top five ENPs utilized as a part of customer items such as paints and additives in pharmaceuticals.339 Commercial products such as sunscreens and self-cleaning window coatings consist of anatase TiO2 ENPs.340,341 Sunlight-illuminated TiO2 catalyses DNA damage, both in vitro and in vivo , since exposure to such nanoparticles is mainly through skin and inhalation342 . In another investigation, it was demonstrated that exposure to nanoparticles affects the brain Murine microglial cells. For this study, commercial anatase and rutile titania of average crystalline size 30nm were selected and they displayed extracellular release of H2O2 and the superoxide radical and furthermore caused hyper-polarization of mitochondrial membrane potential.343 For example, TiO2 absorbs substantial UV radiation yielding in aqueous media hydroxyl species. These species may cause substantial damage to DNA, resulting in additional environmental hazards. 344,



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Hu et al., (2009) investigated the in vitro cytotoxicity of different metal oxide nanoparticles Viz. ZnO, CuO, Al2O3, La2O3, Fe2O3, SnO2 and TiO2 to the test organisms, Escherichia coli.345 It was observed that metal oxide (TiO2) with higher cation charge showed lower cytotoxicity. Among all metal oxides, ZnO was found to be most toxic. Wang et al.,(2007) reported that nano-sized TiO2 can

produce free radicals and exert a strong oxidizing

ability.346 In an another study, Daphnia magna exhibited higher mortality when exposed to TiO2 nanoparticles with an average diameter of 30 nm than those exposed to 100–500 NM.

299

A

significant increase in inflammation signs was observed during the administration of 20 nm TiO2 particles in comparison with the same mass of 250 nm particles.347,348 TiO2 exists in three main crystallographic structures, e.g. anatase, rutile and brookite of which the first two are usually considered the most important in the environment.349 Each of these forms presents different properties and therefore have different applications and environmental impacts. In addition, microorganisms in the presence of light are adversely affected by TiO2 nanoparticles due to the production of ROS.350 This experimental evidence suggests that these nanoparticles can produce oxidative stress in aquatic organisms. Inflammatory injury and respiratory distress were observed after the exposure to TiO2 nanoparticles in rainbow trout.351,352 Musee et al.(2010) selected freshwater snail Physa acuta (Draparnaud) to demonstrate the effect of different nanoparticles viz. γalumina,α-alumina, modified TiO2 (M-TiO2), and commercial TiO2.353 They

reported

that

increases

of

γ-alumina,

α-alumina

concentrations

caused a

vital reduction in the embryo hatchability and the embryo growth rate. It was also observed that these nanoparticles induced

developmental deformities of the embryos. In addition,

available ecotoxicological studies of TiO2 nanoparticles

in aquatic organisms such as

Daphnia magna, algae, fish as well as nematodes were more prominent.299, 354-363


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Wang et al. (2011) reported the combined effect of TiO2 nanoparticles (n-TiO2) and As(V) on Ceriodaphnia. dubia.364 Result showed that n-TiO2 presence of n-TiO2 increase the toxicity of As(V).The Mortality increases with increasing n-TiO2. In 2006, the International Agency for Research on Cancer (IARC) classified TiO2 as a possible carcinogen to humans and animals also.365,366 Different phase compositions of TiO2 nanoparticles (e.g., anatase and rutile, or a mixture of the two) affect cytotoxicity and inflammatory response in lung cells. It was observed that anatase TiO2 nanoparticles were 100 times more toxic than an equivalent sample of rutile TiO2 which revealed that oxidative damage in human lung epithelial cells is strongly dependent on the crystal phase composition of nanoparticles.251 Ecotoxicity study of TiO2

nanoparticles

on microalgae species, Scenedesmus sp. and

Chlorella sp.were investigated by Sadique.(2011) and they noticed a decrease in the chlorophyll content of the treated algae species in comparison to the untreated species367. In 2011, Lapied et al., reported the ecotoxicological effects of an aged TiO2 nanocompositeon the earthworm Lumbricus terrestris for 7 days

Results showed an

enhanced apoptotic frequency which was higher in the cuticule, intestinal epithelium and chloragogenous tissue.368 Noel et al. (2012) investigated the effect of inhaled nano-TiO2 aerosols

on rat lungs and

reported that the dimensions and concentrations of ENPs

agglomerates affected the biological responses.369 The effect of sub-acute exposure to nano-TiO2 on oxidative stress and histopathological changes in juvenile Carp (Cyprinus carpio) were investigated by Linhua et al. (2009).370 This study revealed that a higher concentration of nanoparticles of TiO2 may show abnormal physiological and behavioral changes on carp and also may produce cytotoxic effects. Sha et al. (2011) demonstrated the effect of TiO2 nanoparticles on liver cells from humans and rats. It was found that cell viability exhibited is dose-dependent and time-dependent. 371 Mild cytotoxic response of TiO2 ENPs was determined as clear by the MTT and NR uptake



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measures after 48 h of exposure within the human epidermal cells (A431).372 Mice exposed to TiO2 ENPs with a primary size of 2–5 nm was found to have a significant but moderate inflammatory response in the lungs.373 Effect of size, surface and crystalline structure of titanium dioxide nanoparticles on pelagic filter feeder Daphnia magna and the benthic amphipod Gammarus

Fossarum were

demonstrated by Seitz et al.,(2014) and it was reported that nanoparticle toxicity depends on particle characteristics.378 Effect of two forms of TiO2 viz. anatase (TA) and an anatase/rutile mixture (TM) embryo of the fish Danio rerio were studied and it was demonstrated that TM can induce greater mortality of the larvae in comparison to TA.379 The toxicity of various nanoparticles such as nano-TiO2, ZnO, CuO and Co3O4 on channel catfish hepatocytes and human HepG2 cells were investigated(Wang ,Y., 2011).380 HepG2 cells were found to be more sensitive for nanoparticles than catfish primary hepatocytes. The toxicity of nanoparticles on test species was due to ROS-induced cell death, cell damage and mitochondrial membrane damage. Several types of metal oxide nanoparticles affected mitochondrial functions and induced lactate dehydrogenase (LDH) leakage at concentrations as low as 50–100 g/ L.381, 382 4.5. Toxicity of silver nanoparticles Because of their antiseptic properties, silver nanoparticles are widely used in creams, textiles, surgical prosthesis,

cosmetics and as bacteriocides in fabrics and

other consumer

products.383 Both silver nanoparticles and dissolved silver, are known to have significant antibacterial properties. A higher antimicrobial activity is expected due to its larger specific surface area. The silver nanoparticles inhibit the enzymes for the P, S, and N cycles of nitrifying bacteria. They are also reported to block DNA transcription, and adenosine


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triphosphate (ATP) production. The silver nano particles react with proteins by combining the –SH groups of enzymes, which leads to the inactivation of the proteins and interrupt bacterial respiration.104,384 Due to their widespread applications, silver nanoparticles and their products have emerged as a massive source of ENPs to the surroundings. Potential toxicity from dermal exposure was shown with silver nanoparticles, that diminished human epidermal keratinocyte viability.385 In an in vitro study of cultured human keratinocytes, Lam et al. (2004) observed a substantial decrease in cell viability (0 to 9% cell viability after 30 minutes of incubation) and concluded cytotoxicity of silver nanocrystals (released by Acticoat™). Acticoat™

has been used for several years to heal wounds.386 Phytotoxicity of silver

nanoparticles on Phaseolus radiatus and Sorghum bicolor crop plants were investigated.387 It was observed that seedling growth was adversely affected due to silver nanoparticles. In another study, toxicity of silver nanoparticles on nematode, Caenorhabditis elegans were investigated.388

The

toxicity of

bare silver nanoparticles and PVP-coated silver

nanoparticles were compared and it was observed that coatings on the silver nanoparticles surface increase the toxicity of silver nanoparticles. Larese et al. (2009) demonstrated that polyvinyl pirrolidon coated silver nanoparticles was able to affect the damaged skin in an in vitro diffusion cell system.389

Transmission electron microscope (TEM)

was used for

verification of human skin penetration due to nanoparticles. In another study, toxicity of silver nanoparticles and ionic silver (Ag+) to photosynthesis in Chlamydomonas reinhardtii.390 Silver nanoparticle also showed significant effects on rice (Oryza sativa L.) seedlings391. To investigate the effect of silver nanoparticles on seedllings of rice species, different concentration of silver nanoparticles was taken and it was noted that there was significant 23 ACS Paragon Plus Environment


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reduction in root elongation, total chlorophyll and carotenoids contents. Toxicity of citrate (cit-Ag NPs) or humic acid (HA-Ag NPs) capped silver nanoparticles (Ag NPs) and Ag were demonstrated on. eggs, larvae, juveniles and adults of Platynereis dumerilii and it was observed that cit-Ag NPs and HA-Ag NPs were more toxic than Ag.392 Effect of different sized silver nanoparticles on the seed germination and seedling growth in jasmine rice (Oryza sativa L. cv. KDML 105) were investigated and it was observed that due to silver nanoparticle s showed negative effects on seed germination and seedling growth.393 Vannini et al. reported that 10mg/L AgNPs dose affected the seedling growth and also showed adverse effects on root tip cells.394 Genomic and proteomic changes were observed during AgNps exposure. Silver nanoparticles also induced morphological modification in root tip cells(Vannini et al 2014)”

Silver

nanoparticles were found to induce cytotoxic effects on fish Catla catla and Labeo rohita.395 It was observed that copper and silver nanoparticle(AgNPs) may reduce adult longevity in Drosophila and decreased sperm competition.396 The effects of AgNPs in rainbow trout (Oncorhynchus mykiss) hepatocytes were explored.397 This study proposed that AgNPs could influence hormone-regulated cell signaling pathways.On the exposure of Ag or Al2O3, nanoparticles cyotoxic effects were observed in soil bacteria viz. Pseudomonas stutzeri and Bacillus cereus. A decrease in bacterial transcriptional response was detected in NPs -treated soils.398 In another study it was reported that silver nanoparticles may inhibit the activities of erythrocyte acetylcholinestrase(AChE) and Na+/K+-ATPase and also affected the plasma biochemistry in adult zebrafish (Danio rerio).399 A comparative study was carried out for the toxicity of bare and

polyvinylpyrrolidone (PVP) coated silver nanoparticles


on the

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nematode, Caenorhabditis elegans. Coated silver nanoparticles were found to be more toxic in comparison to bare silver nanoparticles.400 4.6. Toxicity of zinc oxide nanoparticles (ZnO nanoparticles) The remarkable properties of zinc oxide(ZnO) nanoparticles have attracted the interest of many researchers in the past few years.98-100,401 One of the most remarkable commercial applications of 20–100 nm ZnO nanoparticles is their use in the production of sunscreens and cosmetics.402 At nanoscale, ZnO possesses novel electronic and optoelectronic properties and frequently utilized in biosensors, sunscreens, and additionally in medicinal applications like dental filling materials and wound healing.403-406 Due to widespread applications, ZnO nanoparticles are finally reaching the environment unintentionally. Recently, research has been carried out on the potential toxicity of ZnO nanoparticles in particular and other metal oxide

nanoparticles.79,407-411 and it has been

concluded that these nanoparticles ultimately pose threat to fauna, flora and humans. Because of the indiscriminate use of ZnO nanoparticles, it is compulsory to observe their fate in the ecosystem and biocompatibility with biological system. Nations et al., (2011) studied the effect of nano ZnO on Xenopus. laevis. X. laevis exposed to high ZnO nanoparticles concentrations died or displayed slower growth.412 Presence of ZnO nanoparticles in aquatic ecosystems could also reduce food resources such as algae. They reported that ZnO nanoparticles released into aquatic ecosystems in high concentrations could have detrimental effects on aquatic organisms such as amphibians. Exposure to ZnO nanomaterial significantly increased mortality and decreased hatchability of zebrafish at 1mg/L.413



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Zebrafish also had an increased incidence of tissue ulceration beginning 72 hour post fertilization (hpf) and reaching 100% tissue ulceration by 108 hpf. A recent study on ZnO nanoparticles reported that it impels much more noteworthy cytotoxicity than non-metal nanoparticles primary mouse embryo fibroblast cells.414 and prompts apoptosis in neural stem cell.415 The revealed reports have demonstrated that ZnO nanoparticles restrains the seed germination and root development.416 Adams et al.,(2006) studied the antibacterial properties of nanoparticles on Bacillus subtilis and Escherichia. coli.417 Inhalation of ZnO may cause disturbance in pulmonary function(lung disorders) in pigs while in case of inhalation in humans, it may responsible for metal fume fever in humans.418,419 ZnO nanoparticles have been reported to be most toxic nanoparticle among the engineered metal oxide nanoparticles. ZnO nanoparticles have the lowest LD50 value.420 On the other hand, it was additionally reported that ZnO was not discovered to be cytotoxic to human dermal fibroblasts.421 It has reported that ZnO nanoparticles cause membrane damage in the E. coli, possibly due to oxidative stress mechanisms.422 SEM images of E. coli treated with ZnO nanoparticles showed considerable damage to some E. coli and it was due to the breakdown of the bacterial membrane. Huang et al. (2008) investigated the possible interactions that govern the bactericidal activity of 60–100 nm polyvinyl alcohol (PVA)-ZnO nanoparticles against Streptococcus agalactiae and Staphylococcus aureus.423 They observed the cellular damage when the PVA coated ZnO nanoparticle concentrations were higher (> 0.016M) in the ethylene glycol (EG) medium containing the cells. A significant change in the ZnO nanoparticle crystal structure was observed after these cells established contact with PVA-coated ZnO nanoparticles,


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However, in their preliminary studies, contrary to previous studies, they reported that low concentrations of ZnO nanoparticles did not induce any cellular damage.424 Heinlaan et al. (2008) investigated the toxicity of ZnO, Cuo and TiO2nanoparticles to bacteria Vibrio. fischeri ,crustaceans Daphnia magna and Thamnocephalus platyurus.and studied the toxic effects of metal oxides and solubilised metal ions.425 They showed that the metal oxide nanoparticles do not necessarily have to enter to the cells to cause damage in the cell membrane. In fact, the contact between the particle and the cell wall may increase the solubilisation of metals.It has also been demonstrated that the release of metal ions from the ZnO NPs i.e. from the solubilisation, was responsible for toxicity in lung cell lines.426 While under realistic environmental conditions, similar results on algae have been reported.79 Further, Lin and Xing (2008) explored that ZnO nanoparticles enter apoplast and protoplast of the root endodermis and stele.427 Recently, Neuro behavioral toxicity of ZnO nanoparticles was investigated in developing fish. Hatching delay and larval hyperactivity was observed in of embryo–larval zebrafish when exposed to ZnO nanoparticles.428 Changes in plant growth, bioaccumulation and antioxidative enzyme activity in Brassica juncea in the presence of ZnO nanoparticles were studied by Rao et al.,(2014). Nanoparticles of ZnO showed negative effects on plant. Bioaccumulation of ZnO nanoparticles were also reported.429 Yoon et al., (2014) reported long term effect of ZnO nanoparticles on the soybean [Glycine max (L.) Merrill]. It was reported that ZnO nanoparticles adversely influenced the formative stages and reproduction of soybean plants.430 Cytotoxic effects of ZnO nanoparticles on mouse dermal fibroblast cells (mDFs) and human periodontal ligament fibroblast cells (hPDLFs) were investigated.431



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4.7. Toxicity of Iron/ iron oxide nanoparticles Nanostructured iron oxides allow a wide range of potential applications in nanotechnology related fields, including bio-medical imaging, magnetic target drug delivery, environmental catalysis, magnetic storage and so forth. Nanostructured iron has broad applications in the field of environment and biomedical and so it is necessary to study its environmental fate and biocompatibility. Iron oxide NPs, Fe3O4 and Fe2O3 have been synthesized with a number of methods involving different compositions and phases.4 Yan and Zhang (2011) showed the in-vitro cytotoxicity of nanoparticles utilizing Hek 293 cell culture system with diverse dosages.432 Nanoparticle of hematite were found to reduce cell viability. Due to such cytotoxicity, decrease in the activity of antioxidative enzymes induced by oxidative stress in cells may occur. Some researchers studied the comparative toxicity of nano- and micro particles of some metal oxides like Fe2O3, Fe3O4, TiO2 and CuO.433 They reported cell death, mitochondrial damage, DNA damage and oxidative DNA lesions when the human cell line A549 was exposed to these nano and microparticles. This study showed that CuO nanoparticles were much more toxic when compared to the CuO microparticles. Similarly, the microparticles of TiO2 caused more DNA damage compared to the nanoparticles. Toxicity of Au, Ag and Fe3O4 nanoparticles were studied on plants and microorganisms.434 Zero-valent iron nanoparticles have attracted more attention in the field of water and waste water remediation due to their sorbing efficiency, but at the same time it is also becoming a source of nanoparticles contamination in the environment.435-440 Zero valent iron nanoparticles reduced high concentrations of solvents to nearly zero within days but at the


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same time, oxygen levels were reduced causing the groundwater anoxic and pH levels changed significantly.71 Toxicity of four different iron oxide nanoparticles viz. polyvinyl pyrrolidone (PVP-IONP) ,ascorbate (ASC-IONP), dextran (DEX-IONP) and citrate (CIT-IONP), on Daphnia magna were investigated.441 The benefits of different nanoparticles have been demonstrated in several scientific fields, but reports on their capability to penetrate the skin are rare. Bregoli et al.(2009)442 explored the poisonous quality of seven NPs viz. Sb2O3, TiO2,Fe2O3, Fe3O4, Au, Co, and Ag on primary cultures of human hematopoietic progenitor cells with colony forming unit(CFU) assays. Among all seven nanoparticles, Sb2O3 and cobalt (Co) NPs were found to be more toxic than others. They also concluded that Sb2O3 NPs impair the proliferation of erythroid progenitors. 4.8. Toxicity of CuO nanoparticles Toxicity of aggregated zero valent copper nanoparticles on E. coli was studied by that by applying the centroid mixture design of experiment.443 Various

parameters

viz. pH,

temperature, concentration of nanoparticles, aeration rate and concentration of bacteria were taken under consideration. On the basis of linear and quadratic regression model it was concluded that toxicity of copper nanoparticles depends on both primary and interactive effects. The lethal effects of CuO nanoparticles on freshwater shredder Allogamus ligonifer was studied in 2012 by Pradhan et al (2010).444 Different concentration of CuO nano suspension was prepared(0,50,100,250,500 and 1000mg/L) to observe its effect on the shredder. It was observed that nanoparticle exposure led to lethal effects at high concentrations. Metal oxide nanoparticles have been found to adversely affect mammalian cells and some aquatic



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organisms. For example, CuO nanoparticles induced death of H4 and SH-SY5Y cells in a dose dependent manner.445 A comparative study on the toxicity of CuO NPs, core shell(CS-CuO) NPs and ionic copper were compared in the aquatic macrophyte Lemna gibba. The study revealed that the polymer coating changes the mechanism of toxicity due to changes in surface characteristics. CS-CuO nanoparticles showed 50% decreased growth in plant in comparison to CuO nanoparticles.446 Jośko et al., (2014) demonstrated that nanoparticles (ZnO, Cr2O3, CuO and Ni )may also affect the enzymatic activity of the soils.447 In another study it was observed Cu-nanoparticles may cause oxidative stress in the liver, gills and muscles of juvenile Epinephelus coioides.448 4.9. Toxicity of cerium oxide nanoparticles (CeO2) It was observed that due to the presence of cerium oxide nanoparticles in the root vascular tissues and aerial parts of plants, the activity of root antioxidant enzyme were significantly reduced. nCeO2 particles reached into the kidney been root via the gap in the Casparian strip.449 Impact of CeO2 nanoparticles on two amphibian larvae viz. Pleurodeles waltl and Xenopus laevis were reported by Bour et al.,(2014).They also demonstrated its effect on the invertebrate Chironomus riparius and the Diatoms Nitzschia palea. Genotoxic effect was noted in amphibian larvae and Pleurodeles.450 Nanoparticle can also pose toxic effect on fungal population. The toxicity of nano-CeO2 was not ascribed to the dissolved Ce2+ ions, but to the entrapment of algal cells into the aggregates of nanoparticles.451


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Phytotoxicity of four rare earth oxide nanoparticles were explored by Ma et al. (2009). Phytotoxic behaviour of CeO2, La2O3, Gd2O3, and Yb2O3 nanoparticles were studied on seven higher plant species (radish, rape, tomato, lettuce, wheat, cabbage, and cucumber).452 Their effects on root growth varied greatly between different nanoparticles and plant species. It was observed that inhibitory effects different nanoparticles also differed in the different growth process of plants. 4.10. Toxicity of chitosan/gold/silica nanoparticles Loh et al., (2010) studied the effect of chitosan nanoparticles (1% w/v) into the cell nucleus and after 4 h exposure, it was observed that the chitosan NPswere responsible for necrotic or autophagic cell death.453 It may be due to damage of cell membrane and resultant enzyme leakage.The acute toxicity of gold nanoparticles was explored by Cho et al., in 2009 by carrying out an in vivo study using 13 nm-sized gold nanoparticles oxidative stress and membrane damage coated with PEG (MW 5000)454. It was found that the PEG-coated NPs can induce acute inflammation and apoptosis in the liver.

It was reported that the PEG-coated NPs were trapped in liver Kupffer cells and

spleen macrophages. Monodisperse

polypyrrole (PPy) nanoparticles with five different

diameters (20, 40, 60, 80, and 100 nm) were fabricated via chemical oxidation polymerization in order to evaluate size-dependent cytotoxicity .455 Fent et al. (2010) demonstrated the toxic effects of fluorescent silica nanoparticles (FSNP) on early life stages of zebrafish.456 It was observed that the ∼60 and ∼200 nm-sized FSNP were adsorbed on the chorion of eggs. The effect of some nanoparticles on humans is summarized in Table 6.457-466 Now, researchers are focusing not only on the new synthesis method of nanoparticles but also showing their concern

regarding the fate of nanoparticles and their

effects on humans.



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Table 6 5. CONCLUSIONS In the new century, with increasing demands for nanotechnology, there has been a drastic rise in the synthesis as well as application of nanoparticles in widespread areas. This has led to rapid development of commercial applications which involve the utilization of a wide variety of engineered nanoparticles (ENPs). Moreover, the recent use of nanoparticles in biomedical applications has made it even more demanding owing to their specific properties. Further, the application of coated nanoparticles for various purposes is surprisingly showing a great increase. Production of varieties of nanoparticles at astronomically immense scale and their utilization for the benefits of humans will conclusively lead to its entrance in the environment by direct or indirect processes such as disposal, accumulation, etc. The production, utilization and disposal of manufactured nanoparticles will inevitably lead to unintentional discharges in air, soil and aquatic systems which have put forward to the potential risk to the environment. Nanotoxicity of any nanoparticle is greatly influenced by its shape, size, variety, coating material and composition. Sometimes, toxicity can be due to the toxicity of precursors which have been used for its preparation. Toxicity may be increased/decreased depending upon the environment like physical and chemical nature of contacting species, test organism species, and test methods. It was also reviewed that same nanoparticles may pose different extent of toxicity on plants, human beings and animals. Toxicity also depends upon the nature of functional groups. There may be a good correlation in the results for in vitro and in vivo studies, depending upon the experimental conditions and the sensitivity of specimens. So there arises a need for better understanding and assessment of the toxicity and eco-toxicity of engineered nanoparticles to the key ecosystem organisms like algae, plants, and fungi which are


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continuously being exposed to these new materials. Increasing demands of nanoparticles cannot be neglected, due to their vast applications and therefore, for better and safe use of nanoparticles in future we have to be very attentive in understanding the fate, demeanor and toxicity of nanoparticles. Development of some predictive models should be encouraged in order to establish the safety of engineered nanoparticles. Academic Researcher and industries should fixate on reuse/ regeneration of exhausted nanoparticles to obviate its ingression into the environment.

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Table 1. Applications of engineered nanoparticles.8,58,72, 76-112

Nanoparticles CNTs

SnO2 Al2O3 CeO2 SiO2 TiO2

Carbon nanohorns ZnO

Products/applications References Electronic devices, field emission devices, and composite materials , numerous biological and medical applications, As adsorbent material (76-81) for the removal of pollutants from water Transparent conducting coating of glass, gas sensors, solar cell, and (82,83) heat mirror, gas sensors, catalyst supports Batteries, adsorbent, grinding, catalysis; polishing abrasives (84) Abrasive materials of chemical–mechanical polishing (CMP) oxygen (85- 87) sensor , polishing materials, gas sensors, fuel additive; Pharmaceutical products, vegetable oil refining, ceramics, detergents, adhesives, electronics, chromatography , fire proof glass; fillers , (88-91) catalysts Food colouring, photocatalyst; pigments, additive in pharmaceuticals and cosmetics ,paints, antibacterial and self-cleaning materials, (84, 92-94) sunscreen, cosmetics, uv-protection, catalysis, self-cleaning window coating; fillers, catalyst supports, and photocatalysts; Catalyst supports; and drug delivery (95-97)

Electrostatic dissipative coating ,semiconductor material , chemical sensors and solar cells paints, sunscreen, cosmetics, electrical and optical devices ,uv-protection, catalysis; diode lasers , chemical absorbent , pigments, optical materials, Fe3O4 Removal of contaminants, sensors, magnetic resonance imaging , bio manipulation; magnetic storage media magnetic refrigeration magnetic resonance imaging (MRI) DNA detection and drug delivery system and cancer therapy Metallic copper Applications in catalysis nanoparticles Ag Dental resin composites ,coatings of medical equipments Paints, textiles, antibacterial agent, Magnesium– Sensors ,catalysis aluminum oxide, MgAl2O4 CdS Photodetectors, optoelectronics, and for solar cell applications

zero-valent

(84,98-100)

(8,72,101103) (58) (88,89, 104) (105)

(106) (107)

iron Water remediation;

0

(Fe ) Gold nanoparticles(II) ZnS Fullerene

Drug delivery applications

(108)

Electroluminescent (109) devices, solar cells and phosphors Superconductors and for drug delivery; sensors, cosmetics catalyst, (110-112)


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(C60)

polymer modifications optical and electronic devices, sporting goods polymers, and biological and medical applications ,lubricants,

Table 2.Removal of pollutant species from effluents by various nanoparticles pollutants (inorganic/organic).113-153 Pollutants (inorganic/organic) As(III) and As(V) Diethylchloro phosphate (DEClP) Cr(VI) Congo red Cu

Food dyes Fluoride Gold(III) ions Uranium Acridine orange Rhodamine 6G (R6G) Organic dyes Methylene blue and crystal violet dyes Cd(II) Pb(II) Co(II) Zn(II) Cu(II) Mo(VI) Hg(II) Arsenate Acid Black-24 Methyl red Acid Orange 7and Acid Orange 10 Rhodamine 13 Acid Green 27 Trichloroetheneand chlorobenzene TiO2 nanoparticles Flouride

Nanoparticles used for the removal of pollutants Cupric oxide nanoparticles Al2O3nanoparticles Carboxymethyl 2 cellulose-stabilized zero-valent iron nanoparticles; multiwalled carbon nanotubes Maghemite nanoparticles; alumina nanoparticles

Reference s (113) (114) (115,116) (117,118) (119-121)

Multiwalled carbon nanotubes; kaolinite-supported zerovalent iron nanoparticles ;chitosan-coated magnetic nanoparticles modified with α-ketoglutaric acid Spirulina platensis nanoparticles Fe3O4@Al(OH)3magnetic nanoparticles Chitosan coated magnetic nano-adsorbent zero-valent iron nanoparticles Magnetic nanoparticles (gamma -Fe2O3) Fe3O4@PAA nanoparticles; hexadecyl functionalized magnetic silica nanoparticles Fe3O4@C nanoparticles Cadmium sulfide nanoparticles

(122) (123) (124) (125) (126) (127,128)

Akaganeite nanocrystals ; Fe2O3 nanoparticles

(131,132)

Silica–alumina nanoparticles Magnetic chitoson nanoparticles ;zero-valent iron nanoparticles Akaganeite nanocrystals Iron phosphate nanoparticles multiwalled carbon nanotubes Maghemite FeS Nanostructured ZrO2 TiO2 and Fe0 Silica nanoparticles

(133) (134,135)

(129) (130)

(136) (137,138) (139) (140) (141) (142)

Magnetic chitosan nanoparticles

(143) (144)

BiFeO3 Chitosan nanoparticles Palladium/magnetite

(145) (146) (147)

Acid Blue 92 ; Basic Blue 3

(148)

Mg-doped nano ferrihydrite

(149,150)



1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Anthracene-9-carbonxylic acid Methyl orange Cadmium and lead

Fe3O4@Al(OH)3 magnetic nanoparticles CdSe bentonite-supported nanoscale zero-valent iron NiO nanoparticles

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(151) (152) (153)

Table 3.Application of coated nanoparticles154-178 Nanoparticles

Coating materials

Aim

Refernces

Alginate coated chitosan core shell nanoparticles oleic acid-coated Fe3O4 nanoparticles β-lactoglobulin-coated gold nanoparticles

Alginate

Oral delivery of enoxaparin

(154)

Oleic acid-

pH-responsive Pickering emulsions

(155)

β-lactoglobulin

(156)

Citrate-coated silver nanoparticles

Citrate

3-mercaptopropanoic acid-coated superparamagnetic iron oxide nanoparticles Carbon-coated SnSb nanoparticles Polymerized-glucose coated Fe3O4 magnetic nanoparticles Citrate-coated magnetic nanoparticles Polypyrrole-coated magneticnanoparticles Superparamagneticsodium alginate-coated Fe3O4nanoparticles Gold coated ferric oxide nanoparticles Carbon-coated titanium dioxide coreeshell nanoparticles Hyaluronic acid-coated solid lipid nanoparticles Imidazole and imine coated ZnO nanoparticles Citric acid coated Fe3O4 magnetic nanoparticles Nickel oxide coated carbon nanoparticles Salicylic acid-coated magnetic nanoparticles Phosphomolybdate-dopedpoly(3,4ethylenedioxythiophene) coatedgold nanoparticles Sulfate‐A‐coated magnetite

3-mercaptopropanoic acid-

Controlling the gastrointestinal fate of ingested inorganic nanoparticles. Transformation of the morphology, dissolution behavior and reaction product of AgNPs in different amno acid -containing systems in human body Arsenate removal

Carbon

lithium-ion battery anodes

(159)

Polymerized-glucose

Delivery of aspirin

Citrate

For forward osmosis Extraction of nitrophenols

Polypyrrole

(157)

(158)

(160) (161) (162)

Malachite green

(163)

Detection of DNA hybridization processes Microbial fuel cells

(164)

(166)

Imidazole and imine

Targeted deliveryof vorinostat to CD44 overexpressing cancer cells detection of Al(III) and Zn(II)

Citric acid

Biomedical applications

(168)

Nickel oxide

Temperature sensing materials Genomic DNA extraction

(169)

Electrocatalyticreduction of bromate

(171)

Biomedical applications

(172)

Gold Carbon-

Hyaluronic acid

Salicylic acid Phosphomolybdatedoped-poly(3,4ethylenedioxythiophe ne)


(165)

(167)

(170)

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nanoparticles PEG-coated silica nanoparticles Pyrolytic carbon-coated Si nanoparticles Polyethylene glycol (PEG) coated Fe3O4 nanoparticles Sulfonated-mercaptopropanoic acid coated Fe3O4 nanoparticles Oleic acid-coatedFe3O4 nanoparticles Chitosan coated magnetic nanoparticles

Poly ethylene glycol Pyrolytic carbon-

Oral insulin delivery As anode materials for high-performance lithium-ion batteries Biomedical application

(173) (174)

Sulfonatedmercaptopropanoic acid Oleic acid-

Magnetic catalyst for Biginelli reaction

(176)

Biomedical application

(177)

Chitosan

As a support for bio ligands binding

(178)

Polyethylene glycol (PEG)


(175)


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Page 72 of 76

Table 4. In vitro/in vivo studies of Single walled carbon nanotubes(SWCNTs) and multiwalled carbon nanotubes (MWCNTs). 226-266 SWCNT /MWCNT MWCNT

Effects Nitrative DNA damage

Test species human lung epithelial cells

Reference (226)

SWCNT

Acute toxicity on central and peripheral nervous system of chicken

(227)

Multi-walled carbon nanotubes

In vitro cyto toxicity

chicken embryonic spinal cord (SPC) or dorsal root ganglia (DRG) C6 rat glioma cells

marine alga, Dunaliella tertiolecta

(229)

rainbow trout, Oncorhynchus mykiss Mice

(230)

human embryonic kidney (HEK293) cells Human lungs

(232)

Water dispersible oxidized multiwalled carbon nanotubes Carbon nanotube SWCNT Multi wall carbon nanotubes Carbon nanotubes Single-wall carbon nanotubes Water soluble multi-walled Carbon nanotubes MWCNT Carbon nanotubes With impurities Multi-walled carbon nanotubes Carbon nanotubes Single-walled carbon nanotubes dispersed in aqueous media Via non-covalent functionalization Double-walled nanotubescontaminated food Multi-walled carbon nanotube

Cytotoxicity effects

IImmunotoxcity Subcutaneous implantation Induce oxidative stress and cytotoxicity Respiratory toxicity

(228)

(231)

Inflammation on Human macrophages Cells Splenic toxicity

human

(233) (234)

mice

(235)

Intratracheal instillation In vivo immunological toxicity Reactive oxygen species (ROS) increased and cell viability decreased Genotoxicity

Guinea pigs mice

(236)

Cytotoxicity, and epigenetic toxicity of nanotube suspensions

prokaryotic and eukaryotic cell systems

(240)

Lethal and sub-lethal toxicity Soil microbial activity decreased??

Eisenia veneta earthworms

(241)

microorganisms

(242)

(237)

suspension rice cells

Bacteria in vitro and in vivo assays


(238) (239)

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SWCNT

Intratracheal instillation inhibited fecundity and growth in daphnia

Mice

(243)

Daphnids.

(244)

Carbon nanotubes

Dermal and eye irritation and skin sensitization

Male Kbl:NZW rabbits and male Slc:Hartley guinea pigs

(245)

Carbon nanotubes

In vitro cytotoxicity on lung epithelial cells DNA damage in plant and mammalian cells

human lungs

(246)

Allium cepa, human lymphocytes, mouse bone marrow cells and pBR322 plasmid DNA rodent macrophage cells

(247)

Rats, inflammatoryand fibrotic responses

(249)

Suspended multi-walled carbon nanotube

Multi-walled carbon nanotubes (MWCNT)

Functionalized multiwalled carbon nanotubes MWCNT

Cytotoxic and inflammatory responses Intratracheal instillation

(248)

release of the proinflammatory cytokine interleukin 8 from HEKs Single-walled carbon in vitro

Human epidermal keratinocytes (HEK)

(250)

Multi-walled carbon nanotube Single-walled Carbon nanotubes

cultured human dermal fibroblasts (HDF).

(251)

Single walled carbon nanotubes

Depletion in A549 lung cells

252)

SWCNT

SWCNT caused a dosedependentincrease in ROS Idecreasing glutathione (GSH) level, increasing malondialdehyde (MDA), inflammatory cell infiltration Inflammatory response of immortalised and primary human lung epithelial cells (a549 and nhbe) increase in DNA damage , genotoxicity in human bronchial epithelial

human alveolar carcinoma epithelial cell line A549 (ATCC, CCL-185) FE1Muta Mouse lung epithelial cell line mice

human lung epithelium

(255)

Human bronchial epithelial cell line exhibiting an epithelial Phenotype human Wbroblasts

(256)

The Caco-2 cell line from a human caucasian colon adenocarcinoma

(258)

HEK293 cells (human

(259)

Single-walled Carbon nanotubes

SWCNT

Genotoxicity carbon Nanotubes

Single-wall carbon nanotubes Carboxylic acid functionalized single wall carbon nanotubes

Cytotoxicity on human Wbroblasts Cytotoxic effect on on the Caco-2 cells

SWCNT

inhibit HEK293 cell


(253)

(254)

(257)


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Single-wall carbon nanotubes Single-walled carbon nanotubes

Multiwalled Carbon Nanotubes Multi-wall carbon nanotubes Single and multi walled carbon nanotubes Multi-walled carbon nanotube Multi-walled carbon nanotubes

proliferation and decrease cell adhesive ability Human cells of the oral cavity Cytotoxicity of singlewalled carbon nanotubes on human hepatoma Hepg2 cells In Vitro evaluation of cytotoxicity and oxidative Stress Cytotoxic and genotoxic effects Cyto and genotoxicity on macrophages Alteration of protein expression in a target epithelial cell Inhibition of Lactate dehydrogenase activity

Page 74 of 76

embryo kidney cells)

Human gingival fibroblast (HGF) Human hepatoma HepG2 cell

Murine RAW 264.7 macrophages and human A549 lungcells Human umbilical vein endothelial cells mouse macrophages

(260) (261)

(262)

(263) (264)

human keratinocytes

(265)

LDH activity

(266)


Page 75 of 76

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Table 5 Toxicity of Fullerenes300-317 Fullerene/modified Fullerene Fullerene C60 C60 fullerene Fullerene C60 and fullerol C60(OH)18– 22

C60 fullerene particles Fullerene (C60)

Carbon fullerene

Hydroxylated fullerene nanoparticles Fullerene water suspensions Fullerene, C60 Fullerene C60 nanoparticles Fullerene nanoparticles Functionalized fullerene Fullerene C60 nanoparticles

Hydroxylated fullerenes Fullerene (C60)

Fullerene (C60)

Test species

Effects

References

Embryonic zebrafish; Mytilus hemocytes Gills of fish Cyprinus carpio (Cyprinidae)

Induce oxidative stress Immune system Increase in lipid peroxidation, decrease in GCL activity, and the depletion of GSH stock Pulmonary toxicity

(300) (301) (302)

Delay in molting and significantly reduced offspring production

(304)

Increased in malformations, pericardial edema, and mortality Decreased survival rate, shortened lifespan, apoptotic cell death Mortality and glutathione (GSH) induction of embryos

(305)

Rat lung after inhalation aquatic organisms(Daphnia magna and Hyalella azteca) In vivo toxicity; embryonic zebrafish Soil nematode caenorhabditis elegans Japanese medaka(Oryzias latipes) embryos

(303)

(306)

(307)

Aquatic species, Daphnia and Fathead minnow lung cells of rats

Increased LPO in fish

(308)

Genotoxicity

(309)

Escherichia coli K12

Inhibits microbial respiratory activity Decrease in ATP and glutathione Genotoxicity

(310)

Cytotoxicity, Decrease in viability, Antioxidant and oxidative damage

(313)

Oxidative damage,lipid

(315)

Human cells Ames Salmonella typhimurium TA98, TA100, TA1535, and TA1537 strains and Escherichia coli strain; cultured Chinese hamster CHL/IU cells Human epidermal keratinocytes (HEK) Polychaeta Laeonereis acuta (Nereididae) Rat liver


(311) (312)

(314)


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Fullerene (C60) nanoparticles Arsenium and an organic nanomaterial (fullerene, C60)

peroxidation Clastogenicity and phototoxicity Lipid damage

Mammalian cells Zebrafish hepatocytes;In vitro study

Page 76 of 76

(316) (317)

Table 6. Recent studies on toxicity of nanoparticles to humans. 457-467 Nanoparticles ZnO nanoparticles

TiO2 nanoparticles Silver nanoparticles Nickel oxide nanoparticles Fullerenol Nanoparticles ZnO nanoparticles

Silver nanoparticles

Bare titanium dioxide, zinc oxide, magnesium oxide, silver, gold nanoparticles and their Triglyceride-coated form Metal oxide NPs (ZnO, CeO2, TiO2 and Al2O3)

Titanium dioxide nanoparticles

Test Organs /Species Human pulmonary adenocarcinoma cell line LTEP-a-2 Human peripheral blood Mononuclear cells Human colon carcinoma cells Human pulmonary epithelial celllines: BEAS-2B and A549 Cultured human lung fibroblasts Human polymorpho nuclear neutrophil(pmns) Human umbilical vein endothelial cells (huvecs) Suspensions of Balb/c skin cells

Human peripheral blood Lymphocytes (pbls).

Human gastric epithelial cells

Toxic Effects Cytotoxicity on human pulmonary adenocarcinoma cell line LTEP-a-2

References (457)

Suppressed IDO activity and IFN-c production

(458)

Oxidative stress and cytotoxicity

(459)

Inflammation and genotoxic effect in lung epithelial cells

(460)

Cytotoxicity and Genotoxicity Delay in human neutrophil apoptosis

(461) (462)

Endothelial cell injury and dysfunction

(463)

Cytotoxicity

(464)

Induced changes in the expression levels of adhesion molecules and The c-x-c chemokine receptor type 4 (cxcr4) in these cells, T-cell proliferation upon cell exposure to TiO2 and Al2O3 nps Oxidative stress, DNAdamage

(465)


(466)

Critical Review on the Toxicity of Some Widely Used Engineered

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