Minimizing the potential of cancer recurrence and metastasis by the use of graphene oxide nano-flakes released from smart fiducials during image-guided radiation therapy.

An increasing number of studies show that cancer stem cells become more invasive and may escape into blood stream and lymph nodes before they have received a lethal dose during radiation therapy. Recently, it has been found that graphene oxide (GO) can selectively inhibit the proliferative expansion of cancer stem cells across multiple tumor types. In this study, we investigate the feasibility of using GO during radiotherapy to synergistically inhibit cancer stem cells, and lower the risk of cancer metastasis and recurrence. We hypothesize that graphene oxide nano-flakes (GONFs) released from newly-designed radiotherapy biomaterials (fiducial) can reach targeted tumor cells within 14-21 days. These are the typical time periods between the implantation of the fiducial and the start of image-guided radiation therapy. To test this hypothesis, the spatial-temporal diffusion of GONFs in soft tissue is investigated as a function of different particle sizes. Toxicity of GONFs to normal (HUVEC) and cancer (A549) cells has been assessed using the MTT assay. In addition, the survival fraction of A549 cells treated with GONFs is determined via clonogenic assay during radiotherapy. The diffusion study shows that only GONFs sizes of 50 and 200 nm could achieve the desired concentration of 50 µg/mL for 2 cm diameter tumor after 14 and 21 days respectively. The clonogenic and the MTT assay confirm the additional benefit of GONFs in killing lung cancer cells during radiotherapy. This work avails ongoing in vivo studies that use GONFs to enhance the treatment outcome for cancer patients during radiation therapy.

Synthesis and in vitro studies of PLGA-DTX nanoconjugate as potential drug delivery vehicle for oral cancer.

Advances in nanotechnology have led to the design of multifunctional nanoparticles capable of cellular imaging, targeted drug delivery, and diagnostics for early cancer detection. We synthesized poly(lactic-co-glycolic acid) nanoparticles encapsulating a model radiosensitizing drug docetaxel accomplishing localized in situ delivery of the sensitizer to the tumor site. The synthesized nanoparticles have been characterized for their physicochemical properties. The in vitro cytotoxicity of drug-loaded nanoparticles has been studied on human tongue carcinoma cell line SCC-9 (ATCC-CRL-1629).

Synthesis of biocompatible iron oxide nanoparticles as a drug delivery vehicle.

Over the last decade, there has been growing interest in developing novel nanoparticles (NPs) for biomedical applications. A safe-by-design approach was used in this study to synthesize biocompatible iron oxide NPs. The size of the particles obtained was ~100 nm. Although these NPs were significantly (P<0.05) internalized in MCF-7 (human breast adenocarcinoma cell line) cells, no adverse effect was observed in the cells as assessed by cytotoxicity assays (neutral red uptake and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) and cell cycle analysis. Our data demonstrate the potential of iron oxide NPs as a biocompatible carrier for targeted drug delivery.

Synthesis and characterization of multifunctional gold nanoclusters for application in radiation therapy.

Gold nanoparticles, because of their high radiation absorption coefficient and efficient generation of secondary photoelectrons, have been predicted to enhance therapeutic efficacy in radiation therapy. However, high dose for effective treatment limits their use. We have synthesized multifunctional gold nanoclusters (GNCs) that can be used for imaging and radiation therapy. The designed GNCs have been characterized for their physicochemical properties, biocompatibility, and their radiation dose enhancement potential on PC3 cell lines.

Monitoring characteristics and genotoxic effects of engineered nanoparticle-protein corona.

Engineered nanoparticles (ENPs) possess different physical and chemical properties compared to their bulk counterparts. These unique properties have found application in various products in the area of therapeutics, consumer goods, environmental remediation, optical and electronic fields. This has also increased the likelihood of their release into the environment thereby affecting human health and ecosystem. ENPs, when in contact with the biological system have various physical and chemical interactions with cellular macromolecules including proteins. These interactions lead to the formation of protein corona around the ENPs. Consequently, living systems interact with the protein-coated ENP rather than with a bare ENP. This ENP-protein interaction influences uptake, accumulation, distribution and clearance and thereby affecting the cytotoxic and genotoxic responses. Although there are few studies which discussed the fate of ENPs, there is a need for extensive research in the field of ENPs, to understand the interaction of ENPs with biological systems for their safe and productive application.

Heteroagglomeration of zinc oxide nanoparticles with clay mineral modulates the bioavailability and toxicity of nanoparticle in Tetrahymena pyriformis.

The extensive use of zinc oxide nanoparticles (ZnO NPs) in cosmetics, sunscreens and healthcare products increases their release in the aquatic environment. The present study explored the possible interaction of ZnO NPs with montmorillonite clay minerals in aqueous conditions. An addition of ZnO NPs on clay suspension significantly (p<0.05) increases the hydrodymic size of clay particles from 1652±90nm to 2158±13nm due to heteroagglomeration. The electrokinetic measurements showed a significant (p<0.05) difference in the electrophoretic mobilities of bare (-1.80±0.03µmcm/Vs) and ZnO NPs-clay association (-1.37±0.03µmcm/Vs) that results to the electrostatic interaction between ZnO NPs and clay particles. The attenuated total reflectance Fourier transform infrared spectroscopy analysis of ZnO NPs-clay association demonstrated the binding of ZnO NPs with the Si-O-Al region on the edges of clay particles. The increase in size of ZnO NPs-clay heteroagglomerates further leads to their sedimentation at 24h. Although, the stability of ZnO NPs in the clay suspension was decreased due to heteroagglomeration, but the bioavailability and toxicity of ZnO NPs-clay heteroagglomerates in Tetrahymena pyriformis was enhanced. These observations provide an evidence on possible mechanisms available in natural environment that can facilitate nanoparticles entry into the organisms present in lower trophic levels of the food web.

Laboratory Scale Microbial Food Chain To Study Bioaccumulation, Biomagnification, and Ecotoxicity of Cadmium Telluride Quantum Dots.

The increasing applications of engineered nanomaterials (ENMs) in consumer products warrant a careful evaluation of their trophic transfer and consequent ecological impact. In the present study, a laboratory scale aquatic microbial food chain was established using bacteria (Escherichia coli (E. coli)) as a prey and ciliated protozoan (Paramecium caudatum) as a predator organism to determine the impact of cadmium telluride quantum dots (CdTe QDs). We observed that 29% of bacterivory potential of paramecium was lost, including an ∼12 h delay in doubling time on exposure to 25 mg/L CdTe QD (∼4 nm) as compared to control. The fluorescence based stoichiometric analysis revealed that 65% of the QDs bioaccumulated when paramecia were exposed to 25 mg/L QDs at 24 h. There was a significant (p < 0.05) increase in cellular cadmium (Cd) concentration at 24 h (306 ± 192 mg/L) as compared to 1 h (152 ± 50 mg/L). Moreover, the accumulation of Cd in E. coli (147 ± 25 mg/L) at 1 h of exposure to 25 mg/L QDs transferred 1.4 times higher Cd (207 ± 24 mg/L; biomagnification factor = 1.4) to its predator, paramecium.

Zinc oxide nanoparticle induced age dependent immunotoxicity in BALB/c mice.

Zinc oxide (ZnO) nanoparticles (NPs) have potential applications in cosmetics, food packaging and biomedicine but concerns regarding their safety need to be addressed. In the present study, the immunotoxic potential of ZnO NPs was evaluated in different ages of BALB/c mice after sub-acute exposure. The cytokine release, immunophenotyping, distribution of ZnO NPs and ultrastructural changes were assessed. A significant (p < 0.05) change in the CD4- and CD8-cells, levels of IL-6, IFN-γ and TNF-α and reactive oxygen species were observed in aged mice. In juvenile mice, increase in reactive oxygen species and IL-6 and TNF-α levels was observed with no significant changes in adult mice. A significant (p < 0.05) increase in the expression levels of mitogen activated protein kinase (MAPK) cascade proteins such as phospho-ERK1/2, phospho-JNK and phospho-p38 were also induced in aged mice. Collectively, our results indicate that the aged mice are more susceptible to ZnO NP induced immunotoxicity.

Can CuO nanoparticles lead to epigenetic regulation of antioxidant enzyme system?

Copper has been used from ancient time in various applications. Scientists have exploited its means of exposure and consequences to living organisms. The peculiar property of nanomaterials that is a high surface to volume ratio has increased the range of application in products. Copper oxide nanoparticles (CuO NPs) are widely used in industrial applications such as semiconductor devices, gas sensor, batteries, solar energy converter, microelectronics, heat transfer fluids and consumer products. In contrast, acute toxicity of CuO NPs has also been reported. Subsequently, human and environmental health may be at a high risk. Their frequent use can also contaminate ecosystems. Therefore, the toxicity of CuO NPs needs to be thoroughly understood. In this review, we have tried to discuss the recent facts and mechanism that have been explored for CuO NPs-induced toxicity at a cellular, in vivo and ecotoxicological level. Accordingly, the main cause for induction of toxicity by CuO NPs is the generation of reactive oxygen species (ROS) followed by the mitochondrial destruction that leads to apoptosis via the intrinsic pathway or under the condition such as hypoxia cell on exposure to CuO NPs may commit to necrosis. Moreover, CuO NPs also result in activation of MAPK pathways, ERKs and JNK/SAPK thus play an important role in the activation of AP-1. Furthermore, CuO NPs also leads to up-regulation of p53 and caspase three genes. Therefore, careful measures are required to explore omic technology to understand the molecular mechanism of the deleterious effects caused by CuO NPs. Copyright © 2016 John Wiley & Sons, Ltd.

Montmorillonite clay alters toxicity of silver nanoparticles in zebrafish (Danio rerio) eleutheroembryo.

An exponential development in the use of silver nanoparticles (AgNPs) in consumer products has accelerated their release in aquatic environment. As the AgNPs enters into the aquatic systems, their fate may change due to interactions with abiotic (e.g. clay particles) or biotic factors. The abundantly present clay particles are expected to more prone for interaction with nanoparticles in aquatic systems. In the present study, it is demonstrated that AgNPs interacts with clay particles and forms heteroagglomerates. Furthermore, an impact on toxicity potential of AgNPs after interactions with clay particles was assessed by using zebrafish eleutheroembryos (72 h post hatching) as an in vivo model. The mortality rate of zebrafish eleutheroembryos was higher in case of exposure to AgNPs-clay complexes (pH 4.0 and 7.0) as compared to bare AgNPs. In addition, at earlier time points, the eleutheroembryos expressed higher levels of morphological changes in tail, yolk and pericardia, but the edema in yolk sac was followed by cell death. It can be concluded from the observations made in the present study that the inorganic colloids in the aquatic matrices can alter the fate and toxicity potential of nanoparticles.

Assessment of agglomeration, co-sedimentation and trophic transfer of titanium dioxide nanoparticles in a laboratory-scale predator-prey model system.

Nano titanium dioxide (nTiO2) is the most abundantly released engineered nanomaterial (ENM) in aquatic environments. Therefore, it is prudent to assess its fate and its effects on lower trophic-level organisms in the aquatic food chain. A predator-and-prey-based laboratory microcosm was established using Paramecium caudatum and Escherichia coli to evaluate the effects of nTiO2. The surface interaction of nTiO2 with E. coli significantly increased after the addition of Paramecium into the microcosm. This interaction favoured the hetero-agglomeration and co-sedimentation of nTiO2. The extent of nTiO2 agglomeration under experimental conditions was as follows: combined E. coli and Paramecium > Paramecium only > E. coli only > without E. coli or Paramecium. An increase in nTiO2 internalisation in Paramecium cells was also observed in the presence or absence of E. coli cells. These interactions and nTiO2 internalisation in Paramecium cells induced statistically significant (p < 0.05) effects on growth and the bacterial ingestion rate at 24 h. These findings provide new insights into the fate of nTiO2 in the presence of bacterial-ciliate interactions in the aquatic environment.

Multiplex PCR based genotypic characterization of pathogenic vancomycin resistant Enterococcus faecalis recovered from an Indian river along a city landscape.

BACKGROUND: Enterococci are normal commensals of human gut, but vancomycin-resistant enterococci (VRE) are a severe threat to human health. Antimicrobial-resistant enterococci have been reported previously from Indian surface waters. However, the presence of antimicrobial resistance and virulence markers in Enterococcus faecalis, the most dominant enterococci is yet to be investigated. OBJECTIVES: The goal of this study was to analyse concentration of enterococci and distribution of antimicrobial resistance and virulence markers in E. faecalis isolates from river waters along an important north Indian city landscape. METHODS: We enumerated enterococci in river water samples (n = 60) collected from five sites across the Lucknow city landscape using the most probable number and membrane-filtration methods. The antimicrobial sensitivity profile of E. faecalis isolate was generated with the Kirby-Bauer antimicrobial disc diffusion assay. The multiplex PCR was used for genotypic characterization of vancomycin-resistance and virulence in E. faecalis isolates. RESULTS: Enterococci density (p < 0.0001) increased from up-to-down-stream sites. Multiplex PCR based genotypic characterization has shown a significant distribution of virulence-markers gelE, ace or efaA in the E. faecalis isolates (p < 0.05). The range of antimicrobial-resistance varied from 5 to 12 in the landscape with the frequency of vancomycin-resistant E. faecalis (VRE) ranging from 22 to 100 %. CONCLUSION: The occurrence of pathogenic VRE in river Gomti surface water is an important health concern. The observed high background pool of resistance and virulence in E. faecalis in river waters has the potential to disseminate more alarming antimicrobial resistance in the environment and poses serious health risk in developing countries like India as VRE infections could lead to increased cost of healthcare.

Natural water as the test medium for Ag and CuO nanoparticle hazard evaluation: An interlaboratory case study.

Engineered nanoparticles (NPs) have realistic potential of reaching natural waterbodies and of exerting toxicity to freshwater organisms. The toxicity may be influenced by the composition of natural waters as crucial NP properties are influenced by water constituents. To tackle this issue, a case study was set up in the framework of EU FP7 NanoValid project, performing an interlaboratory hazard evaluation of NPs in natural freshwater. Ag and CuO NPs were selected as model NPs because of their potentially high toxicity in the freshwater. Daphnia magna (OECD202) and Danio rerio embryo (OECD236) assays were used to evaluate NP toxicity in natural water, sampled from Lake Greifen and Lake Lucerne (Switzerland). Dissolution of the NPs was evaluated by ultrafiltration, ultracentrifugation and metal specific sensor bacteria. Ag NP size was stable in natural water while CuO NPs agglomerated and settled rapidly. Ag NP suspensions contained a large fraction of Ag(+) ions and CuO NP suspensions had low concentration of Cu(2+) ions. Ag NPs were very toxic (48 h EC50 1-5.5 µg Ag/L) to D. magna as well as to D. rerio embryos (96 h EC50 8.8-61 µg Ag/L) in both standard media and natural waters with results in good agreement between laboratories. CuO NP toxicity to D. magna differed significantly between the laboratories with 48 h EC50 0.9-11 mg Cu/L in standard media, 5.7-75 mg Cu/L in Lake Greifen and 5.5-26 mg Cu/L in Lake Lucerne. No toxicity of CuO NP to zebrafish embryos was detected up to 100 mg/L independent of the medium used. The results show that Ag and CuO NP toxicity may be higher in natural water than in the standard media due to differences in composition. NP environmental hazard evaluation can and should be carried out in natural water to obtain more realistic estimates on the toxicity.

Exploring the potential reservoirs of non specific TEM beta lactamase (bla(TEM)) gene in the Indo-Gangetic region: A risk assessment approach to predict health hazards.

The emergence of antimicrobial resistant bacteria is an important public health and environmental contamination issue. Antimicrobials of ß-lactam group accounts for approximately two thirds, by weight, of all antimicrobials administered to humans due to high clinical efficacy and low toxicity. This study explores ß-lactam resistance determinant gene (blaTEM) as emerging contaminant in Indo-Gangetic region using qPCR in molecular beacon format. Quantitative Microbial Risk Assessment (QMRA) approach was adopted to predict risk to human health associated with consumption/exposure of surface water, potable water and street foods contaminated with bacteria having blaTEM gene. It was observed that surface water and sediments of the river Ganga and Gomti showed high numbers of blaTEM gene copies and varied significantly (p<0.05) among the sampling locations. The potable water collected from drinking water facility and clinical settings exhibit significant number of blaTEM gene copies (13±0.44-10200±316 gene copies/100mL). It was observed that E.crassipes among aquatic flora encountered in both the rivers had high load of blaTEM gene copies. The information on prevalence of environmental reservoirs of blaTEM gene containing bacteria in Indo-Gangetic region and risk associated will be useful for formulating strategies to protect public from menace of clinical risks linked with antimicrobial resistant bacteria.

Cell cycle dependent cellular uptake of zinc oxide nanoparticles in human epidermal cells.

Metal oxide nanoparticles (NPs), including zinc oxide (ZnO) NPs have shown success for use as vehicles for drug delivery and targeting gene delivery in many diseases like cancer. Current anticancer chemotherapeutics fail to effectively differentiate between cancerous and normal cells. There is an urgent need to develop novel drug delivery system that can better target cancer cells while sparing normal cells and tissues. Particularly, ZnO NPs exhibit a high degree of cancer cell selectivity and induce cell death, oxidative stress, interference with the cell cycle progression and genotoxicity in cancerous cells. In this scenario, effective cellular uptake of NP seems to be crucial, which is shown to be affected by cell cycle progression. In the present study, the cytotoxic potential of ZnO NPs and the effect of different cell cycle phases on the uptake of ZnO NPs were examined in A431 cells. It is shown that the ZnO NPs led to cell death and reactive oxygen species generation and were able to induce cell cycle arrest in S and G2/M phase with the higher uptake in G2/M phase compared with other phases.

Q-PCR Based Culture-Independent Enumeration and Detection of Enterobacter: An Emerging Environmental Human Pathogen in Riverine Systems and Potable Water.

The availability of safe and pristine water is a global challenge when large numbers of natural and anthropogenic water resources are being depleted with faster rate. The remaining water resources are severely contaminated with various kinds of contaminants including microorganisms. Enterobacter is one of the fecal coliform bacteria of family Enterobacteriaceae. Enterobacter was earlier used as an indicator bacterium along with other fecal Coliforms namely Escherichia coli, Citrobacter, and Klebsiella, but it is now known to cause various diseases in human beings. In this study, we have collected 55 samples from potable water and riverine system and proved their presence using their conserved sequences of 16S rRNA and 23S rRNA genes with the help of SYBR green real-time PCR, which showed very high specificity for the detection of Enterobacter. The Enterobacter counts in potable water were found to 1290 ± 32.89 to 1460 ± 39.42 cfu/100 ml. The Enterobacter levels in surface water were 1.76 × 10(4) ± 492, 1.33 × 10(4) ± 334, 1.15 × 10(4) ± 308, 2.56 × 10(4) ± 802, 2.89 × 10(4) ± 962, 8.16 × 10(4) ± 3443 cfu/100 ml; the levels of Enterobacter contamination associated with hydrophytes were 4.80 × 10(4) ± 1804, 3.48 × 10(4) ± 856, 8.50 × 10(4) ± 2074, 8.09 × 10(4) ± 1724, 6.30 × 10(4) ± 1738, 3.68 × 10(4) ± 949 cfu/10 g and the Enterobacter counts in sediments of the river, were 2.36 × 10(4) ± 703, 1.98 × 10(4) ± 530, 9.92 × 10(4) ± 3839, 6.80 × 10(4) ± 2230, 8.76 × 10(4) ± 3066 and 2.34 × 10(4) ± 732 cfu/10 g at the sampling Site #1, Site #2, Site #3, Site #4, Site #5, and Site #6, respectively. The assay could be used for the regular monitoring of potable water and other water reservoirs to check waterborne outbreaks.

Nanotechnology in Disease Diagnostic Techniques.

Currently the major research highlights of bioengineering and medical technology are directed towards development of improved diagnostic techniques to screen complex diseases. Screening requirements are to identify the cause of illnesses, monitor improvement or progression of the state of diseases such as cancer, cardiovascular or neurodegenerative diseases. Nanotechnology enables the manipulation of materials at nanoscale and has shown potential to enhance sensitivity, selectivity and lower the cost of a diagnosis. The causative biomolecules (DNA, proteins) can be detected by red-shifted absorbance of gold nanoparticles or alteration in the conductance of a nanowire or nanotubes, and deflection of a micro or nano-cantilever. Several types of nanomaterials such as metals, metal-oxides and quantum dots have shown ample advantages over traditional diagnosis, intracellular labeling and visualization of target cells/tissues. Nanotechnology has also opened several avenues which could be further developed to enable enhanced visualization of tissues, cells, DNA and proteins over a point-of-care device. Protein or gene chips created using nanomaterials could further be integrated into a convenient nano-fluidic device for better disease diagnosis.

Surface functionalization of quantum dots for biological applications.

Quantum dots are a group of inorganic nanomaterials exhibiting exceptional optical and electronic properties which impart distinct advantages over traditional fluorescent organic dyes in terms of tunable broad excitation and narrow emission spectra, signal brightness, high quantum yield and photo-stability. Aqueous solubility and surface functionalization are the most common problems for QDs employed in biological research. This review addresses the recent research progress made to improve aqueous solubility, functionalization of biomolecules to QD surface and the poorly understood chemistry involved in the steps of bio-functionalization of such nanoparticles.