Vacancy engineered ceria nanostructures for protection from radiation-induced cellular damage

The ability of engineered cerium oxide nanoparticles to confer radioprotection was examined. Human normal and tumor cells were treated with nanoceria and irradiated, and cell survival was measured. Treatment of normal cells conferred almost 99% protection from radiation-induced cell death, whereas the same concentration showed almost no protection of tumor cells. For the first time, nanoceria is shown to confer radioprotection to a normal human breast line but not to a human breast tumor line, MCF-7.     

Anti‐inflammatory Properties of Cerium Oxide Nanoparticles

The valence and oxygen defect properties of cerium oxide nanoparticles (nanoceria) suggest that they may act as auto‐regenerative free radical scavengers. Overproduction of the free radical nitric oxide (NO) by the enzyme inducible nitric oxide synthase (iNOS) has been implicated as a critical mediator of inflammation. NO is correlated with disease activity and contributes to tissue destruction. The ability of nanoceria to scavenge free radicals, or reactive oxygen species (ROS), and inhibit inflammatory mediator production in J774A.1 murine macrophages is investigated. Cells internalize nanoceria, the treatment is nontoxic, and oxidative stress and pro‐inflammatory iNOS protein expression are abated with stimulation. In vivo studies show nanoceria deposition in mouse tissues with no pathogenicity. Taken together, it is suggested that cerium oxide nanoparticles are well tolerated in mice and are incorporated into cellular tissues. Furthermore, nanoceria may have the potential to reduce ROS production in states of inflammation and therefore serve as a novel therapy for chronic inflammation.     

Nanoceria coating imperfections and their effect on the high-temperature oxidation resistance of a 304 stainless steel

The microemulsion method was employed in this work for synthesizing nanocrystalline cerium oxide particles. Average nanoparticle sizes of 3.45 nm were produced by these means. XPS determinations indicated that both Ce³⁺ and Ce⁴⁺ are present in the synthesized nanoceria particles, with an average amount of 22.8 % of Ce³⁺ ions. It was found that the nanocrystalline cerium oxide coatings lead to significant improvements (of 1–2 orders of magnitude) in the high-temperature oxidation resistance of 304 SS. In addition, it was found that by decreasing the nanoparticle mean size from 10 to 3.45 nm, the effect of the coating protection was drastically improved. The experimentally determined parabolic rate constants k _p at 1073 and 1273 K for 304 SS indicated a reduction of up to two orders of magnitude when nanoceria coatings with 3.45 nm in mean size were applied. Also, the scale thickness was reduced from 15 to 5 μm when oxidized at 1073 K for 442 h. Coatings with purchased nanoceria particles (10 nm) were not as effective as the coatings with synthesized nanoceria. Apparently, at increasing nanoceria sizes, the oxidation protection is significantly reduced. In addition, it was found that the method of dipping for coating 304 SS does not provide a uniform coverage with nanoceria particles. Fe-rich ‘islands’ develop during high-temperature oxidation indicating that some regions do not exhibit the protection that nanoceria can provide. In contrast, relatively thick-coating regions on the steel substrate exhibit minimal oxidation, and the resultant scale is fine grained and uniform.     

TEM/AFM investigation of size and surface properties of nanocrystalline ceria

A series of ceria nanoparticles were synthesized by using a microemulsion method. The effect of relative concentration of surfactant/water on the size and the surface roughness of ceria nanoparticles was examined using transmission electron microscopy (TEM) and atomic force microscopy (AFM) respectively. The investigation confirmed a relationship between the size and the roughness properties of the nanoceria as a function of the water to surfactant ratio. With increasing dilution of the surfactant, the size distribution became narrow such that average particle size decreased linearly as the ratio increased without affecting lower size threshold of particles (approximately 10 nm). The surface roughness, on the other hand was found to increase with increasing water to surfactant ratio implying diluted surfactant would provide rougher surface of ceria nanoparticles. The information can be used to tailor the adhesion properties of nanoceria by optimizing the size distribution as well as surface roughness as a function of water to surfactant ratio.     

Polysaccharide Nanofiber-Stabilized Pickering Emulsion Microparticles Induce Pyroptotic Cell Death in Hepatocytes and Kupffer Cells

The intracellular uptake and interaction behavior of emulsion microparticles in liver cells critical to host defense and inflammation is significant to understanding their potential cytotoxicity and biomedical applications. In this study, the cell death responses of fibroblastic, hepatocyte, and Kupffer cells (KCs) induced by four types of emulsion particles that are stabilized by polysaccharide nanofibers (cellulose or chitin), an inorganic nanoparticle (β-tricalcium phosphate), or surfactants are compared. Pickering emulsion (PE) microparticles stabilized by polysaccharide nanofibers or inorganic nanoparticles have a droplet size of 1–3 µm, while the surfactant-stabilized emulsion has a diameter of ≈190 nm. Polysaccharide nanofiber-stabilized PEs (PPEs) markedly induce lactate dehydrogenase release in all cell types. Additionally, characteristic pyroptotic cell death, which is accompanied by cell swelling, membrane blebbing, and caspase-1 activation, occurs in hepatocytes and KCs. These PE microparticles are co-cultured with lipopolysaccharide-primed KCs associated with cytokine interleukin-1β release, and the PPEs demonstrate biological activity as a mediator of the inflammation response. Well-designed PPE microparticles induce pyroptosis of liver cells, which may provide new insight into regulating inflammation-related diseases for designing potent anticancer drugs and vaccine adjuvants.     

pH-tunable oxidase-like activity of cerium oxide nanoparticles achieving sensitive fluorigenic detection of cancer biomarkers at neutral pH

The reliable and sensitive detection of cancer-specific biomarkers is important for the diagnosis and treatment of cancer. Hence, detection of these biomarkers has to be reliably and rapidly performed in diverse settings. A limitation of the conventional biomarker-screening method of enzyme-linked immunosorbent assay (ELISA) is the employment of labile components, such as hydrogen peroxide and horseradish peroxidase. Previously, we reported that nanoceria is able to oxidize various colorimertic dyes at acidic pH, such as 3,3',5,5'-tetramethylbenzydine (TMB) and 2,2-azinobis-(3-ethylbenzothizoline-6-sulfonic acid) (AzBTS), and an assay was designed for screening the folate receptor. Herein, we show that the ability of nanoceria to oxidize a substrate can be tuned by modulating the pH. Results showed that nanoceria can oxidize the nonfluorescent substrate ampliflu, either to the very stable fluorescent product resorufin at pH 7.0 or to the nonfluorescent resazurin at pH 4.0. On the basis of these findings, we conjugated Protein G to immobilize antibodies on the surface of nanoceria, in order to detect the expression of prototypic cancer biomarkers at pH 7.0, such as the folate receptor and EpCAM. We found that within 3 h, nanoceria identified the expression of the folate receptor and EpCAM on lung carcinoma and breast adenocarcinoma cells, respectively. Traditional ELISA had a readout time of 15 h and a higher detection threshold, while requiring multiple washing steps. Considering these results and nanoceria's ability to oxidize ampliflu to its stable fluorescent product at neutral pH, the use of antibody-carrying nanoceria in the lab and point-of-care molecular diagnostics is anticipated.     

Toxicity of nano gamma alumina to neural stem cells

Nano alumina, one of the most important nanomaterials, is widely used in diverse areas. It was reported that nano alumina could cross the blood brain barrier to enter the brain. Considering aluminum accumulation in brain is closely related to many neural diseases. We studied the neural toxicity of four nano gamma-alumina samples by using neural stem cells (NSCs) C17.2 as a model. We find that the toxicity of nano gamma-alumina is pretty low, though these alumina particles are easily internalized by cells. The loss of cell viability and membrane integrity are dose-dependent and sample-dependent after alumina exposure. At concentrations lower than 100 microg/mL, no significant toxicity is observed for all alumina samples. When the concentration reaches 200 microg/mL, alumina treated cells begin to loss their activities. No culture period effect (up to 3 days) is observed. Very tiny soluble aluminum and the absorption of culture medium ingredients onto alumina particles do not affect the cell viability. Intracellular reactive oxygen species generation may contribute to the cytotoxicity of alumina particles at high concentration, but it does not induce the apoptosis of NSCs.     

Adverse Effects of Titanium Dioxide Nanoparticles on Human Dermal Fibroblasts and How to Protect Cells

The effects of exposure of human dermal fibroblasts to rutile and anatase TiO₂ nanoparticles are reported. These particles can impair cell function, with the latter being more potent at producing damage. The exposure to nanoparticles decreases cell area, cell proliferation, mobility, and ability to contract collagen. Individual particles are shown to penetrate easily through the cell membrane in the absence of endocytosis, while some endocytosis is observed for larger particle clusters. Once inside, the particles are sequestered in vesicles, which continue to fill up with increasing incubation time till they rupture. Particles coated with a dense grafted polymer brush are also tested, and, using flow cytometry, are shown to prevent adherence to the cell membrane and hence penetration of the cell, which effectively decreases reactive oxygen species (ROS) formation and protects cells, even in the absence of light exposure. Considering the broad applications of these nanoparticles in personal health care products, the functionalized polymer coating can potentially play an important role in protecting cells and tissue from damage.     

Photo‐inactivation of bacteria in hospital effluent via thiolated iron‐doped nanoceria

Hospital wastewater is a major contributor of disease‐causing microbes and the emergence of antibiotic resistant bacteria. In this study, thiolated iron‐doped nanoceria was synthesised and tested for killing of microbes from hospital effluent. These particles were designed to inhibit the efflux pumps of the bacteria found in hospital effluent with further ability to activate in visible light via iron doping thus generating tunable amount of reactive oxygen species (ROS). The quantum yield of the ROS generated by the nanoceria was 0.67 while the ROS types produced were singlet oxygen (36%), hydroxyl radical (31%) and hydroxyl ions (32%), respectively. The particles were initially synthesised through green route using Foeniculum vulgare seeds extract and were annealed at 200°C and further coated with thiolated chitosan to enhance the solubility and efflux pump inhibition. X‐ray diffraction confirmed the polycrystalline nature of nanoparticles and uniform spherical shape with 30 nm size, confirmed by scanning electron microscope. The nanoparticles exhibited 100% bactericidal activity at 100 µg/mL against all the isolated bacteria. The enhanced bactericidal effect of iron‐doped nanoceria could be attributed to efflux inhibition via thiolated chitosan as well as the production of ROS upon illumination in visible light, causing oxidative stress against microbes found in hospital effluent.Inspec keywords: health and safety, chemical engineering, solubility, renewable materials, annealing, hospitals, antibacterial activity, cerium compounds, nanoparticles, photochemistry, wastewater treatment, iron, nanofabrication, microorganisms, X‐ray diffraction, effluents, scanning electron microscopy, particle size, diseasesOther keywords: antibiotic resistant bacteria, thiolated iron‐doped nanoceria, hospital effluent, visible light, reactive oxygen species, hydroxyl ions, thiolated chitosan, solubility, efflux pump inhibition, disease‐causing microbes, wastewater treatment, singlet oxygen, hydroxyl radical, Foeniculum vulgare seeds extract, annealing process, X‐ray diffraction, scanning electron microscopy, particle size, bactericidal activity, oxidative stress, photoinactivation, size 30.0 nm, temperature 200.0 degC, CeO₂ :Fe     

Endothelial cell death on biomaterials: Theoretical and practical aspects of investigation

The use of endothelial cell seeding has been proposed as a solution to increase the patency of blood-contacting devices. While there has been a great deal of research into both biological and mechanical mechanisms of failure, relatively little work involving the effects which biomaterials have on the function of cells seeded on such devices has been conducted. We hypothesize, based on previously published results, that endothelial cells attached to synthetic biomaterials exhibit increased intracellular levels of superoxide and other reactive oxygen species. Previous reports have linked elevated levels of reactive oxygen species to NF-κB activation and subsequent cell death via anoikis, a form of apoptosis. Because of this, it is critical that an understanding of how biomaterials affect cellular behavior is developed, and that this knowledge is used in the creation of future devices which rely on cell seeding and cellular ingrowth.     

Cytoprotective properties of a fullerene derivative against copper

To delineate the complexity of the response of cells to nanoparticles we have performed a study on HT-29 human colon carcinoma cells exposed first to a fullerene derivative C(60)(OH)(20) and then to physiological copper ions. Our cell viability, proliferation, and intracellular reactive oxygen species (ROS) production assays clearly indicated that C(60)(OH)(20) suppressed cell damage as well as ROS production induced by copper, probably through neutralization of the metal ions by C(60)(OH)(20) in the extracellular space, as well as by adsorption and uptake of the nanoparticles surface-modified by the biomolecular species in the cell medium. This double-exposure study provides new data on the effects of nanoparticles on cell metabolism and may aid the treatment of oxidant-mediated diseases using nanomedicine.     

Size-dependent cytotoxicity of gold nanoparticles

Gold nanoparticles are widely used in biomedical imaging and diagnostic tests. Based on their established use in the laboratory and the chemical stability of Au(0), gold nanoparticles were expected to be safe. The recent literature, however, contains conflicting data regarding the cytotoxicity of gold nanoparticles. Against this background a systematic study of water-soluble gold nanoparticles stabilized by triphenylphosphine derivatives ranging in size from 0.8 to 15 nm is made. The cytotoxicity of these particles in four cell lines representing major functional cell types with barrier and phagocyte function are tested. Connective tissue fibroblasts, epithelial cells, macrophages, and melanoma cells prove most sensitive to gold particles 1.4 nm in size, which results in IC(50) values ranging from 30 to 56 microM depending on the particular 1.4-nm Au compound-cell line combination. In contrast, gold particles 15 nm in size and Tauredon (gold thiomalate) are nontoxic at up to 60-fold and 100-fold higher concentrations, respectively. The cellular response is size dependent, in that 1.4-nm particles cause predominantly rapid cell death by necrosis within 12 h while closely related particles 1.2 nm in diameter effect predominantly programmed cell death by apoptosis.     

Mechanisms of Antibacterial Activity of MgO: Non‐ROS Mediated Toxicity of MgO Nanoparticles Towards Escherichia coli

The toxicity of metal oxide nanomaterials and their antimicrobial activity is attracting increasing attention. Among these materials, MgO is particularly interesting as a low cost, environmentally‐friendly material. The toxicity of MgO, similar to other metal oxide nanomaterials, is commonly attributed to the production of reactive oxygen species (ROS). We investigated the toxicity of three different MgO nanoparticle samples, and clearly demonstrated robust toxicity towards Escherichia coli bacterial cells in the absence of ROS production for two MgO nanoparticle samples. Proteomics data also clearly demonstrate the absence of oxidative stress and indicate that the primary mechanism of cell death is related to the cell membrane damage, which does not appear to be due to lipid peroxidation.     

Oxidative stress and apoptosis induced by iron oxide nanoparticles in cultured human umbilical endothelial cells

Recent epidemiologic researches indicate that exposure to ultrafine particles (nanoparticles) is an independent risk factor for several cardiovascular diseases. The induction of endothelial injuries is hypothesized to be an attractive mechanism involved in these cardiovascular diseases. To investigate this hypothesis, the widely used iron nanomaterials, ferric oxide (Fe2O3) and ferriferrous oxide (Fe3O4) nanoparticles were incubated with human umbilical endothelial cells (ECV304 cells) at different concentrations of 2, 20, 100 microg/mL. The cell viability, the rate of apoptosis, the apoptotic nuclear morphology and the mitochondria membrane potential were measured to estimate the cell necrosis and apoptosis caused by the nanoparticle exposure. The stimulation of superoxide anion (O2*-) and nitric oxide (NO) were examined to evaluate the stress responses of endothelial cells. Our results indicated that both the Fe2O3 and Fe3O4 nanoparticles could generate oxidative stress as well as the significant increase of nitric oxide in ECV304 cells. The loss of mitochondria membrane potential and the apoptotic chromatin condensation in the nucleus were observed as the early signs of apoptosis. It is inferred the stress response might be an important mechanism involving in endothelial cells apoptosis and death, and these injuries in endothelial cells might play a key role in downstream cardiovascular diseases such as atheroscelerosis, hypertension and myocardial infarction (MI).     

From radiation-induced chromosome damage to cell death: modelling basic mechanisms and applications to boron neutron capture therapy

Cell death is a crucial endpoint in radiation-induced biological damage: on one side, cell death is a reference endpoint to characterise the action of radiation in biological targets; on the other side, any cancer therapy aims to kill tumour cells. Starting from Lea's target theory, many models have been proposed to interpret radiation-induced cell killing; after briefly discussing some of these models, in this paper, a mechanistic approach based on an experimentally observed link between chromosome aberrations and cell death was presented. More specifically, a model and a Monte Carlo code originally developed for chromosome aberrations were extended to simulate radiation-induced cell death applying an experimentally observed one-to-one relationship between the average number of 'lethal aberrations' (dicentrics, rings and deletions) per cell and -ln S, S being the fraction of surviving cells. Although such observation was related to X rays, in the present work, the approach was also applied to protons and alpha particles. A good agreement between simulation outcomes and literature data provided a model validation for different radiation types. The same approach was then successfully applied to simulate the survival of cells enriched with boron and irradiated with thermal neutrons at the Triga Mark II reactor in Pavia, to mimic a typical treatment for boron neutron capture therapy.     

In vitro and transdermal penetration of PHBV micro/nanoparticles

The purpose of this study was to develop micro and nano sized drug carriers from poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), and study the cell and skin penetration of these particles. PHBV micro/nanospheres were prepared by o/w emulsion method and were stained with a fluorescent dye, Nile Red. The particles were fractionated by centrifugation to produce different sized populations. Topography was studied by SEM and average particle size and its distribution were determined with particle sizer. Cell viability assay (MTT) was carried out using L929 fibroblastic cell line, and particle penetration into the cells were studied. Transdermal permeation of PHBV micro/nanospheres and tissue reaction were studied using a BALB/c mouse model. Skin response was evaluated histologically and amount of PHBV in skin was determined by gas chromatography-mass spectrometry. The average diameters of the PHBV micro/nanosphere batches were found to be 1.9 μm, 426 and 166 nm. Polydispersity indices showed that the size distribution of micro sized particles was broader than the smaller ones. In vitro studies showed that the cells had a normal growth trend. MTT showed no signs of particle toxicity. The 426 and 166 nm sized PHBV spheres were seen to penetrate the cell membrane. The histological sections revealed no adverse effects. In view of this data nano and micro sized PHBV particles appeared to have potential to serve as topical and transdermal drug delivery carriers for use on aged or damaged skin or in cases of skin diseases such as psoriasis, and may even be used in gene transfer to cells.     

Radiobiological effects of high-let particles: DNA strand breaks

Fundamental studies in radiation biology with high-LET charged particles involve a systematic study of the physical, chemical, molecular and cellular processes. Water molecules and DNA present inside a cell constitute the important targets for energy deposition which eventually lead to either cell death or mutation or transformation. High-LET charged particles are very efficient in causing these types of damages. One of the primary lesions for causing injury to a cell is the production of DNA strand breaks. A good understanding of these breaks is essential before ultimate biological effects of heavy particles can be predicted. Based on known molecular mechanisms of the formation of strand breaks, a theoretical model is presented along with a comparison between the predictions of the model and experimental data. The studies have shown that LET is not a convenient physical parameter to relate the extent of strand breaks and one needs to know the microscopic distribution of energy (track structure). A discussion has also been presented to provide a background on various radiobiological characteristics of high-LET charged particles from the point of view of their uniqueness in damaging cancer cells.     

Cancer‐Cell‐Specific Induction of Apoptosis Using Mesoporous Silica Nanoparticles as Drug‐Delivery Vectors

Targeted delivery of the chemotherapeutic agent methotrexate (MTX) to cancer cells using poly(ethyleneimine)‐functionalized mesoporous silica particles as drug‐delivery vectors is reported. Due to its high affinity for folate receptors, the expression of which is elevated in cancer cells, MTX serves as both a targeting ligand and a cytotoxic agent. Enhanced cancer‐cell apoptosis (programmed cell death) relative to free MTX is thus observed at particle concentrations where nonspecific MTX‐induced apoptosis is not observed in the nontargeted healthy cell line, while corresponding amounts of free drug affect both cell lines equally. The particles remain compartmentalized in endo‐/lysosomes during the time of observation (up to 72 h), while the drug is released from the particle only upon cell entry, thereby inducing selective apoptosis in the target cells. As MTX is mainly attached to the particle surface, an additional advantage is that the presented carrier design allows for adsorption (loading) of additional drugs into the pore network for therapies based on a combination of drugs.     

Controllable self-assembly of nanoparticles for specific delivery of multiple therapeutic molecules to cancer cells using RNA nanotechnology

By utilizing RNA nanotechnology, we engineered both therapeutic siRNA and a receptor-binding RNA aptamer into individual pRNAs of phi29's motor. The RNA building block harboring siRNA or other therapeutic molecules was fabricated subsequently into a trimer through the interaction of engineered right and left interlocking RNA loops. The incubation of the protein-free nanoscale particles containing the receptor-binding aptamer or other ligands resulted in the binding and co-entry of the trivalent therapeutic particles into cells, subsequently modulating the apoptosis of cancer cells and leukemia model lymphocytes in cell culture and animal trials. The use of such antigenicity-free 20-40 nm particles holds promise for the repeated long-term treatment of chronic diseases.     

Classification of Fundus Images Based on Deep Learning for Detecting Eye Diseases

Various techniques to diagnose eye diseases such as diabetic retinopathy (DR), glaucoma (GLC), and age-related macular degeneration (AMD), are possible through deep learning algorithms. A few recent studies have examined a couple of major diseases and compared them with data from healthy subjects. However, multiple major eye diseases, such as DR, GLC, and AMD, could not be detected simultaneously by computer-aided systems to date. There were just high-performance-outcome researches on a pair of healthy and eye-diseased group, besides of four categories of fundus image classification. To have a better knowledge of multi-categorical classification of fundus photographs, we used optimal residual deep neural networks and effective image preprocessing techniques, such as shrinking the region of interest, iso-luminance plane contrast-limited adaptive histogram equalization, and data augmentation. Applying these to the classification of three eye diseases from currently available public datasets, we achieved peak and average accuracies of 91.16% and 85.79%, respectively. The specificities for images from the eyes of healthy, GLC, AMD, and DR patients were 90.06%, 99.63%, 99.82%, and 91.90%, respectively. The better specificity performances may alert patient in an early stage of eye diseases to prevent vision loss. This study presents a possible occurrence of a multi-categorical deep neural network technique that can be deemed as a successful pilot study of classification for the three most-common eye diseases and can be used for future assistive devices in computer-aided clinical applications.