Tamoxifen-loaded solid lipid nanoparticles-induced apoptosis in breast cancer cell lines

An in vitro study was conducted to determine the apoptosis induced by tamoxifen (TAM) and TAM-loaded solid lipid nanoparticles (SLNs) in breast cancer cell lines, MCF-7 and MDA-MB231 cells. The effect of free drug and drug-loaded SLN on the cell lines was characterised by cell morphology and cell cycle distribution using phase contrast microscopy, nuclear morphology and flow cytometry, respectively. The results showed that TAM-loaded SLNs have an equally efficient cytotoxic activity against MCF-7 and MDA-MB231 cells, compared to free TAM, and the half maximal inhibitory concentration (IC₅₀) of TAM-loaded SLNs was generally lower than that of free TAM. In the presence of TAM and TAM-loaded SLN, the viability of the both cells diminishes and the cancer cells lose their normal morphological characteristics, detaches, aggregates and later develops apoptotic bodies. Flow cytometry analysis showed that TAM-loaded SLN like the free TAM caused a dose- and time-dependent apoptosis without cell cycle arrest of human breast cancer cells. Therefore, TAM-loaded SLN has great potential in human medicine for the treatment of breast cancers.     

Preferential killing of cancer cells and activated human T cells using ZnO nanoparticles

Nanoparticles are increasingly being recognized for their potential utility in biological applications including nanomedicine. Here we examine the response of normal human cells to ZnO nanoparticles under different signaling environments and compare it to the response of cancerous cells. ZnO nanoparticles exhibit a strong preferential ability to kill cancerous T cells ( approximately 28-35x) compared to normal cells. Interestingly, the activation state of the cell contributes toward nanoparticle toxicity, as resting T cells display a relative resistance while cells stimulated through the T cell receptor and CD28 costimulatory pathway show greater toxicity in direct relation to the level of activation. Mechanisms of toxicity appear to involve the generation of reactive oxygen species, with cancerous T cells producing higher inducible levels than normal T cells. In addition, nanoparticles were found to induce apoptosis and the inhibition of reactive oxygen species was found to be protective against nanoparticle induced cell death. The novel findings of cell selective toxicity, towards potential disease causing cells, indicate a potential utility of ZnO nanoparticles in the treatment of cancer and/or autoimmunity.     

Cytotoxicity of versatile nano-micro-particles based on hierarchical flower-like ZnO

ZnO (nano)structures remain of great interest in biomedical applications due to their unique properties and possible morphologies. Biocompatibility of typically fabricated ZnO structures remains questionable and they still lack desired biological functions, whence their functionalization is of high interest. In this work, we fabricated micro-sized ZnO hierarchical flower-like structures using facile template-free hydrothermal method to act as carriers for the delivery of gold nanoparticles (Au) and/or Biotin (Vitamin B) to cells. Au nanoparticles (~24 nm), as well as Biotin molecules were successfully deposited on the ZnO surface due to non-covalent physical interactions. We have then cultured two cells lines: SH-SY5Y (human malignant neural) and HEK-293 (human non-malignant) and observed that ZnO hierarchical particles exhibited cell line-dependent cytotoxicity. It appeared that further functionalization of ZnO with Au nanoparticles and subsequently with Biotin led to lower discrepancy between the two cell lines response indicating that cytotoxic pathways of pure ZnO were masked by the available surface adsorbed particles (Au/Biotin). Two-photon immunocytochemistry microscopy further confirmed that Biotin decorated particles affected neuroblastoma cells cytoskeleton. These findings contribute to the understanding of cytotoxic pathways of surface-decorated nano-micro-structures made from ZnO with two molecules typically used in anticancer and regenerative medicine therapies.     

Surface modifications of ZnO nanoparticles and their cytotoxicity

ZnO is a well-known UV absorber. At small particle sizes its absorption efficiency is substantially increased and this property, combined with transparency to visible light, has attracted growing interest in its applications in personal care products such as sunscreens. However, some recent studies suggest that ZnO nanoparticles could induce considerable toxicity to certain cells and microorganisms. Aiming to reduce cytotoxicity of ZnO nanoparticles without impairing their unique properties, this paper examines the influence of surface modifications to ZnO nanoparticles using coatings such as silica (SiO2) and Poly methyl Acrylic Acid (PMAA). It was found that both PMAA and SiO2 coatings were physically attached to the ZnO surface and their presence did not weaken UV absorption of the original nanoparticles. Uncoated ZnO and SiO2-coated ZnO exhibited similar cytotoxicity to human lymphoblastoid cells, while PMAA-coated ZnO nanoparticles had little adverse effect except at high concentrations. The type of coating was also shown to affect the generation of Reactive Oxygen Species (ROS). The correlation between cell viability and ROS level led to conclusions that enhanced oxidative stress could be one of the reasons for cytotoxicity of ZnO nanoparticles.     

Nanoscale ZnO induces cytotoxicity and DNA damage in human cell lines and rat primary neuronal cells

The toxic effects of ZnO nanoparticles (nano-ZnO) (1-100 microg/mL) suspended in DMEM were examined in human A549 cells, HepG2 cells, human skin fibroblast cells, human skin keratinocytes, and rat primary neuronal cells for 24 h. Nano-ZnO induced dose dependent cytotoxicity and damaged cell membranes. Cell death was not mediated by reactive oxygen species (ROS) or apoptosis. Nano-ZnO induced DNA damage in rat primary neuronal cells, human fibroblasts, and A549 cells. The cytotoxicity of nano-ZnO in DMEM supplemented with 10% FBS, instead of serum free DMEM, was also examined in the A549 cells, human skin fibroblast cells, and human skin keratinocytes. The levels of cytotoxicity induced were similar to those tested without FBS; in addition, ROS was observed. These results indicate that the cause of cytotoxicity is medium dependent and imply that cellular growth conditions may play a significant role in induction of cytotoxicity and DNA damage by nano-ZnO.     

Effect of the surface texture and crystallinity of ZnO nanoparticles on their toxicity

We have investigated the correlation between the structural properties of ZnO nanoparticles (NPs) and their toxicity to mesenchymal stem cells (C2C12 cell line) and macrophage-derived cells (RAW 264.7 cell line). Nanopowders of grain size ranging between 5 nm and 50 nm were prepared by chemical route. Their structural properties were characterized extensively by X-ray Diffraction (XRD) and High Resolution Transmission Electron Microscopy (HRTEM). The XRD spectra showed that 50 nm sized NPs are well crystallized and present a preferential orientation along the direction normal to the (001) plane while the HREM observations revealed that most of the large size (50 nm) crystallized nanoparticles have polygonal shape which is consistent with a texture of along [001] direction. The toxicity tests showed that [001] large textured NPs have higher toxicity to inflammatory cells than nanoparticles of low crystallinity and much smaller size (5 nm). In addition, NPs have cytotoxic effects on inflammatory cells at concentration as low as 0.05 mM while ten times higher concentrations did not have significant cytotoxic effects on cells representing mesenchymal tissues. These observations are explained by the enhanced generation of Reactive Oxygen Species (ROS) at the (0001) polar surface of ZnO NP. These results provide a direct evidence of the correlation between the toxicity and the surface texture of the oxide nanoparticles. Similar correlation has been reported for the photocatalytic properties of ZnO nanoparticles.     

In vitro toxicity study of plant latex capped silver nanoparticles in human lung carcinoma cells

Organically modified silver nanoparticles were prepared by biosynthetic route induced by stem latex of a medicinally important plant, Euphorbia nivulia. The reduction and stabilization is assisted by certain peptides and terpenoids present within the latex. The aqueous formulation of latex capped silver nanoparticles (LAgNPs) being completely free of toxic chemicals can be directly used for administration/in vivo delivery of nanoparticles. The in vitro cytotoxicity evaluation of the latex capped nanoparticles was carried out using human lung carcinoma cells (A549) by MTT cell viability assay. Further, possible cytotoxic mechanisms were evaluated using various biomarkers for cytotoxicity and oxidative stress viz. extracellular lactate dehydrogenase (LDH) release, reactive oxygen species (ROS) generation, intracellular reduced glutathione (GSH), malondialdehyde (MDA), superoxide generation and acridine orange/ethedium bromide staining. It can be concluded from the present study that LAgNP formulation is toxic to A549 cells in a dose dependent manner. Thus plant latex solubilizes the AgNPs in water and acts as a biocompatible vehicle for transport of AgNPs to tumor/cancer cells.     

In vitro cytotoxicity of transparent yellow iron oxide nanoparticles on human glioma cells

With rapid development of nanotechnology, concerns about the possible adverse health effects on human beings by using nanomaterials have been raised. Transparent yellow iron oxide (alpha-FeOOH) nanoparticles have been widely used in paints, plastic, rubber, building materials, papermaking, food products and pharmaceutical industry, thus the potential health implications by the exposure should be considered. The purpose of this study is to assess the cytotoxicity of transparent yellow iron oxide nanoparticles on U251 human glioma cells. The alpha-FeOOH nanoparticles are in clubbed shapes with 9 nm in diameter and 43 nm long. The specific surface area is 115.3 m2/g. After physicochemical characterization of the nanoparticles, U251 cells were exposed to a-FeOOH at the doses of 0, 3.75, 15, 60 and 120 microg/mL. The results showed that the alpha-FeOOH nanoparticles reduced the cell viability and induced necrosis and apoptosis in U251 cells. In addition, nanoparticle exposure significantly increased the levels of superoxide anion and nitric oxide in a dose-dependent fashion in the cells. Our results suggest that exposure to alpha-FeOOH nanoparticles induce significant free radical formation and cytotoxic effects. The large surface area that induced high surface reactivity may play an important role in the cytotoxic effect of alpha-FeOOH nanoparticles.     

Light‐Induced ROS Generation and 2‐DG‐Activated Endoplasmic Reticulum Stress by Antitumor Nanosystems: An Effective Combination Therapy by Regulating the Tumor Microenvironment

A multimodal cancer therapeutic nanoplatform is reported. It demonstrates a promising approach to synergistically regulating the tumor microenvironment. The combination of intracellular reactive oxygen species (ROS) generated by irradiation of photosensitizer and endoplasmic reticulum (ER) stress induced by 2‐deoxy‐glucose (2‐DG) has a profound effect on necrotic or apoptotic cell death. Especially, targeting metabolic pathway by 2‐DG is a promising strategy to promote the effect of photodynamic therapy and chemotherapy. The nanoplatform can readily release its cargoes inside cancer cells and combines the advantages of ROS‐sensitive releasing chemotherapeutic drugs, upregulating apoptosis pathways under ER stress, light‐induced generation of cytotoxic ROS, achieving tumor accumulation, and in vivo fluorescence imaging capability. This work highlights the importance of considering multiple intracellular stresses as design parameters for nanoscale functional materials in cell biology, immune response, as well as medical treatments of cancer, Alzheimer's disease, etc.     

Role of size scale of ZnO nanoparticles and microparticles on toxicity toward bacteria and osteoblast cancer cells

The specific role of size scale, surface capping, and aspect ratio of zinc oxide (ZnO) particles on toxicity toward prokaryotic and eukaryotic cells was investigated. ZnO nano and microparticles of controlled size and morphology were synthesized by wet chemical methods. Cytotoxicity toward mammalian cells was studied using a human osteoblast cancer cell line and antibacterial activity using Gram-negative bacteria (Escherichia coli) as well as using Gram-positive bacteria (Staphylococcus aureus). Scanning electron microscopy (SEM) was conducted to characterize any visual features of the biocidal action of ZnO. We observed that antibacterial activity increased with reduction in particle size. Toxicity toward the human cancer cell line was considerably higher than previously observed by other researchers on the corresponding primary cells, suggesting selective toxicity of the ZnO to cancer cells. Surface capping was also found to profoundly influence the toxicity of ZnO nanoparticles toward the cancer cell line, with the toxicity of starch-capped ZnO being the lowest. Our results are found to be consistent with a membrane-related mechanism for nanoparticle toxicity toward microbes.     

Synthesis and characterization of bicalutamide-loaded magnetic nanoparticles as anti-tumor drug carriers

This study examined the optical characteristics of bicalutamide-loaded magnetic/ethylene glycol composite nanoparticles (BMP), as well as their anti-cancer activity against cancer cells. The gamma-Fe2O3 magnetic nanoparticles (MNPs), approximately 20 nm in diameter, were prepared via a chemical co-precipitation method and coated with two surfactants to yield a water-based product. The characteristics of the particles were determined via X-ray diffraction (XRD), field emission scanning electron microscopy, and Raman spectrophotometry. The Raman spectra of the BMP showed peaks at 222, 283, 395, 520, 669 and 1316 cm(-1), with broadened band in comparison to the Raman spectra of the magnetic nanoparticles. The BMP absorbance evidenced a rapid increase, with a broad peak at 409 nm, thus reflecting a good loading of the bicalutamide onto the magnetic nanoparticles. The results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the MNPs were non-toxic against human brain cancer cells (SH-SY5Y), human cervical cancer cells (Hela), human liver cancer cells (HepG2), breast cancer cells (MCF-7), colon cancer cells (CaCO2) and human prostate cancers (Du 145, PC3) tested herein. In particular, BMPs were cytotoxic at 56% against DU145 cells, at 74.37% in SH-SY5Y cells, and at 58% in Hela cells. Our results demonstrated the biological applicability of BMP nanoparticles as anticancer agents and as agents for enhanced drug delivery against human prostate cancer cells. Our results indicated that the MNPs were biostable and that the BMP functioned effectively as drug delivery vehicles.     

Functionalised gold nanoparticles for selective induction of in vitro apoptosis among human cancer cell lines

The interaction of citrate- and polyethylene imine (PEI)-functionalised gold nanoparticles (GNP) with cancer cell lines with respect to the cellular response was studied. It was found that GNP/citrate nanoparticles were able to induce apoptosis in human carcinoma lung cell lines A549, but GNP/PEI did not show any reduction in the viability of the cells in human breast cancer cell line MCF-7 and A549 cell lines. FACS data confirmed that the number of apoptotic cells increased with increase in the concentration of GNP/citrate nanoparticles. Decline in cellular expansion and changes in the nuclear morphology were noted after the treatment of GNP/citrate nanoparticles on A549 cell lines, which itself is a direct response for stress induction. The induction of cellular apoptosis was further confirmed by DNA fragmentation assay. These data confirm the potential of GNP/citrate nanoparticle to evoke cell-specific death response in the A549 cell lines.     

Zinc oxide nanoparticle induced genotoxicity in primary human epidermal keratinocytes

Zinc oxide (ZnO) nanoparticles are widely used in cosmetics and sunscreens. Human epidermal keratinocytes may serve as the first portal of entry for these nanoparticles either directly through topically applied cosmetics or indirectly through any breaches in the skin integrity. Therefore, the objective of the present study was to assess the biological interactions of ZnO nanoparticles in primary human epidermal keratinocytes (HEK) as they are the most abundant cell type in the human epidermis. Cellular uptake of nanoparticles was investigated by scanning electron microscopy using back scattered electrons imaging as well as transmission electron microscopy. The electron microscopy revealed the internalization of ZnO nanoparticles in primary HEK after 6 h exposure at 14 microg/ml concentration. ZnO nanoparticles exhibited a time (6-24 h) as well as concentration (8-20 microg/ml) dependent inhibition of mitochondrial activity as evident by the MTT assay. A significant (p < 0.05) induction in DNA damage was observed in cells exposed to ZnO nanoparticles for 6 h at 8 and 14 microg/ml concentrations compared to control as evident in the Comet assay. This is the first study providing information on biological interactions of ZnO nanoparticles with primary human epidermal keratinocytes. Our findings demonstrate that ZnO nanoparticles are internalized by the human epidermal keratinocytes and elicit a cytotoxic and genotoxic response. Therefore, caution should be taken while using consumer products containing nanoparticles as any perturbation in the skin barrier could expose the underlying cells to nanoparticles.     

Overcoming multidrug resistance of breast cancer cells by the micellar doxorubicin nanoparticles of mPEG-PCL-graft-cellulose

The amphiphilic block copolymer methoxy-poly(ethylene glycol)-poly(epsilon-caprolactone) (mPEG-PCL) was grafted to 2-hydroxyethyl cellulose (HEC) to produce nano-sized micellar nanoparticles. The nanoparticles were loaded with anti-tumor drug, doxorubicin (DOX) and the size of the DOX-loaded nanoparticles were determined by dynamic light scattering (DLS) in aqueous solution to be from 197.4 to 230 nm. The nanoparticles subjected to co-culture with macrophage cells showed that these nanoparticles used as drug carrier are not recognized as foreign bodies. Overexpression of P-glycoprotein (P-gp) is an important factor in the development of multidrug resistance (MDR) in many cancer cells. In this study, Western blot and Rhodamine 123 were used to monitor the relative P-glycoprotein expression in human breast cancer cell lines MCF-7/WT and MCF-7/ADR. The endocytosis of the DOX-loaded nanoparticles by breast cancer cells is more efficient observed under a confocal laser scanning microscopy (CLSM) and a flow cytometry in MCF7/ADR cells, compared to the diffusion of the free drug into the cytoplasm of cells. Based on these findings, we concluded that the nanoparticles made from mPEG-PCL-g-cellulose were effective in overcoming P-gp efflux in MDR breast cancer cells.     

Toxicological Aspects of Long‐Term Treatment of Keratinocytes with ZnO and TiO2 Nanoparticles

Sunscreens containing ZnO and TiO₂ nanoparticles (NPs) are increasingly applied to skin over long time periods to reduce the risk of skin cancer. However, long‐term toxicological studies of NPs are very sparse. The in vitro toxicity of ZnO and TiO₂ NPs on keratinocytes over short‐ and long‐term applications is reported. The effects studied are intracellular formation of radicals, alterations in cell morphology, mitochondrial activity, and cell‐cycle distribution. Cellular response depends on the type of NP, concentration, and exposure time. ZnO NPs have more pronounced adverse effects on keratinocytes than TiO₂. TiO₂ has no effect on cell viability up to 100 μg mL^?1, whereas ZnO reduces viability above 15 μg mL^?1 after short‐term exposure. Prolonged exposure to ZnO NPs at 10 μg mL^?1 results in decreased mitochondrial activity, loss of normal cell morphology, and disturbances in cell‐cycle distribution. From this point of view TiO₂ has no harmful effect. More nanotubular intercellular structures are observed in keratinocytes exposed to either type of NP than in untreated cells. This observation may indicate cellular transformation from normal to tumor cells due to NP treatment. Transmission electron microscopy images show NPs in vesicles within the cell cytoplasm, particularly in early and late endosomes and amphisomes. Contrary to insoluble TiO₂, partially soluble ZnO stimulates generation of reactive oxygen species to swamp the cell redox defense system thus initiating the death processes, seen also in cell‐cycle distribution and fluorescence imaging. Long‐term exposure to NPs has adverse effects on human keratinocytes in vitro, which indicates a potential health risk.     

Bioeffects of CdTe quantum dots on human umbilical vein endothelial cells

Quantum dots (QDs) hold great potential for applications in nanomedicine, however, their health effects are largely unknown. In the present study, the cytotoxicity and genotoxicity of CdTe QDs were examined in human umbilical vein endothelial cells (HUVECs). The QDs exhibited a dose-dependent inhibitory effect on cell growth. It was shown that after a 12 h treatment QDs at 1, 10, and 50 microg x ml(-1) induced formation of yH2AX foci, indicative of DNA damage, in a dose-dependent manner. Moreover, QD treatment clearly induced the generation of reactive oxygen species (ROS). Pre-treatment with N-acetyl-cysteine (NAC), a ROS scavenger, could inhibit the induction of ROS by QDs, as well as the formation of yH2AX foci. Taken together, our data indicate that CdTe QDs have cytotoxic and genotoxic effects on HUVECs, and that ROS generation may be involved in QD induced DNA damage.     

A pH-switched mesoporous nanoreactor for synergetic therapy

Zinc oxide nanoparticles (ZnO NPs),as a new type of pH-sensitive drug carrier,have received much attention.ZnO NPs are stable at physiological pH,but can dissolve quickly in the acidic tumor environment (pH ＜ 6) to generate cytotoxic zinc ions and reactive oxygen species (ROS).However,the protein corona usually causes the non-specific degradation of ZnO NPs,which has limited their application considerably.Herein,a new type of pH-sensitive nanoreactor (ZnO-DOX@F-mSiO2-FA),aimed at reducing the non-specific degradation of ZnO NPs,is presented.In the acidic tumor environment (pH ＜ 6),it can release cytotoxic zinc ions,ROS,and anticancer drugs to kill cancer cells effectively.In addition,the fluorescence emitted from fluorescein isothiocyanate (FITC)-labeled mesoporous silica (F-mSiO2) and doxorubicin (DOX) can be used to monitor the release behavior of the anticancer drug.This report provides a new method to avoid the non-specific degradation of ZnO NPs,resulting in synergetic therapy by taking advantage of ZnO NPs-induced oxidative stress and targeted drug release.     

Antibacterial and anticancer activity of biosynthesised CuO nanoparticles

The present investigation aims for the synthesis of copper oxide nanoparticles (CuO NPs) using Nilgirianthus ciliatus plant extract. The obtained CuO NPs were characterised by X‐ray diffraction, Fourier transform infrared spectrum, ultraviolet–visible spectroscopy, photoluminescence, scanning electron microscopy and transmission electron microscopy analysis. Significant bacterial activity was manifested by CuO nanoparticles against both Gram‐positive (Staphylococcus aureus and Staphylococcus mutans) and Gram‐negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. The synthesised CuO NPs have good cytotoxicity against both human breast cancer cell line (MCF‐7) and lung cancer cell line (A549) with minimum cytotoxic effect on normal L929 (fibroblast) cell lines.Inspec keywords: microorganisms, ultraviolet spectra, nanomedicine, transmission electron microscopy, visible spectra, cellular biophysics, antibacterial activity, nanoparticles, X‐ray diffraction, lung, copper compounds, cancer, toxicology, biomedical materials, scanning electron microscopy, photoluminescence, Fourier transform infrared spectraOther keywords: antibacterial activity, anticancer activity, biosynthesised CuO nanoparticles, copper oxide nanoparticles, Nilgirianthus ciliatus plant, X‐ray diffraction, infrared spectrum, ultraviolet–visible spectroscopy, transmission electron microscopy analysis, bacterial activity, Gram‐negative bacteria, synthesised CuO NPs, human breast cancer cell line, Staphylococcus aureus, Staphylococcus mutans, CuO     

Apoptotic efficacy and antiproliferative potential of silver nanoparticles synthesised from aqueous extract of sumac (Rhus coriaria L.)

Currently, nanotechnology and nanoparticles (NPs) are recognised due to their extensive applications in medicine and the treatment of certain diseases, including cancer. Silver NPs (AgNPs) synthesised by environmentally friendly method exhibit a high medical potential. This study was conducted to determine the cytotoxic and apoptotic effects of AgNPs synthesised from sumac (Anacardiaceae family) fruit aqueous extract (AgSu/NPs) on human breast cancer cells (MCF‐7). The anti‐proliferative effect of AgSu/NPs was determined by MTT assay. The apoptotic properties of AgSu/NPs were assessed by morphological analysis and acridine orange/propidium iodide (AO/PI) and DAPI staining. The mechanism of apoptosis induction in treated cells was investigated using molecular analysis. Overall results of morphological examination and cytotoxic assay revealed that AgSu/NPs exert a concentration‐dependent inhibitory effect on the viability of MCF‐7 cells (IC50 of ∼10 µmol/48 h). AO/PI staining confirmed the occurrence of apoptosis in cells treated with AgSu/NPs. In addition, molecular analysis demonstrated that the apoptosis in MCF‐7 cells exposed to AgSu/NPs was induced via up‐regulation of Bax and down‐regulation of Bcl‐2. These findings suggested the potential use of AgSu/NP as cytotoxic and pro‐apoptotic efficacy and its possible application in modern medicine for treating certain disorders, such as cancer.Inspec keywords: nanoparticles, silver, nanomedicine, biomedical materials, toxicology, cancer, molecular biophysics, proteins, biochemistry, cellular biophysics, nanofabricationOther keywords: Ag, Bcl‐2 down‐regulation, Bax up‐regulation, MCF‐7 cell viability, concentration‐dependent inhibitory effect, cytotoxic assay, molecular analysis, DAPI staining, acridine orange‐propidium iodide staining, morphological analysis, MTT assay, human breast cancer cells, sumac fruit aqueous extract, Anacardiaceae family, cytotoxic effects, drug delivery function, diseases, Rhus coriaria L, silver nanoparticles, antiproliferative potential, apoptotic efficacy