Cytotoxic Effects Of Zinc Oxide Nanoparticles Research Articles

The Cytotoxic Effects of Zinc Oxide Nanoparticles on SW480 Cell Lines and Measurement of Nitric Oxide in Cell Culture Medium

Background: Colorectal cancer (CRC) is one of the most common gastrointestinal cancers worldwide. It is the second leading cause of death in women after breast cancer. Objectives: The present study aimed to determine the cytotoxic effects of zinc oxide (ZnO) nanoparticles on colon cancer (SW480) viability and nitric oxide (NO) level in vitro. Methods: In this experimental study, the values of 15.62, 31.25, 62.5, 125, 250, and 500 μg/mL of ZnO nanoparticles were considered to treat SW480 cells for 24 hours. The MTT method was used to determine cell viability. The amount of NO was also measured using the grease method. An analysis of variance (ANOVA) was used to analyze the data. Results: Treatment of SW480 cells with ZnO nanoparticles resulted in decreased viability in a dose-dependent pattern. NO concentration significantly increased in response to an effective dose of ZnO nanoparticles (108 µg/mL) compared to the control group (P < 0.001). Conclusions: ZnO nanoparticles have cytotoxic effects on colon cancer cells. The cytotoxic effect of ZnO nanoparticles on colon cancer cells is partly mediated by NO in cancer cells.

Synthesis, Characterization, and Assessment of Anti-Cancer Potential of ZnO Nanoparticles in an In Vitro Model of Breast Cancer.

Advanced innovations for combating variants of aggressive breast cancer and overcoming drug resistance are desired. In cancer treatment, ZnO nanoparticles (NPs) have the capacity to specifically and compellingly activate apoptosis of cancer cells. There is also a pressing need to develop innovative anti-cancer therapeutics, and recent research suggests that ZnO nanoparticles hold great potential. Here, the in vitro chemical effectiveness of ZnO NPs has been tested. Zinc oxide (ZnO) nanoparticles were synthesized using Citrullus colocynthis (L.) Schrad by green methods approach. The generated ZnO was observed to have a hexagonal wurtzite crystal arrangement. The generated nanomaterials were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), UV-visible spectroscopy. The crystallinity of ZnO was reported to be in the range 50–60 nm. The NPs morphology showed a strong absorbance at 374 nm with an estimated gap band of 3.20 eV to 3.32 eV. Microscopy analysis proved the morphology and distribution of the generated nanoparticles to be around 50 nm, with the elemental studies showing the elemental composition of ZnO and further confirming the purity of ZnO NPs. The cytotoxic effect of ZnO NPs was evaluated against wild-type and doxorubicin-resistant MCF-7 and MDA-MB-231 breast cancer cell lines. The results showed the ability of ZnO NPs to inhibit the prefoliation of MCF-7 and MDA-MB-231 prefoliation through the induction of apoptosis without significant differences in both wild-type and resistance to doxorubicin.

Cytotoxic effect, apoptotic activity, hematological and histological alterations induced by green synthesized ZnO nanoparticles applying Hyssopus officinalis leaves

The synthesis of zinc oxide nanoparticles (ZnO-NPs) provides interesting biological characteristics and therapeutic applications in a particular method, which evolve the versatile platform for the essential development of the new anticancer agents. Cytotoxic effect of ZnO-NPs was evaluated by MTT assay on A549 cell line. Besides, the anti-apoptotic effects of ZnO-NPs were investigated using fluorescent microscopy and flow cytometry techniques. Histological assessment was carried out using H&E staining. Finally, complete blood count and hematological factors were analyzed on RBC, platelets, and WBC. Obtained data showed the slight cellular viability (IC50 = 37.2 after 24 h) and increased sub-G1 cell cycle arrest of A549 lung carcinoma cells under ZnO-NPs exposure, indicate the significant dose-dependent and time-dependent cytotoxic effects. In vivo analysis demonstrated raise level of MCH, RBC, and WBC count. ZnO-NPs have the sufficient potential used to be as a safe efficient apoptotic inducer especially in lung cancer therapy.

Antifungal, antioxidant and photocatalytic activities of zinc nanoparticles synthesized by Sargassum vulgare extract

Zinc oxide nanoparticles (ZnO-NPs) have attracted considerable interest owing to their physicochemical properties and vigorous biological activity. In this research, ZnO-NPs were synthesized using aqueous extract of Sargassum vulgare (SVE) as a reducing and capping agent. Morphological and physical features were analyzed through Uv–visible, SEM, TEM, XRD, EDX, DLS and zeta potential studies. Subsequently, antifungal, antioxidant, cytotoxicity and photocatalytic activities were evaluated. FTIR and Uv–vis spectra showed absorption peaks of SVE metabolites and biosynthesized ZnO-NPs before and after the formation of NPs. According to the SEM and TEM micrographs, the biosynthesized ZnO-NPs were spherical with a diameter of about 50–150 nm. The XRD Spectrum indicated the existence of eight peaks corresponding to the crystal angle of the ZnO-NPs. The EDX analysis confirms the presence of oxygen and zinc in the ZnO-NPs structure. The DLS and Zeta potential revealed that the hydrodynamic diameter of ZnO-NPs was approximately 200 nm and +12.7 mV, respectively. The ZnO-NPs demonstrated wide-spectrum antifungal action against Aspergillus, Candida and saccharomyces cerevisiae. The antioxidant potential of ZnO-NPs was determined to be around 65% lower than that of ascorbic acid and BHA, but higher than that of SVE. The cytotoxic effect of ZnO-NPs against the HepG2 cell line was estimated to be an IC50 value of 128.0 μg/ml. Given the potent antioxidant and photocatalytic capacity as well as the robust antifungal activity of ZnO-NPs, a broad variety of applications such as the biological control of plant diseases and the removal of pesticide residues may be recommended.

Cultivation of hMSCs in Human Plasma Prevents the Cytotoxic and Genotoxic Potential of ZnO-NP In Vitro

Zinc oxide nanoparticles (ZnO-NPs) are commonly used for industrial applications. Consequently, there is increasing exposure of humans to them. The in vitro analysis of cytotoxicity and genotoxicity is commonly performed under standard cell culture conditions. Thus, the question arises of how the results of genotoxicity and cytotoxicity experiments would alter if human plasma was used instead of cell culture medium containing of fetal calf serum (FCS). Human mesenchymal stem cells (hMSCs) were cultured in human plasma and exposed to ZnO-NPs. A cultivation in expansion medium made of DMEM consisting 10% FCS (DMEM-EM) served as control. Genotoxic and cytotoxic effects were evaluated with the comet and MTT assay, respectively. hMSC differentiation capacity and ZnO-NP disposition were evaluated by histology and transmission electron microscopy (TEM). The protein concentration and the amount of soluble Zn2+ were measured. The cultivation of hMSCs in plasma leads to an attenuation of genotoxic and cytotoxic effects of ZnO-NPs compared to control. The differentiation capacity of hMSCs was not altered. The TEM showed ZnO-NP persistence in cytoplasm in both groups. The concentrations of protein and Zn2+ were higher in plasma than in DMEM-EM. In conclusion, the cultivation of hMSCs in plasma compared to DMEM-EM leads to an attenuation of cytotoxicity and genotoxicity in vitro.

Time-Dependent Toxic and Genotoxic Effects of Zinc Oxide Nanoparticles after Long-Term and Repetitive Exposure to Human Mesenchymal Stem Cells.

Zinc oxide nanoparticles (ZnO-NP) are widely spread in consumer products. Data about the toxicological characteristics of ZnO-NP is still under controversial discussion. The human skin is the most important organ concerning ZnO-NP exposure. Intact skin was demonstrated to be a sufficient barrier against NPs; however, defect skin may allow NP contact to proliferating cells. Within these cells, stem cells are the most important toxicological target for NPs. The aim of this study was to evaluate the genotoxic and cytotoxic effects of ZnO-NP at low-dose concentrations after long-term and repetitive exposure to human mesenchymal stem cells (hMSC). Cytotoxic effects of ZnO-NP were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Furthermore, genotoxicity was evaluated by the comet assay. For long-term observation over 6 weeks, transmission electron microscopy (TEM) was applied. The results of the study indicated cytotoxic effects of ZnO-NP beginning at high concentrations of 50 μg/mL and genotoxic effects in hMSC exposed to 1 and 10 μg/mL ZnO-NP. Repetitive exposure enhanced cyto- but not genotoxicity. Intracellular NP accumulation was observed up to 6 weeks. The results suggest cytotoxic and genotoxic potential of ZnO-NP. Even low doses of ZnO-NP may induce toxic effects as a result of repetitive exposure and long-term cellular accumulation. This data should be considered before using ZnO-NP on damaged skin.

Cytotoxic effect of zinc oxide nanoparticles on murine photoreceptor cells via potassium channel block and Na+ /K+ -ATPase inhibition.

Zinc oxide (ZnO) nanoparticles can exhibit toxicity towards organisms and oxidative stress is often hypothesized to be one of the most important factors. Nevertheless, the detailed mechanism of toxicity-induced by ZnO nanoparticles has not been completely addressed. The present study aimed to investigate the toxic effects of ZnO nanoparticles on the expression and activity of Na+ /K+ -ATPase and on potassium channel block. In the present study, we explored the cytotoxic effect of ZnO nanoparticles on murine photoreceptor cells using lactate dehydrogenase (LDH) release assay, reactive oxygen species (ROS) determination, mitochondrial membrane potential (Δφm) measurement, delayed rectifier potassium current recordings and Na+ /K+ -ATPase expression and activity monitoring. The results indicated that ZnO nanoparticles could increase the LDH release in medium, aggravate the ROS level within cells, collapse the Δφm, block the delayed rectifier potassium current, and attenuate the expressions of Na+ /K+ -ATPase at both mRNA and protein levels and its activity, and thus exert cytotoxic effects on murine photoreceptor cells, finally damaging target cells. Our findings will facilitate the understanding of the mechanism involved in ZnO nanoparticle-induced cytotoxicity in murine photoreceptor cells via potassium channel block and Na+ /K+ -ATPase inhibition.

Zinc-Oxide Nanoparticles Exhibit Genotoxic, Clastogenic, Cytotoxic and Actin Depolymerization Effects by Inducing Oxidative Stress Responses in Macrophages and Adult Mice.

Zinc oxide nanoparticles (ZnO-NPs) have wide biological applications, which have raised serious concerns about their impact on the health and environment. Although, various studies have shown ZnO-NP toxicity on different cells underin vitroconditions, sufficient information is lacking regarding toxicity and underlying mechanisms underin vivoconditions. In this work, we investigated genotoxic, clastogenic, and cytotoxic effects of ZnO-NPs on macrophages and in adult mice. ZnO-NP-treated mice showed signs of toxicity such as loss in body weight, passive behavior and reduced survival. Further mechanistic studies revealed that administration of higher dose caused severe DNA damage in peripheral blood and bone marrow cells as evident by the formation of COMET tail, micronuclei, chromosomal fragmentation, and phosphorylation of H2A histone family member X. Moreover, ZnO-NPs inhibited DNA repair mechanism by downregulating the expression offen-1andpolBproteins. Histopathological examinations showed severe inflammation and damage to liver, lungs, and kidneys. Cell viability and wound healing assays revealed that ZnO-NPs killed macrophages in a dose-dependent manner, caused severe wounds and inhibited cellular migration by irreversible actin depolymerization and degradation. Reduction in the viability of macrophages was due to the arrest of the cell cycle at the G0/G1 phase, inhibition of superoxide dismutase and catalase and eventually reactive oxygen species. Furthermore, treatment with an antioxidant drug N-acetyl cysteine significantly reduced the ZnO-NP induced genotoxicity bothin vitroandin vivo Altogether, this study gives detailed pathological insights of ZnO-NP that impair cellular functions, thus will enable to arbitrate their biological applications.

Irradiation-Enhanced Cytotoxicity of Zinc Oxide Nanoparticles

Zinc oxide (ZnO) nanoparticles (NPs) are being widely utilized in industry due to their versatile properties. The in vitro cytotoxicity findings and the potential for exposures to ZnO NP from different sources via different routes of entry into the body have raised public health concerns. Although recent studies have shown the cytotoxic effects of these NPs, including oxidative stress, apoptosis and necrosis induction, genotoxicity, and others, irradiation-induced cytotoxicity has not been systematically studied. The goal of this study was to determine whether irradiation in the forms of visible light (approximately 400-600 nm), ultraviolet (UV) A (366 nm), and UVC (254 nm) would affect ZnO NPs-induced cytotoxicity. The results of this study demonstrated that the cytotoxicity of 60 to 80 nm ZnO NPs to A549 cells is dosage, time, and wavelength dependent. Nuclear decomposition by ZnO NPs, prior to membrane deformation, was found to be enhanced when exposed to irradiation. This study suggests that this phenomenon may be attributed to the irradiation-induced formation of positively charged sites on the ZnO NPs, which enhances nuclear affinity and generation of reactive oxygen species. Finally, the data demonstrated that while ZnO NPs act preferentially toward nuclear regions, destructions of cell membrane and the cytosol have also been observed. The photocatalytic properties of ZnO NPs play a critical role during the early stages of cell death, and their effects were reduced through the use of an antioxidant, N-acetylcysteine. In conclusion, both visible light and UV irradiations have been found to enhance the cytotoxic effect of ZnO NPs on the A549 cell line. This finding supports the need for further in vivo exposure studies relevant to actual conditions to confirm whether combined irradiation and ZnO NP exposure could enhance the nanotoxicity of ZnO NPs.

Phosphate-enhanced cytotoxicity of zinc oxide nanoparticles and agglomerates

Zinc oxide (ZnO) nanoparticles (NPs) have been found to readily react with phosphate ions to form zinc phosphate (Zn3(PO4)2) crystallites. Because phosphates are ubiquitous in physiological fluids as well as waste water streams, it is important to examine the potential effects that the formation of Zn3(PO4)2 crystallites may have on cell viability. Thus, the cytotoxic response of NIH/3T3 fibroblast cells was assessed following 24h of exposure to ZnO NPs suspended in media with and without the standard phosphate salt supplement. Both particle dosage and size have been shown to impact the cytotoxic effects of ZnO NPs, so doses ranging from 5 to 50μg/mL were examined and agglomerate size effects were investigated by using the bioinert amphiphilic polymer polyvinylpyrrolidone (PVP) to generate water-soluble ZnO ranging from individually dispersed 4nm NPs up to micron-sized agglomerates. Cell metabolic activity measures indicated that the presence of phosphate in the suspension media can led to significantly reduced cell viability at all agglomerate sizes and at lower ZnO dosages. In addition, a reduction in cell viability was observed when agglomerate size was decreased, but only in the phosphate-containing media. These metabolic activity results were reflected in separate measures of cell death via the lactate dehydrogenase assay. Our results suggest that, while higher doses of water-soluble ZnO NPs are cytotoxic, the presence of phosphates in the surrounding fluid can lead to significantly elevated levels of cell death at lower ZnO NP doses. Moreover, the extent of this death can potentially be modulated or offset by tuning the agglomerate size. These findings underscore the importance of understanding how nanoscale materials can interact with the components of surrounding fluids so that potential adverse effects of such interactions can be controlled.

Zinc oxide nanoparticles inhibit Ca2+-ATPase expression in human lens epithelial cells under UVB irradiation

Epidemiological and experimental studies have revealed that lens epithelial cells exposed to ultraviolet B (UVB) light could be induced apoptosis, and lens epithelial cell apoptosis can initiate cataractogenesis. Posterior capsular opacification (PCO), the most frequent complication after cataract surgery, is induced by the proliferation, differentiation, migration of lens epithelial cells. Thus, inhibiting the proliferation of lens epithelial cells could reduce the occurrence of PCO. It is reported that zinc oxide (ZnO) nanoparticles have great potential for the application of biomedical field including cancer treatment. In the present study, we investigated the cytotoxic effect of ZnO nanoparticles on human lens epithelial cell (HLEC) viability. In addition, changes in cell nuclei, apoptosis, reactive oxygen species and intracellular calcium ion levels were also investigated after cells treated with ZnO nanoparticles in the presence and absence of UVB irradiation. Meanwhile, the expression of plasma membrane calcium ATPase 1 (PMCA1) was also determined at gene and protein levels. The results indicate that ZnO nanoparticles and UVB irradiation have synergistic inhibitory effect on HLEC proliferation in a concentration-dependent manner. ZnO nanoparticles can increase the intracellular calcium ion level, disrupt the intracellular calcium homeostasis, and decrease the expression level of PMCA1. UVB irradiation can strengthen the effect of reduced expression of PMCA1, suggesting that both UVB irradiation and ZnO nanoparticles could exert inhibitory effect on HLECs via calcium-mediated signaling pathway. ZnO nanoparticles have great potential for the treatment of PCO under UVB irradiation.

Cytotoxicity of zinc oxide (ZnO) nanoparticles is influenced by cell density and culture format

A parameter that has often been overlooked in cytotoxicity assays is the density and confluency of mammalian cell monolayers utilized for toxicology screening. Hence, this study investigated how different cell seeding densities influenced their response to cytotoxic challenge with ZnO nanoparticles. Utilizing the same volume (1ml per well) and concentration range (5-40μg/ml) of ZnO nanoparticles, contradictory results were observed with higher-density cell monolayers (BEAS-2B cells) obtained either by increasing the number of seeded cells per well (50,000 vs. 200,000 cells per well of 12-well plate) or by seeding the same numbers of cells (50,000) within a smaller surface area (12-well vs. 48-well plate, 4.8 vs. 1.2cm(2), respectively). Further experiments demonstrated that the data may be skewed by inconsistency in the mass/number of nanoparticles per unit area of culture surface, as well as by inconsistent nanoparticle to cell ratio. To keep these parameters constant, the same number of cells (50,000 per well) were seeded on 12-well plates, but with the cells being seeded at the edge of the well for the experimental group (by tilting the plate) to form a dense confluent monolayer, as opposed to a sparse monolayer for the control group seeded in the conventional manner. Utilizing such an experimental set-up for the comparative evaluation of four different cell lines (BEAS-2B, L-929, CRL-2922 and C2C12), it was observed that the high cell density monolayer was consistently more resistant to the cytotoxic effects of ZnO nanoparticles compared to the sparse monolayer for all four different cell types, with the greatest differences being observed above a ZnO concentration of 10μg/ml. Hence, the results of this study demonstrate the need for the standardization of cell culture protocols utilized for toxicology screening of nanoparticles, with respect to cell density and mass/number of nanoparticles per unit area of culture surface.