Investigations of the Toxic Effect of Silver Nanoparticles on Mammalian Cell Lines (original) (raw)
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Toxicity Effect of Silver Nanoparticles on Mice Liver Primary Cell Culture and HepG2 Cell Line
Iranian journal of pharmaceutical research : IJPR, 2014
Nano-silver (AgNP) has biological properties which are significant for consumer products, food technology, textiles and medical applications (e.g. wound care products, implantable medical devices, in diagnosis, drug delivery, and imaging). For their antibacterial activity, silver nanoparticles are largely used in various commercially available products. Thus, the use of nano-silver is becoming more and more widespread in medicine. In this study we investigated the cytotoxic effects of AgNPs on liver primary cells of mice, as well as the human liver HepG2 cell. Cell viability was examined with MTT assay after HepG2 cells exposure to AgNPs at 1, 2, 3, 4, 5, 7.5, 10 ppm compared to mice primary liver cells at 1, 10, 50, 100, 150, 200, 400 ppm for 24h. AgNPs caused a concentration-dependent decrease of cell viability in both cells. IC50 value of 2.764 ppm (µg/mL) was calculated in HepG2 cell line and IC50 value of 121.7 ppm (µg/mL) was calculated in primary liver cells of mice. The resu...
Despite the widespread use of silver nanoparticles (AgNPs) in different fields and the amount of investigations available, to date, there are many contradictory results on their potential toxicity. In the present study, extensively characterized 20-nm AgNPs were investigated using optimized protocols and standardized methods to test several toxicological endpoints in different cell lines. The agglomeration/aggregation state of AgNPs in culture media was measured by dynamic light scattering (DLS). DNA and chromosomal damage on BEAS-2B and RAW 264.7 cells were evaluated
Archives of Toxicology, 2012
Silver nanoparticles (SNPs) are among the most commercialized nanoparticles worldwide. Often SNP are used because of their antibacterial properties. Besides that they possess unique optic and catalytic features, making them highly interesting for the creation of novel and advanced functional materials. Despite its widespread use only little data exist in terms of possible adverse effects of SNP on human health. Conventional synthesis routes usually yield products of varying quality and property. It thus may become puzzling to compare biological data from different studies due to the great variety in sizes, coatings or shapes of the particles applied. Here, we applied a novel synthesis approach to obtain SNP of well-defined colloidal and structural properties. Being stabilized by a covalently linked small peptide, these particles are nicely homogenous, with narrow size distribution, and form monodisperse suspensions in aqueous solutions. We applied these peptide-coated SNP in two different sizes of 20 or 40 nm (Ag20Pep and Ag40Pep) and analyzed responses of THP-1-derived human macrophages while being exposed against these particles. Gold nanoparticles of similar size and coating (Au20Pep) were used for comparison. The cytotoxicity of particles was assessed by WST-1 and LDH assays, and the uptake into the cells was confirmed via transmission electron microscopy. In summary, our data demonstrate that this novel type of SNP is well suited to serve as model system for nanoparticles to be tested in toxicological studies in vitro.
Environmental and Human Health Issues of Silver Nanoparticles Applications
2011
The significant growth in applications of silver nanoparticles across various branches of industry as well as in consumer products has caused concerns that nanosilver may have a toxic effect on the environment and human health and may have implications for eco-terorism. This paper presents research on antimicrobial effects of silver nanoparticles. We studied the cytotoxicity of silver nanoparticles via an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromid) assay that measures cell activity through the mitochondrial cleavage of a molecule that exhibits a change of colour that can be measured spectrophotometrically. NIH-3T3 (Swiss mouse embryo), HEP-G2 (human hepatocellular carcinoma), A-549 (human lung carcinoma), PC-12 (rat adrenal pheochromocytoma), and Colo-320 (human colon adenocarcinoma) cells were chosen in order to study different possible absorption paths of nanoparticles into the organism and various areas of particle accumulation in the body. The obtained MTT test results have shown that silver nanoparticles with concentrations of ~1-10 ppm entering the body from air or liquid suspensions can present a potential risk to human health.
Interactions of silver nanoparticles with primary mouse fibroblasts and liver cells
Toxicology and Applied Pharmacology, 2009
Primary cells are ideal for in vitro toxicity studies since they closely resemble tissue environment. Here, we report a detailed study on the in vitro interactions of 7-20 nm spherical silver nanoparticles (SNP) with primary fibroblasts and primary liver cells isolated from Swiss albino mice. The intended use of silver nanoparticles is in the form of a topical antimicrobial gel formulation for the treatment of burns and wounds. Upon exposure to SNP for 24 h, morphology of primary fibroblasts and primary liver cells remained unaltered up to 25 μg/mL and 100 μg/mL SNP, respectively, although with minor decrease in confluence. IC 50 values for primary fibroblasts and primary liver cells as revealed by XTT assay were 61 μg/mL and 449 μg/mL, respectively. Ultra-thin sections of primary cells exposed to 1/2 IC 50 SNP for 24 h, visualized under Transmission electron microscope showed the presence of dark, electron dense, spherical aggregates inside the mitochondria, and cytoplasm, probably representing the intracellular SNP. When the cells were challenged with ∼1/2 IC 50 concentration of SNP (i.e. 30 μg/mL and 225 μg/mL for primary fibroblasts and primary liver cells, respectively), enhancement of GSH (∼1.2 fold) and depletion of lipid peroxidation (∼1.4 fold) were seen in primary fibroblasts which probably protect the cells from functional damage. In case of primary liver cells; increased levels of SOD (∼1.4 fold) and GSH (∼1.1 fold) as compared to unexposed cells were observed. Caspase-3 activity assay indicated that the SNP concentrations required for the onset of apoptosis were found to be much lower (3.12 μg/mL in primary fibroblasts, 12.5 μg/mL in primary liver cells) than the necrotic concentration (100 μg/mL in primary fibroblasts, 500 μg/mL in primary liver cells). These observations were confirmed by CLSM studies by exposure of cells to 1/2 IC 50 SNP (resulting in apoptosis) and 2× IC 50 ) cells (resulting in necrosis). These results clearly suggest that although silver nanoparticles seem to enter the eukaryotic cells, cellular antioxidant mechanisms protect the cells from possible oxidative damage. This property, in conjunction with the finding that primary cells possess much higher SNP tolerance than the concentration in the gel (∼20 μg/g), indicates preliminary safety of the formulation and warrants further study for possible human application.
Size-dependent cellular toxicity of silver nanoparticles
Journal of Biomedical Materials Research Part A, 2012
Silver nanoparticles (AgNPs) have found a variety of uses including biomedical materials; however, studies of the cytotoxicity of AgNPs by size effects are only in the beginning stage. In this study, we examined the size-dependent cellular toxicity of AgNPs using three different characteristic sizes ($ 10, 50, and 100 nm) against several cell lines including MC3T3-E1 and PC12. The cytotoxic effect determined based on the cell viability, intracellular reactive oxygen species generation, lactate dehydrogenase release, ultrastructural changes in cell morphology, and upregulation of stress-related genes (ho-1 and MMP-3) was fairly size-and dose-dependent. In particular, AgNPs stimulated apoptosis in the MC3T3-E1 cells, but induced necrotic cell death in the PC12 cells. Furthermore, the smallest sized AgNPs (10 nm size) had a greater ability to induce apoptosis in the MC3T3-E1 cells than the other sized AgNPs (50 and 100 nm). These data suggest that the AgNPs-induced cytotoxic effects against tissue cells are particle size-dependent, and thus, the particle size needs careful consideration in the design of the nanoparticles for biomedical uses.
Biochemical Changes Induced by the Toxicity of Variable Sizes of Silver Nanoparticles
British Journal of Pharmaceutical Research, 2014
This project was carried out in collaboration between all authors. Author QJ anchored the literature for the study, performed all experimental work of the project and wrote the first draft of the manuscript. Author AB managed all aspects of the project while authors FO and KB managed the statistical analysis and provided the background for the interpretation of the results. All authors read and approved the manuscript.
With the development of nanotechnology, silver nanoparticles (Ag-NPs) have become one of the most in-demand nanoparticles owing to their exponential number of uses in various sectors. The increased use of Ag-NPs-enhanced products may result in an increased level of toxicity affecting both the environment and living organisms. Several studies have used different model cell lines to exhibit the cytotoxicity of Ag-NPs, and their underlying molecular mechanisms. This review aimed to elucidate different properties of Ag-NPs that are responsible for the induction of cellular toxicity along with the critical mechanism of action and subsequent defense mechanisms observed in vitro. Our results show that the properties of Ag-NPs largely vary based on the diversified synthesis processes. The physiochemical properties of Ag-NPs (e.g., size, shape, concentration, agglomeration, or aggregation interaction with a biological system) can cause impairment of mitochondrial function prior to their penetration and accumulation in the mito-chondrial membrane. Thus, Ag-NPs exhibit properties that play a central role in their use as biocides
Environmental Toxicology, 2013
Silver nanoparticles are increasingly used in various products, due to their antibacterial properties. Despite its wide spread use, only little information on possible adverse health effects exists. Therefore, the aim of this study was to assess the toxic potential of silver nanoparticles (<100 nm) in human lung epithelial (A549) cells and the underlying mechanism of its cellular toxicity. Silver nanoparticles induced dose and time-dependent cytotoxicity in A549 cells demonstrated by MTT and LDH assays. Silver nanoparticles were also found to induce oxidative stress in dose and time-dependent manner indicated by depletion of GSH and induction of ROS, LPO, SOD, and catalase. Further, the activities of caspases and the level of proinflammatory cytokines, namely interleukin-1b (IL-1b) and interleukin-6 (IL-6) were significantly higher in treated cells. DNA damage, as measured by single cell gel electrophoresis, was also dose and time-dependent signicants in A549 cells. This study investigating the effects of silver nanoparticles in human lung epithelial cells has provided valuable insights into the mechanism of potential toxicity induced by silver nanoparticles and warrants more careful assessment of silver nanoparticles before their industrial applications. V C 2013 Wiley Periodicals, Inc. Environ Toxicol 00: 000-000, 2013.
Iranian Red Crescent Medical Journal
Background: Cancer is currently the second leading cause of death worldwide that is originated from cell growth and proliferation without control. Acute lymphoblastic leukemia (ALL) is one of the types of leukemia that affects lymphocyte maturation and it is common among children. Silver nanoparticles are considered one of the targeted chemotherapy methods by creating cytotoxicity. Objectives: In this research, a comparative study of cytotoxic effect of silver nanoparticles was evaluated on human lymphocytes and HPB-ALL cell line as an in vitro study. Methods: In this experimental study, lymphocytes and HPB-ALL cell line were exposed to silver nanoparticles at RPMI 1640 medium culture in order to assess toxicity for 24 hours. To this aim, MTT assay was used to evaluate the toxicity of the silver nanoparticles. DNA fragmentation and apoptosis were evaluated by Gel Electrophoresis and Flow Cytometry, respectively. Moreover, quantitative PCR was performed on bax, bcl-2, and caspase-9 genes. Results: The results of MTT assay showed IC50 values of silver nanoparticles were 5.87 and 2.68 µg/mL for lymphocytes and HPB-ALL cell line, respectively. The results showed that silver nanoparticles could split DNA of the HPB-ALL cell line more than DNA of the lymphocytes during DNA fragmentation. Flow Cytometry results indicated that the early apoptosis was 6.04% and 22.75% in lymphocytes and HPB-ALL cell line, respectively. Moreover, Q-PCR results showed a significant up-regulation of caspase-9 and bax genes and downregulation of bcl-2 gene in comparison to the control group. Conclusions: The silver nanoparticles had cytotoxic effects on the lymphocytes and HPB-ALL cell line. The results showed that the silver nanoparticle had a significant cytotoxic effect on HPB-ALL cell line.