Emodin induces embryonic toxicity in mouse blastocysts through apoptosis

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Previous studies have established that emodin inhibits cell proliferation and induces caspase 3-dependent apoptosis. However, its side-effects, particularly those on embryonic development, have not been well characterized as yet. In the current study, we examined the cytotoxic effects of emodin on mouse embryos at the blastocyst stage, subsequent embryonic attachment and outgrowth in vitro, and in vivo implantation by embryo transfer. Blastocysts treated with 25-75 μM emodin exhibited significantly increased apoptosis and a corresponding decrease in total cell number. Notably, the implantation success rate of blastocysts pretreated with emodin was lower than that of their control counterparts. Moreover, in vitro treatment with 25-75 μM emodin was associated with increased resorption of post-implantation embryos and decreased fetal weight. With the aid of an in vivo mouse model, we showed that consumption of drinking water containing emodin led to apoptosis and decreased cell proliferation, and inhibited early embryonic development to the blastocyst stage. Our findings support a degree of selective inhibition of retinoic acid receptors in blastocysts treated with emodin. In addition, emodin appears to induce injury in mouse blastocysts through intrinsic apoptotic signaling processes to impair sequent embryonic development. These results collectively indicate that emodin has the potential to induce embryonic cytotoxicity.     

Embryonic toxicity of sanguinarine through apoptotic processes in mouse blastocysts

In this study, we examined the cytotoxic effects of sanguinarine, a phytoalexin with antimicrobial, anti-oxidant, anti-inflammatory and pro-apoptotic effects, on the blastocyst stage of mouse embryos, subsequent embryonic attachment and outgrowth in vitro and in vivo implantation via embryo transfer. Blastocysts treated with 0.5-2 μM sanguinarine exhibited significantly increased apoptosis and a corresponding decrease in total cell number. Notably, the implantation success rates of blastocysts pretreated with sanguinarine were lower than that of their control counterparts. Moreover, in vitro treatment with 0.5-2 μM sanguinarine was associated with increased resorption of post-implantation embryos and decreased fetal weight. Our results collectively indicate that sanguinarine induces apoptosis and retards early post-implantation development in vitro and in vivo. In addition, sanguinarine induces apoptotic injury effects on mouse blastocysts through intrinsic and extrinsic apoptotic signaling processes to impair sequent embryonic development. However, the extent to which sanguinarine exerts teratogenic effects on early human development is not known at present, and further studies are required to establish effective protection strategies against its cytotoxic effects.     

PVP-coated silver nanoparticles and silver ions induce reactive oxygen species, apoptosis and necrosis in THP-1 monocytes

The objective of the present study was to investigate the toxicity of silver nanoparticles (Ag NPs) in vitro. Silver ions (Ag+) have been used in medical treatments for decades whereas Ag NPs have been used in a variety of consumer products within recent years. This study was undertaken to compare the effect of well characterized, PVP-coated Ag NPs (69 nm ± 3 nm) and Ag+ in a human monocytic cell line (THP-1). Characterization of the Ag NPs was conducted in both stock suspension and cell media with or without serum and antibiotics. By using the flowcytometric annexin V/propidium iodide (PI) assay, both Ag NPs and Ag+ were shown to induce apoptosis and necrosis in THP-1 cells depending on dose and exposure time. Furthermore, the presence of apoptosis could be confirmed by the TUNEL method. A number of studies have implicated the production of reactive oxygen species (ROS) in cytotoxicity mediated by NPs. We used the fluorogenic probe, 2′,7′-dichlorofluorescein to assess the levels of intracellular ROS during exposure to Ag NPs and Ag+. A drastic increase in ROS levels could be detected after 6–24 h suggesting that oxidative stress is an important mediator of cytotoxicity caused by Ag NPs and Ag+.     

Cellular uptake, intracellular trafficking and cytotoxicity of silver nanoparticles

Silver nanoparticles (Ag NPs) are used in consumer products and wound dressings due to their antimicrobial properties. However, in addition to toxic effects on microbes, Ag NPs can also induce stress responses as well as cytotoxicity in mammalian cells. We observed that Ag NPs are efficiently internalized via scavenger receptor-mediated phagocytosis in murine macrophages. Confocal and electron microscopy analysis revealed that internalized Ag NPs localize in the cytoplasm. Ag NPs cause mitochondrial damage, induce apoptosis and cell death. These effects were abrogated in presence of Ag ion-reactive, thiol-containing compounds suggesting the central of Ag ions in Ag NP toxicity. Quantitative image analysis revealed that intracellular dissolution of Ag NPs occurs about 50 times faster than in water. In conclusion, we demonstrate for the first time that Ag NPs are internalized by scavenger receptors, trafficked to cytoplasm and induce toxicity by releasing Ag ions.     

Interactions of silver nanoparticles with primary mouse fibroblasts and liver cells

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. IC₅₀ 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 IC₅₀ 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 IC₅₀ 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 IC₅₀ SNP (resulting in apoptosis) and 2× IC₅₀) 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.     

Differential transformation and antibacterial effects of silver nanoparticles in aerobic and anaerobic environment

Silver nanoparticles (AgNP) undergo various transformations into different Ag species in the environment, which determines their toxicity in microorganisms. In aerobic condition, AgNPs release Ag⁺ that causes cell inactivation. Limited information is known about the AgNP-cell interaction in oxygen-free environment. Here we compared the transformation and antibacterial effects of AgNPs in aerobic and anaerobic environment. The bacterium Pseudomonas aeruginosa was relatively not susceptible to Ag⁺ or AgNP in anaerobic environment, indicated by near two orders of magnitude greater of anaerobic minimum inhibitory concentration (MIC) than the aerobic counterpart. In anaerobic environment, the dissolved Ag concentration decreased due to the reduction of Ag⁺. Electron microscopy images showed the formation of new AgNPs and aggregates, preferably on cell surface or associated with extracellular polymer substances (EPS) matrix. Accumulating AgNPs onto the cells could cause membrane damage, cytoplasm release or bacterial death. Meanwhile, EPS and cell lysate were very likely to bind AgNPs, facilitating the extensively assembling of AgNPs into large aggregates. This reduced the effective Ag exposure to cells and might contribute to the detoxification in anaerobic environment. Further, flow cytometry analysis quantified that bacterial membrane was largely intact under the treatment of AgNPs in anaerobic condition compared to the dose–response manner in aerobic condition.     

Synthesis of silver and copper oxide nanoparticles using Myristica fragrans fruit extract: Antimicrobial and catalytic applications

The pericarp of Myristica fragrans fruit extract was utilized for a low cost, eco-friendly synthesis of silver (AgNPs) and copper oxide (CuONPs) nanoparticles. The aqueous fruit extract of the plant was used as reducing and stabilizing agents for this preparation. Characterization of the biosynthesized nanoparticles was carried out using UV–Vis spectroscopy, FTIR spectroscopy and X-Ray Diffraction studies. Morphology and size of the particles was observed using Field-Emission Scanning Electron Microscopy (FESEM) and High Resolution Transmission Electron Microscopy (HRTEM). The copper and silver nanoparticles show Surface Plasmon Resonance (SPR) band at 360 and 478 nm respectively in the UV–Vis spectrum. It was observed that size of the synthesized copper oxide and silver nanoparticles are in the range 10–50 nm. The presence of copper and silver elements was confirmed from their respective EDS spectrum. Involvement of phytochemicals in the stabilization and reduction of the nanoparticles was confirmed by FTIR spectroscopy. CuONPs exhibited catalytic activity in 1,3-dipolar cycloaddition reaction between azides and terminal alkynes to form 1,2,3-triazoles. Silver nanoparticle possesses good antibacterial activity against multidrug human pathogens Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis. The present study focuses on the utilization of the less economic part of Myristica fragrans fruit's pericarp for the preparation of copper oxide and silver nanoparticles which have good catalytic and antibacterial activities.     

Differentiation of the toxicities of silver nanoparticles and silver ions to the Japanese medaka (Oryzias latipes) and the cladoceran Daphnia magna

Abstract

Silver nanoparticles (AgNPs) are increasingly used in various fields. However, little is known about the environmental effects of widespread use of products containing AgNPs. The objective of this study was to determine the ecotoxicity caused by AgNPs. The 48-h effective concentration 50 (EC₅₀) values for Daphnia magna of suspensions of 60 nm and 300 nm AgNPs were 1.0 (95% confidence interval [CI] = 0.1–1.3) and 1.4 (95% CI = 0.3–2.1) μg Ag/l, respectively. The 96 h LC₅₀ values for Oryzias latipes of 60 nm and 300 nm AgNP suspensions were 28 (95% CI = 23–34) and 67 (95% CI = 45–108) μg Ag/l, respectively. To show that toxicity is caused only by Ag⁺ and not by AgNPs, Ag⁺ was adsorbed onto the synthesized sorbents packed in a column and D. magna was exposed to the column-passed-300 nm AgNP suspensions. There was no acute toxicity with the AgNP suspensions not containing Ag⁺.     

Cellular responses induced by silver nanoparticles: In vitro studies

A systematic study on the in vitro interactions of 7-20nm spherical silver nanoparticles (SNP) with HT-1080 and A431 cells was undertaken as a part of an on-going program in our laboratory to develop a topical antimicrobial agent for the treatment of burn wound infections. Upon exposure to SNP (up to 6.25mug/mL), morphology of both the cell types remained unaltered. However, at higher concentrations (6.25-50mug/mL) cells became less polyhedral, more fusiform, shrunken and rounded. IC(50) values for HT-1080 and A431 as revealed by XTT assay were 10.6 and 11.6mug/mL, respectively. When the cells were challenged with approximately 1/2 IC(50) concentration of SNP (6.25mug/mL), clear signs of oxidative stress, i.e. decreased GSH ( approximately 2.5-folds in HT-1080, approximately 2-folds in A431) and SOD ( approximately 1.6-folds in HT-1080, 3-folds in A431) as well as increased lipid peroxidation ( approximately 2.5-folds in HT-1080, approximately 2-folds in A431) were seen. Changes in the levels of catalase and GPx in A431 cells were statistically insignificant in both cell types. DNA fragmentation in SNP-exposed cells suggested apoptosis. When the apoptotic thresholds of SNP were monitored with caspase-3 assay the concentrations required for the onset of apoptosis were found to be much lower (0.78mug/mL in HT-1080, 1.56mug/mL in A431) than the necrotic concentration (12.5mug/mL in both cell types). These results can be used to define a safe range of SNP for the intended application as a topical antimicrobial agent after appropriate in vivo studies.     

Liposomal encapsulation of silver nanoparticles enhances cytotoxicity and causes induction of reactive oxygen species‐independent apoptosis

Silver nanoparticles (AgNP) are one of the most widely investigated metallic NPs due to their promising antibacterial activities. In recent years, AgNP research has shifted beyond antimicrobial use to potential applications in the medical arena. This shift coupled with the extensive commercial applications of AgNP will further increase human exposure and the subsequent risk of adverse effects that may result from repeated exposures and inefficient delivery, meaning research into improved AgNP delivery is of paramount importance. In this study, AgNP were encapsulated in a natural biosurfactant, dipalmitoylphosphatidylcholine, in an attempt to enhance the intracellular delivery and simultaneously mediate the associated cytotoxicity of the AgNP. It was noted that because of the encapsulation, liposomal AgNP (Lipo‐AgNP) at 0.625 μg ml^–1 induced significant cell death in THP1 cell lines a notably lower dose than that of the uncoated AgNP induced cytotoxicity. The induced cytotoxicity was shown to result in an increased level of DNA fragmentation resulting in a cell cycle interruption at the S phase. It was shown that the predominate form of cell death upon exposure to both uncoated AgNP and Lipo‐AgNP was apoptosis. However, a reactive oxygen species‐independent activation of the executioner caspases 3/7 occurred when exposed to the Lipo‐AgNP. These findings showed that encapsulation of AgNP enhance AgNP cytotoxicity and mediates a reactive oxygen species‐independent induction of apoptosis.     

Characterisation,cytotoxicity and apoptosis studies of methotrexate-loaded PLGA and PLGA-PEG nanoparticles

Abstract

Methotrexate (MTX) widely used in the treatments of various types of malignancies, but high toxicity and short plasma half-life have limited its use. This study was aimed at developing a polymeric drug delivery system for improving the therapeutic index of this potent drug. To achieve these goals, PLGA and PLGA-PEG nanoparticles were prepared using the emulsification-solvent diffusion technique and were optimized for particle size and entrapment efficiency. The optimum loaded nanoparticles were evaluated by cytotoxicity and their ability to induce apoptosis compared to free drug by examining of caspase-3 activity. The results showed that optimized particles were 182?±?14?nm and 258?±?10?nm in size for PLGA-PEG and PLGA nanoparticles, respectively, with an entrapment efficiency of more than 51%. The cytotoxicity experiment showed that the nanoparticles were more effective than pure MTX and increase the activity of caspase-3 in MCF7 and AGS and A549 cell lines.     

Multi-platform genotoxicity analysis of silver nanoparticles in the model cell line CHO-K1

Investigation of the genotoxic potential of nanomaterials is essential to evaluate if they pose a cancer risk for exposed workers and consumers. The Chinese hamster ovary cell line CHO-K1 is recommended by the OECD for use in the micronucleus assay and is commonly used for genotoxicity testing. However, studies investigating if this cell line is suitable for the genotoxic evaluation of nanomaterials, including induction of DNA adduct and micronuclei formation, are rare and for silver nanoparticles (Ag NPs) missing. Therefore, we here systematically investigated DNA and chromosomal damage induced by BSA coated Ag NPs (15.9 ± 7.6 nm) in CHO-K1 cells in relation to cellular uptake and intracellular localization, their effects on mitochondrial activity and production of reactive oxygen species (ROS), cell cycle, apoptosis and necrosis. Ag NPs are taken up by CHO-K1 cells and are presumably translocated into endosomes/lysosomes. Our cytotoxicity studies demonstrated a concentration-dependent decrease of mitochondrial activity and increase of intracellular reactive oxygen species (ROS) in CHO-K1 cells following exposure to Ag NPs and Ag⁺ (0–20 μg/ml) for 24 h. Annexin V/propidium iodide assay showed that Ag NPs and Ag⁺ induced apoptosis and necrosis, which is in agreement with an increased fraction of cells in subG1 phase of the cell cycle. Genotoxicity studies showed that Ag NPs but also silver ions (Ag⁺) induced bulky-DNA adducts, 8-oxodG and micronuclei formation in a concentration-dependent manner, however, there were quantitative and qualitative differences between the particulate and ionic form of silver. Taken together, our multi-platform genotoxicity and cytotoxicity analysis demonstrates that CHO-K1 cells are suitable for the investigation of genotoxicity of nanoparticles like Ag NPs.     

DNA damage response to different surface chemistry of silver nanoparticles in mammalian cells

Silver nanoparticles (Ag NPs) have recently received much attention for their possible applications in biotechnology and life sciences. Ag NPs are of interest to defense and engineering programs for new material applications as well as for commercial purposes as an antimicrobial. However, little is known about the genotoxicity of Ag NPs following exposure to mammalian cells. This study was undertaken to examine the DNA damage response to polysaccharide surface functionalized (coated) and non-functionalized (uncoated) Ag NPs in two types of mammalian cells; mouse embryonic stem (mES) cells and mouse embryonic fibroblasts (MEF). Both types of Ag NPs up-regulated the cell cycle checkpoint protein p53 and DNA damage repair proteins Rad51 and phosphorylated-H2AX expression. Furthermore both of them induced cell death as measured by the annexin V protein expression and MTT assay. Our observations also suggested that the different surface chemistry of Ag NPs induce different DNA damage response: coated Ag NPs exhibited more severe damage than uncoated Ag NPs. The results suggest that polysaccharide coated particles are more individually distributed while agglomeration of the uncoated particles limits the surface area availability and access to membrane bound organelles.     

Exposure to silver nanoparticles induces size- and dose-dependent oxidative stress and cytotoxicity in human colon carcinoma cells

The antimicrobial properties of silver nanoparticles (AgNPs) have made these particles one of the most frequently utilized nanomaterials in consumer products; therefore, a comprehensive understanding of their toxicity is necessary. In particular, information about the cellular uptake and size dependence of AgNPs is insufficient.In this study, we evaluated the size-dependent effects of AgNPs by treating the human LoVo cell line, an intestinal epithelium model, with spherical AgNPs of well-defined sizes (10, 20, 40, 60 and 100 nm). The cellular uptake was visualized by confocal laser scanning microscopy, and various cytotoxicity parameters were analyzed in a size- and dose-dependent manner. In addition, the cellular proteomic response to 20 and 100 nm AgNPs was investigated to increase the understanding of potential mechanisms of action. Our data indicated that cellular uptake and toxicity were regulated by size; smaller particles easily penetrated the cells, and 100 nm particles did not. It was hypothesized that this size-dependent effect resulted from the stimulation of a signaling cascade that generated ROS and inflammatory markers, leading to mitochondrial dysfunction and subsequently inducing apoptosis. By contrast, the cell proliferation, was independent of AgNPs particle size, indicating a differentially regulated, ROS-independent pathway.     

Effects of citrinin on maturation of mouse oocytes, fertilization, and fetal development in vitro and in vivo

The mycotoxin citrinin (CTN), a natural contaminant in foodstuffs and animal feeds, exerts cytotoxic and genotoxic effects on various mammalian cells. An earlier study by our group shows that CTN has cytotoxic effects on mouse embryonic stem cells and blastocysts, and is associated with defects in their subsequent development, both in vitro and in vivo. Here, we further investigate the effects of CTN on oocyte maturation, and subsequent pre- and postimplantation development in vitro and in vivo. CTN induced a significant reduction in the rate of oocyte maturation, fertilization, and in vitro embryo development. Treatment of oocytes with 5 microM CTN during in vitro maturation (IVM) led to increased resorption of postimplantation embryos, and decreased placental and fetal weight. Using an in vivo mouse model, we show that consumption of drinking water containing 5 microM CTN results in decreased oocyte maturation and in vitro fertilization, as well as early embryonic developmental injury. To our knowledge, this is the first study investigating the impact of CTN on maturation of mouse oocytes, fertilization, and sequential embryonic development.     

Antioxidant,antimicrobial and cytotoxic potential of silver nanoparticles synthesized using flavonoid rich alcoholic leaves extract of Reinwardtia indica

The present work discusses the establishment of a green route for the rapid synthesis of silver nanoparticles (AgNPs) using an alcoholic extract of Reinwardtia indica (AERI) leaves which act as a reducing as well as a capping agent. The change in color from yellowish green to dark brown confirmed the synthesis of AgNPs. A characteristic surface plasmon resonance (SPR) band at 436?nm advocated the presence of AgNPs. The synthesis process was optimized using one factor at a time approach where 1.0?mM AgNO₃ concentration, 5?mL 0.4% (v/v) of AER inoculum dose and 30?min of sunlight exposure were found to be the optimum conditions. The synthesized AgNPs was characterized by several characterizing techniques such as HR- TEM, SAED, HR-SEM, EDX, XRD, FTIR and AFM analysis. For evaluation and comparison of AgNPs with AERI used human pathogen E. coli, P. aeurogenosa, S. aeurus and C. albicans for antimicrobial, for cytotoxicity study SiHa cell line at concentration of (10, 50, 100, 250 and 500?µg mL^?1) and for enzymatic assay superoxide dismutase, catalase, malondialdehyde and glutathione peroxidase method were used. The size of nanoparticle in the range of 3–15?nm was confirmed TEM, spherical shape by SEM and crystal lattice nature by XRD. AFM results revealed the 2?D and 3?D pattern of particle scatter nature on the surface. This protocol as simple, rapid, one step, eco-friendly, nontoxic and AgNPs showed strong antimicrobial activity as well as cytotoxic potential in comparison to AERI.     

Assessment of total silver and silver nanoparticle extraction from medical devices

There is concern over the release of silver nanoparticles (AgNPs) from medical devices due to their potential toxicological consequences inside the body. Towards developing the exposure component of a risk assessment model, the purpose of this study was to determine the amount and physical form of silver released from medical devices. Scanning electron microscopy was used to confirm that three of five marketed medical devices contained nanosilver coatings (mean feature sizes 115–341 nm). Aqueous device extracts (water, saline and human plasma) were analyzed with inductively coupled plasma mass spectrometry, ultraviolet–visible spectroscopy, dynamic light scattering, transmission electron microscopy, and nanoparticle tracking analysis. The amount of silver extracted from the devices ranged from 1 × 10⁻¹ to 1 × 10⁶ ng/cm² (conditions ranged from 37 to 50 °C, over one hour to seven days). The results further indicated that one of the five devices (labeled MD1) released significantly more AgNPs than the other devices. This data suggests that some but not all devices that are formulated with nanosilver may release detectable levels of AgNPs upon extraction. Further work is underway to quantitate the proportion of silver released as AgNPs and to incorporate this data into a risk assessment for AgNP exposure from medical devices.     

The acute toxic effects of silver nanoparticles on myocardial transmembrane potential,INa and IK1 channels and heart rhythm in mice

This study focused on the potential toxicity of silver nanoparticles (AgNPs) on cardiac electrophysiology which is rarely investigated. We found that AgNPs (10^?9–10^?6?g/ml) concentration-dependently depolarized the resting potential, diminished the action potential, and finally led to loss of excitability in mice cardiac papillary muscle cells in vitro. In cultured neonatal mice cardiomyocytes, AgNPs (10^?9–10^?7?g/ml) concentration-dependently decreased the Na⁺ currents (I_Na), accelerated the activation, and delayed the inactivation and recovery of Na⁺ channels from inactivation within 5?min. AgNPs at 10^?8?g/ml also rapidly decreased the inwardly rectifying K⁺ currents (I_K1) and delayed the activation of I_K1 channels. Intravenous injection of AgNPs at 3?mg/kg only decreased the heart rate, while at ≥4?mg/kg sequentially induced sinus bradycardia, complete atrio-ventricular conduction block, and cardiac asystole. AgNPs at 10^?10–10^?6?g/ml did not increase reactive oxygen species (ROS) generation and only at 10^?6?g/ml mildly induced lactate dehydrogenase (LDH) release in the cardiomyocytes within 5?min. Endocytosis of AgNPs by cardiomyocytes was not observed within 5?min, but was observed 1?h after exposing to AgNPs. Comparative Ag⁺ (≤0.02% of the AgNPs) could not induce above toxicities. We conclude that AgNPs exert rapid toxic effects on myocardial electrophysiology and induce lethal bradyarrhythmias. These acute toxicities are likely due to direct effects of AgNPs on ion channels at the nano-scale level, but not caused by Ag⁺, ROS, and membrane injury. These findings provide warning to the nanomedical practice using AgNPs.     

Multivariate optimization of mebeverine analysis using molecularly imprinted polymer electrochemical sensor based on silver nanoparticles

Thin film of a moleculary imprinted polymer (MIP) based on electropolymerization method with sensitive and selective binding sites for mebeverine (MEB) was developed. This film was cast on pencil graphite electrode (PGE) by electrochemical polymerization in solution of pyrrole (PY) and template MEB via cyclic voltammetry scans and further electrodeposition of silver nanoparticles (AgNPs). Several parameters controlling the performance of the silver nano particles MIP pencil graphite electrode (AgNPs-MIP-PGE) including concentration of PY(mM) concentration of mebeverine (mM), number of cycles in electropolymerization, scan rate of CV process (mV. s⁻¹), deposition time of AgNPs on to the MIP surface (s), stirring rate of loading solution (rpm), electrode loading time (min), pH of Britton–Robinson Buffer (BRB) solution were examined and optimized using multivariate optimization methods such as Plackett–Burman design (PBD) and central composite design (CCD). Two dynamic linear ranges of concentration for the MIP sensor were obtained as. 1 × 10⁻⁸ to 1 × 10⁻⁶ and 1 × 10⁻⁵ to 1 × 10⁻³ M with the limit of detection (LOD) of 8.6 × 10⁻⁹ M (S/N = 3). The proposed method was successfully intended for the determination of MEB in real samples (serum, capsule). The sensor was showed highly reproducible response (RSD 1.1%) to MEB concentration.     

Silver nanoparticles induce oxidative cell damage in human liver cells through inhibition of reduced glutathione and induction of mitochondria-involved apoptosis

Silver nanoparticles (AgNPs), which have well-known antimicrobial properties, are extensively used in various medical and general applications. Despite the widespread use of AgNPs, relatively few studies have been undertaken to determine the cytotoxic effects of AgNPs exposure. This study investigates possible molecular mechanisms underlying the cytotoxic effects of AgNPs. Here, we show that AgNPs-induced cytotoxicity was higher compared than that observed when AgNO₃ was used as a silver ion source. AgNPs induced reactive oxygen species (ROS) generation and suppression of reduced glutathione (GSH) in human Chang liver cells. ROS generated by AgNPs resulted in damage to various cellular components, DNA breaks, lipid membrane peroxidation, and protein carbonylation. Upon AgNPs exposure, cell viability decreased due to apoptosis, as demonstrated by the formation of apoptotic bodies, sub-G₁ hypodiploid cells, and DNA fragmentation. AgNPs induced a mitochondria-dependent apoptotic pathway via modulation of Bax and Bcl-2 expressions, resulting in the disruption of mitochondrial membrane potential (Δψ_m). Loss of Δψ_m was followed by cytochrome c release from the mitochondria, resulting in the activation of caspases 9 and 3. The apoptotic effect of AgNPs was exerted via the activation of c-Jun NH₂-terminal kinase (JNK) and was abrogated by the JNK-specific inhibitor, SP600125 and siRNA targeting JNK. In summary, the results suggest that AgNPs cause cytotoxicity by oxidative stress-induced apoptosis and damage to cellular components.