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Chlorpromazine (CPZ) is a first-generation neuroleptic with well-established antitumor and antiviral properties. Currently, numerous studies are focused on developing new methods for CPZ delivery; however, the knowledge regarding its conjugates with metal nanoparticles remains limited. The aim of this study was to prepare CPZ conjugates with gold nanoparticles (AuNPs) and evaluate their biological activity on human lymphocytes (HUT-78 and COLO 720L), as well as human (COLO 679) and murine (B16-F0) melanoma cells, in comparison to the effects induced by unconjugated CPZ molecules and AuNPs with well-defined properties. During the treatment of cells with CPZ, AuNPs, and CPZ-AuNP conjugates, changes in mitochondrial activity, membrane integrity, and the secretion of lipid peroxidation mediators were studied using standard biological assays such as MTT, LDH, and MDA assays. It was found that positively charged CPZ-AuNP conjugates more effectively reduced cell viability compared to AuNPs alone. The dose-dependent membrane damage was correlated with oxidative stress resulting from exposure to CPZ-AuNP conjugates. The activity of the conjugates depended on their composition and the size of the AuNPs. It was concluded that conjugating CPZ to AuNPs reduced its biological activity, while the cellular response to the treatment varied depending on the specific cell type.
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The toxicity of silver nanoparticles (AgNPs) depends on their physicochemical properties. The ongoing research aims to develop effective methods for modifying AgNPs using molecules that enable control over the processes induced by nanoparticles in both normal and cancerous cells. Application of amino acid-stabilized nanoparticles appears promising, exhibiting tunable electrokinetic properties. Therefore, this study focused on determining the influence of the surface charge of cysteine (CYS)-stabilized AgNPs on their toxicity towards human normal B (COLO-720L) and T (HUT-78) lymphocyte cell lines. CYS-AgNPs were synthesized via the chemical reduction. Transmission electron microcopy (TEM) imaging revealed that they exhibited a quasi-spherical shape with an average size of 18 ± 3 nm. CYS-AgNPs remained stable under mild acidic (pH 4.0) and alkaline (7.4 and 9.0) conditions, with an isoelectric point observed at pH 5.1. Following a 24 h treatment of lymphocytes with CYS-AgNPs, concentration-dependent alterations in cell morphology were observed. Positively charged CYS-AgNPs notably decreased lymphocyte viability. Furthermore, they exhibited grater genotoxicity and more pronounced disruption of biological membranes compared to negatively charged CYZ-AgNPs. Despite both types of AgNPs interacting similarly with fetal bovine serum (FBS) and showing comparable profiles of silver ion release, the biological assays consistently revealed that the positively charged CYS-AgNPs exerted stronger effects at all investigated cellular levels. Although both types of CYS-AgNPs have the same chemical structure in their stabilizing layers, the pH-induced alterations in their surface charge significantly affect their biological activity.
Assuntos
Cisteína , Nanopartículas Metálicas , Prata , Prata/química , Cisteína/química , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Humanos , Sobrevivência Celular/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linhagem Celular , Propriedades de Superfície , Concentração de Íons de Hidrogênio , Tamanho da PartículaRESUMO
Scientific studies have demonstrated that conjugates of anticancer drugs with metal nanoparticles (MeNPs) lead to a more effective deactivation of tumor cells compared to free drugs. Similarly, it has been established that conjugates of antibiotics with MeNPs exhibit higher biocidal activity against bacteria than their unbound counterparts. However, limited information is available regarding conjugates formed from drugs other than anticancer and antibiotics. Therefore, our research aims to develop synthesis methods for conjugates of chlorpromazine (CPZ), a neuroleptic, with gold nanoparticles (AuNPs). CPZ-AuNP conjugates were prepared through a ligand exchange reaction conducted on the surface of quasi-spherical, negatively charged citrate-stabilized TC-AuNPs with an average size of 55 ± 5 nm. UV-vis spectroscopy was employed to determine the stability range of the conjugates under controlled conditions of pH and ionic strength. Based on electrokinetic measurements, it was observed that the zeta potential of CPZ-AuNP conjugates strongly depends on the amount of CPZ adsorbed on the TC-AuNP surface. Additionally, the conjugates exhibited an isoelectric point at pH 8.8. Surface-enhanced Raman spectroscopy (SERS) and surface-enhanced infrared absorption spectroscopy (SEIRA) were employed to elucidate the adsorption structure of CPZ on TC-AuNPs. The interpretation of the spectra was conducted based on the Raman and FTIR spectra of CPZ, along with calculations performed using Density Functional Theory (DFT). The results indicated that CPZ primarily interacts with the TC-AuNP surface through the angularly oriented phenothiazine ring and the propylene bridge. Furthermore, it was demonstrated that the C-N-C fragment is perpendicular to the surface of the TC-AuNP with which it interacts. The findings from this analysis suggest the potential for further research on the use of these conjugates in biomedical applications.
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Clorpromazina , Ouro , Nanopartículas Metálicas , Espectrofotometria Ultravioleta , Análise Espectral Raman , Ouro/química , Clorpromazina/química , Clorpromazina/farmacologia , Nanopartículas Metálicas/química , Concentração de Íons de Hidrogênio , Antipsicóticos/química , Antipsicóticos/farmacologia , AdsorçãoRESUMO
This study focuses on the adsorption process of L-cysteine (Cys), a sulfur-containing amino acid, onto monolayers of gold nanoparticles (AuNPs) prepared through distinct protocols on mica substrates. Two types of AuNPs were prepared using two different methods: the first employed a physical approach, which combined the Inert Gas Condensation (IGC) technique with the magnetron sputtering method, while the second utilized a chemical method involving the reduction of tetrachloroauric acid with trisodium citrate (TC). The characterization of AuNPs was performed using transmission electron microscopy (TEM) and atomic force microscopy (AFM), of up to 5 ± 1.3 nm for bare AuNPs obtained through vacuum techniques, and up to 12 ± 5 nm for negatively charged, citrate-stabilized TCAuNPs(-). The application of spectroscopic techniques based on the surface-enhanced effects allows for describing the adsorption process in both micro- and nanoscale systems: Cys/bare AuNPs and Cys/ TCAuNPs(-). The commonly used surface-enhanced Raman spectroscopy (SERS) technique provided insights into adsorption behaviours at the microscale level. In the case of TCAuNPs(-), an interaction involving the lone electron pair of sulfur (S) atom and metal surface, while on the bare AuNPs, S is adsorbed on the surface, but the cleavage of the SH group is not discernible. Nanoscale analysis was complemented using AFM combined with the surface-enhanced infrared absorption spectroscopy (AFM-SEIRA) technique. AFM-SEIRA map indicated the formation of hot spot which were predominantly located between aggregated TCAuNPs(-) and on specific NPs surfaces (area between NPs and gold-coated tip). Results from the SERS and AFM-SEIRA techniques were in good agreement, underscoring the comprehensive understanding achieved through the chosen experimental approach regarding the Cys interactions with layers of AuNPs.
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The study investigated the effects of different silver nanoparticles (AgNPs) on the soil microbiome and wheat growth. For comparison purposes, a commercial fungicide and silver nitrate (AgNO3) were used. The results revealed three distinct groups of nanoparticles based on their impacts. Small-size AgNPs (10 nm) with a negative charge, as well as fungicide had limited effects on the microbiome, similar to the no-treatment control. Bigger in size (30-60 nm) and a negative charge AgNPs showed the most beneficial effects on soil microbiota shifts. These AgNPs increased the abundance of bacteria with beneficial traits such as nitrogen-fixing, urease, protease, and lignin degradation bacteria. The third type of AgNPs had a positive charge of nanostructure and influenced specific microbial populations, increasing the abundance of anaerobic and autotrophic groups of microorganisms, which could be assessed as a harmful shift for plants growth promotions and was similar to the AgNO3 treatment. Overall, the study emphasized the potential of AgNPs in agriculture not only as biocidal. The conducted study proved that AgNPs with bigger size/negative charge, used in low concentration can have a surprisingly stimulating effect on the positive characteristics of the rhizosphere microbiome. Moreover, the surface charge of AgNPs is a significant factor affecting microbial activity of wheat rhizosphere soil, which in this treatment is significantly similar to the AgNO3 treatment.
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Fungicidas Industriais , Nanopartículas Metálicas , Microbiota , Rizosfera , Triticum , Nanopartículas Metálicas/toxicidade , Prata , Solo/química , Microbiologia do SoloRESUMO
Vimentin, a protein that builds part of the cytoskeleton and is involved in many aspects of cellular function, was recently identified as a cell surface attachment site for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The present study investigated the physicochemical nature of the binding between the SARS-CoV-2 S1 glycoprotein receptor binding domain (S1 RBD) and human vimentin using atomic force microscopy and a quartz crystal microbalance. The molecular interactions of S1 RBD and vimentin proteins were quantified using vimentin monolayers attached to the cleaved mica or a gold microbalance sensor as well as in its native extracellular form present on the live cell surface. The presence of specific interactions between vimentin and S1 RBD was also confirmed using in silico studies. This work provides new evidence that cell-surface vimentin (CSV) functions as a site for SARS-CoV-2 virus attachment and is involved in the pathogenesis of Covid-19, providing a potential target for therapeutic countermeasures.
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COVID-19 , Humanos , SARS-CoV-2/metabolismo , Glicoproteína da Espícula de Coronavírus/química , Vimentina/metabolismo , Enzima de Conversão de Angiotensina 2/metabolismo , Ligação ProteicaRESUMO
In this study for the first time, surface-enhanced Raman spectroscopy (SERS) and tip-enhanced infrared (TEIRA) nanospectrocopy together with a quartz crystal microbalance (QCM) are postulated as powerful tools for comprehensive qualitative and quantitative analyses of drug/metal nanocarrier conjugates. The development of efficient drug/carrier systems requires that the stability of the drug/carrier connection be estimated and the number of drug molecules immobilized on the carrier surface be determined. Thus, such a characterization study is highly desirable. Here, the SERS technique was applied to identify how erlotinib, a drug applied in non-small cell lung cancer (NSCLC) therapy, interacts with silver nanoparticles (AgNPs) that are considered as drug carriers. These investigations indicate that in the erlotinib/AgNP suspension, the drug strongly connects with the NPs mainly through the phenylacetylene moiety. The QCM was used to prepare an AgNP monolayer with a monitored degree of coverage and to perform controlled erlotinib adsorption as a next step. The results indicate that the drug forms a stable layer on the AgNP monolayer and also show the amount of the erlotinib molecules which underwent immobilization on the metal nanosurface. Simultaneously, it was identified how the erlotinib layer adsorbs on the AgNP monolayer using TEIRA nanospectroscopy with ultra-high spatial resolution. The obtained results show that the phenylacetylene, ethoxy, and methoxy moieties are mainly responsible for the drug/AgNP monolayer connection. Additionally, the performed studies also try to explain the surface-enhanced phenomena that occur during the TEIRA experiments and attempt to prove the statement that the "tip-enhanced" effect plays a crucial role in the detection of the thin erlotinib layer deposited on the AgNP monolayer.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Nanopartículas Metálicas , Humanos , Técnicas de Microbalança de Cristal de Quartzo , Cloridrato de Erlotinib , Prata/química , Nanopartículas Metálicas/química , Adsorção , Análise Espectral RamanRESUMO
The aim of the study was to evaluate the effect of two types of negatively charged quasi-spherical silver nanoparticles (AgNPs) at concentrations of 10, 20 and 30mgL-1 and silver ions at a concentration of 30mgL-1 on the growth, selected physiological aspects and yielding of wheat (Triticum aestivum L.) cv. Tybalt, and on plant resistance to seedling blight. Seed germination, α-amylase activity in seeds, morphology and infestation of seedlings by pathogens were assessed in a hydroponic treatment. Growth rate, PSII efficiency, heading and yield of the same plants were then analysed in pot culture. Results showed that the AgNPs and silver ions had a negative effect on roots, but reduced seedling blight and improved leaf area compared to the control. In addition, the AgNPs reduced with sodium borohydride in the presence of trisodium citrate at concentrations of 10 and 20mgL-1 stimulated germination, α-amylase activity and shoot length, which was not observed in the case of silver ions and the AgNPs reduced with sodium hypophosphite in the presence of sodium hexametaphosphate. In a pot experiment, the AgNPs improved plant growth, PSII efficiency, accelerated heading and increased yield-related parameters compared with the control. Results revealed the interaction strength in the following order: TCSB-AgNPs>SHSH-AgNPs>silver ions. TCSB-AgNPs in the lowest concentration had the most favourable effect, indicating their great potential for use in improving wheat cultivation.
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Nanopartículas Metálicas , Plântula , Germinação , Triticum , Prata/farmacologia , Plantas , alfa-Amilases/farmacologiaRESUMO
The microbiome's significance in chronic rhinosinusitis (CRS) is unclear. Antimicrobials are recommended in acute exacerbations of the disease (AECRS). Increasing rates of antibiotic resistance have stimulated research on alternative therapeutic options, including silver nanoparticles (AgNPs). However, there are concerns regarding the safety of silver administration. The aim of this study was to assess the biological activity of tannic acid-prepared AgNPs (TA-AgNPs) towards sinonasal pathogens and nasal epithelial cells (HNEpC). The minimal inhibitory concentration (MIC) for pathogens isolated from patients with AECRS was approximated using the well diffusion method. The cytotoxicity of TA-AgNPswas evaluated using an MTT assay and trypan blue exclusion. A total of 48 clinical isolates and 4 reference strains were included in the study (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Klebsiellaoxytoca, Acinetobacter baumannii, Serratia marcescens, Enterobacter cloacae). The results of the studies revealed that the MIC values differed between isolates, even within the same species. All the isolates were sensitive to TA-AgNPs in concentrations non-toxic to human cells during 24 h exposition. However, 48 h exposure to TA-AgNPs increased toxicity to HNEpC, narrowing their therapeutic window and enabling 19% of pathogens to resist the TA-AgNPs' biocidal action. It was concluded that TA-AgNPs are non-toxic for the investigated eukaryotic cells after short-term exposure and effective against most pathogens isolated from patients with AECRS, but sensitivity testing may be necessary before application.
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Nanopartículas Metálicas , Prata , Humanos , Prata/farmacologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Taninos/farmacologia , Escherichia coliRESUMO
In this work, we have assessed the exposure of soil bacteria from potato monoculture to three types of silver nanoparticles (AgNPs) as well as silver ions (Ag+ ions) delivered in the form of silver nitrate and a commercially available fungicide. The diversity of the soil microbial community, enzymatic activity, and carbon source utilization were evaluated. It was found that only the fungicide significantly limited the abundance and activity of soil bacteria. Silver ions significantly reduced bacterial metabolic activity. In turn, one type of AgNPs prepared with the use of tannic acid (TA) increased bacterial load and activity. There was found in all AgNPs treated soils (1) a greater proportion of all types of persistent bacteria, i.e., Bacillus, Paenibacillus, and Clostridium; (2) a visible decrease in the proportion of Nocardioides, Arthrobacter, and Candidatus Solibacter; (3) almost complete depletion of Pseudomonas; (4) increase in the number of low-frequency taxa and decrease in dominant taxa compared to the control soil. Despite the general trend of qualitative changes in the bacterial community, it was found that the differences in the chemical structure of the AgNP stabilizing layers had a significant impact on the specific metabolic activity resulting from qualitative changes in the microbiome.
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Fungicidas Industriais , Nanopartículas Metálicas , Microbiota , Solo/química , Microbiologia do Solo , Nanopartículas Metálicas/química , Prata , Bactérias , ÍonsRESUMO
Silver nanoparticles (AgNPs) exhibit unusual biocidal properties thanks to which they find a wide range of applications in diverse fields of science and industry. Numerous research studies have been devoted to the bactericidal properties of AgNPs while less attention has been focused on their fungicidal activity. Our studies were therefore oriented toward determining the impact of AgNPs characterized by different physicochemical properties on Fusarium avenaceum and Fusarium equiseti. The main hypothesis assumed that the fungicidal properties of AgNPs characterized by comparable morphology can be shaped by stabilizing agent molecules adsorbed on nanoparticle surfaces. Two types of AgNPs were prepared by the reduction of silver ions with sodium borohydride (SB) in the presence of trisodium citrate (TC) or cysteamine hydrochloride (CH). Both types of AgNPs exhibited a quasi-spherical shape. Citrate-stabilized AgNPs (TCSB-AgNPs) of an average size of 15 ± 4 nm were negatively charged. Smaller (12 ± 4 nm), cysteamine-capped AgNPs (CHSB-AgNPs) were characterized by a positive surface charge and higher silver ion release profile. The phytopathogens were exposed to the AgNPs in three doses equal to 2.5, 5 and 10 mg L-1 over 24 and 240 h. Additionally, the impact of silver ions delivered in the form of silver nitrate and the stabilizing agents of AgNPs on the fungi was also investigated. The response of phytopathogens to these treatments was evaluated by determining mycelial growth, sporulation and changes in the cell morphology. The results of our studies showed that CHSB-AgNPs, especially at a concentration of 10 mg L-1, strongly limited the vegetative mycelium growth of both species for short and long treatment times. The cell imaging revealed that CHSB-AgNPs damaged the conidia membranes and penetrated into the cells, while TCSB-AgNPs were deposited on their surface. The fungistatic (lethal) effect was demonstrated only for silver ions at the highest concentration for the F. equiseti species in the 240 h treatment. The number of spores of both Fusarium species was significantly reduced independently of the type of silver compounds used. Generally, it was found that the positively charged CHSB-AgNPs were more fungicidal than negatively charged TCSB-AgNPs. Thereby, it was established that the stabilizing agents of AgNPs and surface charge play a crucial role in the shaping of their fungicidal properties.
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Antifúngicos , Nanopartículas Metálicas , Antifúngicos/farmacologia , Nanopartículas Metálicas/química , Excipientes , Prata/farmacologia , Propriedades de Superfície , ÍonsRESUMO
DNA double-strand breaks (DSBs) are typical DNA lesions that can lead to cell death, translocations, and cancer-driving mutations. The repair process of DSBs is crucial to the maintenance of genomic integrity in all forms of life. However, the limitations of sensitivity and special resolution of analytical techniques make it difficult to investigate the local effects of chemotherapeutic drugs on DNA molecular structure. In this work, we exposed DNA to the anticancer antibiotic bleomycin (BLM), a damaging factor known to induce DSBs. We applied a multimodal approach combining (i) atomic force microscopy (AFM) for direct visualization of DSBs, (ii) surface-enhanced Raman spectroscopy (SERS) to monitor local conformational transitions induced by DSBs, and (iii) multivariate statistical analysis to correlate the AFM and SERS results. On the basis of SERS results, we identified that bands at 1050 cm-1 and 730 cm-1 associated with backbone and nucleobase vibrations shifted and changed their intensities, indicating conformational modifications and strand ruptures. Based on averaged SERS spectra, the PLS regressions for the number of DSBs caused by corresponding molar concentrations of bleomycin were calculated. The strong correlation (R2 = 0.92 for LV = 2) between the predicted and observed number of DSBs indicates, that the model can not only predict the number of DSBs from the spectra but also detect the spectroscopic markers of DNA damage and the associated conformational changes.
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Quebras de DNA de Cadeia Dupla , Reparo do DNA , Bleomicina/farmacologia , DNA/química , Dano ao DNARESUMO
Silver nanoparticles (AgNPs) prepared and stabilized by diverse biologically active substances seem to be especially useful in diverse biological and medical applications. The combination of AgNPs with bioactive substances, such as antioxidants, can lead to the development of new systems of desired anticancer properties. In this research, AgNPs were prepared with the use of diverse antioxidant combinations including gallic acid (GA), (-)-epicatechin-3-gallate (EGCG), and caffeine (CAF). The insightful physicochemical characteristic revealed that each type of AgNPs exhibited spherical shape, comparable size distribution and negative surface charge. Surface-enhanced Raman spectroscopy (SERS) delivered the information about the chemistry of AgNP stabilizing layers, which turned out to be a crucial factor tuning toxicity of AgNPs toward murine B16 melanoma cells (B16-F0) and human skin melanoma (COLO 679) cells. EGCGAgNPs were the most cytotoxic among all the investigated AgNPs. They strongly reduced the activity of mitochondria, damaged cell membrane integrity, and penetrated inside the cells causing DNA damage. In turn, the toxicity of GAAgNPs strongly manifested via the induction of oxidative stress in the cells. It was found that CAFGAAgNPs exhibited the lowest toxicity toward the melanoma cells, which proved that a proper combination of antioxidants enable to prepare AgNPs of differentiated toxicity. It was established that human skin melanoma cells were significantly more sensitive to AgNPs than the murine melanoma cells.
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Antineoplásicos , Melanoma , Nanopartículas Metálicas , Animais , Antineoplásicos/farmacologia , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Humanos , Melanoma/tratamento farmacológico , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Camundongos , Prata/química , Prata/toxicidade , Análise Espectral RamanRESUMO
The biocidal properties of silver nanoparticles (AgNPs) prepared with the use of biologically active compounds seem to be especially significant for biological and medical application. Therefore, the aim of this research was to determine and compare the antibacterial and fungicidal properties of fifteen types of AgNPs. The main hypothesis was that the biological activity of AgNPs characterized by comparable size distributions, shapes, and ion release profiles is dependent on the properties of stabilizing agent molecules adsorbed on their surfaces. Escherichia coli and Staphylococcus aureus were selected as models of two types of bacterial cells. Candida albicans was selected for the research as a representative type of eukaryotic microorganism. The conducted studies reveal that larger AgNPs can be more biocidal than smaller ones. It was found that positively charged arginine-stabilized AgNPs (ARGSBAgNPs) were the most biocidal among all studied nanoparticles. The strongest fungicidal properties were detected for negatively charged EGCGAgNPs obtained using (-)-epigallocatechin gallate (EGCG). It was concluded that, by applying a specific stabilizing agent, one can tune the selectivity of AgNP toxicity towards desired pathogens. It was established that E. coli was more sensitive to AgNP exposure than S. aureus regardless of AgNP size and surface properties.
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Antibacterianos/farmacologia , Antifúngicos/farmacologia , Excipientes/farmacologia , Prata/farmacologia , Antioxidantes/farmacologia , Bactérias/efeitos dos fármacos , Boroidretos/farmacologia , Citratos/farmacologia , Fungos/efeitos dos fármacos , Glucose/farmacologia , Testes de Sensibilidade Microbiana , Propriedades de SuperfícieRESUMO
The properties of silver nanoparticles (AgNPs) synthesized using compounds exhibiting biological activity seem to constitute an interesting issue worthy of examination. In these studies, two types of AgNPs were synthesized by a chemical reduction method using well-known antioxidants: gallic acid (GA) and ascorbic acid (AA). Transmission electron microscopy (TEM) and atomic force microscopy (AFM) revealed that the AgNPs were spherical. The average size was equal to 26 ± 6 nm and 20 ± 7 nm in the case of ascorbic acid-silver nanoparticles (AAgNPs) and gallic acid-silver nanoparticles (GAAgNPs), respectively. Surface-enhanced Raman spectroscopy (SERS) confirmed that the AgNPs were not stabilized by pure forms of applied antioxidants. Changes in mitochondrial activity and secretion of inflammatory and apoptosis mediators after the exposure of human promyelocytic (HL-60) and histiocytic lymphoma (U-937) cells to the AgNPs were studied to determine the impact of stabilizing layers on nanoparticle toxicity. The GAAgNPs were found to be more toxic for the cells than the AAgNPs. Their toxicity was manifested by a strong reduction in mitochondrial activity and induction of the secretion of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and caspase-9. The addition of pure antioxidants to the AgNP suspensions was found to influence their toxicity. There was a significant positive effect in the case of the mixture of AA with AAgNPs and GA with GAAgNPs. The results obtained suggest that the presence of stabilizing agents adsorbed on the surface of AgNPs is the main factor in shaping their toxicity. Nevertheless, the toxic effect can be also tuned by the introduction of free antioxidant molecules to the AgNP suspensions.
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Antioxidantes/metabolismo , Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Células HL-60 , Humanos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Prata/química , Análise Espectral Raman , Células U937RESUMO
In this work, we clearly focus on the comparative cytotoxicity investigations of several protein-stabilized gold nanoclusters (Au NCs) towards lymphocytes B (COLO-720â¯L) and lymphocytes T (HUT-78) cells. For synthesis, the one-pot template-assisted method was carried out using lysozyme (LYZ), human (HSA) and bovine (BSA) serum albumins, and gamma globulin (γG) as stabilizing agents. Regardless of the type of proteins, all synthesized Au NCs possess intense red emission (λem â¼ 650â¯nm) and have similar size of a metal core (ca. 1.4â¯nm) with negative surface charge at pHâ¯=â¯7.4. During the treatment of cells with clusters, changes in mitochondrial activity, membrane integrity, secretion of inflammatory and apoptosis mediators of the lymphocytes were studied to determine the potential effect of protein layers on the toxicity of clusters. It was found that γG-Au NCs induced the highest disorders in mitochondrial activity, but the influence of other NCs on the cell viability was minor. Besides, all Au NCs caused oxidative stress by peroxidation of membrane lipids. The secretion of malonic dialdehyde (MDA) was enhanced by LYZ- and γG-Au NCs. Apart from LYZ-Au NCs, the clusters did not exhibit strong proinflammatory and apoptotic properties. The enhanced secretion of tumor necrosis factor (TNF-α) by lymphocytes B, in comparison to control, was independent of the clusters type. Despite the lack of significant influence of the Au NCs on the viability of the lymphocytes, they can stimulate undesirable cellular processes, which clearly depends on the stabilizing proteins.
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Ouro , Nanopartículas Metálicas , Animais , Bovinos , Sobrevivência Celular , Corantes , Corantes Fluorescentes , Humanos , Linfócitos , Nanopartículas Metálicas/toxicidadeRESUMO
In this study, surface - enhanced Raman spectroscopy (SERS) was applied at the first time for estimation of how pH, temperature, and nanoparticle (NP) stabilizer affect an adsorption behavior of erlotinib (drug approved in a non-small cell lung cancer therapy) onto citrate-stabilized silver nanoparticles (AgNPs). Novel approach to improve cancer therapy assumes application of NPs as an efficient drug delivery system. This strategy requires designing stable drug/nanocarrier conjugates that can effectively interact in the target site. It is also important to perform deeply characterization of a drug orientation on the potential carrier surface and estimation how stable the appeared interaction is. Performed analysis, indicates that pH, temperature, presence of NP stabilizers, and time of incubation have an influence on the occurring adsorption geometry of the drug. However, the observed erlotinib/AgNP interaction remains stable regardless of the applied conditions. These considerations were supported by insightful physicochemical characteristics of the AgNPs and the erlotinib/AgNP conjugates by conducting transmission electron microscopy (TEM) imaging, determination of colloid stability conducted with the use of dynamic light scattering technique (DLS) and measurements of electrophoretic mobility. Such complex approach allows a better understanding of the stability of the erlotinib/AgNP conjugates and provides information how the investigated interaction is affected by the induced perturbations.
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Antineoplásicos/química , Cloridrato de Erlotinib/química , Nanopartículas Metálicas/química , Prata/química , Ácido Cítrico/química , Concentração de Íons de Hidrogênio , Modelos Moleculares , Análise Espectral Raman , TemperaturaRESUMO
The kinetics of positively charged gold nanoparticle self-assembly on oxidized silicon substrates (wafers) under diffusion-controlled transport was studied using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The latter technique allowed the roughness parameters of the monolayer (root mean square) to be determined as a function of the particle coverage. These results were adequately interpreted in terms of a theoretical model developed for surfaces covered by features of spherical shape considering the tip convolution effect. The stability and the electrokinetic characteristics (zeta potential) of the monolayers were also acquired using streaming potential measurements. It was shown that the inversion of the negative zeta potential of the bare substrate (overcharging) occurs at the particle coverage equal to 0.15, and for larger coverages positive zeta potential values were asymptotically attained. Additionally, the desorption kinetics of the particles was investigated by the streaming potential method, which confirmed the stability of the monolayers for a broad range of pHs. It was argued that these results enable to develop an efficient method for the preparation of gold sensors exhibiting a well-controlled surface roughness and electrostatic charge comprising both negative and positive values.
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Silver and titanium dioxide nanoparticles (AgNPs and TiO2NPs) are highly useful, but they are also a significant reason for concern as they exert toxicity. The goal of research was to assess the role of three kinds of NPs in concentrations of 100 mg L-1 on early growth plants (wheat, flax) and bacterial community in rhizoplane. Titanium (IV) oxide anatase (TiO2NPs1) and titanium (IV) oxide nanopowder (TiO2NPs2) are commercial products. A suspension of AgNPs was prepared via a procedure of reduction with tannic acid. The response of Monocot and Dicot growth form plants to the tested NPs was different. Germination and seedling growth of wheat treated with TiO2NPs1 was better. The response of flax to NPs was noted as an increase of chlorophyll content. The bacterial community in wheat rhizoplane was not significantly modified, but there was a declining trend. In turn, a difference in the surface charge of NPs had an influence on the total bacterial community in Dicot rhizoplane. Positively charged TiO2NPs2 significantly decreased the quantity of total bacteria in contrast to negatively charged AgNPs and TiO2NPs1 which increased it. A qualitative analysis did not confirm the influence of the surface charge of NPs on an increase/decrease in the quantity of Pseudomonas and Bacillus bacteria, but did show that there was no toxicity of the tested NPs to the plant growth-promoting bacteria community. The rhizoplane microbiome was dependent on the species of plant, and the bacteria found in the communities are sensitive to NPs to a varying degree.
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Linho/efeitos dos fármacos , Nanopartículas/toxicidade , Prata/toxicidade , Titânio/toxicidade , Triticum/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Linho/crescimento & desenvolvimento , Germinação/efeitos dos fármacos , Microbiota/efeitos dos fármacos , Nanopartículas/química , Rizosfera , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Microbiologia do Solo , Poluentes do Solo/toxicidade , Especificidade da Espécie , Triticum/crescimento & desenvolvimentoRESUMO
Formation of positively charged gold nanoparticle layers on polystyrene microparticles (PSMs600) was studied using the electrokinetic and the concentration depletion methods based on atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging. Primarily, the dependence of electrophoretic mobility of microparticles on the gold nanoparticle concentration in the suspension was measured. These results were quantitatively interpreted in terms of the three-dimensional electrokinetic model. This allowed to derive a formula for calculating the coverage of nanoparticles under in situ conditions whose validity was confirmed by direct SEM imaging of deposited gold nanoparticles (AuNPs). Additionally, the maximum coverage of gold nanoparticles for various ionic strengths was determined using a concentration depletion method based on AFM imaging of residual particles deposited on the silica substrate. The maximum coverage increased with ionic strength attaining a value of 0.35 for the ionic strength of 3 × 10-3 M. This effect was attributed to the decreasing range of lateral electrostatic interactions among deposited particles. The electrokinetic properties of the gold nanoparticle layers were also evaluated in pH cycling experiments that confirmed their stability. Beyond significance to basic science, the new data acquired in this work confirm the feasibility of preparing gold nanoparticle layers on polymer microparticles characterized by a controlled structure, coverage, and electrokinetic properties.