RESUMEN
This study investigated the effect of AgNPs and AgNO3, at concentrations equivalent, on the production of primary and secondary metabolites on transgenic soybean plants through an NMR-based metabolomics. The plants were cultivated in a germination chamber following three different treatments: T0 (addition of water), T1 (addition of AgNPs), and T2 (addition of AgNO3). Physiological characteristics, anatomical analyses through microscopic structures, and metabolic profile studies were carried out to establish the effect of abiotic stress on these parameters in soybean plants. Analysis of the 1H NMR spectra revealed the presence of amino acids, organic acids, sugars, and polyphenols. The metabolic profiles of plants with AgNP and AgNO3 were qualitatively similar to the metabolic profile of the control group, suggesting that the application of silver does not affect secondary metabolites. From the PCA, it was possible to differentiate the three treatments applied, mainly based on the content of fatty acids, pinitol, choline, and betaine.
Asunto(s)
Glycine max , Espectroscopía de Resonancia Magnética , Metabolómica , Nanopartículas del Metal , Plantas Modificadas Genéticamente , Plata , Glycine max/metabolismo , Glycine max/genética , Glycine max/química , Glycine max/efectos de los fármacos , Glycine max/crecimiento & desarrollo , Plata/metabolismo , Plata/química , Nanopartículas del Metal/química , Espectroscopía de Resonancia Magnética/métodos , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Plantas Modificadas Genéticamente/química , Aminoácidos/metabolismo , Ácidos Grasos/metabolismo , Ácidos Grasos/químicaRESUMEN
The interaction between silver nanoparticles (AgNPs) and molecules producing coronas plays a key role in cytotoxicity mechanisms. Once adsorbed coronas determine the destiny of nanomaterials in vivo, their effective deployment in the biomedical field requires a comprehensive understanding of the dynamic interactions of biomolecules with nanoparticles. In this work, we characterized 40 nm AgNPs in three different nutritional cell media at different molar concentrations and incubation times to study the binding mechanism of molecules on surface nanoparticles. In addition, their cytotoxic effects have been studied in three cell lineages used as tissue regeneration models: FN1, HUV-EC-C, RAW 264.7. According to the data, when biomolecules from DMEM medium were in contact with AgNPs, agglomeration and precipitation occurred. However, FBS medium proteins indicated the formation of coronas over the nanoparticles. Nonetheless, little adsorption of molecules around the nanoparticles was observed when compared to DMEM supplemented with 10% FBS. These findings indicate that when nanoparticles and bioproteins from supplemented media interact, inorganic salts from DMEM contribute to produce large bio-coronas, the size of which varies with the concentration and time. The static quenching mechanism was shown to be responsible for the fluorescence quenching of the bioprotein aggregates on the AgNPs surface. The calculated bioprotein-nanoparticle surface binding constants were on the order of 105 M-1 at 37 °C, with hydrophobic interactions driven by enthalpy and entropy playing a role, as confirmed by thermodynamic analysis. Cytotoxicity data showed a systematic degrowth in the viable cell population as the number of nanoparticles increased and the diameter of coronas decreased. Cytotoxic intervals associated with half decrease of cell population were established for AgNPs molar concentration of 75 µM for 24 h and 50 µM for 48 h. In summary, through the cytotoxicity mechanism of bio-coronas we are able to manipulate cells' expansion rates to promote specific processes, such inflammatory mechanisms, at different time instants.
RESUMEN
Metallic nanoparticles, such as gold (Au, Z = 79) and silver (Ag, Z = 47) nanoparticles (AuNPs and AgNPs, respectively), possess strong surface plasmonic resonance (SPR) and high atomic number, which makes them ideal candidates for enhancing dosimeter sensitivity. In this study, we have inserted different mass percentages (from 0 to 0.015 wt%) of AuNPs into a gelatinous Fricke-xylenol-orange (FXO-f) gel matrix and irradiated it with doses ranging from 2 to 32 Gy, using a source of x-ray of low energy with an effective energy of 42 keV. Optical absorption increased significantly; sensitivity gains of up to 50% were achieved for the FXO-f gel matrix containing 0.011 wt% AuNPs. To elucidate the mechanism underlying this increased sensitivity, we also evaluated FXO-f gel matrixes containing AgNPs. AgNPs insertion into the FXO-f gel matrix did not enhance sensitivity, which suggested that the AgNPs plasmonic absorption band and the FXO-f gel matrix absorption band at 441 nm overlapped, to increase absorption even after the gel matrix was irradiated. To visualize the dose distribution, we recorded optical tomography and acquired 3D reconstruction maps. In addition, we analyzed the dose enhancement factor (DEF) by using magnetic resonance images. AuNPs insertion into the FXO-f gel matrix resulted in a DEF gain of 1.37, associated with the photoelectric effect originating from the increased number of free radicals.
Asunto(s)
Oro , Nanopartículas del Metal , Radiometría/métodos , Imagen por Resonancia MagnéticaRESUMEN
Chitosan (CHI) and carboxymethyl cellulose (CMC) are naturally sourced materials with excellent physical, chemical, and biological properties, which make them a promising tool for the development of different medical devices. In this research, CHI-CMC wound dressings were manufactured, by using different colloidal suspensions of silver nanoparticles (AgNPs) synthesized from the ligninolytic fungus Anamorphous Bjerkandera sp. R1, called CS and SN. Transmission electron microscopy (TEM), UV-Vis spectroscopy, and dynamic light scattering (DLS) analysis were used to characterize AgNPs. The wound dressings were characterized, by scanning electron microscopy (SEM), optical microscopy and their mechanical, antimicrobial, and biological properties were evaluated. The results of the different characterizations revealed the formation of spherical AgNPs with a mean size between 10 and 70 nm for the different mixtures worked. The mechanical properties of CHI-CMS-AgNPs doped with CS and SN suspensions showed superior mechanical properties with respect to CHI-CMC wound dressings. Compared to the latter, CHI-CMC-AgNPs wound dressings yielded better antibacterial activity against the pathogen Escherichia coli. In biological assays, it was observed that manufactured CHI-CMC-AgNPs wound dressings were not toxic when in contact with human skin fibroblasts (Detroit). This study, then, suggests that this type of wound dressings with a chitosan matrix and carboxymethyl cellulose doped with biologically synthesized nanoparticles from the fungus Bjerkandera sp., may be an ideal alternative for the manufacture of new wound dressings.
RESUMEN
Salmonella spp. is an important causal agent of salmonellosis in humans. Controlling Salmonella spp. in eggs is important as the bacterium passes through the shell to an embryo and remains in the terrain. Disinfection is usually performed by using several sanitizers. However, novel, more efficient ways of controlling this agent have been studied with advances in nanotechnology, including nanoparticles. Preliminary studies of nanoparticles have shown they are successful in controlling such microorganisms. Standardizing the ideal concentration of this nanocomposite is fundamental for optimum efficiency in the control of Salmonella spp. In this study, eggs from commercial laying chickens were purchased from local trade and treated in laboratory with silver and zinc nanoparticles in different concentrations. Biofilm was formed 24 hours after that; then, the eggs were washed for the removal of free bacteria. Conventional microbiology was performed to isolate Salmonella spp., and PCR was performed to identify colonies. The effectiveness of using nanocomposite of silver oxide with silver-doped zinc oxide (ZnO:Ag-AgO) was evaluated in different concentrations to prevent the formation of eggshell biofilms.
As Salmonellas spp. são importantes agentes causadores de salmonelose em humanos. O controle da Salmonella spp. é importante, pois a bactéria ultrapassa a barreira da casca atingindo o embrião e infecta lotes de aves que podem levar a infecção ao ser humano. A desinfecção costuma ser feita por vários sanitizantes; porém, com os avanços da nanotecnologia, formas novas e mais eficientes de controle desse agente estão sendo estudadas, como as nanopartículas. Estudos preliminares dessas nanopartículas têm mostrado o sucesso de seu uso no controle de microrganismos. A padronização da concentração ideal de uso desse nanocomposto é fundamental para a máxima eficiência no controle de Salmonella spp. Ovos vermelhos oriundos de postura comercial foram comprados no comércio local e tratados em laboratório com as nanopartículas em diferentes concentrações; após 24 horas, formaram o biolfilme. Os ovos foram lavados para a retirada das bactérias livres. Realizaram-se exame microbiológico convencional, para isolamento de Salmonella spp., e PCR, para identificação das colônias. O objetivo deste artigo foi avaliar a eficácia da utilização de nanocompostos de óxido de prata com óxido de zinco dopado com óxido de prata (ZnO: Ag-Ago) em diferentes concentrações na prevenção da formação de biofilmes na casca dos ovos.
Asunto(s)
Biopelículas , Cáscara de Huevo/microbiología , Compuestos de Plata , Nanopartículas del Metal , Huevos/análisis , Salmonella/aislamiento & purificación , Óxido de Zinc/administración & dosificación , Pollos , Infecciones por Salmonella/prevención & control , Nanocompuestos , Reacción en Cadena de la PolimerasaRESUMEN
Salmonella spp. is an important causal agent of salmonellosis in humans. Controlling Salmonella spp. in eggs is important as the bacterium passes through the shell to an embryo and remains in the terrain. Disinfection is usually performed by using several sanitizers. However, novel, more efficient ways of controlling this agent have been studied with advances in nanotechnology, including nanoparticles. Preliminary studies of nanoparticles have shown they are successful in controlling such microorganisms. Standardizing the ideal concentration of this nanocomposite is fundamental for optimum efficiency in the control of Salmonella spp. In this study, eggs from commercial laying chickens were purchased from local trade and treated in laboratory with silver and zinc nanoparticles in different concentrations. Biofilm was formed 24 hours after that; then, the eggs were washed for the removal of free bacteria. Conventional microbiology was performed to isolate Salmonella spp., and PCR was performed to identify colonies. The effectiveness of using nanocomposite of silver oxide with silver-doped zinc oxide (ZnO:Ag-AgO) was evaluated in different concentrations to prevent the formation of eggshell biofilms.(AU)
As Salmonellas spp. são importantes agentes causadores de salmonelose em humanos. O controle da Salmonella spp. é importante, pois a bactéria ultrapassa a barreira da casca atingindo o embrião e infecta lotes de aves que podem levar a infecção ao ser humano. A desinfecção costuma ser feita por vários sanitizantes; porém, com os avanços da nanotecnologia, formas novas e mais eficientes de controle desse agente estão sendo estudadas, como as nanopartículas. Estudos preliminares dessas nanopartículas têm mostrado o sucesso de seu uso no controle de microrganismos. A padronização da concentração ideal de uso desse nanocomposto é fundamental para a máxima eficiência no controle de Salmonella spp. Ovos vermelhos oriundos de postura comercial foram comprados no comércio local e tratados em laboratório com as nanopartículas em diferentes concentrações; após 24 horas, formaram o biolfilme. Os ovos foram lavados para a retirada das bactérias livres. Realizaram-se exame microbiológico convencional, para isolamento de Salmonella spp., e PCR, para identificação das colônias. O objetivo deste artigo foi avaliar a eficácia da utilização de nanocompostos de óxido de prata com óxido de zinco dopado com óxido de prata (ZnO: Ag-Ago) em diferentes concentrações na prevenção da formação de biofilmes na casca dos ovos.(AU)
Asunto(s)
Huevos/análisis , Biopelículas , Nanopartículas del Metal , Salmonella/aislamiento & purificación , Óxido de Zinc/administración & dosificación , Compuestos de Plata , Cáscara de Huevo/microbiología , Nanocompuestos , Pollos , Infecciones por Salmonella/prevención & control , Reacción en Cadena de la PolimerasaRESUMEN
Photochemical method was used to synthesize silver nanoparticles (AgNPs) in the presence of citrate or clay (SWy-1, SYn-1, and Laponite B) as stabilizers and Lucirin TPO as photoinitiator. During the photochemical synthesis, an appearance of the plasmon absorption band was seen around 400 nm, indicating the formation of AgNPs. X-ray diffraction results suggested that AgNPs prepared in SWy-1 were adsorbed into interlamellar space, and moreover, showed some clay exfoliation. In the case of SYn-1, AgNPs was not intercalated. For the AgNP/Lap B sample, the formation of an exfoliated structure occurred. Transmission electron microscopy revealed the spherical shape of AgNPs for all samples. The particle sizes obtained for AgNP/SWy-1, AgNP/SYn-1, and AgNP/Lap B were 2.6, 5.1, and 3.8 nm, respectively. AgNPs adsorbed on SYn-1 reveal nonuniform size and aggregation of some particles. However, AgNP/SWy-1 and AgNP/Lap B samples are more uniform and have diameters smaller than those prepared with SYn-1. This behavior is due to the ability to exfoliate these clays. The antibacterial activities of pure clays, AgNP/citrate, and AgNP/clays were investigated against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). AgNPs in the presence of clays (AgNPs/SYn-1 and AgNPs/SWy-1) showed a lower survival index percentage compared to those obtained for pure clays and AgNPs. The AgNP/SWy-1 sample showed good antibacterial activity against both tested species and the lowest survival index of 3.9 and 4.3 against E. coli and S. aureus, respectively. AgNPs are located in the interlayer region of the SWy-1, which has acid sites. These acidic sites may contribute to the release of Ag(+) ions from the surface of AgNPs. On the other hand, Laponite B and AgNP/Lap B samples did not demonstrate any bactericidal activity.
Asunto(s)
Nanopartículas del Metal , Antibacterianos , Escherichia coli , Plata , Staphylococcus aureusRESUMEN
Silver nanoparticles (AgNPs) have been broadly used as antibacterial and antiviral agents. Further, interests for green AgNP synthesis have increased in recent years and several results for AgNP biological synthesis have been reported using bacteria, fungi and plant extracts. The understanding of the role and nature of fungal proteins, their interaction with AgNPs and the subsequent stabilization of nanosilver is yet to be deeply investigated. Therefore, in an attempt to better understand biogenic AgNP stabilization with the extracellular fungal proteins and to describe these supramolecular interactions between proteins and silver nanoparticles, AgNPs, produced extracellularly by Aspergillus tubingensis-isolated as an endophytic fungus from Rizophora mangle-were characterized in order to study their physical characteristics, identify the involved proteins, and shed light into the interactions among protein-NPs by several techniques. AgNPs of around 35 nm in diameter as measured by TEM and a positive zeta potential of +8.48 mV were obtained. These AgNPs exhibited a surface plasmon resonance (SPR) band at 440 nm, indicating the nanoparticles formation, and another band at 280 nm, attributed to the electronic excitations in tryptophan, tyrosine, and/or phenylalanine residues in fungal proteins. Fungal proteins were covalently bounded to the AgNPs, mainly through S-Ag bonds due to cysteine residues (HS-) and with few N-Ag bonds from H2N- groups, as verified by Raman spectroscopy. Observed supramolecular interactions also occur by electrostatic and other protein-protein interactions. Furthermore, proteins that remain free on AgNP surface may perform hydrogen bonds with other proteins or water increasing thus the capping layer around the AgNPs and consequently expanding the hydrodynamic diameter of the particles (~264 nm, measured by DLS). FTIR results enabled us to state that proteins adsorbed to the AgNPs did not suffer relevant secondary structure alteration upon their physical interaction with the AgNPs or when covalently bonded to them. Eight proteins in the AgNP dispersion were identified by mass spectrometry analyses. All these proteins are involved in metabolic pathways of the fungus and are important for carbon, phosphorous and nitrogen uptake, and for the fungal growth. Thereby, important proteins for fungi are also involved in the formation and stabilization of the biogenic AgNPs.