Nail Lacquer Containing Origanum vulgare and Rosmarinus officinalis Essential Oils and Biogenic Silver Nanoparticles for Onychomycosis: Development, Characterization, and Evaluation of Antifungal Efficacy.

Onychomycosis is a common fungal nail infection for which new antifungals are needed to overcome antimicrobial resistance and the limitations of conventional treatments. This study reports the development of antifungal nail lacquers containing oregano essential oil (OEO), rosemary essential oil (REO), and biogenic silver nanoparticles (bioAgNPs). The formulations (F) were tested against dermatophytes using agar diffusion, ex vivo nail infection, and scanning electron microscopy techniques. They were evaluated for their pharmacotechnical characteristics and by FTIR-PAS to assess permeation across the nail. F-OEO and F-OEO/bioAgNPs were promising candidates for the final nail lacquer formulation, as they permeated through the nail and showed antifungal efficacy against dermatophytes-contaminated nails after 5 days of treatment. Treated nails exhibited decreased hyphae and spores compared to the untreated control; the hyphae were atypically flattened, indicating loss of cytoplasmic content due to damage to the cytoplasmic membrane. The formulations were stable after centrifugation and thermal stress, maintaining organoleptic and physicochemical characteristics. Both F-OEO and F-OEO/bioAgNPs had pH compatible with the nail and drying times (59-90 s) within the reference for nail lacquer. For the first time, OEO and bioAgNPs were incorporated into nail lacquer, resulting in a natural and nanotechnological product for onychomycosis that could combat microbial resistance.

Integrating Natural Deep Eutectic Solvents into Nanostructured Lipid Carriers: An Industrial Look.

The industries are searching for greener alternatives for their productions due to the rising concern about the environment and creation of waste and by-products without industrial utility for that specific line of products. This investigation describes the development of two stable nanostructured lipid carriers (NLCs): one is the formulation of a standard NLC, and the other one is the same NLC formulation associated with a natural deep eutectic solvent (NaDES). The research presents the formulation paths of the NLCs through completeness, which encompass dynamic light scattering (DLS), zeta potential tests, and pH. Transmission electron microscopy (TEM) and confocal microscopy were performed to clarify the morphology. Cytotoxicity tests with zebrafish were realized, and the results are complementary to the in vitro outcomes reached with fibroblast L132 tests by the MTT technique and the zymography test. Infrared spectroscopy and X-ray diffractometry tests elucidated the link between the physicochemical characteristics of the formulation and its behavior and properties. Different cooling techniques were explored to prove the tailorable properties of the NLCs for any industrial applications. In conclusion, the compiled results show the successful formulation of new nanocarriers based on a sustainable, eco-friendly, and highly tailorable technology, which presents low cytotoxic potential.

Tiny Green Army: Fighting Malaria with Plants and Nanotechnology.

Malaria poses a global threat to human health, with millions of cases and thousands of deaths each year, mainly affecting developing countries in tropical and subtropical regions. Malaria's causative agent is Plasmodium species, generally transmitted in the hematophagous act of female Anopheles sp. mosquitoes. The main approaches to fighting malaria are eliminating the parasite through drug treatments and preventing transmission with vector control. However, vector and parasite resistance to current strategies set a challenge. In response to the loss of drug efficacy and the environmental impact of pesticides, the focus shifted to the search for biocompatible products that could be antimalarial. Plant derivatives have a millennial application in traditional medicine, including the treatment of malaria, and show toxic effects towards the parasite and the mosquito, aside from being accessible and affordable. Its disadvantage lies in the type of administration because green chemical compounds rapidly degrade. The nanoformulation of these compounds can improve bioavailability, solubility, and efficacy. Thus, the nanotechnology-based development of plant products represents a relevant tool in the fight against malaria. We aim to review the effects of nanoparticles synthesized with plant extracts on Anopheles and Plasmodium while outlining the nanotechnology green synthesis and current malaria prevention strategies.

Novel Microsynthesis of High-Yield Gold Nanoparticles to Accelerate Research in Biosensing and Other Bioapplications.

Gold nanoparticles (AuNPs) exhibit unique properties that make them appealing for applications in biosensing and other emerging fields. Despite the availability of numerous synthesis methods, important questions remain to be addressed regarding the volume effect on the synthesis yield and quality of AuNPs in the light of biosensing research. The present study addresses these issues by developing a novel microvolumetric citrate-reduction method to improve the synthesis of AuNPs, which were characterized by electronic microscopy, energy dispersive spectroscopy, zeta potential and colorimetric analysis. A comparison of the novel microsynthesis method with the standard Turkevich method demonstrated its superior performance in terms of yield, monodispersity, rapidity (in one step), reproducibility, and stability. The analytical behavior of AuNPs-based aptasensors prepared by microsynthesis was investigated using kanamycin detection and showed higher reproducibility and improved detection limits (3.4 times) compared to those of Turkevich AuNPs. Finally, the effect of pH was studied to demonstrate the suitability of the method for the screening of AuNP synthesis parameters that are of direct interest in biosensing research; the results showed an optimal pH range between 5.0 and 5.5. In summary, the approach described herein has the potential to improve research capabilities in biosensing, with the added benefits of lowering costs and minimizing waste generation in line with current trends in green nanotechnology.

Hydrogel-Containing Biogenic Silver Nanoparticles: Antibacterial Action, Evaluation of Wound Healing, and Bioaccumulation in Wistar Rats.

Wound infections are feared complications due to their potential to increase healthcare costs and cause mortality since multidrug-resistant bacteria reduce treatment options. This study reports the development of a carbomer hydrogel containing biogenic silver nanoparticles (bioAgNPs) and its effectiveness in wound treatment. This hydrogel showed in vitro bactericidal activity after 2 h, according to the time-kill assay. It also reduced bacterial contamination in rat wounds without impairing their healing since the hydrogel hydrophilic groups provided hydration for the injured skin. The high number of inflammatory cells in the first days of the skin lesion and the greater degree of neovascularization one week after wound onset showed that the healing process occurred normally. Furthermore, the hydrogel-containing bioAgNPs did not cause toxic silver accumulation in the organs and blood of the rats. This study developed a bioAgNP hydrogel for the treatment of wounds; it has a potent antimicrobial action without interfering with cicatrization or causing silver bioaccumulation. This formulation is effective against bacteria that commonly cause wound infections, such as Pseudomonas aeruginosa and Staphylococcus aureus, and for which new antimicrobials are urgently needed, according to the World Health Organization's warning.

Antibiofilm Effect of Biogenic Silver Nanoparticles Combined with Oregano Derivatives against Carbapenem-Resistant Klebsiella pneumoniae.

Resistant bacteria may kill more people than COVID-19, so the development of new antibacterials is essential, especially against microbial biofilms that are reservoirs of resistant cells. Silver nanoparticles (bioAgNP), biogenically synthesized using Fusarium oxysporum, combined with oregano derivatives, present a strategic antibacterial mechanism and prevent the emergence of resistance against planktonic microorganisms. Antibiofilm activity of four binary combinations was tested against enteroaggregative Escherichia coli (EAEC) and Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC): oregano essential oil (OEO) plus bioAgNP, carvacrol (Car) plus bioAgNP, thymol (Thy) plus bioAgNP, and Car plus Thy. The antibiofilm effect was accessed using crystal violet, MTT, scanning electron microscopy, and Chromobacterium violaceum anti-quorum-sensing assays. All binary combinations acted against preformed biofilm and prevented its formation; they showed improved antibiofilm activity compared to antimicrobials individually by reducing sessile minimal inhibitory concentration up to 87.5% or further decreasing biofilm metabolic activity and total biomass. Thy plus bioAgNP extensively inhibited the growth of biofilm in polystyrene and glass surfaces, disrupted three-dimensional biofilm structure, and quorum-sensing inhibition may be involved in its antibiofilm activity. For the first time, it is shown that bioAgNP combined with oregano has antibiofilm effect against bacteria for which antimicrobials are urgently needed, such as KPC.

Green nanotechnology for the development of nanoparticles based on alginate associated with essential and vegetable oils for application in fruits and seeds protection.

Aiming to highlight the valorization of the natural products and the green synthesis processes, this work describes the development of a nanoscale system based on the use of alginate to encapsulate a blend of oils (vegetable and essential oils), not previously reported, with antibacterial and antioxidant actions. The study shows the influence of the polymer and surfactant concentrations on the physicochemical properties of the nanoparticles. The formulations were characterized by DLS, zeta potential, efficiency of encapsulation and stability. In addition, the antioxidant and antimicrobial properties of the systems were evaluated using the DPPH method and disk diffusion assays, respectively. The shelf life was studied by coating fruits and seeds. The results showed that the nanostructured system was stable, the efficiency of encapsulation was high and the nanoparticles size range was about 200-400 nm. The coating of fruits and seeds showed that the system was capable of inhibiting the growth of microorganisms and delaying the fruit maturation, indicating its potential for prolonging the shelf-life of fresh food.

Photo-catalytic and biomedical applications of one-step, plant extract-mediated green-synthesized cobalt oxide nanoparticles.

In the present work, for the first time, green chemically synthesized and stabilized Co3O4 nanoparticles were employed for catalytic conversion of isopropyl alcohol to acetone by dehydrogenation of IPA. Plant extract of Rosmarinus officinalis was used as a reducing and stabilizing agent for this synthesis. The biosynthesized Co3O4 nanoparticles were annealed at 450℃ followed by their physiochemical characterizations through XRD, SEM, AFM, and FTIR. Size distribution information collected through XRD and AFM back each other, and it was found to be 6.5 nm, having the highest number of nanoparticles in this size range. While SEM confirms the self-arranging property of synthesized nanoparticles due to their magnetic nature, furthermore, the biogenic Co3O4 nanoparticles were studied for their catalytic potential to convert isopropyl alcohol to acetone with the help of a UV-Visible spectrophotometer. The highest photocatalytic conversion of 99% was obtained in time period of 48 s. For the first time ever, nanoparticles were used for 5 cycles to evaluate their recyclable nature and conversion fell from 99 to 86% and the end of the 5th cycle. Later anti-bacterial activity against 3 Gram-positive and 3 Gram-negative strains gave the highest inhibition value of 99% against Streptococcus pneumoniae at 500 µg/mL. Finally, a cytotoxicity study on synthesized nanomaterials was carried out by exposing freshly drawn human macrophages to them. It was found that even at the highest concentration of 500 µg/mL, the nanoparticles showed only 28% lysis.

Biogenic Silver Nanoparticles Strategically Combined With Origanum vulgare Derivatives: Antibacterial Mechanism of Action and Effect on Multidrug-Resistant Strains.

Multidrug-resistant bacteria have become a public health problem worldwide, reducing treatment options against several pathogens. If we do not act against this problem, it is estimated that by 2050 superbugs will kill more people than the current COVID-19 pandemic. Among solutions to combat antibacterial resistance, there is increasing demand for new antimicrobials. The antibacterial activity of binary combinations containing bioAgNP (biogenically synthesized silver nanoparticles using Fusarium oxysporum), oregano essential oil (OEO), carvacrol (Car), and thymol (Thy) was evaluated: OEO plus bioAgNP, Car plus bioAgNP, Thy plus bioAgNP, and Car plus Thy. This study shows that the mechanism of action of Thy, bioAgNP, and Thy plus bioAgNP involves damaging the membrane and cell wall (surface blebbing and disruption seen with an electron microscope), causing cytoplasmic molecule leakage (ATP, DNA, RNA, and total proteins) and oxidative stress by enhancing intracellular reactive oxygen species and lipid peroxidation; a similar mechanism happens for OEO and Car, except for oxidative stress. The combination containing bioAgNP and oregano derivatives, especially thymol, shows strategic antibacterial mechanism; thymol disturbs the selective permeability of the cell membrane and consequently facilitates access of the nanoparticles to bacterial cytoplasm. BioAgNP-treated Escherichia coli developed resistance to nanosilver after 12 days of daily exposition. The combination of Thy and bioAgNP prevented the emergence of resistance to both antimicrobials; therefore, mixture of antimicrobials is a strategy to extend their life. For antimicrobials alone, minimal bactericidal concentration ranges were 0.3-2.38 mg/ml (OEO), 0.31-1.22 mg/ml (Car), 0.25-1 mg/ml (Thy), and 15.75-31.5 µg/ml (bioAgNP). The time-kill assays showed that the oregano derivatives acted very fast (at least 10 s), while the bioAgNP took at least 30 min to kill Gram-negative bacteria and 7 h to kill methicillin-resistant Staphylococcus aureus (MRSA). All the combinations resulted in additive antibacterial effect, reducing significantly minimal inhibitory concentration and acting faster than the bioAgNP alone; they also showed no cytotoxicity. This study describes for the first time the effect of Car and Thy combined with bioAgNP (produced with F. oxysporum components) against bacteria for which efficient antimicrobials are urgently needed, such as carbapenem-resistant strains (E. coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa) and MRSA.

Green nanotechnology: only the final product that matters?

Nanotechnology is increasingly showing interest in eco-friendly nanotechnology products. However, the final product does not always establish a connection with the principles established by green chemistry. Thus, much confusion is present in the literature, where nanostructures are synthesized with aggressive protocols to the environment but are subsequently exposed to natural products. The confusion established is associated with the natural products related to these nanostructures. Thus, the final nanotechnological product is considered green, even though it undergoes too drastic processes related to toxic precursors, the formation of by-products, high energy consumption. In this article, the importance of strengthening green chemistry principles to discuss green nanotechnology will be discussed.

Silver nanoparticles from residual biomass: Biosynthesis, characterization and antimicrobial activity.

The industrial effluent contaminated with organic pollutants has been causing an increase in the toxicity of the ecosystem, causing a great environmental impact. Thus, the present work aims the green synthesis of silver nanoparticles (AgNPs) from Aloe vera, its characterization and antimicrobial activity against Pseudomonas aeruginosa (ATCC 27853) and Staphylococcus aureus (ATCC 25923). AgNPs were characterized by X-ray diffraction (XRD), Scanning Electronic Microscopy with Energy Dispersive Spectroscopy (SEM-EDS), Zeta Potential (ZP) and N2 porosimetry (BET/BJH method). Antimicrobial activity were carried out by Minimal Inhibitory Concentration (MIC) method. The XRD demonstrated characteristic peaks of AgNPs at 38.29°; 44.55° and 64.81°, and SEM-EDS micrographs showed that AgNPs produced by biomolecules of Aloe vera extract resulted in a weight concentration around 92.59% silver, 7.15% oxygen and 0.26% chlorine. Regarding zeta potential, all samples showed negative electric charge (around -35.3 mV), while N2 porosimetry resulted in a surface specific area of 6.09 m2 g-1, with a volume and diameter pore of 0.032 cm³ g-1 and 33.47, respectively. Antimicrobial activity was observed at 15.62 µg mL-1 and 31.25 µg mL-1 for P. aeruginosa and S. aureus, respectively. Thus, AgNPs can be considered a promising nanoparticle for degradation of organic pollutants in aqueous solution as well as an adjuvant for treatment of microbial infections.

Protein corona on biogenic silver nanoparticles provides higher stability and protects cells from toxicity in comparison to chemical nanoparticles.

The development of environmentally friendly new procedures for the synthesis of metallic nanoparticles is one of the main goals of nanotechnology. Proteins and enzymes from plants, filamentous fungi, yeast, and bacteria to produce nanoparticles are both valuable and viable alternatives to conventional synthesis of nanomaterials due to their high efficiency and the low cost to scale up and generate large quantities. The aim of this work is to compare biogenic silver nanoparticles (AgNPs) obtained from cell-free filtrates from the fungus Macrophomina phaseolina to conventional chemical AgNPs, in biocidal activity and toxicity. Our results show that bio-AgNPs displayed similar bactericidal activity than chemical AgNPs, but less toxicity in the model organism Caenorhabditis elegans. We employed biochemical and proteomic techniques to profile the unique surface chemistry of the capping in the bio-AgNPs and therefore to identify the proteins involved in their synthesis and stability. These results not only suggest that the proteins involved in the synthesis of the nanoparticles and corona formation in the bio-AgNPs are responsible for keeping the silver core preserved making them more stable in time, but also masking and protecting eukaryotic cells from metal toxicity.

Green-tea-synthesized silver nanoparticles as a sensing platform for determination of tetracycline in honey samples.

BACKGROUND: Silver nanoparticles (AgNPs) can be easily obtained in aqueous solution by chemical reduction using appropriate reducing agents and stabilizers. The development of environmentally friendly methods using non-toxic solvents and reagents has become an alternative for the synthesis of these particles and their future application as sensor probes for agricultural products. In this work, a straightforward method based on green tea extracts as reducing and capping agent is proposed for the synthesis of AgNPs, followed by their evaluation as a sensing platform for determination of tetracycline in honey samples. RESULTS: Highly stable nanoparticles were easily obtained by combining green tea aqueous extracts and ultrasound irradiation for 2 min. The as-synthesized AgNPs, spherical in shape and with average size of 8.5 nm, were evaluated for determination of tetracycline by following the changes on the localized surface plasmon resonance band at 450 nm induced by the presence of this antibiotic at pH 5.8. The method was successfully applied in the concentration range between 200 and 800 µg L-1 with R2 > 0.996 and limit of detection of 52.7 µg L-1 . Multiple honey samples were analyzed, and the recovery values obtained ranged between 82.8% and 116%, with relative standard deviation values lower than 6.69%. CONCLUSION: The results obtained demonstrate that the AgNPs synthesized using just green tea extracts represent a promising and sustainable alternative tool for the cost-effective determination of tetracycline antibiotics in honey. © 2021 Society of Chemical Industry.

Synthesis and characterization of iron oxide nanoparticles/carboxymethyl cellulose core-shell nanohybrids for killing cancer cells in vitro.

Novel core-shell superparamagnetic nanofluids composed of magnetic iron oxide (Fe3O4, MION) and cobalt-doped (CoxFe3-xO4, Co-MION) nanoparticles functionalized with carboxymethyl cellulose (CMC) ligands were designed and produced via green colloidal aqueous process. The effect of the degree of substitution (DS = 0.7 and 1.2) and molecular mass (Mw) of CMC and cobalt doping concentration on the physicochemical and magnetic properties of these nanoconjugates were comprehensively investigated using Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction, transmission electron microscopy (TEM) with selected area electron diffraction, X-ray fluorescence, dynamic light scattering (DLS), zeta potential (ZP) analysis, vibrating sample magnetometry (VSM) and electron paramagnetic resonance spectroscopy (EPR). The results demonstrated the effect of concentration of carboxylate groups and Mw of CMC on the hydrodynamic dimension, zeta potential, and generated heat by magnetic hyperthermia of MION nanoconjugates. Co-doping of MION showed significant alteration of the electrostatic balance of charges of the nanoconjugates interpreted as effect of surface interactions. Moreover, the VSM and EPR results proved the superparamagnetic properties of these nanocolloids, which were affected by the presence of CMC and Co-doping of iron oxide nanoparticles. These magnetic nanohybrids behaved as nanoheaters for killing brain cancer cells in vitro with prospective future applications in oncology and nanomedicine.

Green synthesis of silver nanoparticles using tomato leaf extract and their entrapment in chitosan nanoparticles to control bacterial wilt.

BACKGROUND: Silver nanoparticles (AgNPs), particularly those entrapped in polymeric nanosystems, have arisen as options for managing plant bacterial diseases. Among the biopolymers useful for the entrapment of AgNPs, chitosan is promising because of its low cost, good biocompatibility, antimicrobial properties and biodegradability. The present study aimed: (i) to greenly-synthesize AgNPs using different concentrations of aqueous extract of tomato leaves followed by entrapment of AgNPs with chitosan (CH-AgNPs); (ii) to characterize the optical, structural and biological properties of the nanosystems produced; (iii) to evaluate the antimicrobial activities of AgNPs and nanomaterials; and (iv) to assess the effectiveness of AgNPs and nanomaterials for controlling tomato bacterial wilt caused by Ralstonia solanacearum. RESULTS: Spherical and oval AgNPs had incipient colloidal instability, although the concentration of the tomato leaf extract influenced both size (< 87 nm) and the polydispersity index. Nanomaterials (< 271 nm in size) were characterized by a highly stable matrix of chitosan containing polydisperse AgNPs. Free AgNPs and CH-AgNPs were stable for up to 30 days, with no significant alteration in physicochemical parameters. The AgNPs and nanomaterials had antibacterial activity and decreased bacterial growth at micromolar concentrations after 48 h. Morphological changes in R. solanacearum cells were observed after treatment with CH-AgNPs. The application of CH-AgNPs at 256 µmol L-1 reduced the incidence of bacterial wilt in a partially resistant tomato genotype but not in the susceptible line. CONCLUSION: Greenly-synthesized chitosan-derived nanomaterials containing AgNPs produced with leaf extracts from their own species appear to comprise a promising and sustainable alternative in an integrated management approach aiming to reduce the yield losses caused by bacterial wilt. © 2019 Society of Chemical Industry.

Gold nanoparticle-carboxymethyl cellulose nanocolloids for detection of human immunodeficiency virus type-1 (HIV-1) using laser light scattering immunoassay.

It is estimated that over 100 million people have been infected with human immunodeficiency virus (HIV-1) resulting in approximately 30 million deaths globally. Herein, we designed and developed novel nano-immunoconjugates using gold nanoparticles (AuNPs) and carboxymethylcellulose (CMC) biopolymer, which performed simultaneously as an eco-friendly in situ reducing agent and surface stabilizing ligand for the aqueous colloidal process. These AuNPs-CMC nanocolloids were biofunctionalized with the gp41 glycoprotein receptor (AuNPs-CMC-gp41) or HIV monoclonal antibodies (AuNPs-CMC_PolyArg-abHIV) for detection using the laser light scattering immunoassay (LIA). These AuNPs-CMC bioengineered nanoconjugates were extensively characterized by morphological and physicochemical methods, which demonstrated the formation of spherical nanocrystalline colloidal AuNPs with the average size from 12 to 20 nm and surface plasmon resonance peak at 520 nm. Thus, stable nanocolloids were formed with core-shell nanostructures composed of AuNPs and biomacromolecules of CMC-gp41, which were cytocompatible based on in vitro cell viability results. The AuNPs-CMC-gp41 nanoconjugates were tested against HIV monoclonal antibodies conjugates (AuNPs-CMC_PolyArg-abHIV) using the light scattering immunoassay (LIA) where they behaved as active nanoprobes for the detection at nM level of HIV-1 antigenic proteins. This strategy offers a novel nanoplatform for creating bioprobes using green nanotechnology for the detection of HIV-1 and other virus-related diseases.

Application of poly(epsilon-caprolactone) nanoparticles containing atrazine herbicide as an alternative technique to control weeds and reduce damage to the environment.

Nanoparticles of poly(epsilon-caprolactone) containing the herbicide atrazine were prepared, characterized, and evaluated in terms of their herbicidal activity and genotoxicity. The stability of the nanoparticles was evaluated over a period of three months, considering the variables: size, polydispersion index, pH, and encapsulation efficiency. Tests on plants were performed with target (Brassica sp.) and non-target (Zea mays) organisms, and the nanoparticle formulations were shown to be effective for the control of the target species. Experiments using soil columns revealed that the use of nanoparticles reduced the mobility of atrazine in the soil. Application of the Allium cepa chromosome aberration assay demonstrated that the nanoparticle systems were able to reduce the genotoxicity of the herbicide. The formulations developed offer a useful means of controlling agricultural weeds, while at the same time reducing the risk of harm to the environment and human health.