A scalable route to quaternary ammonium-functionalized AgCl colloidal antimicrobials inhibiting food pathogenic bacteria and biofilms.

This study explores how a simple argentometric titration-like approach could be evolved into a versatile, scalable, fast, and robust strategy for the production of AgCl/quaternary ammonium compounds (QACs) colloidal nanoantimicrobials (NAMs). These systems, which are green, stable, cost-effective, and reproducible are found to be effective against a wide range of food pathogenic bacteria and biofilms. The option of a large-scale production for such colloidal suspensions was explored via the use of a peristaltic pump. The utilization of various types of biosafe QACs and a wide range of solvents including aqueous and organic ones renders this system green and versatile. Nanocolloids (NCs) were characterized using UV-Vis, X-ray photoelectron and Fourier transform infrared (FTIR) spectroscopies. Their morphology and crystalline nature were investigated by transmission electron microscopy (TEM) and selected area diffraction pattern (SAED). Nanoparticle (NP) size distribution and hydrodynamic radius were measured by dynamic light scattering (DLS), while the ζ-potential was found to be highly positive, thus indicating significant colloidal stability and antimicrobial activity. In fact, the higher the NP surface charge, the stronger was their bioactivity. Furthermore, the antibacterial and antibiofilm effects of the as-prepared NCs were tested against Gram-positive bacteria, such as Staphylococcus aureus (ATCC 29213) and Listeria monocytogenes 46, and Gram-negative bacteria, such as Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). The results clearly indicate that AgCl/QACs provide pronounced antibiofilm activity with long-term bacteriostatic effects against foodborne pathogenic bacteria rendering them an ideal choice for active food packaging systems.

A Review on Montmorillonite-Based Nanoantimicrobials: State of the Art.

One of the crucial challenges of our time is to effectively use metal and metal oxide nanoparticles (NPs) as an alternative way to combat drug-resistant infections. Metal and metal oxide NPs such as Ag, Ag2O, Cu, Cu2O, CuO, and ZnO have found their way against antimicrobial resistance. However, they also suffer from several limitations ranging from toxicity issues to resistance mechanisms by complex structures of bacterial communities, so-called biofilms. In this regard, scientists are urgently looking for convenient approaches to develop heterostructure synergistic nanocomposites which could overcome toxicity issues, enhance antimicrobial activity, improve thermal and mechanical stability, and increase shelf life. These nanocomposites provide a controlled release of bioactive substances into the surrounding medium, are cost effective, reproducible, and scalable for real life applications such as food additives, nanoantimicrobial coating in food technology, food preservation, optical limiters, the bio medical field, and wastewater treatment application. Naturally abundant and non-toxic Montmorillonite (MMT) is a novel support to accommodate NPs, due to its negative surface charge and control release of NPs and ions. At the time of this review, around 250 articles have been published focusing on the incorporation of Ag-, Cu-, and ZnO-based NPs into MMT support and thus furthering their introduction into polymer matrix composites dominantly used for antimicrobial application. Therefore, it is highly relevant to report a comprehensive review of Ag-, Cu-, and ZnO-modified MMT. This review provides a comprehensive overview of MMT-based nanoantimicrobials, particularly dealing with preparation methods, materials characterization, and mechanisms of action, antimicrobial activity on different bacterial strains, real life applications, and environmental and toxicity issues.

Preparation of Laser-Ablated Ag Nanoparticle-MMT Clay-Based Beeswax Antibiofilm Coating.

Unlike other antimicrobial agents, Ag-based composites are stable and currently widely used as broad spectral additives, fighting microbial biofilms and other biological threats. The goal of the present study is to develop a green, multifunctional, and robust antibiofilm water-insoluble coating, inhibiting histamine-producing Lentilactobacillus parabuchneri biofilms. Herein, laser-ablated Ag NPs (L-Ag NPs) were incorporated into and onto a montmorillonite (MMT) surface layer with a simple wet chemical method, provided that the electrostatic interaction between L-Ag NPs and MMT clay led to the formation of L-Ag/MMT nanoantimicrobials (NAMs). The use of MMT support can facilitate handling Ag NPs in industrial applications. The Ag/MMT composite was characterized with transmission electron microscopy (TEM) and scanning electron microscopy (SEM), which confirmed the entrapment of L-Ag NPs into MMT clay. The surface chemical composition was assessed with X-ray photoelectron spectroscopy, proving that Ag NPs were in contact with and deposited onto the surface of MMT. The characteristic L-Ag/MMT band was investigated with UV-vis spectroscopy. Following that, the L-Ag/MMT composite was embedded into a biosafe water-insoluble beeswax agent with a spin coating technique. The antimicrobial ion release kinetic profile of the L-Ag/MMT/beeswax coating through an electrothermal atomic absorption spectroscopy (ETAAS) study supported the controlled release of Ag ions, reaching a plateau at 420 ± 80 nM, which is safe from the point of view of Ag toxicity. Microbial biofilm growth inhibition was assessed with real-time in situ Fourier transform infrared attenuated total reflection spectroscopy (FTIR-ATR) in a flow cell assembly over 32 h. The study was further supported by optical density (OD) measurements and SEM on bacteria incubated in the presence of the L-Ag/MMT/beeswax coating.

Green Synthesis and Analytical Characterization of Core-Shell Copper Sub-Microparticles.

This study demonstrates a simple and reproducible approach to synthesize green core-shell copper sub-microparticles stabilized by poly(n-vinyl)pyrrolidone (PVP). Cu@PVP colloids were here prepared using copper sulfate pentahydrate as precursor and glucose as reducing agent. The presence of PVP in the synthetic medium eliminates the need for an inert atmosphere during the process, thus simplifying the whole method. Both the morphology and the spectroscopic properties of Cu@PVP colloids were investigated by transmission electron microscopy, and infrared, UV-Vis and X-ray photoelectron spectroscopies. Size distributions and average shell thickness were obtained by statistical analysis on TEM micrographs, and spectroscopies demonstrated the formation of a PVP layer around the copper core. The produced colloids were employed in composite thin films for potential antimicrobial application, in association with a highly-recyclable polymer: polycarbonate (4,4'-(1-methylethylidene)bis(phenol)).

Self-Standing Bioinspired Polymer Films Doped with Ultrafine Silver Nanoparticles as Innovative Antimicrobial Material.

Thin self-standing films with potential antimicrobial synergistic activity have been produced by a simple green chemical synthesis with overnight thermal treatment. Their properties have been studied by scanning electron microscopy, X-ray photoelectron spectroscopy and other techniques to understand their potential range of applications. In this work, the focus was set on the development of a potential novel and effective alternative to conventional antimicrobial materials. By creating an antimicrobial polymer blend, and using it to develop and immobilize fine (~25 nm) silver nanophases, we further aimed to exploit its film-forming properties and create a solid composite material. The resulting polymer matrix showed improved water uptake percentage and better stability in the presence of water. Moreover, the antimicrobial activity of the films, which is due to both organic and inorganic components, has been evaluated by Kirby-Bauer assay against common foodborne pathogens (Staphylococcus aureus and Salmonella enterica) and resulted in a clear inhibition zone of 1.2 cm for the most complex nanocomposition. The excellent performance against bacteria of fresh and 6-month-old samples proves the prospects of this material for the development of smart and biodegradable food packaging applications.

Highly Stable Core-Shell Nanocolloids: Synergy between Nano-Silver and Natural Polymers to Prevent Biofilm Formation.

Active investment in research time in the development and study of novel unconventional antimicrobials is trending for several reasons. First, it is one of the ways which might help to fight antimicrobial resistance and bacterial contamination due to uncontrolled biofilm growth. Second, minimizing harmful environmental outcomes due to the overuse of toxic chemicals is one of the highest priorities nowadays. We propose the application of two common natural compounds, chitosan and tannic acid, for the creation of a highly crosslinked polymer blend with not only intrinsic antimicrobial properties but also reducing and stabilizing powers. Thus, the fast and green synthesis of fine spherically shaped Ag nanoparticles and further study of the composition and properties of the colloids took place. A positively charged core-shell nanocomposition, with an average size in terms of the metal core of 17 ± 4 nm, was developed. Nanoantimicrobials were characterized by several spectroscopic (UV-vis and FTIR) and microscopic (transmission and scanning electron microscopies) techniques. The use of AgNPs as a core and an organic polymer blend as a shell potentially enable a synergistic long-lasting antipathogen effect. The antibiofilm potential was studied against the food-borne pathogens Salmonella enterica and Listeria monocytogenes. The antibiofilm protocol efficiency was evaluated by performing crystal violet assay and optical density measurements, direct visualization by confocal laser scanning microscopy and morphological studies by SEM. It was found that the complex nanocomposite has the ability to prevent the growth of biofilm. Further investigation for the potential application of this stable composition in food packaging will be carried out.

Green Synthesis and Characterization of Antimicrobial Synergistic AgCl/BAC Nanocolloids.

All over the world, one of the major challenges is the green synthesis of potential materials against antimicrobial resistance and viruses. This study demonstrates a simple method like chemistry lab titration to synthesize green, facile, scalable, reproducible, and stable synergistic silver chloride/benzyldimethylhexadecyl-ammonium chloride (AgCl/BAC) colloidal Nanoantimicrobials (NAMs). Nanocolloidal dispersions of AgCl in an aqueous medium are prepared by using silver nitrate (AgNO3) as precursor and BAC as both sources of chloride and stabilizer, holding an asymmetric molecular structure. The synthetic approach is scalable and green. Both the morphology and stability of AgCl/BAC nanocolloids (NCs) were investigated as a function of different molar fractions of the reagents. AgCl/BAC NCs were characterized by transmission electron microscopy (TEM) and X-ray photoelectron and UV-vis spectroscopies. Zeta potential measurements revealed increasing positive potential values at every stage of the synthesis. Size distribution and hydrodynamic diameter of the particles were measured by dynamic light scattering (DLS), which predicted the formation of BAC layered structures associated with the AgCl nanoparticles (NPs). Small-angle X-ray scattering (SAXS) experiments verify the thickness of the BAC bilayer around AgCl. The produced AgCl/BAC NCs probably have synergistic antimicrobial properties from the AgCl core and the biocide BAC shell. AgCl/BAC NCs stability over months was investigated. The experimental evidence supports the morphological stability of the AgCl/BAC NCs, while higher positive zeta potential values anticipate a long-term antimicrobial effect: a higher surface charge causes NPs to be potentially more lethal to bacteria. AgCl/BAC antimicrobial aqueous colloidal suspensions will be used as additives for the industrial production of antimicrobial coatings.

Silver-fluoropolymer (Ag-CFX) films: Kinetic study of silver release, and spectroscopic-microscopic insight into the inhibition of P. fluorescens biofilm formation.

Silver-fluoropolymer (Ag-CFX) composed of encapsulated bioactive nanophases within a thin polymer coating are promising antimicrobial films with excellent bioactivity. In this contribution, we report on Ag-CFX thin films obtained by ion beam co-sputtering, accurately tuning film thickness, and inorganic loading. The Ag-CFX films were characterized by spectroscopic and scanning probe microscopy techniques with respect to composition and swelling behavior. Next to electrothermal atomic absorption spectroscopy (ETAAS) studies, scanning electrochemical microscopy (SECM) experiments in combination with anodic stripping voltammetry (ASV) were carried out to study the release mechanism of silver(I) from the embedded silver nanoparticles (AgNPs). Silver(I) concentration profiles at the Ag-CFX films in contact with water resulted in a release of 1310 ± 50 µg L-1 (n = 3) after 27 h of immersion and corresponded well to the swelling of the films. The antimicrobial properties towards biofilm formation of P. fluorescens were studied by attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy during a period of 48 h. The obtained IR data revealed biofilm inhibition due to the presence of the antimicrobial layer but also indicated potential surface re-colonization after 30 h of contact with the bacteria-containing solution. The occurrence of cyclic changes in the characteristic IR bands correlated with apparent stress of bottom-layered bacteria, along with re-colonization on top of dead biomass, indicative of potential cannibalism events.

On the Efficacy of ZnO Nanostructures against SARS-CoV-2.

In 2019, the new coronavirus disease (COVID-19), related to the severe acute respiratory syndrome coronavirus (SARS-CoV-2), started spreading around the word, giving rise to the world pandemic we are still facing. Since then, many strategies for the prevention and control of COVID-19 have been studied and implemented. In addition to pharmacological treatments and vaccines, it is mandatory to ensure the cleaning and disinfection of the skin and inanimate surfaces, especially in those contexts where the contagion could spread quickly, such as hospitals and clinical laboratories, schools, transport, and public places in general. Here, we report the efficacy of ZnO nanoparticles (ZnONPs) against SARS-CoV-2. NPs were produced using an ecofriendly method and fully characterized; their antiviral activity was tested in vitro against SARS-CoV-2, showing a decrease in viral load between 70% and 90%, as a function of the material's composition. Application of these nano-antimicrobials as coatings for commonly touched surfaces is envisaged.

Recent advances on the spectroscopic characterization of microbial biofilms: A critical review.

Biofilms are a major cause of health and environmental issues. Bacteria organized in biofilms are much more resistant to biocides than their equivalents in the planktonic state. In this context, spectroscopic techniques have significantly contributed to a more fundamental understanding of biofilm formation, which is crucial to prevent and limit their generation, spreading, and maturation. In this review, recent progress on the main analytical approaches enabling the spectroscopic characterization of microbial biofilms is comparatively discussed. In addition, less commonly used techniques, facilitating biofilm studies, will be also presented. Advantages and drawbacks of each discussed technique will be underlined, thus providing an overview on spectroscopic approaches for studying biofilms.

Sensing nanoparticle-protein corona using nanoparticle enhanced Laser Induced Breakdown Spectroscopy signal enhancement.

Recently nanoparticle enhanced Laser Induced Breakdown Spectroscopy (NELIBS) is getting a growing interest as an effective alternative method for improving the analytical performance of LIBS. On the other hand, the plasmonic effect during laser ablation can be used for a different task rather than elemental analysis. In this paper, the dependence of NELIBS emission signal enhancement on nanoparticle-protein solutions dried on a reference substrate (metallic titanium) was investigated. Two proteins were studied: Human Serum Albumin (HSA) and Cytochrome C (CytC). Both proteins have a strong affinity for the gold nanoparticles (AuNPs) due to the bonding between the single free exterior thiol (associated with a cysteine residue) and the gold surface to form a stable protein corona. Then, since the protein sizes are vastly different, a different number of protein units is needed to cover AuNP surface to form a protein layer. The NP-protein solution was dropped and dried onto the titanium substrate. Then the NELIBS signal enhancement of Ti emission lines was correlated to the solution characteristics as determined with Dynamic Light Scattering (DLS), Surface Plasmon Resonance (SPR) spectroscopy and Laser Doppler Electrophoresis (LDE) for ζ-potential determination. Moreover, the dried solutions were studied with TEM (Transmission Electron Microscopy) for the inspection of the inter-particle distance. The structural effect of the NP-protein conjugates on the NELIBS signal reveals that NELIBS can be used to determine the number of protein units required to form the nanoparticle-protein corona with good accuracy. Although the investigated NP-protein systems are simple cases in biological applications, this work demonstrates, for the first time, a different use of NELIBS that is beyond elemental analysis and it opens the way for sensing the nanoparticle protein corona.

A New Nanocomposite Packaging Based on LASiS-Generated AgNPs for the Preservation of Apple Juice.

Designing bioactive materials, with controlled metal ion release, exerting a significant biological action and associated to low toxicity for humans, is nowadays one of the most important challenges for our community. The most looked-for nanoantimicrobials are capable of releasing metal species with defined kinetic profiles, either by slowing down or inhibiting bacterial growth and pathogenic microorganism diffusion. In this study, laser ablation synthesis in solution (LASiS) has been used to produce bioactive Ag-based nanocolloids, in isopropyl alcohol, which can be used as water-insoluble nano-reservoirs in composite materials like poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Infrared spectroscopy was used to evaluate the chemical state of pristine polymer and final composite material, thus providing useful information about synthesis processes, as well as storage and processing conditions. Transmission electron microscopy was exploited to study the morphology of nano-colloids, along with UV-Vis for bulk chemical characterization, highlighting the presence of spheroidal particles with average diameter around 12 nm. Electro-thermal atomic absorption spectroscopy was used to investigate metal ion release from Ag-modified products, showing a maximum release around 60 ppb, which ensures an efficient antimicrobial activity, being much lower than what recommended by health institutions. Analytical spectroscopy results were matched with bioactivity tests carried out on target microorganisms of food spoilage.

Ag-Based Synergistic Antimicrobial Composites. A Critical Review.

The emerging problem of the antibiotic resistance development and the consequences that the health, food and other sectors face stimulate researchers to find safe and effective alternative methods to fight antimicrobial resistance (AMR) and biofilm formation. One of the most promising and efficient groups of materials known for robust antimicrobial performance is noble metal nanoparticles. Notably, silver nanoparticles (AgNPs) have been already widely investigated and applied as antimicrobial agents. However, it has been proposed to create synergistic composites, because pathogens can find their way to develop resistance against metal nanophases; therefore, it could be important to strengthen and secure their antipathogen potency. These complex materials are comprised of individual components with intrinsic antimicrobial action against a wide range of pathogens. One part consists of inorganic AgNPs, and the other, of active organic molecules with pronounced germicidal effects: both phases complement each other, and the effect might just be the sum of the individual effects, or it can be reinforced by the simultaneous application. Many organic molecules have been proposed as potential candidates and successfully united with inorganic counterparts: polysaccharides, with chitosan being the most used component; phenols and organic acids; and peptides and other agents of animal and synthetic origin. In this review, we overview the available literature and critically discuss the findings, including the mechanisms of action, efficacy and application of the silver-based synergistic antimicrobial composites. Hence, we provide a structured summary of the current state of the research direction and give an opinion on perspectives on the development of hybrid Ag-based nanoantimicrobials (NAMs).

Can Nanotechnology and Materials Science Help the Fight against SARS-CoV-2?

Since 2004, we have been developing nanomaterials with antimicrobial properties, the so-called nanoantimicrobials. When the coronavirus disease 2019 (COVID-19) emerged, we started investigating new and challenging routes to nanoantivirals. The two fields have some important points of contact. We would like to share with the readership our vision of the role a (nano)materials scientist can play in the fight against the COVID-19 pandemic. As researchers specifically working on surfaces and nanomaterials, in this letter we underline the importance of nanomaterial-based technological solutions in several aspects of the fight against the virus. While great resources are understandably being dedicated to treatment and diagnosis, more efforts could be dedicated to limit the virus spread. Increasing the efficacy of personal protection equipment, developing synergistic antiviral coatings, are only two of the cases discussed. This is not the first nor the last pandemic: our nanomaterials community may offer several technological solutions to challenge the ongoing and future global health emergencies. Readers' feedback and suggestions are warmly encouraged.

ZnO Nanostructures with Antibacterial Properties Prepared by a Green Electrochemical-Thermal Approach.

Zinc oxide (ZnO) nanostructures are widely applied materials, and are also capable of antimicrobial action. They can be obtained by several methods, which include physical and chemical approaches. Considering the recent rise of green and low-cost synthetic routes for nanomaterial development, electrochemical techniques represent a valid alternative to biogenic synthesis. Following a hybrid electrochemical-thermal method modified by our group, here we report on the aqueous electrosynthesis of ZnO nanomaterials based on the use of alternative stabilizers. We tested both benzyl-hexadecyl-dimetylammonium chloride (BAC) and poly-diallyl-(dimethylammonium) chloride (PDDA). Transmission electron microscopy images showed the formation of rod-like and flower-like structures with a variable aspect-ratio. The combination of UV-Vis, FTIR and XPS spectroscopies allowed for the univocal assessment of the material composition as a function of different thermal treatments. In fact, the latter guaranteed the complete conversion of the as-prepared colloidal materials into stoichiometric ZnO species without excessive morphological modification. The antimicrobial efficacy of both materials was tested against Bacillus subtilis as a Gram-positive model microorganism.

Electrochemical Preparation of Synergistic Nanoantimicrobials.

The rapid spreading of resistance among common bacterial pathogens towards the misused antibiotics/disinfectant agents has drawn much attention worldwide to bacterial infections. In light of this, the present work aimed at the realization of core-shell nanoparticles possessing remarkable antimicrobial properties thanks to the synergistic action of the metal core and the disinfectant shell. Copper nanoparticles stabilized by benzalkonium chloride were prepared, characterized, and implemented in poly-vinyl-methyl ketone to obtain nanoantimicrobial composite coatings. Bioactivity tests are reported, proving the excellent disinfectant properties of the proposed nanomaterials, as compared to one of the well-known and strongest silver-based nanoantimicrobials. Applications are also briefly described.

The Pros and Cons of the Use of Laser Ablation Synthesis for the Production of Silver Nano-Antimicrobials.

Silver nanoparticles (AgNPs) are well-known for their antimicrobial effects and several groups are proposing them as active agents to fight antimicrobial resistance. A wide variety of methods is available for nanoparticle synthesis, affording a broad spectrum of chemical and physical properties. In this work, we report on AgNPs produced by laser ablation synthesis in solution (LASiS), discussing the major features of this approach. Laser ablation synthesis is one of the best candidates, as compared to wet-chemical syntheses, for preparing Ag nano-antimicrobials. In fact, this method allows the preparation of stable Ag colloids in pure solvents without using either capping and stabilizing agents or reductants. LASiS produces AgNPs, which can be more suitable for medical and food-related applications where it is important to use non-toxic chemicals and materials for humans. In addition, laser ablation allows for achieving nanoparticles with different properties according to experimental laser parameters, thus influencing antibacterial mechanisms. However, the concentration obtained by laser-generated AgNP colloids is often low, and it is hard to implement them on an industrial scale. To obtain interesting concentrations for final applications, it is necessary to exploit high-energy lasers, which are quite expensive. In this review, we discuss the pros and cons of the use of laser ablation synthesis for the production of Ag antimicrobial colloids, taking into account applications in the food packaging field.

Catalytic Activity of Silicon Nanowires Decorated with Gold and Copper Nanoparticles Deposited by Pulsed Laser Ablation.

Silicon nanowires (SiNWs) decorated by pulsed laser ablation with gold or copper nanoparticles (labeled as AuNPs@SiNWs and CuNPs@SiNWs) were investigated for their catalytic properties. Results demonstrated high catalytic performances in the Caryl-N couplings and subsequent carbonylations for gold and copper catalysts, respectively, that have no precedents in the literature. The excellent activity, attested by the very high turn over number (TON) values, was due both to the uniform coverage along the NW length and to the absence of the chemical shell surrounding the metal nanoparticles (MeNPs). A high recyclability was also observed and can be ascribed to the strong covalent interaction at the Me-Si interface by virtue of metal "silicides" formation.

Spectroscopic Characterization and Nanosafety of Ag-Modified Antibacterial Leather and Leatherette.

The development of antibacterial coatings is of great interest from both industry and the consumer's point of view. In this study, we characterized tanned leather and polyurethane leatherette, typically employed in the automotive and footwear industries, which were modified by photo-deposition of antibacterial silver nanoparticles (AgNPs). Material surface chemical composition was investigated in detail by X-ray photoelectron spectroscopy (XPS). The material's antibacterial capability was checked against Escherichia coli and Staphylococcus aureus, as representative microorganisms in cross transmissions. Due to the presence of silver in a nanostructured form, nanosafety issues were considered, as well. Ionic release in contact media, as well as whole nanoparticle release from treated materials, were quantitatively evaluated, thus providing specific information on potential product nanotoxicity, which was further investigated through cytocompatibility MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, also after surface abrasion of the materials. The proved negligible nanoparticle release, as well as the controlled release of antibacterial ions, shed light on the materials' potentialities, in terms of both high activity and safety.

Combined Approach for the Development of Efficient and Safe Nanoantimicrobials: The Case of Nanosilver-Modified Polyurethane Foams.

Silver nanophases are routinely used as bioactive additives in commercial products. Besides their antimicrobial activity, nanosafety issues regarding the application of (silver-based) nanoantimicrobials should be considered, as well. In this study, we modified polyurethane foams, typically employed in air filtration and stuffing, by photodeposited silver nanoparticles for preparing hybrid materials (Ag-PU) with antibacterial properties. The composite materials were characterized in terms of morphology, surface chemical composition, ionic release in contact media, bioactivity, as well as whole nanoparticle release. Cytocompatibility was also assessed on 3T3 mouse fibroblasts. The proposed systematic approach allows for defining suitable composite final properties, in terms of bioactivity and safety, by properly tuning the deposition parameters.