Green silver nanoparticles: Prospective nanotools against neurodegenerative cell line model.

Neurodegenerative diseases represent an increasingly burdensome challenge of the past decade, primarily driven by the global aging of the population. Ongoing efforts focus on implementing diverse strategies to mitigate the adverse effects of neurodegeneration, with the goal of decelerating the pathology progression. Notably, in recent years, it has emerged that the use of nanoparticles (NPs), particularly those obtained through green chemical processes, could constitute a promising therapeutic approach. Green NPs, exclusively sourced from phytochemicals, are deemed safer compared to NPs synthetized through conventional chemical route. In this study, the effects of green chemistry-derived silver NPs (AgNPs) were assessed in neuroblastoma cells, SHSY-5Y, which are considered a pivotal model for investigating neurodegenerative diseases. Specifically, we used two different concentrations (0.5 and 1 µM) of AgNPs and two time points (24 and 48 h) to evaluate the impact on neuroblastoma cells by observing viability reduction and intracellular calcium production, especially using 1 µM at 48 h. Furthermore, investigation using atomic force microscopy (AFM) unveiled an alteration in Young's modulus due to the reorganization of cortical actin following exposure to green AgNPs. This evidence was further corroborated by confocal microscopy acquisitions as well as coherency and density analyses on actin fibers. Our in vitro findings suggest the potential efficacy of green AgNPs against neurodegeneration; therefore, further in vivo studies are imperative to optimize possible therapeutic protocols.

Nanoparticle delivery through the BBB in central nervous system tuberculosis.

Recent advances in Nanotechnology have revolutionized the production of materials for biomedical applications. Nowadays, there is a plethora of nanomaterials with potential for use towards improvement of human health. On the other hand, very little is known about how these materials interact with biological systems, especially at the nanoscale level, mainly because of the lack of specific methods to probe these interactions. In this review, we will analytically describe the journey of nanoparticles (NPs) through the brain, starting from the very first moment upon injection. We will preliminarily provide a brief overlook of the physicochemical properties of NPs. Then, we will discuss how these NPs interact with the body compartments and biological barriers, before reaching the blood-brain barrier (BBB), the last gate guarding the brain. Particular attention will be paid to the interaction with the biomolecular, the bio-mesoscopic, the (blood) cellular, and the tissue barriers, with a focus on the BBB. This will be framed in the context of brain infections, especially considering central nervous system tuberculosis (CNS-TB), which is one of the most devastating forms of human mycobacterial infections. The final aim of this review is not a collection, nor a list, of current literature data, as it provides the readers with the analytical tools and guidelines for the design of effective and rational NPs for delivery in the infected brain.

Gold and silver nanoparticles in Alzheimer's and Parkinson's diagnostics and treatments.

Neurodegenerative diseases (NDs) impose substantial medical and public health burdens on people worldwide and represent one of the major threats to human health. The prevalence of these age-dependent disorders is dramatically increasing over time, a process intrinsically related to a constantly rising percentage of the elderly population in recent years. Among all the NDs, Alzheimer's and Parkinson's are considered the most debilitating as they cause memory and cognitive loss, as well as severely affecting basic physiological conditions such as the ability to move, speak, and breathe. There is an extreme need for new and more effective therapies to counteract these devastating diseases, as the available treatments are only able to slow down the pathogenic process without really stopping or resolving it. This review aims to elucidate the current nanotechnology-based tools representing a future hope for NDs treatment. Noble metal nano-systems, that is, gold and silver nanoparticles (NPs), have indeed unique physicochemical characteristics enabling them to deliver any pharmacological treatment in a more effective way within the central nervous system. This can potentially make NPs a new hope for reversing the actual therapeutic strategy based on slowing down an irreversible process into a more effective and permanent treatment.

Shape-Driven Response of Gold Nanoparticles to X-rays.

Radiotherapy (RT) involves delivering X-ray beams to the tumor site to trigger DNA damage. In this approach, it is fundamental to preserve healthy cells and to confine the X-ray beam only to the malignant cells. The integration of gold nanoparticles (AuNPs) in the X-ray methodology could be considered a powerful tool to improve the efficacy of RT. Indeed, AuNPs have proven to be excellent allies in contrasting tumor pathology upon RT due to their high photoelectric absorption coefficient and unique physiochemical properties. However, an analysis of their physical and morphological reaction to X-ray exposure is necessary to fully understand the AuNPs' behavior upon irradiation before treating the cells, since there are currently no studies on the evaluation of potential NP morphological changes upon specific irradiations. In this work, we synthesized two differently shaped AuNPs adopting two different techniques to achieve either spherical or star-shaped AuNPs. The spherical AuNPs were obtained with the Turkevich-Frens method, while the star-shaped AuNPs (AuNSs) involved a seed-mediated approach. We then characterized all AuNPs with Transmission Electron Microscopy (TEM), Uv-Vis spectroscopy, Dynamic Light Scattering (DLS), zeta potential and Fourier Transform Infrared (FTIR) spectroscopy. The next step involved the treatment of AuNPs with two different doses of X-radiation commonly used in RT, namely 1.8 Gy and 2 Gy, respectively. Following the X-rays' exposure, the AuNPs were further characterized to investigate their possible physicochemical and morphological alterations induced with the X-rays. We found that AuNPs do not undergo any alteration, concluding that they can be safely used in RT treatments. Lastly, the actin rearrangements of THP-1 monocytes treated with AuNPs were also assessed in terms of coherency. This is a key proof to evaluate the possible activation of an immune response, which still represents a big limitation for the clinical translation of NPs.

Silver-Gold Alloy Nanoparticles (AgAu NPs): Photochemical Synthesis of Novel Biocompatible, Bimetallic Alloy Nanoparticles and Study of Their In Vitro Peroxidase Nanozyme Activity.

Facile synthesis of metal nanoparticles with controlled physicochemical properties using environment-friendly reagents can open new avenues in biomedical applications. Nanomaterials with controlled physicochemical properties have opened new prospects for a variety of applications. In the present study, we report a single-step photochemical synthesis of ~5 nm-sized silver (Ag) and gold (Au) nanoparticles (NPs), and Ag-Au alloy nanoparticles using L-tyrosine. The physicochemical and surface properties of both monometallic and bimetallic NPs were investigated by analytical, spectroscopic, and microscopic techniques. Our results also displayed an interaction between L-tyrosine and surface atoms that leads to the formation of AgAu NPs by preventing the growth and aggregation of the NPs. This method efficiently produced monodispersed NPs, with a narrow-sized distribution and good stability in an aqueous solution. The cytotoxicity assessment performed on breast cancer cell lines (MCF-7) revealed that the biofriendly L-tyrosine-capped AgNPs, AuNPs, and bimetallic AgAu NPs were biocompatible. Interestingly, AgAu NPs have also unveiled controlled cytotoxicity, cell viability, and in vitro peroxidase nanozyme activity reliant on metal composition and surface coating.

Cancer Treatment Using Different Shapes of Gold-Based Nanomaterials in Combination with Conventional Physical Techniques.

The conventional methods of cancer treatment and diagnosis, such as radiotherapy, chemotherapy, and computed tomography, have developed a great deal. However, the effectiveness of such methods is limited to the possible failure or collateral effects on the patients. In recent years, nanoscale materials have been studied in the field of medical physics to develop increasingly efficient methods to treat diseases. Gold nanoparticles (AuNPs), thanks to their unique physicochemical and optical properties, were introduced to medicine to promote highly effective treatments. Several studies have confirmed the advantages of AuNPs such as their biocompatibility and the possibility to tune their shapes and sizes or modify their surfaces using different chemical compounds. In this review, the main properties of AuNPs are analyzed, with particular focus on star-shaped AuNPs. In addition, the main methods of tumor treatment and diagnosis involving AuNPs are reviewed.

Investigation of the Effects of Pulse-Atomic Force Nanolithography Parameters on 2.5D Nanostructures' Morphology.

In recent years, Atomic Force Microscope (AFM)-based nanolithography techniques have emerged as a very powerful approach for the machining of countless types of nanostructures. However, the conventional AFM-based nanolithography methods suffer from low efficiency, low rate of patterning, and high complexity of execution. In this frame, we first developed an easy and effective nanopatterning technique, termed Pulse-Atomic Force Lithography (P-AFL), with which we were able to pattern 2.5D nanogrooves on a thin polymer layer. Indeed, for the first time, we patterned nanogrooves with either constant or varying depth profiles, with sub-nanometre resolution, high accuracy, and reproducibility. In this paper, we present the results on the investigation of the effects of P-AFL parameters on 2.5D nanostructures' morphology. We considered three main P-AFL parameters, i.e., the pulse's amplitude (setpoint), the pulses' width, and the distance between the following indentations (step), and we patterned arrays of grooves after a precise and well-established variation of the aforementioned parameters. Optimizing the nanolithography process, in terms of patterning time and nanostructures quality, we realized unconventional shape nanostructures with high accuracy and fidelity. Finally, a scanning electron microscope was used to confirm that P-AFL does not induce any damage on AFM tips used to pattern the nanostructures.

Pile-Ups Formation in AFM-Based Nanolithography: Morpho-Mechanical Characterization and Removal Strategies.

In recent decades, great efforts have been made to develop innovative, effective, and accurate nanofabrication techniques stimulated by the growing demand for nanostructures. Nowadays, mechanical tip-based emerged as the most promising nanolithography technique, allowing the pattern of nanostructures with a sub-nanometer resolution, high reproducibility, and accuracy. Unfortunately, these nanostructures result in contoured pile-ups that could limit their use and future integration into high-tech devices. The removal of pile-ups is still an open challenge. In this perspective, two different AFM-based approaches, i.e., Force Modulation Mode imaging and force-distance curve analysis, were used to characterize the structure of pile-ups at the edges of nanogrooves patterned on PMMA substrate by means of Pulse-Atomic Force Lithography. Our experimental results showed that the material in pile-ups was less stiff than the pristine polymer. Based on this evidence, we have developed an effective strategy to easily remove pile-ups, preserving the shape and the morphology of nanostructures.

Inorganic Nanomaterials versus Polymer-Based Nanoparticles for Overcoming Neurodegeneration.

Neurodegenerative disorders (NDs) affect a great number of people worldwide and also have a significant socio-economic impact on the aging population. In this context, nanomedicine applied to neurological disorders provides several biotechnological strategies and nanoformulations that improve life expectancy and the quality of life of patients affected by brain disorders. However, available treatments are limited by the presence of the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (B-CSFB). In this regard, nanotechnological approaches could overcome these obstacles by updating various aspects (e.g., enhanced drug-delivery efficiency and bioavailability, BBB permeation and targeting the brain parenchyma, minimizing side effects). The aim of this review is to carefully explore the key elements of different neurological disorders and summarize the available nanomaterials applied for neurodegeneration therapy looking at several types of nanocarriers. Moreover, nutraceutical-loaded nanoparticles (NPs) and synthesized NPs using green approaches are also discussed underling the need to adopt eco-friendly procedures with a low environmental impact. The proven antioxidant properties related to several natural products provide an interesting starting point for developing efficient and green nanotools useful for neuroprotection.

AFM Characterization of Halloysite Clay Nanocomposites' Superficial Properties: Current State-of-the-Art and Perspectives.

Natural halloysite clay nanotubes (HNTs) are versatile inorganic reinforcing materials for creating hybrid composites. Upon doping HNTs with polymers, coating, or loading them with bioactive molecules, the production of novel nanocomposites is possible, having specific features for several applications. To investigate HNTs composites nanostructures, AFM is a very powerful tool since it allows for performing nano-topographic and morpho-mechanical measurements in any environment (air or liquid) without treatment of samples, like electron microscopes require. In this review, we aimed to provide an overview of recent AFM investigations of HNTs and HNT nanocomposites for unveiling hidden characteristics inside them envisaging future perspectives for AFM as a smart device in nanomaterials characterization.

Impact of Nanomaterials in Biological Systems and Applications in Nanomedicine Field.

The increasingly widespread use of engineered nanomaterials in many applications increases the need to understand the mechanisms behind their toxicity [...].

Pulse-Atomic Force Lithography: A Powerful Nanofabrication Technique to Fabricate Constant and Varying-Depth Nanostructures.

The widespread use of nanotechnology in different application fields, resulting in the integration of nanostructures in a plethora of devices, has addressed the research toward novel and easy-to-setup nanofabrication techniques to realize nanostructures with high spatial resolution and reproducibility. Owing to countless applications in molecular electronics, data storage, nanoelectromechanical, and systems for the Internet of Things, in recent decades, the scientific community has focused on developing methods suitable for nanopattern polymers. To this purpose, Atomic Force Microscopy-based nanolithographic techniques are effective methods that are relatively less complex and inexpensive than equally resolute and accurate techniques, such as Electron Beam lithography and Focused Ion Beam lithography. In this work, we propose an evolution of nanoindentation, named Pulse-Atomic Force Microscopy, to obtain continuous structures with a controlled depth profile, either constant or variable, on a polymer layer. Due to the modulation of the characteristics of voltage pulses fed to the AFM piezo-scanner and distance between nanoindentations, it was possible to indent sample surface with high spatial control and fabricate highly resolved 2.5D nanogrooves. That is the real strength of the proposed technique, as no other technique can achieve similar results in tailor-made graded nanogrooves without the need for additional manufacturing steps.

High Doses of Silica Nanoparticles Obtained by Microemulsion and Green Routes Compromise Human Alveolar Cells Morphology and Stiffness Differently.

Among all the inorganic nanomaterials used in commercial products, industry, and medicine, the amorphous silica nanoparticles (SiO2 NPs) appeared to be often tolerated in living organisms. However, despite several toxicity studies, some concerns about the exposure to high doses of SiO2 NPs with different sizes were raised. Then, we used the microemulsion method to obtain stable SiO2 NPs having different sizes (110 nm, 50 nm, and 25 nm). In addition, a new one-pot green synthetic route using leaves extract of Laurus nobilis was performed, obtaining monodispersed ultrasmall SiO2 NPs without the use of dangerous chemicals. The NPs achieved by microemulsion were further functionalized with amino groups making the NPs surface positively charged. Then, high doses of SiO2 NPs (1 mg/mL and 3 mg/mL) achieved from the two routes, having different sizes and surface charges, were used to assess their impact on human alveolar cells (A549), being the best cell model mimicking the inhalation route. Cell viability and caspase-3 induction were analyzed as well as the cellular uptake, obtaining that the smallest (25 nm) and positive-charged NPs were more able to induce cytotoxicity, reaching values of about 60% of cell death. Surprisingly, cells incubated with green SiO2 NPs did not show strong toxicity, and 70% of them remained vital. This result was unusual for ultrasmall nanoobjects, generally highly toxic. The actin reorganization, nuclear morphology alteration, and cell membrane elasticity analyses confirmed the trend achieved from the biological assays. The obtained data demonstrate that the increase in cellular softness, i.e., the decrease in Young's modulus, could be associated with the smaller and positive NPs, recording values of about 3 kPa. On the contrary, green NPs triggered a slight decrease of stiffness values (c.a. 6 kPa) compared to the untreated cells (c.a. 8 kPa). As the softer cells were implicated in cancer progression and metastasization, this evidence strongly supported the idea of a link between the cell elasticity and physicochemical properties of NPs that, in turn, influenced the interaction with the cell membrane. Thus, the green SiO2 NPs compromised cells to a lesser extent than the other SiO2 NPs types. In this scenario, the elasticity evaluation could be an interesting tool to understand the toxicity of NPs with the aim of predicting some pathological phenomena associated with their exposure.

Green Silver Nanoparticles Promote Inflammation Shutdown in Human Leukemic Monocytes.

The use of silver nanoparticles (Ag NPs) in the biomedical field deserves a mindful analysis of the possible inflammatory response which could limit their use in the clinic. Despite the anti-cancer properties of Ag NPs having been widely demonstrated, there are still few studies concerning their involvement in the activation of specific inflammatory pathways. The inflammatory outcome depends on the synthetic route used in the NPs production, in which toxic reagents are employed. In this work, we compared two types of Ag NPs, obtained by two different chemical routes: conventional synthesis using sodium citrate and a green protocol based on leaf extracts as a source of reduction and capping agents. A careful physicochemical characterization was carried out showing spherical and stable Ag NPs with an average size between 20 nm and 35 nm for conventional and green Ag NPs respectively. Then, we evaluated their ability to induce the activation of inflammation in Human Leukemic Monocytes (THP-1) differentiated into M0 macrophages using 1 µM and 2 µM NPs concentrations (corresponded to 0.1 µg/mL and 0.2 µg/mL respectively) and two-time points (24 h and 48 h). Our results showed a clear difference in Nuclear Factor κB (NF-κb) activation, Interleukins 6-8 (IL-6, IL-8) secretion, Tumor Necrosis Factor-α (TNF-α) and Cyclooxygenase-2 (COX-2) expression exerted by the two kinds of Ag NPs. Green Ag NPs were definitely tolerated by macrophages compared to conventional Ag NPs which induced the activation of all the factors mentioned above. Subsequently, the exposure of breast cancer cell line (MCF-7) to the green Ag NPs showed that they exhibited antitumor activity like the conventional ones, but surprisingly, using the MCF-10A line (not tumoral breast cells) the green Ag NPs did not cause a significant decrease in cell viability.

Marine Macroalgae Display Bioreductant Efficacy for Fabricating Metallic Nanoparticles: Intra/Extracellular Mechanism and Potential Biomedical Applications.

The application of hazardous chemicals during nanoparticle (NP) synthesis has raised alarming concerns pertaining to their biocompatibility and equally to the environmental harmlessness. In the recent decade, nanotechnological research has made a gigantic shift in order to include the natural resources to produce biogenic NPs. Within this approach, researchers have utilized marine resources such as macroalgae and microalgae, land plants, bacteria, fungi, yeast, actinomycetes, and viruses to synthesize NPs. Marine macroalgae (brown, red, and green) are rich in polysaccharides including alginates, fucose-containing sulfated polysaccharides (FCSPs), galactans, agars or carrageenans, semicrystalline cellulose, ulvans, and hemicelluloses. Phytochemicals are abundant in phenols, tannins, alkaloids, terpenoids, and vitamins. However, microorganisms have an abundance of active compounds ranging from sugar molecules, enzymes, canonical membrane proteins, reductase enzymes (NADH and NADPH), membrane proteins to many more. The prime reason for using the aforesaid entities in the metallic NPs synthesis is based on their intrinsic properties to act as bioreductants, having the capability to reduce and cap the metal ions into stabilized NPs. Several green NPs have been verified for their biocompatibility in human cells. Bioactive constituents from the above resources have been found on the green metallic NPs, which has demonstrated their efficacies as prospective antibiotics and anti-cancer agents against a range of human pathogens and cancer cells. Moreover, these NPs can be characterized for the size, shapes, functional groups, surface properties, porosity, hydrodynamic stability, and surface charge using different characterization techniques. The novelty and originality of this review is that we provide recent research compilations on green synthesis of NPs by marine macroalgae and other biological sources (plant, bacteria, fungi, actinomycetes, yeast, and virus). Besides, we elaborated on the detailed intra- and extracellular mechanisms of NPs synthesis by marine macroalgae. The application of green NPs as anti-bacterial, anti-cancer, and popular methods of NPs characterization techniques has also been critically reviewed.

Improvement of PMMA Dental Matrix Performance by Addition of Titanium Dioxide Nanoparticles and Clay Nanotubes.

Over the last decades, several materials have been proposed for the fabrication of dental and mandibular prosthetic implants. Today, the poly(methyl-methacrylate) (PMMA) resin is the most spread material, due to its ease of processing, low cost, aesthetic properties, low weight, biocompatibility, and biostability in the oral cavity. However, the porous surface (which favors the adhesion of microorganisms) and the weak mechanical properties (which lead to wear or fracture) are the major concerns. The inclusion of engineered nanomaterials in the acrylic matrix could improve the performances of PMMA. In this study, we added two different kind of nanomaterials, namely titanium dioxide nanoparticles (TiO2NPs) and halloysite clay nanotubes (HNTs) at two concentrations (1% and 3% w/w) in PMMA. Then, we assessed the effect of nanomaterials inclusion by the evaluation of specific physical parameters: Young's modulus, roughness, and wettability. In addition, we investigated the potential beneficial effects regarding the Candida albicans (C. albicans) colonization reduction, the most common yeast responsible of several infections in oral cavity. Our experimental results showed an improvement of PMMA performance, following the addition of TiO2NPs and HNTs, in a dose dependent manner. In particular, the presence of TiO2NPs in the methacrylate matrix induced a greater increase in PMMA stiffness respect to HNTs addition. On the other hand, HNTs reduced the rate of C. albicans colonization more significantly than TiO2NPs. The results obtained are of great interest for the improvement of PMMA physico-chemical properties, in view of its possible application in clinical dentistry.

Synergistic Effect Induced by Gold Nanoparticles with Polyphenols Shell during Thermal Therapy: Macrophage Inflammatory Response and Cancer Cell Death Assessment.

BACKGROUND: In recent decades, gold nanoparticle (Au NP)-based cancer therapy has been heavily debated. The physico-chemical properties of AuNPs can be exploited in photothermal therapy, making them a powerful tool for selectively killing cancer cells. However, the synthetic side products and capping agents often induce a strong activation of the inflammatory pathways of macrophages, thus limiting their further applications in vivo. METHODS: Here, we described a green method to obtain stable polyphenol-capped AuNPs (Au NPs@polyphenols), as polyphenols are known for their anti-inflammatory and anticancer properties. These NPs were used in human macrophages to test key inflammation-related markers, such as NF-κB, TNF-α, and interleukins-6 and 8. The results were compared with similar NPs obtained by a traditional chemical route (without the polyphenol coating), proving the potential of Au NPs@polyphenols to strongly promote the shutdown of inflammation. This was useful in developing them for use as heat-synergized tools in the thermal treatment of two types of cancer cells, namely, breast cancer (MCF-7) and neuroblastoma (SH-SY5Y) cells. The cell viability, calcium release, oxidative stress, HSP-70 expression, mitochondrial, and DNA damage, as well as cytoskeleton alteration, were evaluated. RESULTS: Our results clearly demonstrate that the combined strategy markedly exerts anticancer effects against the tested cancer cell, while neither of the single treatments (only heat or only NPs) induced significant changes. CONCLUSIONS: Au NP@polyphenols may be powerful agents in cancer treatment.

Physico-Chemical Properties of Inorganic NPs Influence the Absorption Rate of Aquatic Mosses Reducing Cytotoxicity on Intestinal Epithelial Barrier Model.

Noble metals nanoparticles (NPs) and metal oxide NPs are widely used in different fields of application and commercial products, exposing living organisms to their potential adverse effects. Recent evidences suggest their presence in the aquifers water and consequently in drinking water. In this work, we have carefully synthesized four types of NPs, namely, silver and gold NPs (Ag NPs and Au NPs) and silica and titanium dioxide NPs (SiO2 NPs and TiO2 NPs) having a similar size and negatively charged surfaces. The synthesis of Ag NPs and Au NPs was carried out by colloidal route using silver nitrate (AgNO3) and tetrachloroauric (III) acid (HAuCl4) while SiO2 NPs and TiO2 NPs were achieved by ternary microemulsion and sol-gel routes, respectively. Once the characterization of NPs was carried out in order to assess their physico-chemical properties, their impact on living cells was studied. We used the human colorectal adenocarcinoma cells (Caco-2), known as the best representative intestinal epithelial barrier model to understand the effects triggered by NPs through ingestion. Then, we moved to explore how water contamination caused by NPs can be lowered by the ability of three species of aquatic moss, namely, Leptodictyum riparium, Vesicularia ferriei, and Taxiphyllum barbieri, to absorb them. The experiments were conducted using two concentrations of NPs (100 µM and 500 Μm as metal content) and two time points (24 h and 48 h), showing a capture rate dependent on the moss species and NPs type. Then, the selected moss species, able to actively capture NPs, appear as a powerful tool capable to purify water from nanostructured materials, and then, to reduce the toxicity associated to the ingestion of contaminated drinking water.

Engineering Polymeric Nanosystems against Oral Diseases.

Nanotechnology and nanoparticles (NPs) are at the forefront of modern research, particularly in the case of healthcare therapeutic applications. Polymeric NPs, specifically, hold high promise for these purposes, including towards oral diseases. Careful optimisation of the production of polymeric NPs, however, is required to generate a product which can be easily translated from a laboratory environment to the actual clinical usage. Indeed, considerations such as biocompatibility, biodistribution, and biodegradability are paramount. Moreover, a pre-clinical assessment in adequate in vitro, ex vivo or in vivo model is also required. Last but not least, considerations for the scale-up are also important, together with an appropriate clinical testing pathway. This review aims to eviscerate the above topics, sourcing at examples from the recent literature to put in context the current most burdening oral diseases and the most promising polymeric NPs which would be suitable against them.