Mariafrancesca Cascione

The growing request for more sustainable materials and environmentally friendly nanofabrication methods in the electronics field has recently driven the scientific community in the development of bio-derived materials as an alternative to conventional lithographic resists. In this work, we used chitosan, a biodegradable and biocompatible polysaccharide, as a green direct-write resist material for Atomic Force Microscopy-based nanolithography. Chitosan thin layers were obtained by spin coating and systematically characterized, in terms of thickness and surface roughness, demonstrating nanoscale smoothness and tunable film thickness. Three Pulse–Atomic Force Lithography (P-AFL) approaches, i.e., Constant Pulse, Gradient Pulse, and Raster Pulse AFL methods, were used to pattern nanostructures with constant-depth nanogrooves, variable-depth (2.5D) profile, and three-dimensional nanoholes on chitosan films. The results reveal high pattern fidelity, reproducibility, and tunability of feature dimensions as a function of applied force and scanning direction. Moreover, the RP-AFL technique enabled the fabrication of well-defined 3D nanostructures with depths matching the film thickness, which is a prerequisite for subsequent pattern transfer. This experimental work provided a first proof-of-concept to adopt chitosan as a more sustainable alternative with respect to conventional resists. Moreover, the results highlight P-AFL methods as a versatile and low-impact nanofabrication strategy, contributing to the development of greener micro- and nano-manufacturing technologies.

Epigallocatechin gallate (EGCG), the predominant catechin in green tea, displays strong antioxidant and cytoprotective properties; however, its poor stability and limited oral bioavailability significantly restrict its therapeutic potential, prompting the need for new therapeutic strategies. Here, we reported synthesis and physicochemical characterization of chitosan nanoparticles (Ch-NPs) encapsulating EGCG (Ch-EGCG-NPs) by the ionic gelation method. The Ch-EGCG-NPs displayed tunable particle size (184-236 nm), narrow polydispersity, and positive zeta potential, supporting colloidal stability and efficient EGCG loading. NPs uptake was followed by confocal microscopy and flow cytometry, using FITC-labeled Ch-NPs demonstrating efficient internalization in pancreatic and hepatic cancer cell models, i.e HepG2 and PANC-1 respectively. In vitro assays revealed that Ch-EGCG-NPs preserved cell viability under hydrogen peroxide-induced oxidative stress and significantly modulated intracellular ROS levels. The oxidative treatment induced a 1.89-fold increase in ROS production compared to the control (corresponding to an 89% up-regulation), representing the largest percentage change observed. Conversely, Ch-EGCG-NPs markedly reduced ROS accumulation compared to free EGCG, lowering ROS levels from 2.24 to 1.06 (≈52.7% reduction) and from 2.24 to 0.66 (≈70.5% reduction), with reductions ranging from 52% to 70.5% in PANC-1 cells and up to 60.3% in HepG2 cells.

Chitosan (CS), a naturally abundant biopolymer mainly sourced from marine crustacean waste, has emerged as a sustainable alternative to conventional, nonbiodegradable synthetic polymers in food packaging due to its intrinsic biocompatibility, nontoxicity, higher sustainability, biodegradability, and excellent film-forming ability. Furthermore, CS exhibits a remarkable dual role as both a reducing and capping agent in the green synthesis of silver nanoparticles (AgNPs). In the present work, CS-based nanocomposite films embedded with green AgNPs were produced via a straightforward one-step UV photoreduction method. CS simultaneously acted as a reducing and capping agent for AgNPs obtained in situ and as a rigid matrix for their confinement. By varying the UV exposure times (15, 45, and 90 min), the morphology and size of the NPs were characterized using transmission electron microscopy (TEM), revealing a predominantly spherical shape with an average diameter of ∼60 nm. In parallel, each resulting film was thoroughly analyzed using different techniques to evaluate the impact of UV radiation and in situ AgNP formation on the polymers’ physical and structural properties, including wettability, moisture content, swelling degree, water solubility, surface morphology, roughness, and mechanical behavior. In addition, antibacterial efficacy was assessed against both Escherichia coli and Staphylococcus aureus, demonstrating inhibition of both Gram-positive and Gram-negative strains. Moreover, the silver ion release in aqueous media (pH 7) was quantified via ICP-OES. The simplicity, scalability, and effectiveness of the proposed method underscore the potential of the AgNPs@CS film as a sustainable antibacterial material for next-generation food packaging solutions.

Carbon dots (CDs) derived from renewable biomass are emerging as sustainable alternatives to traditional nanomaterials for applications in bioimaging, sensing, and photonics. In this study, we reported a one-step synthesis of photoluminescent CDs from Laurus nobilis leaves particularly spread in the Mediterranean area. The resulting nanoparticles (NPs) exhibited average diameters of 3–5 nm and high colloidal stability in water. Structural analysis by X-Rays Diffraction revealed the presence of amorphous graphitic domains, while infrared spectroscopy confirmed oxygenated functional groups on the CD surface. Spectrofluorimetric analysis showed excitation-dependent blue–green emission with a maximum at 490 nm that can be applied also as label agents for cells. The environmental sustainability of the synthetic procedure was evaluated through a Life Cycle Assessment (LCA), highlighting that the current impacts were primarily associated with electricity consumption, due to the laboratory-scale nature of the process. These impacts are expected to decrease significantly with future scale-up and process optimization.

Carbon dots (CDs) derived from renewable biomass are emerging as sustainable alternatives to traditional nanomaterials for applications in bioimaging, sensing, and photonics. In this study, we report a one-step hydrothermal synthesis of photoluminescent CDs from Laurus nobilis leaves particularly spread in the Mediterranean area. The resulting nanoparticles exhibited average diameters of 3–5 nm and high colloidal stability in water. Structural analysis by XRD revealed the presence of amorphous graphitic domains, while FTIR spectroscopy confirmed oxygenated functional groups on the CD surface. Spectrofluorimetric analysis showed excitation-dependent blue–green emission with a maximum at 490 nm that can be applied also as label agents for cells. The environmental sustainability of the synthesis was evaluated through a Life Cycle Assessment (LCA), highlighting that the current impacts are primarily associated with electricity consumption, due to the laboratory-scale nature of the process. These impacts are expected to decrease significantly with future scale-up and process optimization.

The implementation of novel analytic methodologies in cancer and biomedical research has enabled the quantification of parameters that were previously disregarded only a few decades ago. A notable example of this paradigm shift is the widespread integration of atomic force microscopy (AFM) into biomedical laboratories, significantly advancing our understanding of cancer cell biology and treatment response. AFM allows for the meticulous monitoring of different parameters at the molecular and nanoscale levels, encompassing critical aspects such as cell morphology, roughness, adhesion, stiffness, and elasticity. These parameters can be systematically investigated in correlation with specific cell treatment, providing important insights into morpho-mechanical properties during normal and treated conditions. The resolution of this system holds the potential for its systematic adoption in clinics; its application could produce useful diagnostic information regarding the aggressiveness of cancer and the efficacy of treatment. This review endeavors to analyze the current literature, underscoring the pivotal role of AFM in biomedical research, especially in cancer cases, while also contemplating its prospective application in a clinical context.

Melanoma is an aggressive cancer with rising incidence and high mortality rates, largely due to chemotherapy resistance and molecular dysregulation. Nanotechnology, particularly silver nanoparticles (AgNPs), has emerged as a promising therapeutic avenue because of the nanoparticles’ ability to induce oxidative stress and apoptosis in cancer cells. However, conventional colloidal AgNPs lack selectivity, often causing significant damage to healthy cells. In this study, we introduce a green synthesis of AgNPs using plant extracts, providing an eco-friendly alternative with improved antitumor selectivity compared to traditional colloidal AgNPs. Leveraging label-free Data-Independent Acquisition/Sequential Window Acquisition of All Theoretical Mass Spectrometry (DIA/SWATH MS) quantitative proteomics, we investigated the antitumor effects of green-synthesized versus traditional AgNPs on A375 melanoma cells at 24 and 48 h. Our findings reveal that green AgNPs selectively reduced melanoma cell viability while sparing healthy keratinocytes (HaCaT), a benefit not observed with colloidal AgNPs. Proteomic analysis highlighted that green AgNPs significantly downregulated oncogenes, enhanced carbohydrate metabolism, and disrupted copper homeostasis in melanoma cells. This marks the first study to explore the differential effects of green and traditional AgNPs on melanoma using an integrated proteomic approach, underscoring the molecular potential of green AgNPs as a targeted and sustainable option for cancer therapy.

Melanoma is an aggressive cancer with increasing incidence and high mortality, driven by chemotherapy resistance and molecular dysregulation. Nanotechnology, specifically silver nanoparticles (AgNPs), shows promise as a treatment due to their ability to cause oxidative stress and apoptosis in cancer cells. However, traditional colloidal AgNPs lack selectivity, causing significant harm to normal cells. In this study, we present a green synthesis of AgNPs using plant extracts, which offers an eco-friendly alternative with a more targeted antitumor effect than traditional colloidal AgNPs. Using label-free DIA/SWATH MS quantitative proteomics, we compared the antitumor effects of AgNPs synthesized via traditional and green methods on A375 melanoma cells at 24 and 48 hours. Green AgNP exposure selectively reduced melanoma cell viability while sparing healthy keratinocytes (HaCaT), unlike Colloidal AgNPs. Proteomic analysis revealed that Green AgNPs exposure significantly decreased a panel of oncogenes and increasing carbohydrate metabolism, and dysregulating copper homeostasis. This is the first study to report the effects of differently synthesized AgNPs on melanoma cells using an integrated proteomic approach. It emphasizes the potential of green AgNPs in cancer therapy at a molecular level.