Electrospun films incorporating humic substances of application interest in sustainable active food packaging.

Sustainable active food packaging is essential to reduce the use of plastics, preserve food quality and minimize the environmental impact. Humic substances (HS) are rich in redox-active compounds, such as quinones, phenols, carboxyl, and hydroxyl moieties, making them functional additives for biopolymeric matrices, such as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Herein, composites made by incorporating different amounts of HS into PHBV were developed using the electrospinning technology and converted into homogeneous and continuous films by a thermal post-treatment to obtain a bioactive and biodegradable layer which could be part of a multilayer food packaging solution. The morphology, thermal, optical, mechanical, antioxidant and barrier properties of the resulting PHBV-based films have been evaluated, as well as the antifungal activity against Aspergillus flavus and Candida albicans and the antimicrobial properties against both Gram (+) and Gram (-) bacterial strains. HS show great potential as natural additives for biopolymer matrices, since they confer antioxidant, antimicrobial, and antifungal properties to the resulting materials. In addition, barrier, optical and mechanical properties highlighted that the obtained films are suitable for sustainable active packaging. Therefore, the electrospinning methodology is a promising and sustainable approach to give biowaste a new life through the development of multifunctional materials suitable in the active bio-packaging.

Humic substance/metal-oxide multifunctional nanoparticles as advanced antibacterial-antimycotic agents and photocatalysts for the degradation of PLA microplastics under UVA/solar radiation.

As a result of the accumulation of plastic in the environment, microplastics have become part of the food chain, boosting the resistance of fungi and bacteria which can frequently encounter human beings. Employing photocatalytic degradation is a possible route towards the removal of chemical and biological pollutants, such as plastics and microplastic wastes as well as microorganisms. Using biowaste materials to design hybrid nanoparticles with enhanced photocatalytic and antimicrobial features would uphold the principles of the circular bioeconomy. Here, two unexpensive semiconductors-namely titanium dioxide (TiO2) and zinc oxide (ZnO) - were synthetized through solvothermal synthesis and combined with humic substances deriving from agrifood biomass. The preparation led to hybrid nanoparticles exhibiting enhanced ROS-generating properties for simultaneous applications as antimicrobial agents against different bacterial and fungal strains and as photoactive catalysts to degrade polylactic acid (PLA) microplastics under UVA and solar irradiation. In comparison to bare nanoparticles, hybrid nanoparticles demonstrated higher antibacterial and antimycotic capabilities toward various pathogenic microorganisms as well as advanced photocatalytic activity in the degradation of PLA with a carbonyl index reduction in the range of 15-23%, thus confirming a noteworthy ability in microplastics photodegradation under UVA and solar irradiation.

Humic acids on fire? Physico-chemical, thermal, flammability features and extraction process of different humic acids in support of their possible applications.

Humic acids (HA) consist in a multitude of heterogeneous organic molecules surviving the biological and chemical degradation of both vegetal and animal biomasses. The great abundance and chemical richness of these residues make their valorisation one of the most promising approaches to move towards a circular economy. However, the heterogeneity of the biomass from which HA are extracted, as well as the production process, significantly affects the nature and the relative content of functional groups (i.e. quinones, phenols and carboxylic and hydroxyl moieties), eventually changing HA reactivity and ultimately determining their application field. Indeed, depending on their properties, these substances can be used as flame retardants in the case of pronounced resilience degree (i.e., absent or low reactivity), or as antioxidant or antimicrobial agents in the case of pronounced reactivity, thanks to their redox behaviour. In this work we investigated the flammable, the thermal and the physico-chemical features of HA extracted from different composted biomasses to identify the reactivity or the resiliency of these moieties. Several techniques, including flammability characterization (LIT and MIE), laser diffraction granulometry, TG, XRD analyses, FTIR spectroscopy on both solid and gaseous phases, and Raman spectroscopy were integrated to investigate the correlation among the safety parameters, the distributions of particle sizes, as well as the thermal, the chemical properties of HA powders and the influence of post-extraction processes on HA final properties.

Waste to Wealth Approach: Improved Antimicrobial Properties in Bioactive Hydrogels through Humic Substance-Gelatin Chemical Conjugation.

Exploring opportunities for biowaste valorization, herein, humic substances (HS) were combined with gelatin, a hydrophilic biocompatible and bioavailable polymer, to obtain 3D hydrogels. Hybrid gels (Gel HS) were prepared at different HS contents, exploiting physical or chemical cross-linking, through 1-ethyl-(3-3-dimethylaminopropyl)carbodiimide (EDC) chemistry, between HS and gelatin. Physicochemical features were assessed through rheological measurements, X-ray diffraction, attenuated total reflectance (ATR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and scanning electron microscopy (SEM). ATR and NMR spectroscopies suggested the formation of an amide bond between HS and Gel via EDC chemistry. In addition, antioxidant and antimicrobial features toward both Gram(-) and Gram(+) strains were evaluated. HS confers great antioxidant and widespread antibiotic performance to the whole gel. Furthermore, the chemical cross-linking affects the viscoelastic behavior, crystalline structures, water uptake, and functional performance and produces a marked improvement of biocide action.

Shall We Tune? From Core-Shell to Cloud Type Nanostructures in Heparin/Silica Hybrids.

Heparin plays multiple biological roles depending on the availability of active sites strongly influenced by the conformation and the structure of polysaccharide chains. Combining different components at the molecular scale offers an extraordinary chance to easily tune the structural organization of heparin required for exploring new potential applications. In fact, the combination of different material types leads to challenges that cannot be achieved by each single component. In this study, hybrid heparin/silica nanoparticles were synthesized, and the role of silica as a templating agent for heparin supramolecular organization was investigated. The effect of synthesis parameters on particles compositions was deeply investigated by Fourier Transform Infrared Spectroscopy (FTIR) and Thermogravimetric Analysis (TGA). Transmission Electron Microscopy (TEM) reveals a different supramolecular organization of both components, leading to amazing organic-inorganic nanoparticles with different behavior in drug encapsulation and release. Furthermore, favorable biocompatibility for healthy human dermal fibroblasts (HDF) and tumor HS578T cells has been assessed, and a different biological behavior was observed, ascribed to different surface charge and morphology of synthesized nanoparticles.

A study on structural evolution of hybrid humic Acids-SiO2 nanostructures in pure water: Effects on physico-chemical and functional properties.

Humic acids (HA) are considered a promising and inexpensive source for novel multifunctional materials for a huge range of applications. However, aggregation and degradation phenomena in aqueous environment prevent from their full exploitation. A valid strategy to address these issues relies on combining HA moieties at the molecular scale with an inorganic nanostructured component, leading to more stable hybrid nanomaterials with tunable functionalities. Indeed, chemical composition of HA can determine their interactions with the inorganic constituent in the hybrid nanoparticles and consequently affect their overall physico-chemical properties, including their stability and functional properties in aqueous environment. As a fundamental contribution to HA materials-based technology, this study aims at unveiling this aspect. To this purpose, SiO2 nanoparticles have been chosen as a model platform and three different HAs extracted from composted biomasses, manure (HA_Man), artichoke residues (HA_Art) and coffee grounds (HA_Cof), were employed to synthetize hybrid HA-SiO2 nanoparticles through in-situ sol-gel synthesis. Prepared samples were submitted to aging in water to assess their stability. Furthermore, antioxidant properties and physico-chemical properties of both as prepared and aged samples in aqueous environment were assessed through Scanning Electron Microscopy (SEM), N2 physisorption, Simultaneous Thermogravimetric (TGA) and Differential Scanning Calorimetric (DSC) Analysis, Fourier Transform Infrared (FT-IR), Nuclear Magnetic Resonance (NMR), Electron Paramagnetic Resonance (EPR) spectroscopies. The experimental results highlighted that hybrid HA-SiO2 nanostructures acted as dynamic systems which exhibit structural supramolecular reorganization during aging in aqueous environment with marked effects on physico-chemical and functional properties, including improved antioxidant activity. Obtained results enlighten a unique aspect of interactions between HA and inorganic nanoparticles that could be useful to predict their behavior in aqueous environment. Furthermore, the proposed approach traces a technological route for the exploitation of organic biowaste in the design of hybrid nanomaterials, providing a significant contribution to the development of waste to wealth strategies based on humic substances.

Adding Humic Acids to Gelatin Hydrogels: A Way to Tune Gelation.

Exploring the chance to convert biowaste into a valuable resource, this study tests the potential role of humic acids (HA), a class of multifunctional compounds obtained by oxidative decomposition of biomass, as physical agents to improve gelatin's mechanical and thermal properties. To this purpose, gelatin-HA aqueous samples were prepared at increasing HA content. HA/gelatin concentrations changed in the range 2.67-26.67 (wt/wt)%. Multiple techniques were employed to assess the influence of HA content on the gel properties and to unveil the underlying mechanisms. HAs increased gel strength up to a concentration of 13.33 (wt/wt)% and led to a weaker gel at higher concentrations. FT-IR and DSC results proved that HAs can establish noncovalent interactions through H-bonding with gelatin. Coagulation phenomena occur because of HA-gelatin interactions, and at concentrations greater than 13.33 (wt/wt)%, HAs established preferential bonds with water molecules, preventing them from coordinating with gelatin chains. These features were accompanied by a change in the secondary structure of gelatin, which lost the triple helix structure and exhibited an increase in the random coil conformation. Besides, higher HA weight content caused swelling phenomena due to HA water absorption, contributing to a weaker gel. The current findings may be useful to enable a better control of gelatin structures modified with composted biowaste, extending their exploitation for a large set of technological applications.

Hybrid humic acid/titanium dioxide nanomaterials as highly effective antimicrobial agents against gram(-) pathogens and antibiotic contaminants in wastewater.

Humic acids (HAs) provide an important bio-source for redox-active materials. Their functional chemical groups are responsible for several properties, such as metal ion chelating activity, adsorption ability towards small molecules and antibacterial activity, through reactive oxygen species (ROS) generation. However, the poor selectivity and instability of HAs in solution hinder their application. A promising strategy for overcoming these disadvantages is conjugation with an inorganic phase, which leads to more stable hybrid nanomaterials with tuneable functionalities. In this study, we demonstrate that hybrid humic acid/titanium dioxide nanostructured materials that are prepared via a versatile in situ hydrothermal strategy display promising antibacterial activity against various pathogens and behave as selective sequestering agents of amoxicillin and tetracycline antibiotics from wastewater. A physicochemical investigation in which a combination of techniques were utilized, which included TEM, BET, 13C-CPMAS-NMR, EPR, DLS and SANS, shed light on the structure-property-function relationships of the nanohybrids. The proposed approach traces a technological path for the exploitation of organic biowaste in the design at the molecular scale of multifunctional nanomaterials, which is useful for addressing environmental and health problems that are related to water contamination by antibiotics and pathogens.

Adsorption of Cellulase on Wrinkled Silica Nanoparticles with Enhanced Inter-Wrinkle Distance.

Mesoporous silica materials offer a unique opportunity for enzyme immobilization thanks to their properties, such as tuneable pore size, large surface area and easy functionalization. However, a significant enhancement of cellulase enzyme activity entrapped inside the silica pores still represents a challenge. In this work, we immobilized cellulase by adsorption on wrinkled silica nanoparticles (WSNs), obtaining an active and stable biocatalyst. We used pentanol as co-solvent to synthesize WSNs with enhanced inter-wrinkle distance in order to improve cellulase hosting. The physical-chemical and morphological characterization of WSNs and cellulase/WSNs was performed by thermogravimetric (TG), Fourier transform infrared (FT-IR), and transmission electron microscopy (TEM) analyses. The obtained results showed that this matrix generates a favourable microenvironment for hosting cellulase. The results of the catalytic assays and operational stability confirmed the key role of size, morphology and distribution of the pores in the successful outcome of the cellulase immobilization process. The immobilization procedure used allowed preserving most of the secondary structure of the enzyme and, consequently, its catalytic activity. Moreover, the same value of glucose yield was observed for five consecutive runs, showing a high operational stability of the biocatalyst.

CFD Simulations of Microreactors for the Hydrolysis of Cellobiose to Glucose by ß-Glucosidase Enzyme.

The enzymatic hydrolysis of lignocellulosic biomass-derived compounds represents a valid strategy to reduce the dependence on fossil fuels, with geopolitical and environmental benefits. In particular, ß-glucosidase (BG) enzyme is the bottleneck in the degradation of cellulose because it catalyzes the hydrolysis of cellobiose, a known inhibitor of the other cellulolytic enzymes. However, free enzymes are unstable, expensive and difficult to recover. For this reason, the immobilization of BG on a suitable support is crucial to improve its catalytic performance. In this paper, computational fluid dynamics (CFD) simulations were performed to test the hydrolysis reaction in a monolith channel coated by BG adsorbed on a wrinkled silica nanoparticles (WSNs) washcoat. We initially defined the physical properties of the mixture, the parameters related to kinetics and mass transfers and the initial and boundary conditions thanks to our preliminary experimental tests. Numerical simulation results have shown great similarity with the experimental ones, demonstrating the validity of this model. Following this, it was possible to explore in real time the behavior of the system, varying other specified parameters (i.e., the mixture inlet velocity or the enzymatic load on the reactor surface) without carrying out other experimental analyses.

Tuning Functional Behavior of Humic Acids through Interactions with Stöber Silica Nanoparticles.

Humic acids (HA) exhibit fascinating multifunctional features, yet degradation phenomena as well as poor stability in aqueous environments strongly limit their use. Inorganic nanoparticles are emerging as a powerful interface for the development of robust HA bio-hybrid materials with enhanced chemical stability and tunable properties. Hybrid organic-inorganic SiO2/HA nanostructures were synthesized via an in-situ sol-gel route, exploiting both physical entrapment and chemical coupling. The latter was achieved through amide bond formation between carboxyl groups of HA and the amino group of 3-aminopropyltriethoxysilane (APTS), as confirmed by Fourier-Transform Infrared (FTIR) and Nuclear Magnetic Resonance (NMR) spectroscopy. Monodisperse hybrid nanoparticles about 90 nm in diameter were obtained in both cases, yet Electron Paramagnetic Resonance (EPR) spectroscopy highlighted the different supramolecular organization of HA. The altered HA conformation was reflected in different antioxidant properties of the conjugated nanoparticles that, however, resulted in being higher than for pure HA. Our findings proved the key role of both components in defining the morphology of the final system, as well as the efficacy of the ceramic component in templating the HA supramolecular organization and consequently tuning their functional features, thus defining a green strategy for bio-waste valorization.

Improving Interaction at Polymer-Filler Interface: The Efficacy of Wrinkle Texture.

One of the main issues in preparing polymer-based nanocomposites with effective properties is to achieve a good dispersion of the nanoparticles into the matrix. Chemical interfacial modifications by specific coupling agents represents a good way to reach this objective. Actually, time consuming compatibilization procedures strongly compromise the sustainability of these strategies. In this study, the role of particles' architectures in their dispersion into a poly-lactic acid matrix and their subsequent influences on physical-chemical properties of the obtained nanocomposites were investigated. Two kinds of silica nanoparticles, "smooth" and "wrinkled," with different surface areas (≈30 and ≈600 m2/g respectively) were synthesized through a modified Stöber method and used, without any chemical surface pre-treatments, as fillers to produce poly-lactic acid based nanocomposites. The key role played by wrinkled texture in modifying the physical interaction at the polymer-filler interface and in driving composite properties, was investigated and reflected in the final bulk properties. Detailed investigations revealed the presence of wrinkled nanoparticles, leading to (i) an enormous increase of the chain relaxation time, by almost 30 times compared to the neat PLA matrix; (ii) intensification of the shear-thinning behavior at low shear-rates; and (iii) slightly slower thermal degradation of polylactic acid.

Covalent Immobilization of ß-Glucosidase into Mesoporous Silica Nanoparticles from Anhydrous Acetone Enhances Its Catalytic Performance.

An immobilization protocol of a model enzyme into silica nanoparticles was applied. This protocol exploited the use of the bifunctional molecule triethoxysilylpropylisocyanate (TEPI) for covalent binding through a linker of suitable length. The enzyme ß-glucosidase (BG) was anchored onto wrinkled silica nanoparticles (WSNs). BG represents a bottleneck in the conversion of lignocellulosic biomass into biofuels through cellulose hydrolysis and fermentation. The key aspect of the procedure was the use of an organic solvent (anhydrous acetone) in which the enzyme was not soluble. This aimed to restrict its conformational changes and thus preserve its native structure. This approach led to a biocatalyst with improved thermal stability, characterized by high immobilization efficiency and yield. It was found that the apparent KM value was about half of that of the free enzyme. The Vmax was about the same than that of the free enzyme. The biocatalyst showed a high operational stability, losing only 30% of its activity after seven reuses.