Designing an antimicrobial film for wound applications incorporating bacteriophages and ε-poly-l-lysine.

Alginate-based dressings have been shown to promote wound healing, leveraging the unique properties of alginate. This work aimed to develop and characterize flexible individual and bilayered films to deliver bacteriophages (phages) and ε-Poly-l-lysine (ε-PLL). Films varied in different properties. The moisture content, swelling and solubility increased with higher alginate concentrations. The water vapour permeability, crucial in biomedical films to balance moisture levels for effective wound healing, reached optimal levels in bilayer films, indicating these will be able to sustain an ideal moist environment. The bilayer films showed improved ductility (lower tensile strength and increased elongation at break) compared to individual films. The incorporated phages maintained viability for 12 weeks under vacuum and refrigerated conditions, and their release was sustained and gradual. Antibacterial immersion tests showed that films with phages and ε-PLL significantly inhibited Pseudomonas aeruginosa PAO1 growth (>3.1 Log CFU/cm2). Particle release was influenced by the swelling degree and diffusional processes within the polymer network, providing insights into controlled release mechanisms for particles of varying size (50 nm to 6 µm) and charge. The films developed, demonstrated modulated release capabilities for active agents, and may show potential as controlled delivery systems for phages and wound healing adjuvants.

Active Low-Density Polyethylene-Based Films by Incorporating α-Tocopherol in the Free State and Loaded in PLA Nanoparticles: A Comparative Study.

In this work, alpha-tocopherol (α-TOC) was encapsulated in poly(lactic acid) nanoparticles (PLA NPs) and added to low-density polyethylene (LDPE) films with the aim of producing an active film for food packaging applications. PLA NPs loaded with α-TOC were produced through nanoprecipitation and dried using two methods (freeze-dryer and oven). LDPE-based films with final polymeric matrix concentrations of 10 and 20 g/kg were then produced through blow extrusion. The results showed that LDPE-based films loaded with α-TOC can be produced using blow extrusion, and a good distribution of PLA NPs can be obtained within the LDPE matrix as observed using scanning electron microscopy (SEM). The mechanical properties were affected by the incorporation of α-TOC and PLA NPs loaded with α-TOC, with the observation of a decrease in tensile strength and Young's Modulus values and an increase in elongation at break. Regarding water vapor permeability, the films showed a reduction in the values with the addition of α-TOC and PLA NPs loaded with α-TOC compared to the LDPE film (control). Films with α-TOC in the free state and loaded in PLA NPs showed antioxidant activity, but their behavior was affected by the encapsulation process.

Rhamnolipids: A biosurfactant for the development of lipid-based nanosystems for food applications.

Biosurfactants (surfactants synthesized by microorganisms) are produced by microorganisms and are suitable for use in different areas. Among biosurfactants, rhamnolipids are the most studied and popular, attracting scientists, and industries' interest. Due to their unique characteristics, the rhamnolipids have been used as synthetic surfactants' alternatives and explored in food applications. Besides the production challenges that need to be tackled to guarantee efficient production and low cost, their properties need to be adjusted to the final application, where the pH instability needs to be considered. Moreover, regulatory approval is needed to start being used in commercial applications. One characteristic of interest is their capacity to form oil-in-water nanosystems. Some of the most explored have been nanoemulsions, solid-lipid nanoparticles and nanostructured lipid carriers. This review presents an overview of the main properties of rhamnolipids, asserts the potential and efficiency of rhamnolipids to replace the synthetic surfactants in the development of nanosystems, and describes the rhamnolipids-based nanosystems used in food applications. It also discusses the main characteristics and methodologies used for their characterization and in the end, some of the main challenges are highlighted.

Resveratrol-loaded octenyl succinic anhydride modified starch emulsions and hydroxypropyl methylcellulose (HPMC) microparticles: Cytotoxicity and antioxidant bioactivity assessment after in vitro digestion.

Hydroxypropyl methylcellulose (HPMC)-based microparticles and modified starch emulsions (OSA-MS) were loaded with resveratrol and characterized regarding their physicochemical and thermal properties. Both delivery systems were subject to an in vitro gastrointestinal digestion to assess the bioaccessibility of resveratrol. In addition, cell-based studies were conducted after in vitro digestion and cytotoxicity and oxidative stress were assessed. HPMC-based microparticles displayed higher average sizes (d) and lower polydispersity index (PDI) (d = 948 nm, PDI < 0.2) when compared to OSA-MS-based emulsions (d = 217 nm, PDI < 0.3). Both proved to protect resveratrol under digestive conditions, leading to an increase in bioaccessibility. Resveratrol-loaded HPMC-microparticles showed a higher bioaccessibility (56.7 %) than resveratrol-loaded emulsions (19.7 %). Digested samples were tested in differentiated co-cultures of Caco-2 and HT29-MTX, aiming at assessing cytotoxicity and oxidative stress, and a lack of cytotoxicity was observed for all samples. Results displayed an increasing antioxidant activity, with 1.6-fold and 1.4-fold increases over the antioxidant activity of free resveratrol, for HPMC-microparticles and OSA-MS nanoemulsions, respectively. Our results offer insight into physiological relevancy due to assessment post-digestion and highlight the protection that the use of micro-nano delivery systems can confer to resveratrol and their potential to be used as functional food ingredients capable of providing antioxidant benefits upon consumption.

Recent advances in oral delivery systems of resveratrol: foreseeing their use in functional foods.

Herein, we review the current state-of-the-art on the use of micro- and nano-delivery systems, a possible solution to some of the drawbacks associated with the incorporation of resveratrol in foods. Specifically, we present an overview of a wide range of micro-nanostructures, namely, lipidic and polymeric, used for the delivery of resveratrol. Also, the gastrointestinal fate of resveratrol-loaded micro-nanostructures, as a critical parameter for their use as functional food, is explored in terms of stability, bioaccessibility, and bioavailability. Different micro-nanostructures are of interest for the development of functional foods given that they can provide different advantages and properties to these foods and even be tailor-made to address specific issues (e.g., controlled or targeted release). Therefore, we discuss a wide range of micro-nanostructures, namely, lipidic and polymeric, used to deliver resveratrol and aimed at the development of functional foods. It has been reported that the use of some production methodologies can be of greater interest than others, for example, emulsification, solvent displacement and electrohydrodynamic processing (EHDP) enable a greater increase in bioaccessibility. Additionally, the use of coatings facilitates further improvements in bioaccessibility, which is likely due to the increased gastric stability of the coated micro-nanostructures. Other properties, such as mucoadhesion, can also help improve bioaccessibility due to the increase in gut retention time. Additionally, cytotoxicity (e.g., biocompatibility, antioxidant, and anti-inflammatory) and possible sensorial impact of resveratrol-loaded micro- and nano-systems in foods are highlighted.

Characterization of Sodium Alginate-Based Films Blended with Olive Leaf and Laurel Leaf Extracts Obtained by Ultrasound-Assisted Technology.

Due to environmental concerns, there is an increasing need to reduce the use of synthetic and non-renewable packaging materials to reduce waste and increase sustainability. This study aimed to characterise sodium alginate edible-based films (SA) incorporated with laurel leaf extract (LLE) and olive leaf extract (OLE) obtained by ultrasound-assisted extraction. Determination of total phenolic content, antioxidant, and antimicrobial activity was performed for the extracts and films. Also, thickness, tensile strength, elongation at break, modulus of elasticity, opacity and colour, moisture content, water vapour permeability (WVP), Fourier-transform infrared spectroscopy (FTIR) spectra, and surface morphology by scanning electron microscope (SEM) analyses were performed for the films. LLE yielded better results in terms of phenolic content (195 mg GAE/g), antioxidant (2.1 TE/g extract) and antimicrobial activity (MIC at 1% for Listeria monocytogenes and Staphylococcus aureus, and 1.8% for Enterococcus faecalis). For the films, the simultaneous incorporation of LLE 1% (w/v) and OLE 1% (w/v) resulted in a significant reduction of approximately 2 log CFU/g against S. aureus. The addition of LLE and OLE extracts also proved to improve barrier properties (lower WVP for SA films with LLE 1% + OLE 1%, 3.49 × 10-11 g m-1 s-1 Pa-1) and promoted changes in resistance and flexibility. The results demonstrated that active alginate-based films can be valuable for enhancing food preservation.

Evaluation of the Antimicrobial Activity of Chitosan Nanoparticles against Listeria monocytogenes.

Chitosan is obtained from the deacetylation of chitin, and it is known to possess antimicrobial activity. It has attracted attention as it may be used for treating infections caused by different types of microorganisms due to its broad spectrum. Its application in the form of micro- or nanoparticles (CM/CN) has expanded its usage, as in this form, it retains its activity, and remain stable in aqueous solutions. However, inconsistencies in the results reported by different authors have been identified. In this communication, the antimicrobial activity of CN produced from different starting materials was tested against Listeria monocytogenes. It was observed that, even though all the starting materials were reported to have a molecular weight (MW) below 200 kDa and degree of deacetylation (DD) > 75%, the size of the CNs were significantly different (263 nm vs. 607 nm). Furthermore, these differences in sizes exerted a direct effect on the antimicrobial properties of the particles, as when testing the ones with the smallest size, i.e., 263 nm, a lower Minimum Inhibitory Concentration (MIC) was achieved, i.e., 0.04 mg/mL. Even though the largest particles, i.e., 607 nm, in individual experiments were able to achieve an MIC of 0.03 mg/mL, the results with CN presented great variation among replicates and up to 0.2 mg/mL were needed in other replicates. The starting material has a critical impact on the properties of the CN, and it must be carefully characterized and selected for the intended application, and MW and DD solely do not fully account for these properties.

Encapsulation of vitamin D3 using rhamnolipids-based nanostructured lipid carriers.

This work had as main objective to encapsulate vitamin D3 (VD3) into nanostructured lipid carriers (NLCs) using rhamnolipids as surfactant. Glycerol monostearate and medium chain triglycerides with 2.625 % of VD3 were used as lipid materials. The three formulations of NLCs with VD3 (NLCs + VD3) were composed by 99 % of aqueous phase, 1 % of lipid phase and 0.05 % of surfactant. The difference between them was the ratio of solid:liquid in lipid phase. The NLCs + VD3 sizes ranged between 92.1 and 108.1 nm. The most stable formulation maintaining their caracteristics for 60 days at 4 °C. The NLCs + VD3 cytotoxicity demonstrated that concentrations of 0.25 mg/mL or lower up had a good biocompatibility in vitro. During the in vitro digestion, formulations with lower sizes and higher content on solid lipid had higher lipolysis rate and consequently higher VD3 bioaccessibility. The rhamnolipids-based NLCs are a good option for the encapsulation of VD3.

Bacteriophage Delivery Systems for Food Applications: Opportunities and Perspectives.

Currently, one-third of all food produced worldwide is wasted or lost, and bacterial contamination is one of the main reasons. Moreover, foodborne diseases are a severe problem, causing more than 420,000 deaths and nearly 600 million illnesses yearly, demanding more attention to food safety. Thus, new solutions need to be explored to tackle these problems. A possible solution for bacterial contamination is using bacteriophages (phages), which are harmless to humans; these natural viruses can be used to prevent or reduce food contamination by foodborne pathogens. In this regard, several studies showed the effectiveness of phages against bacteria. However, when used in their free form, phages can lose infectivity, decreasing the application in foods. To overcome this problem, new delivery systems are being studied to incorporate phages and ensure prolonged activity and controlled release in food systems. This review focuses on the existent and new phage delivery systems applied in the food industry to promote food safety. Initially, an overview of phages, their main advantages, and challenges is presented, followed by the different delivery systems, focused in methodologies, and biomaterials that can be used. In the end, examples of phage applications in foods are disclosed and future perspectives are approached.

Edible alginate-based films with anti-SARS-CoV-2 activity.

The viability of SARS-CoV-2 on food surfaces and its propagation through the food chain has been discussed by several stakeholders, as it may represent a serious public health problem, bringing new challenges to the food system. This work shows for the first time that edible films can be used against SARS-CoV-2. Sodium alginate-based films containing gallic acid, geraniol, and green tea extract were evaluated in terms of their antiviral activity against SARS-CoV-2. The results showed that all these films have strong in vitro antiviral activity against this virus. However, a higher concentration of the active compound (1.25%) is needed for the film containing gallic acid to achieve similar results to those obtained for lower concentrations of geraniol and green tea extract (0.313%). Furthermore, critical concentrations of the active compounds in the films were used to evaluate their stability during storage. Results showed that gallic acid-loaded films lose their activity from the second week of storage, while films with geraniol and green tea extract only show a drop in activity after four weeks. These results highlight the possibility of using edible films and coatings as antiviral materials on food surfaces or food contact materials, which may help to reduce the spreading of viruses through the food chain.

Postharvest Quality Improvement of Tomato (Solanum lycopersicum L.) Fruit Using a Nanomultilayer Coating Containing Aloe vera.

The effectiveness of an alginate/chitosan nanomultilayer coating without (NM) and with Aloe vera liquid fraction (NM+Av) was evaluated on the postharvest quality of tomato fruit at 20 °C and 85% relative humidity (RH) to simulate direct consumption. Both nanomultilayer coatings had comparable effects on firmness and pH values. However, the NM+Av coating significantly reduced weight loss (4.5 ± 0.2%) and molds and yeasts (3.5-4.0 log CFU g-1) compared to uncoated fruit (16.2 ± 1.2% and 8.0 ± 0.0 log CFU g-1, respectively). It notably lowered O2 consumption by 70% and a 52% decrease in CO2 production, inhibiting ethylene synthesis. Visual evaluation confirmed NM+Av's efficacy in preserving the postharvest quality of tomato. The preservation of color, indicated by the Minolta color (a*/b*) values, demonstrated NM+Av's ability to keep the light red stage compared to uncoated fruit. The favorable effects of NM+Av coating on enhancing postharvest quality indicates it as a potential alternative for large-scale tomato fruit preservation.

Assessment of the Antibiofilm Performance of Chitosan-Based Surfaces in Marine Environments.

Marine biofouling is a natural process often associated with biofilm formation on submerged surfaces, creating a massive economic and ecological burden. Although several antifouling paints have been used to prevent biofouling, growing ecological concerns emphasize the need to develop new and environmentally friendly antifouling approaches such as bio-based coatings. Chitosan (CS) is a natural polymer that has been widely used due to its outstanding biological properties, including non-toxicity and antimicrobial activity. This work aims to produce and characterize poly (lactic acid) (PLA)-CS surfaces with CS of different molecular weight (Mw) at different concentrations for application in marine paints. Loligo opalescens pens, a waste from the fishery industry, were used as a CS source. The antimicrobial activity of the CS and CS-functionalized surfaces was assessed against Cobetia marina, a model proteobacterium for marine biofouling. Results demonstrate that CS targets the bacterial cell membrane, and PLA-CS surfaces were able to reduce the number of culturable cells up to 68% compared to control, with this activity dependent on CS Mw. The antifouling performance was corroborated by Optical Coherence Tomography since PLA-CS surfaces reduced the biofilm thickness by up to 36%, as well as the percentage and size of biofilm empty spaces. Overall, CS coatings showed to be a promising approach to reducing biofouling in marine environments mimicked in this work, contributing to the valorization of fishing waste and encouraging further research on this topic.

Alginate Particles for Encapsulation of Phenolic Extract from Spirulina sp. LEB-18: Physicochemical Characterization and Assessment of In Vitro Gastrointestinal Behavior.

Encapsulation can be used as a strategy to protect and control the release of bioactive extracts. In this work, an extract from Spirulina sp. LEB-18, rich in phenolic compounds, was encapsulated in biopolymeric particles (i.e., composed of alginate) and characterized concerning their thermal behavior using differential scanning calorimetry (DSC), size, morphology, swelling index (S), and encapsulation efficiency (EE%); the release profile of the phenolic compounds at different pHs and the particle behavior under in vitro gastrointestinal digestion were also evaluated. It was shown that it is possible to encapsulate the phenolic extract from Spirulina sp. LEB-18 in alginate particles with high encapsulation efficiency (88.97%). It was also observed that the particles are amorphous and that the encapsulated phenolic compounds were released at a pH 7.2 but not at pH 1.5, which means that the alginate particles are able to protect the phenolic compounds from the harsh stomach conditions but lose their integrity under intestinal pH conditions. Regarding bioaccessibility, it was observed that the encapsulated phenolic compounds showed higher bioaccessibility compared to phenolic compounds in free form. This work increases the knowledge about the behavior of alginate particles encapsulating phenolic compounds during in vitro gastrointestinal digestion. It also provides essential information for designing biopolymeric particle formulations encapsulating phenolic compounds for application in pharmaceutical and food products.

Oleogels and Organogels: A Promising Tool for New Functionalities.

Growing awareness concerning human health and sustainability has been continually driving the need to change consumers' habits and develop new bio-based and environmentally friendly materials that could be used in new product formulations [...].

Zn and Zn-Fe Nanostructures with Multifunctional Properties as Components for Food Packaging Materials.

Metallic and bimetallic nanostructures have shown interesting chromatic and antibacterial properties, and they can be used in various applications. In this work, zinc (Zn) and iron (Fe) nanostructures were produced with different morphologies: (i) pure Zn; (ii) Zn-Fe nanoalloys; (iii) Zn-Fe nanolayers (Zn-Fe NLs); and (iv) Zn nanolayers combined with Fe nanoparticles (Zn NLs + Fe NPs). The aim was to produce components for food packaging materials with active and intelligent properties, including oxygen absorption capacity, chromatic properties, and antibacterial properties. Thus, the morphology, structure, and chemical composition of the samples were characterized and correlated with their oxidation, chromatic, and antibacterial properties. The results revealed a relevant reduction in the coating's opacity after oxidation varying from 100 to 10% depending on the morphology of the system. All coatings exhibited significant antibacterial activity against S. aureus, revealing a direct correlation with Zn content. The incorporation of Fe for all atomic arrangements showed a negative impact on the antibacterial effect against E. coli, decreasing to less than half the zone of inhibition for Zn-Fe NLs and Zn NLs + Fe NPs and suppressing the antibacterial effect for Zn-Fe alloy when compared with the pure Zn system.

Active Flexible Films for Food Packaging: A Review.

Active food packaging is a dynamic area where the scientific community and industry have been trying to find new strategies to produce innovative packaging that is economically viable and compatible with conventional production processes. The materials used to develop active packaging can be organized into scavenging and emitting materials, and based on organic and inorganic materials. However, the incorporation of these materials in polymer-based flexible packaging is not always straightforward. The challenges to be faced are mainly related to active agents' sensitivity to high temperatures or difficulties in dispersing them in the high viscosity polymer matrix. This review provides an overview of methodologies and processes used in the production of active packaging, particularly for the production of active flexible films at the industrial level. The direct incorporation of active agents in polymer films is presented, focusing on the processing conditions and their effect on the active agent, and final application of the packaging material. Moreover, the incorporation of active agents by coating technologies and supercritical impregnation are presented. Finally, the use of carriers to help the incorporation of active agents and several methodologies is discussed. This review aims to guide academic and industrial researchers in the development of active flexible packaging, namely in the selection of the materials, methodologies, and process conditions.

Gelation Behavior and Stability of Multicomponent Sterol-Based Oleogels.

Novel fat mimetic materials, such as oleogels, are advancing the personalization of healthier food products and can be developed from low molecular weight compounds such as γ-oryzanol and ß-sitosterol. Following molecular assembly, the formation of a tubular system ensues, which seems to be influenced by elements such as the oleogelators' concentration and ratio, cooling rates, and storage periods. Sterol-based oleogels were formulated under distinct environmental conditions, and a comprehensive study aimed to assess the effects of the mentioned factors on oleogel formation and stability, through visual observation and by using techniques such as small-angle X-ray scattering, X-ray diffraction, confocal Raman spectroscopy, rheology, and polarized microscopy. The long, rod-like conformations, identified by small-angle X-ray scattering, showed that different cooling rates influence oleogels' texture. Raman spectra showed that the stabilization time is associated with the interfibrillar aggregation, which occurred differently for 8 and 10 wt%, with a proven relationship between ferulic acid and the tubular formation. This report gives fundamental insight into the critical point of gelation, referring to the time scale of the molecular stabilization. Our results verify that understanding the structuring mechanisms of oleogelation is decisive for the processing and manufacturing of novel foods which integrate oleogels in their structure.

Performance of Gelatin Films Reinforced with Cloisite Na+ and Black Pepper Essential Oil Loaded Nanoemulsion.

The concern about consuming eco-friendly products has motivated research in the development of new materials. Therefore, films based on natural polymers have been used to replace traditional polymers. This study consists of a production of films based on gelatin reinforced with black pepper essential oil-loaded nanoemulsions and Cloisite Na+. The films were characterized by water vapor permeability, mechanical and thermal properties, surface contact angle, X-ray diffraction and scanning electron microscopy. It was observed that the films containing the nanoemulsion have higher permeability values and an increase in their mechanical resistance. The addition of nanoclay contributed to an increase in the surface hydrophobicity of the film and an increase in the tensile strength, at break, by about 150%. The addition of essential oil nanoemulsions led to an increase in thermal stability. The presence of clay dispersion contributed to the formation of a surface that was slightly rougher and grainier. The addition of the black pepper essential oil nanoemulsion resulted in an increase in porosity of the gelatin matrix. Through X-ray diffraction analysis, it was possible to conclude that both the polymeric gelatin matrix and the essential oils nanoemulsion are intercalated with the clay dispersion.

The Effect of Molecular Weight on the Antimicrobial Activity of Chitosan from Loligo opalescens for Food Packaging Applications.

The growing requirement for sustainable processes has boosted the development of biodegradable plastic-based materials incorporating bioactive compounds obtained from waste, adding value to these products. Chitosan (Ch) is a biopolymer that can be obtained by deacetylation of chitin (found abundantly in waste from the fishery industry) and has valuable properties such as biocompatibility, biodegradability, antimicrobial activity, and easy film-forming ability. This study aimed to produce and characterize poly(lactic acid) (PLA) surfaces coated with ß-chitosan and ß-chitooligosaccharides from a Loligo opalescens pen with different molecular weights for application in the food industry. The PLA films with native and depolymerized Ch were functionalized through plasma oxygen treatment followed by dip-coating, and their physicochemical properties were assessed by Fourier-transform infrared spectroscopy, X-ray diffraction, water contact angle, and scanning electron microscopy. Their antimicrobial properties were assessed against Escherichia coli and Pseudomonas putida, where Ch-based surfaces reduced the number of biofilm viable, viable but nonculturable, and culturable cells by up to 73%, 74%, and 87%, respectively, compared to PLA. Biofilm growth inhibition was confirmed by confocal laser scanning microscopy. Results suggest that Ch films of higher molecular weight had higher antibiofilm activity under the food storage conditions mimicked in this work, contributing simultaneously to the reuse of marine waste.

Active Carboxymethylcellulose-Based Edible Films: Influence of Free and Encapsulated Curcumin on Films' Properties.

Carboxymethylcellulose (CMC)-based films can act as a protective barrier in food surfaces and a carrier of bioactive compounds, such as curcumin. However, incorporating curcumin in hydrophilic matrixes can be a challenge, and new strategies need to be explored. In this work, CMC-based films containing free curcumin and curcumin-loaded nanohydrogels (composed of lactoferrin and glycomacropeptide) were produced and characterized. The incorporation of curcumin-loaded nanohydrogels showed a significant decrease in films' thickness (from 0.0791 to 0.029 mm). Furthermore, the water vapor permeability of CMC-based films was significantly decreased (62%) by incorporating curcumin-loaded nanohydrogels in the films. The water affinity's properties (moisture, solubility, and contact angle) of films were also affected by incorporating encapsulated curcumin. The addition of nanohydrogels to CMC-based films reduced the tensile strength values from 16.46 to 9.87 MPa. Chemical interactions were analyzed using Fourier transform infrared spectroscopy. The release profile of curcumin from CMC-based films was evaluated at 25 °C using a hydrophilic food simulant and suggests that the release mechanism of the curcumin happens by Fick's diffusion and Case II transport. Results showed that protein-based nanohydrogels can be a good strategy for incorporating curcumin in edible films, highlighting their potential for use in food applications.