Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides.

In this study, multilayer microcapsules (two-layer and four-layer) based on furcellaran (FUR) and chitosan (CHIT) were produced, enclosing a tripeptide with an antioxidant effect-glutathione-in different concentrations. In addition, for the first time, an empty, four-layer microcapsule based on CHIT and FUR (ECAPS) was obtained, which can be used to contain sensitive, active substances of a hydrophobic nature. Layering was monitored using zeta potential, and the presence of the resulting capsules was confirmed by SEM imaging. In the current study, we also investigated whether the studied capsules had any effect on the Hep G2 cancer cell line. An attempt was also made to identify the possible molecular mechanism(s) by which the examined capsules suppressed the growth of Hep G2 cells. In this report, we demonstrate that the capsules suppressed the growth of cancer cells. This mechanism was linked to the modulation of the AKT/PI3K signaling pathway and the induction of the G2/M arrest cell cycle. Furthermore, the results indicate that the tested multilayer microcapsules induced cell death through an apoptotic pathway.

Recent progress on Pickering emulsion stabilized essential oil added biopolymer-based film for food packaging applications: A review.

Nowadays food safety and protection are a growing concern for food producers and food industry. The stability of food-grade materials is key in food processing and shelf life. Pickering emulsions (PEs) have gained significant attention in food regimes owing to their stability enhancement of food specimens. PE can be developed by high and low-energy methods. The use of PE in the food sector is completely safe as it uses solid biodegradable particles to stabilize the oil in water and it also acts as an excellent carrier of essential oils (EOs). EOs are useful functional ingredients, the inclusion of EOs in the packaging film or coating formulation significantly helps in the improvement of the shelf life of the packed food item. The highly volatile nature, limited solubility and ease of oxidation in light of EOs restricts their direct use in packaging. In this context, the use of PEs of EOs is suitable to overcome most of the challenges, Therefore, recently there have been many papers published on PEs of EOs including active packaging film and coatings and the obtained results are promising. The current review amalgamates these studies to inform about the chemistry of PEs followed by types of stabilizers, factors affecting the stability and different high and low-energy manufacturing methods. Finally, the review summarizes the recent advancement in PEs-added packaging film and their application in the enhancement of shelf life of food.

Influence of Curcuma Longa extract in citral addition on functional properties of thin films with triple-layer structure based on furcellaran and gelatin.

In this study, we obtained triple-layer films based on furcellaran and gelatin, in which the middle layer was enriched with extract of Curcuma longa in citral. This newly developed material underwent a comprehensive characterisation process to identify significant improvements in its functional properties. Both SEM, XRD and FTIR analyzes indicated the formation of interactions not only between the components but also between the film layers. Notably, the incorporation of the natural extract led to a significant reduction in solubility, decreasing it from 74.79 % to 57.25 %, while enhancing thermal stability expressed as a melting point elevating it from 147.10 °C in the control film to 158.80 °C in the film with the highest concentration of the active ingredient. Simultaneously, the addition of this active ingredient resulted in decreased water contact angle (WCA) values, rendering the film more hydrophilic. The produced films exhibit great promise as packaging materials, particularly within the food industry, and the conducted research is marked by its forward-looking and developmental approach.

Application of antimicrobial chitosan-Furcellaran-hydrolysate gelatin edible coatings enriched with bioactive peptides in shelf-life extension of pork loin stored at 4 and -20 °C.

In this study, the authors investigate the preservative capabilities of edible coatings comprising a blend of chitosan, furcellaran and gelatin hydrolysate enhanced with the bioactive peptides RW4 and LL37. The preservative effects on pork samples stored for 21 days at 4 °C and 6 months at -20 °C were studied, while examining changes in microbiological contamination, pH levels, water activity and sensory attributes. Microbiological analyses reveal the coatings' antimicrobial efficacy against aerobic bacteria, microscopic fungi and yeasts, particularly during the initial storage period, when coated samples exhibit microbial reductions of 0.5-2 log CFU/g compared to the controls. The coatings have no discernible impact on water activity during storage in refrigerated or freezer conditions. Notably, differences in pH development can be observed between the coated and control samples, potentially attributable to the antimicrobial action of the coatings. Sensory analysis allows to highlight the inhibition of deterioration related to sensory attributes through the use of edible coatings. In conclusion, employing bioactive peptide-enriched edible coatings holds promise for extending the shelf-life of perishable foods.

The use of active compounds to shape the quality of active double-layer films based on furcellaran intended for packaging salad-dressing - Assessment of utilitarian and storage properties.

In order to obtain innovative, high-quality biodegradable packaging, double-layer films based on furcellaran and gelatin, enriched with plant extracts were created. The films were assessed considering their potential utility, applications and environmental impact. The mechanical properties over a period of nine months were studied and it was noted that the passing of time had a beneficial effect on these parameters. The antioxidant properties was also examined, with the highest results obtained using the DPPH and metal chelating activity methods for GE (76.64 % and 9.85 % respectively), while this film showed the lowest FRAP value (5.99 %) compared to the highest obtained for DTE (52.62 %). For the first time, the possibility of using the double-layer active FUR/GEL film as packaging for salad-dressing was evaluated, but no improvement in parameters was observed regardless of the extract used. The environmental impact analysis showed the ability to completely decomposed in vermicompost within several days.

Valorisation of prawn/shrimp shell waste through the production of biologically active components for functional food purposes.

BACKGROUND: The aim of the work was to develop a technology for using waste from prawn and shrimp processing as a source of active ingredients that could be used in the promotion of healthy foods. From fresh and freeze-dried prawn and shrimp shells, protein hydrolysates (carotenoproteins) were obtained using two different enzymes, Flavourzyme and Protamex. RESULTS: The obtained hydrolysates were characterised in terms of protein content, degree of hydrolysis, and antioxidant and antimicrobial activity. The hydrolysate with the best antioxidant properties (FRAP value of 2933.33 µmol L-1 TE; ORAC value of 115.58 µmol L-1 TE) was selected and tested for its possible use as a component of functional foods. Molecular weight distribution, amino acid profile and free amino acids, the solubility of the hydrolysate in different pH ranges as well as foaming ability were determined. It was found that this hydrolysate was characterised by an amino acid profile with high nutritional value, flavour enhancement properties and excellent solubility in a wide pH range (from 97.06% to 100%). Afterward, the possibility of using carotenoproteins from prawn waste as a component of an emulsion with furcellaran and a lipid preparation of astaxanthin, taken from post-hydrolysate production waste, was investigated. The obtained complexes were stable as proved by the measurement of zeta potential (ζ = -23.87 and -22.32 to -27.79 mV). CONCLUSION: It is possible to produce stable complexes of the hydrolysate with furcellaran and to emulsify a lipid preparation of astaxanthin, obtained from waste following production of the hydrolysate, in them. © 2023 Society of Chemical Industry.

Replacement of milk fat by rapeseed oil stabilised emulsion in commercial yogurt.

The incorporation of lipid droplets and further characterization of matrices within dairy products may be possible using such adjacent particles as protein complexes/lipids. Among the range of varied emulsions and their functionalities, great attention has recently focused on the fabrication of high internal phase types. Feasibly, stable alternatives structured with health-beneficial lipids like those derived from plants could replace saturated fatty acids. As a fat replacement strategy, the fate of incorporated HIPE would require some adjustments either with storage stability and/or structural feat for the food matrix. Therefore, the replacement of milk fat by rapeseed oil stabilised emulsion in commercial yogurt was investigated. This involved 25%, 50% and 75% rapeseed oil respectively assigned as low (LIPE), medium (MIPE), and high internal phase emulsion (HIPE). Specifically, emulsions were examined by droplet size, encapsulation, pH, zeta potential, phase separation, and rheology. The fat free yogurt supplemented by HIPE were examined by droplet size, zeta potential, pH, color, sensory, texture and microbiological aspects against positive (regular milk fat) and negative (fat free) yogurt controls. Results showed increasing rapeseed oil contents would form smaller droplet-like emulsions. Within the yogurt matrix however, incorporating HIPE would seemingly reduce oil droplet size without much compromise to bacterial viability, sensory, or texture. Overall, this simple method of lipid alternation shows promise in dairy products.

Sugar-Free, Vegan, Furcellaran Gummy Jellies with Plant-Based Triple-Layer Films.

Increasing consumer awareness of the impact of nutrition on health and the growing popularity of vegan diets are causing a need to look for new plant-based formulations of standard confectionery products with high energy density and low nutritional value, containing gelatin. Therefore, the aim of this study was to develop vegan and sugar-free gummy jellies based on an algae-derived polysaccharide-furcellaran (FUR). Until now, FUR has not been used as a gel-forming agent despite the fact that its structure-forming properties show high potential in the production of vegan confectionery. The basic formulation of gummy jellies included the addition of soy protein isolate and/or inulin. The final product was characterized regarding its rheological, antioxidant, mechanical and physicochemical properties. Eco-friendly packaging for the jellies composed of a three-layer polymer film has also been developed. It was observed that the highest values of textural parameters were obtained in jellies containing the addition of soy protein isolate, whose positive effect was also found on antioxidant activity. Before drying, all furcellaran-based gel systems showed G' and G" values characteristic of strong elastic hydrogels. Storing jellies for a week under refrigeration resulted in an increase in hardness, a decrease in moisture content and reduced water activity values. Overall, our study indicates the high potential of furcellaran both as a gelling agent in confectionery products and as a base polymer for their packaging.

Improving the quality of multi-layer films based on furcellaran by immobilising active ingredients and impact assessment of the use of a new packaging material.

To improve the quality of multi-layer film, four-layer films based on furcellaran and active ingredients: gelatin hydrolysate, curcumin, capsaicin, montmorillonite and AgNPs, were produced in an innovative manner. The films were characterised by SEM and AFM analysis. Along with an increase in the concentration of active ingredients, the structure of the film becomes less homogeneous, which may affect the functional properties. The objective of the study was to analyse changes in the functional properties of the newly-obtained films and to verify their potential as packaging materials for fish products. With the increase in active ingredient concentration, water properties also improved, but there were no noticeable significant effects on mechanical properties. For antioxidant properties, the obtained values were within 1.04-2.74 mM Trolox/mg (FRAP) and 7.67-40.49% (DPPH). The obtained multi-layer films were examined with regard to the shelf-life of salmon. For this purpose, salmon fillets were packed in films having good antioxidant and functional properties. The films were effective in microorganism growth inhibition responsible for fillet spoilage during storage. The microorganism number in the active film-stored samples was lower by 0.13 log CFU/g on day 12 versus the control. However, film application did not retard lipid oxidation in the salmon fillets. Nonetheless, the films show great potential as active packaging materials, extending the shelf-life of the packed foods.

The influence of aqueous butterfly pea (Clitoria ternatea) flower extract on active and intelligent properties of furcellaran Double-Layered films - in vitro and in vivo research.

Innovative, intelligent and active double-layer films, based on furcellaran and with the addition of gelatin hydrolysates, have been obtained for the first time. An aqueous extract of clitoria flower in 3 concentrations was included in the 1st FUR layer. The films demonstrated strong antimicrobial effects, but did not exhibit fungicidal properties. The antioxidant properties of the films were within the range of 2.27-3.92 mM Trolox/mg (FRAP method) and 36.67-61.24 % (DPPH method). The films were used as active packaging materials in salmon fillets, which were stored for a period of 12 days in 4 °C. Analysis concerning microbiological properties of the stored fillets showed the possibility of extending their shelf-life by 6 days. Lipid oxidation, determined by TBARS has delayed. The obtained films are a promising material for the packaging industry. This is an important aspect within the context of global food waste and also the need to reduce synthetic materials.

Biological potential of polysaccharides extracted from Nostoc colonies for film production - Physical and biological properties.

Cyanobacteria of the Nostoc genus secrete a number of biologically active compounds, including polysaccharides, which may exhibit antioxidant, antimicrobial, anticancer, and anti-inflammatory properties. The aim of the study was to investigate the biological properties of Nostoc polysaccharides (NPs) (antioxidative and antimicrobial) and the possibility of using NPs addition in the production of biofoils. Our results allow to indicate that NPs were compatible with the used biopolymer matrix (furcellaran and chitosan) and showed antioxidant properties. The tested polysaccharide extracts (0.14%) exhibited the ability to neutralize free radicals - 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) at a level of 4.46% and 10.14%, respectively. NP extracts demonstrated reducing properties of 15.35 and 30.07 mg Trolox equivalents (FRAP and CUPRAC methods, respectively) and 2.64 mg chlorogenic acid equivalents (tested with Folin's reagent). NP extracts showed: a growth-stimulating effect (Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae), no effect (Penicillum sp.), or a slight inhibitory effect (Streptomyces sp.) on the tested microorganisms. The enrichment of the film with NPs influenced the physic-chemical properties of the obtained biofoils. The addition of polysaccharides to furcellaran and chitosan films decreased their water solubility (by approximately 40% and 9%, respectively, compared to the control) and, at the same time increased, their water absorption.

Incorporation of curcumin extract with lemongrass essential oil into the middle layer of triple-layered films based on furcellaran/chitosan/gelatin hydrolysates - In vitro and in vivo studies on active and intelligent properties.

Triple-layered, pH-sensitive and active films based on furcellaran (FUR), chitosan (CHIT) and gelatin hydrolysates (HGEL) were obtained for the first time. Curcumin ethanol extract (CUR) enriched with lemongrass essential oil (LEO) in three concentrations was added to the middle layer of the film. SEM and AFM analysis confirmed the formation of 3 layers, and CUR-LEO was evenly dispersed in the 2nd layer. The addition of CUR-LEO influenced the color of the surface and improved the UV blocking properties of the film. However, the mechanical and water vapor barrier properties remained unchanged. The CUR-LEO demonstrated antibacterial properties against bacteria and fungi. The antioxidant activity of the films increased along with the increasing concentrations of CUR-LEO. The presented films were able to effectively extend the storage life of tomatoes. Moreover, the colored films had the monitoring freshness effect with a distinct color change during carp storage.

The impact of nano/micro-plastics toxicity on seafood quality and human health: facts and gaps.

Contamination of the food and especially marine environment with nano/micro-plastic particles has raised serious concern in recent years. Environmental pollution and the resulting seafood contamination with microplastic (MP) pose a potential threat to consumers. The absorption rate of the MP by fish is generally considered low, although the bioavailability depends on the physical and chemical properties of the consumed MP. The available safety studies are inconclusive, although there is an indication that prolonged exposure to high levels of orally administered MP can be hazardous for consumers. This review details novel findings about the occurrence of MP, along with its physical and chemical properties, in the marine environment and seafood. The effect of processing on the content of MP in the final product is also reviewed. Additionally, recent findings regarding the impact of exposure of MP on human health are discussed. Finally, gaps in current knowledge are underlined, and the possibilities for future research are indicated in the review. There is an urgent need for further research on the absorption and bioavailability of consumed MP and in vivo studies on chronic exposure. Policymakers should also consider the implementation of novel legislation related to MP presence in food.

Antioxidant edible double-layered film based on waste from soybean production as a vegan active packaging for perishable food products.

Edible vegan films were obtained on the basis of furcellaran and protein hydrolysates from soybean bran. The aim of the study was to investigate the possibility of using innovative single- and double-layer films with antioxidant properties as packaging materials for vegetarian products. A further objective of the study was to evaluate the degree of biodegradation and potential toxicity with regard to the developed films. The tofu was analysed for changes in physicochemical parameters (weight loss, dry weight texture and colour), microbiological quality (total microbial count, yeast and moulds, psychotropic bacteria), and lipid oxidation (peroxidase and acid values, TBARS, fatty acid profile). It was found that films undergo almost complete biodegradation within 10 days and do not demonstrate any toxicity for cress seeds during their growth. Films are not effective in inhibiting the growth of microorganisms in tofu samples. Furthermore, they are not efficient in inhibiting the oxidation of tofu lipids. Compared to the control, the tofu samples covered with the innovative biopolymer films showed significantly lower moisture content and higher weight loss as well as hardness during the whole storage period.

Double-Layered Films Based on Furcellaran, Chitosan, and Gelatin Hydrolysates Enriched with AgNPs in Yerba Mate Extract, Montmorillonite, and Curcumin with Rosemary Essential Oil.

Double-layered active films based on furcellaran (1st layer-FUR), chitosan, and gelatin hydrolysates (2nd layer-CHIT+HGEL) were successfully prepared. Bioactive ingredients were added to the 1st film layer: AgNPs, which were synthesized in situ with yerba mate extract; montmorillonite clay (MMT); and different loads of ethanolic curcumin (CUR) extract enriched with rosemary essential oil (REO). SEM images confirmed the presence of AgNPs with a size distribution of 94.96 ± 3.33 nm throughout the films, and AFM and SEM photos indicated that the higher substance concentrations had rougher and more porous film microstructures. However, the water vapor transmission rate was reduced only at the lowest load of this ingredient. Despite the tensile strength of the films having decreased, the incorporation of the compounds showed a tendency towards reducing the modulus of elasticity, resulting in a lower stiffness of the composites. The addition of CUR and AgNPs improved the UV light barrier properties of the materials. The presented films showed quick reactions to changes in the pH value (from orange to red along with an increase in pH from 2 to 10), which indicates their potential use as indicators for monitoring the freshness of food products. Composite No. 2 showed the highest antimicrobial potential, while none of the presented films showed an antifungal effect. Finally, the antioxidant activities of the films increased dramatically at higher AgNP and CUR loads, suggesting an outstanding potential for active food packaging applications.

Biopolymer-Based Films from Sodium Alginate and Citrus Pectin Reinforced with SiO2.

Blend films based on sodium alginate (SA) and citrus pectin (P) reinforced with different concentrations of SiO2 (0-10% w/w) were developed in this study. From the morphological (SEM) and structural (FT-IR) evaluation, it was verified that the incorporation of the reinforcing agent did not drastically modify the microstructure of the films, nor did new chemical bonds form. However, the XRD results suggested a slight reduction in the crystallinities of the blends by the incorporation of SiO2. Among the formulations prepared, the addition of a 5% reinforcing agent was responsible for the simultaneous improvement of mechanical and barrier properties. Comparing the control sample (SA/P) with the SA/P/5.0%SiO2 film, the tensile strength increased from 27.7 ± 3.7 to 40.6 ± 4.5 MPa, and the water-vapor transmission rate decreased from 319.8 ± 38.7 to 288.9 ± 23.5 g m-2 day-1. Therefore, SiO2, as a reinforcing agent in SA/P blends, represents a simple and effective strategy for improving the properties of biopolymer-based films in applications, such as packaging.

Attempt to Extend the Shelf-Life of Fish Products by Means of Innovative Double-Layer Active Biodegradable Films.

In this study, we aimed to produce, innovative and, at the same time, environmentally-friendly, biopolymer double-layer films with fish processing waste and active lingonberry extract as additives. These double-layered films were based on furcellaran (FUR) (1st layer) and carboxymethyl cellulose (CMC) with a gelatin hydrolysate (HGEL) (2nd layer). The aim of the study was to assess their impact on the durability of perishable salmon fillets during storage, and to evaluate their degree of biodegradation. The fillets were analyzed for changes in microbiological quality (total microbial count, yeast and molds, and psychrotrophic bacteria), biogenic amine content (HPLC), and lipid oxidation (peroxidase and acid values, TBARS). The degree of biodegradation includes analysis of film and compost chemical composition solubility, respiratory activity, and ecotoxicity testing. The obtained results allow to suggest that active films are not only bacteriostatic, but even bactericidal when they used to coat fish fillets. Concerning the group of samples covered with the double-layer films, a 19.42% lower total bacteria count was noted compared to the control samples. Furthermore, it can be observed that the applied double-layer films have a potentially strong inhibitory effect on the accumulation of biogenic amines in fish, which is correlated with its antimicrobial effect (the total biogenic amine content for control samples totaled 263.51 mg/kg, while for the double-layer samples, their value equaled: 164.90 mg/kg). The achieved results indicate a high biodegradation potential, however, a too low pH of the film results in limiting seed germination and growth. Despite that, of these, double-layer films are a technology that has applicative potential.

Utilisation of soybean post-production waste in single- and double-layered films based on furcellaran to obtain packaging materials for food products prone to oxidation.

Single-layered films, based on furcellaran and protein hydrolysates from soybean bran (1st layer), were obtained. Moreover, a procedure for the preparation of double-layered films was developed, in which an ethanol extract from soybean bran was deposited onto the 1st layer. It was checked how the addition of the 2nd layer affects the functional properties of the film. The addition of the 2nd layer increased the thermal properties, modulus of elasticity and antioxidant activity, while decreases were noted for tensile strength and elongation at break parameters. The films were used as packaging materials for storing butter and the active films did not extend the quality of butter during storage, however, they behaved in the same way as synthetic films. Therefore they have the potential to be used as packaging material instead of a synthetic film.

Shelf-life extension of salmon using active total biodegradable packaging with tea ground waste and furcellaran-CMC double-layered films.

In this study, active biopolymer trays, being part of the biodegradable packaging, were developed and characterised. The aim of our research was to determine how active packaging (trays + films) affects the quality of salmon storage. The trays had high antioxidant potential and were biodegradable, however, they limited germination and seed growth, which may have been caused by the low pH of the material. Furthermore, the applied packaging demonstrated a potential possible inhibitory effect on the accumulation of biogenic amines and the growth of microorganisms responsible for the spoilage of salmon fillets. Compared to the control group, fillets stored in the tested pack had a 19% lower total bacteria count on the 6th day of storage. The innovative packing is easily biodegradable and prolongs the shelf-life of salmon fillets, therefore, it shows promise as a packaging material for perishable food products.

Developing Technology for the Production of Innovative Coatings with Antioxidant Properties for Packaging Fish Products.

In this study, we investigated the effects of furcellaran−gelatine (FUR/GEL) coatings incorporated with herb extracts on the quality retention of carp fish during refrigeration. Nutmeg, rosemary, thyme, milfoil, marjoram, parsley, turmeric, basil and ginger were subjected to water and ethanol extraction methods (10% concentration of herbs). The water extractions of the rosemary and thyme (5%) were used for the further development of coatings due to their high 2,2-Diphenyl-1-picrylhydrazyl (DPPH: 85.49 and 83.28%) and Ferric Reducing Antioxidant Power Assay values (FRAP: 0.46 and 0.56 mM/L) (p < 0.05), respectively. A new, ready-to-cook product with the coatings (carp fillets) was evaluated regarding quality in terms of colour parameters, texture profile, water activity, Thiobarbituric Acid Reactive Substances (TBARSs) and sensory analyses during 12 days of storage at 4 °C. The results show that the colour of the carp fillets treated with the rosemary and thyme extracts became slightly darker and had a propensity towards redness and yellowness. In contrast to the control group, the carp fillets stored in the coatings with the rosemary extract effectively slowed the lipid oxidation processes. Therefore, the innovative coatings produced from carp processing waste may have high potential as components in convenience food products and could extend the shelf-life of carp fillets during refrigerated storage. However, further research is needed to assess the microbiological stability of the obtained food products.