Examining the Impact of Squaric Acid as a Crosslinking Agent on the Properties of Chitosan-Based Films.

Hydrogels based on chitosan are very versatile materials which can be used for tissue engineering as well as in controlled drug delivery systems. One of the methods for obtaining a chitosan-based hydrogel is crosslinking by applying different components. The objective of the present study was to obtain a series of new crosslinked chitosan-based films by means of solvent casting method. Squaric acid-3,4-dihydroxy-3-cyclobutene-1,2-dione-was used as a safe crosslinking agent. The effect of the squaric acid on the structural, mechanical, thermal, and swelling properties of the formed films was determined. It was established that the addition of the squaric acid significantly improved Young's modulus, tensile strength, and thermal stability of the obtained materials. Moreover, it should be stressed that the samples consisting of chitosan and squaric acid were characterized by a higher swelling than pure chitosan. The detailed characterization proved that squaric acid could be used as a new effective crosslinking agent.

Comparison of Polylactide-Based Active Films Containing Berberine and Quercetin as Systems for Maintaining the Quality and Safety of Blueberries.

Polymeric thin films based on polylactide with an addition of poly(ethylene glycol) as a plasticizer and flavonoids in the form of quercetin and berberine were subjected to tests that were particularly relevant from the point of view of contact with food. A comparative analysis of the effect of individual flavonoids on the antioxidative properties of tested films and blueberry storage was carried out. The influence of active compounds on the water vapor permeability, as well as UV protection, of the obtained materials was investigated. Also, the specific migration of individual flavonoids from obtained materials to food simulants in the form of acetic acid and ethyl alcohol was determined. The crucial point of this study is the storage of blueberries. The obtained results indicate that the selection of packaging, containing individual active compounds, depends on the purpose and requirements that the packaging must meet for particular types of food.

The adenine-modified edible chitosan films containing choline chloride and citric acid mixture.

A series of biopolymeric chitosan-based (Ch) films were prepared with choline chloride and citric acid plasticizer (deep eutectic solvent, DES). An effect of adenine (A, vitamin B4) addition on the functional properties of these films was evaluated. Several physicochemical and mechanical properties were tested: Fourier-transformed infrared spectra proved DES's plasticizing and crosslinking effect, while scanning electron microscopy and atomic force microscopy techniques confirmed the possible phase separation after adenine addition. These changes affected the mechanical characteristics and the water vapor and oxygen permeability. The prepared materials are not water soluble because the CA acts as a crosslinker. The adenine addition on antioxidative and antimicrobial properties was also checked. It was found that Ch-DES materials with A exhibit improved antioxidative properties (55.8-66.1% of H2O2 scavenging activity) in contrast to the pristine chitosan-DES material (51.1% of H2O2 scavenging activity), while the material is still non-mutagenic (lack of growth of Salmonella typhimurium) and possesses antimicrobial features (no E. coli observed for all the tested films and inhibition zones noted for S. aureus). The mentioned properties, reduced oxygen transmission (1.6-2.1 g m-2 h-1), and mechanical characteristics within the range of typical food packaging plastics proved the potential of Ch-DES-A films in the packaging sector. Moreover, the antioxidative properties, usage of substrates being allowed as food additives, and the presence of adenine create the advantage of the Ch-DES-A materials as edible coatings, being also a source of Vitamin B4.

Microplastics Derived from Food Packaging Waste-Their Origin and Health Risks.

Plastics are commonly used for packaging in the food industry. The most popular thermoplastic materials that have found such applications are polyethylene (PE), polypropylene (PP), poly(ethylene terephthalate) (PET), and polystyrene (PS). Unfortunately, most plastic packaging is disposable. As a consequence, significant amounts of waste are generated, entering the environment, and undergoing degradation processes. They can occur under the influence of mechanical forces, temperature, light, chemical, and biological factors. These factors can present synergistic or antagonistic effects. As a result of their action, microplastics are formed, which can undergo further fragmentation and decomposition into small-molecule compounds. During the degradation process, various additives used at the plastics' processing stage can also be released. Both microplastics and additives can negatively affect human and animal health. Determination of the negative consequences of microplastics on the environment and health is not possible without knowing the course of degradation processes of packaging waste and their products. In this article, we present the sources of microplastics, the causes and places of their formation, the transport of such particles, the degradation of plastics most often used in the production of packaging for food storage, the factors affecting the said process, and its effects.

Development and characterization of active packaging films based on chitosan, plasticizer, and quercetin for repassed oil storage.

Novel chitosan (Ch) films containing choline chloride and citric acid mixture as plasticizer (deep eutectic solvent, DES) and different amounts of quercetin (QUE) as antioxidant additive were prepared. Physicochemical and mechanical characteristics of the developed Ch/DES/QUE films were studied using FTIR, SEM, and AFM techniques. FTIR spectra revealed the possible interactions between all the components. The surface of the films was dense and rough. The addition of quercetin caused an increase in the tensile strength (TS) and Young's modulus, but significantly decreased the elongation at break. The films containing quercetin showed improved antioxidant activity in relation to Ch/DES film. Finally, the oxidation phenomena of rapeseed oils with and without chitosan films were evaluated as amounts of primary and secondary oxidation products and total oxidation index. The addition of Ch/DES films with quercetin to oil samples successfully retarded secondary lipid oxidation processes and improved its antioxidant activity under the accelerated storage condition.

The Use of Chitin from the Molts of Mealworm (Tenebrio molitor) for the Removal of Anionic and Cationic Dyes from Aqueous Solutions.

The possibility of using chitin from the molts of an insect-ealworm (Tenebrio molitor) to remove anionic (RB5, RY84) and cationic dyes (BV10, BR46) from aqueous solutions was investigated. The scope of the research included, among others: Characteristics of chitin from mealworms (FTIR, SEM, pHPZC), the effect of pH on sorption efficiency, sorption kinetics (pseudo-first, pseudo-second order, intramolecular diffusion models) and the determination of the maximum sorption capacity (Langmuir and Freundlich models). The sorption efficiency of anionic dyes on chitin from mealworm was the highest at pH 2-3, and for cationic dyes at pH 6. The equilibrium time of sorption of anionic dyes was 240-300 min and for cationic dyes it was 180-240 min. The experimental data on dye sorption kinetics was best described by the pseudo-second order model. The maximum sorption capacity of chitin from the mealworm for the anionic dyes RB5 and RY84 was 121.15 mg/g and 138.55 mg/g, respectively, and was higher than with some carbon-based materials (literature data). In the case of cationic dyes, the sorption capacity of the tested chitin was lower and reached 3.22 mg/g and 59.56 mg/g for BV10 and BR46, respectively.

Chitosan/Phenolic Compounds Scaffolds for Connective Tissue Regeneration.

Chitosan-based scaffolds modified by gallic acid, ferulic acid, and tannic acid were fabricated. The aim of the experiment was to compare the compatibility of scaffolds based on chitosan with gallic acid, ferulic acid, or tannic acid using the in vivo method. For this purpose, materials were implanted into rabbits in the middle of the latissimus dorsi muscle length. A scaffold based on unmodified chitosan was implanted by the same method as a control. Moreover, the Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) spectra and scanning electron microscope (SEM) observations were made to study the interactions between chitosan and phenolic acids. Additionally, antioxidant properties and blood compatibility were investigated. The results showed that all studied materials were safe and non-toxic. However, chitosan scaffolds modified by gallic acid and tannic acid were resorbed faster and, as a result, tissues were organized faster than those modified by ferulic acid or unmodified.

Carboxymethyl cellulose based films enriched with polysaccharides from mulberry leaves (Morus alba L.) as new biodegradable packaging material.

The objective of this study was to determine the properties of a new active packaging film developed by the addition of mulberry leaves polysaccharides (MLP) into carboxymethyl cellulose (CMC). Biodegradable CMC-MLP films were fabricated by casting method with various concentrations of MLP (1, 5 and 10 % w/w). The addition of MLP into the CMC matrix resulted increased thickness (0.126 to 0.163 mm) and roughness of the films. Also, the decline in moisture content from 27.91 to 14.12 %, water vapor permeability from 8.95 to 5.21 × 10-10 g-1 s-1 Pa-1, and a swelling degree from 59.11 to 37.45 % were observed. With the increasing concentration of MLP, the mechanical properties of the films were improved and higher dispersion of UV light were noted. Fourier transform - infrared spectroscopy (FT-IR) and X-ray diffraction revealed good inter-molecular interaction between CMC matrix and MLP. The prepared films showed excellent thermal stability, antioxidant and antibacterial properties as well as susceptibility to biodegradation in the soil environment. Moreover, it was proved that the films have ability to retard oil oxidation. Overall, it was concluded that CMC-MLP films constitute a promising biomaterial that may be applied as active food packaging.

Polylactide-Based Films Incorporated with Berberine-Physicochemical and Antibacterial Properties.

A series of new polymeric materials consisting of polylactide (PLA), polyethylene glycol (PEG) and berberine chloride (B) was evaluated. PEG was incorporated into the polymer matrix with the aim of obtaining a plasticizing effect, while berberine was added in order to obtain antibacterial properties in formed packaging materials. Materials were formed using the solvent-casting procedure. Fourier transform infrared spectroscopy and scanning electron microscopy were used so as to establish the structural changes resulting from the introduction of berberine. Thermogravimetry and differential scanning calorimetry were applied to study the thermal properties. Further, mechanical properties and differences in colour and transparency between the control sample and films containing berberine were also studied. The recorded data indicates that berberine formed a network on the surface of the PLA-based materials. Introduction of an active compound significantly improved thermal stability and greatly affected the Young's modulus values of the studied polymeric films. Moreover, it should be stressed that the addition of the studied active compound leads to an improvement of the antibacterial properties, resulting in a significant decrease in growth of E. coli and the S. aureus bacteria cultures.

Polylactide-Based Films with the Addition of Poly(ethylene glycol) and Extract of Propolis-Physico-Chemical and Storage Properties.

Polymeric films based on polylactide (PLA) with the addition of poly(ethylene glycol) (PEG) and a chloroformic extract of propolis were obtained. In the case of the studied films, polylactide (PLA) played the role of polymeric matrix and poly(ethylene glycol) was used as a plasticizer, while the extract of propolis was incorporated as a compound that could significantly affect the properties of the obtained materials, especially the water vapour permeation rate and the stability of the food products. Moreover, changes in structure, morphology, mechanical and storage properties as well as differences in colour, thickness and transparency after introducing propolis into the PLA-PEG system were determined. Based on the obtained results, it was established that the addition of the chloroformic extract of propolis significantly influences the most important properties taken into account during food packaging. It was also noticed that films with incorporated propolis were characterised by a significant improvement in the water vapour barrier property. Moreover, the obtained results prove that packaging containing a chloroformic propolis extract allow for the maintenance of the quality of the fruit stored for an extended period of time. To summarise, the application of a chloroformic propolis extract enables the formation of packaging materials that extend the shelf life of stored food products.

Polylactide Films with the Addition of Olive Leaf Extract-Physico-Chemical Characterization.

The aim of this work was to obtain and characterize polylactide films (PLA) with the addition of poly(ethylene glycol) (PEG) as a plasticizer and chloroformic olive leaf extract (OLE). The composition of OLE was characterized by LC-MS/MS techniques. The films with the potential for using in the food packaging industry were prepared using a solvent evaporation method. The total content of the phenolic compounds and DPPH radical scavenging assay of all the obtained materials have been tested. Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (FTIR-ATR) allows for determining the molecular structure, while Scanning Electron Microscopy (SEM) indicated differences in the films' surface morphology. Among other crucial properties, mechanical properties, thickness, degree of crystallinity, water vapor permeation rate (WVPR), and color change have also been evaluated. The results showed that OLE contains numerous active substances, including phenolic compounds, and PLA/PEG/OLE films are characterized by improved antioxidant properties. The OLE addition into PLA/PEG increases the material crystallinity, while the WVPR values remain almost unaffected. From these studies, significant insight was gained into the possibility of the application of chloroform as a solvent for both olive leaf extraction and for the preparation of OLE, PLA, and PEG-containing film-forming solutions. Finally, evaporation of the solvent from OLE can be omitted.

The role of a deep eutectic solvent in changes of physicochemical and antioxidative properties of chitosan-based films.

In this work deep eutectic solvent (DES), based on the mixture of choline chloride and lactic acid, were suggested as chitosan films plasticizers. The molecular structure and properties of films obtained using chitosan, with different degree of deacetylation and 0-80 wt.% DES content (ωDES), were studied by means of FTIR spectroscopy, SEM and AFM microscopy (films' surface properties) together with optical characteristics, water vapor transmission rate (WVTR), water vapor permeability (WVP), tensile strength (TS) and elongation at break (Eb). Scanning electron micrographs revealed that all chitosan-DES films were smooth and uniform. DES significantly improves the film flexibility (Eb increases of ca. 160 % after incorporation of 80 wt.% DES), slightly decreases tensile strength and also improves antioxidative properties while simultaneously, increasing water vapor permeability (WVP). Films prepared in this study exhibit characteristics that qualify them for potential use as an active packaging material.

Antibacterial Films Based on Polylactide with the Addition of Quercetin and Poly(Ethylene Glycol).

A series of new films with antibacterial properties has been obtained by means of solvent casting method. Biodegradable materials including polylactide (PLA), quercetin (Q) acting as an antibacterial compound and polyethylene glycol (PEG) acting as a plasticizer have been used in the process. The effect of quercetin as well as the amount of PEG on the structural, thermal, mechanical and antibacterial properties of the obtained materials has been determined. It was found that an addition of quercetin significantly influences thermal stability. It should be stressed that samples containing the studied flavonoid are characterized by a higher Young modulus and elongation at break than materials consisting only of PLA and PEG. Moreover, the introduction of 1% of quercetin grants antibacterial properties to the new materials. Recorded results showed that the amount of plasticizer did not influence the antibacterial properties; it does, however, cause changes in physicochemical properties of the obtained materials. These results prove that quercetin could be used as an antibacterial compound and simultaneously improve mechanical and thermal properties of polylactide-based films.

Polycistronic Expression System for Pichia pastoris Composed of Chitino- and Chitosanolytic Enzymes.

Chitin is one of the most abundant biopolymers. Due to its recalcitrant nature and insolubility in accessible solvents, it is often considered waste and not a bioresource. The products of chitin modification such as chitosan and chitooligosaccharides are highly sought, but their preparation is a challenging process, typically performed with thermochemical methods that lack specificities and generate hazardous waste. Enzymatic treatment is a promising alternative to these methods, but the preparation of multiple biocatalysts is costly. In this manuscript, we biochemically characterised chitin deacetylases of Mucor circinelloides IBT-83 and utilised one of them for the construction of the first eukaryotic, polycistronic expression system employing self-processing 2A sequences. The three chitin-processing enzymes; chitin deacetylase of M. circinelloides IBT-83, chitinase from Thermomyces lanuginosus, and chitosanase from Aspergillus fumigatus were expressed under the control of the same promoter in methylotrophic yeast Pichia pastoris and characterised for their synergistic action towards their respective substrates.

Influence of Tea Tree Essential Oil and Poly(ethylene glycol) on Antibacterial and Physicochemical Properties of Polylactide-Based Films.

The aim of the study was to establish the influence of poly(ethylene glycol) (PEG) on the properties of potential biodegradable packaging materials with antibacterial properties, based on polylactide (PLA) and tea tree essential oil (TTO). The obtained polymeric films consisted of PLA, a natural biocide, and tea tree essential oil (5-20 wt. %) was prepared with or without an addition of 5 wt. % PEG. The PLA-based materials have been tested, taking into account their morphology, and their thermal, mechanical and antibacterial properties against Staphylococcus aureus and Escherichia coli. It was established that the introduction of a plasticizer into the PLA-TTO systems leads to an increase in tensile strength, resistance to deformation, as well an increased thermal stability, in comparison to films modified using only TTO. The incorporation of 5 wt. % PEG in the PLA solution containing 5 wt. % TTO allowed us to obtain a material exhibiting a satisfactory antibacterial effect on both groups of representative bacteria. The presented results indicated a beneficial effect of PEG on the antibacterial and functional properties of materials with the addition of TTO.

Antibacterial Films Based on PVA and PVA-Chitosan Modified with Poly(Hexamethylene Guanidine).

In this study, thin, polymeric films consisting of poly(vinyl alcohol) (PVA) and chitosan (Ch) with the addition of poly(hexamethylene guanidine) (PHMG) were successfully prepared. The obtained materials were analyzed to determine their physicochemical and biocidal properties. In order to confirm the structure of PHMG, nuclear magnetic resonance spectroscopy (1H NMR) was applied, while in the case of the obtained films, attenuated total reflectance infrared spectroscopy with Fourier transform (FTIR-ATR) was used. The surface morphology of the polymer films was evaluated based on atomic force microscopy. Furthermore, the mechanical properties, color changes, and thermal stability of the obtained materials were determined. Microbiological tests were performed to evaluate the biocidal properties of the new materials with and without the addition of PHMG. These analyses confirmed the biocidal potential of films modified by PHMG and allowed for comparisons of their physicochemical properties with the properties of native films. In summary, films consisting of PVA and PHMG displayed higher antimicrobial potentials against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria in comparison to PVA:Ch-based films with the addition of PHMG.

Development and Characterization of Polyamide-Supported Chitosan Nanocomposite Membranes for Hydrophilic Pervaporation.

An experimental protocol of preparation of homogeneous and nanocomposite chitosan (Ch) based membranes supported on polyamide-6 (PA6) films was developed and described in detail. Montmorillonite (MMT) and Cloisite 30B (C30B) nanoclays were used as nanofillers to improve mechanical properties of chitosan films. The surface, mechanical, and transport properties of PA6 supported Ch, Ch/MMT and Ch/C30B membranes were studied and compared with a pristine, non-supported chitosan membrane. Implementation of advanced analytical techniques e.g., SEM reveal the clays nanoparticles are well dispersed in the chitosan matrix. According to AFM images, composite chitosan/nanoclay membranes possess higher roughness compared with unfilled ones. On the other hand, an incorporation of clay particles insignificantly changed the mechanical and thermal properties of the membranes. It was also found that all membranes are hydrophilic and water is preferentially removed from EtOH/H2O and iPrOH/H2O mixtures by pervaporation. Supporting of chitosan and chitosan/nanoclay thin films onto PA6 porous substrate enhanced permeate flux and pervaporation separation index, in comparison to the pristine Ch membrane. Concerning separation factor (ß), the highest value equal to 4500 has been found for a chitosan composite membrane containing Cloisite 30B contacting 85/15 wt % iPrOH/H2O mixture. The mentioned membrane was characterized by the normalized flux of 0.5 µm·kg·m-2·h-1. Based on the established data, it was possible to conclude that chitosan membranes are meaningful material in dehydration of azeotropic mixtures. Nevertheless, to boost up the membrane efficiency, the further modification process is required.

The Influence of the Morphology and Mechanical Properties of Polymer Inclusion Membranes (PIMs) on Zinc Ion Separation from Aqueous Solutions.

The transport of Zn(II) ions across polymer inclusion membranes (PIMs) with acetylacetone (ACAC) or di(2-ethylhexyl)phosphoric acid (D2EHPA) as carriers was studied. Polymeric membranes consisting of polyvinylchloride (PVC) as the support, bis(2-ethylhexyl)adipate (DAO) as plasticizer, and ACAC or D2EHPA as ion carriers were investigated. The highest recovery factors for Zn(II) ions were observed in the case of a membrane containing 20% acac (99.6%) and 60% D2EHPA (56.3%). The prepared PIMs were examined using atomic force microscopy (AFM) techniques. Their mechanical properties were also determined. The influence of membrane morphology and mechanical properties on the zinc transport process was discussed.

Hsp78 chaperone functions in restoration of mitochondrial network following heat stress.

Under physiological conditions mitochondria of yeast Saccharomyces cerevisiae form a branched tubular network, the continuity of which is maintained by balanced membrane fusion and fission processes. Here, we show using mitochondrial matrix targeted green fluorescent protein that exposure of cells to extreme heat shock led to dramatic changes in mitochondrial morphology, as tubular network disintegrated into several fragmented vesicles. Interestingly, this fragmentation did not affect mitochondrial ability to maintain the membrane potential. Cells subjected to recovery at physiological temperature were able to restore the mitochondrial network, as long as an active matrix chaperone, Hsp78, was present. Deletion of HSP78 gene did not affect fragmentation of mitochondria upon heat stress, but significantly inhibited ability to restore mitochondrial network. Changes of mitochondrial morphology correlated with aggregation of mitochondrial proteins. On the other hand, recovery of mitochondrial network correlated with disappearance of protein aggregates and reactivation of enzymatic activity of a model thermo-sensitive protein: mitochondrial DNA polymerase. Since protein disaggregation and refolding is mediated by Hsp78 chaperone collaborating with Hsp70 chaperone system, we postulate that effect of Hsp78 on mitochondrial morphology upon recovery after heat shock is mediated by its ability to restore activity of unknown protein(s) responsible for maintenance of mitochondrial morphology.