Effect of Chemical Degradation of Sodium Alginate on Capsaicin Encapsulation.

Capsaicin is known as an oily extract of paprika that is characterized by pungent taste and bioactivity. It also may cause irritation to the mouth and stomach which is why is so important to immobilize capsaicin on a carrier to prevent it. The usage of alginate oligomers, which has an antioxidant potential compared to alginate, is of benefit because it may be used in the immobilization of bioactive substances that are fragile to oxidation. The purpose of this study was to use sodium alginate oligomers as a coating material in the encapsulation process of paprika oleoresin. Alginate oligomers were produced by chemical degradation with hydrogen peroxide. The characteristics of the samples were obtained by measuring the viscosity, the contact angle of the surface, and the surface tension of solutions. The obtained solution of alginate oligomers served as the carrier material for the immobilization of capsaicin. Capsules were prepared by ionic gelation using a calcium chloride solution as a crosslinking agent. In this way, capsules without and with the core (capsaicin) were prepared and their ability to scavenge free radicals (DPPH) and iron-reducing properties (FRAP) were determined. The stability of the capsules was examined by thermal decomposition and under conditions of the gastric and small intestine, and capsaicin content was detected using high-performance liquid chromatography. It was found that alginate oligomers could be used in the encapsulation of bioactive compounds and the efficiency was above 80%. Capsule production from alginate oligomers affected their thermal stability. The use of alginate derivatives as a carrier increased the antioxidant properties of the finished product, as well as its ability to reduce iron ions. The use of alginate oligomers as a coating material prevented the active substance from being released too early in the conditions of the small intestine, prolonged the stability of the capsules, and supported their durability in gastric conditions.

Functional Properties of Chitosan Oligomers Obtained by Enzymatic Hydrolysis.

The aims of this study were to obtain chitooligosaccharides (COS) from chitosan (CH) with improved functional properties and comparison of the use of two different enzymes: commercial cellulase (CL) and the dedicated enzyme chitosanase (CS). After enzymatic reaction, chitosan oligomers (NFs) were isolated by methanol into two fractions: precipitate (HMF) and supernatant (LMF). The occurrence of a hydrolysis reaction was confirmed by an increased reducing sugar content and viscosity reduction of chitosan oligomers. CPMAS 13C NMR analysis confirmed the dissimilar cleavage mechanism of the enzymes used. LMF and NF fractions were characterised by improved solubility in water (94.56%) compared to the HMF and CH samples (70.64%). Thermogravimetric analysis (TGA) showed that the HMF decomposed in two-stage process while CH, NF, and LMF decomposed in a three-stage process. The greatest mass loss of LMF samples (58.35%) suggests their sensitivity to high-temperature treatments. COS were a mixture of DP (degrees of polymerisation) from 3 to 18 hetero-chitooligomers, with an average Mw of <3 kDa. CL consisted of more low-DP products (DP 3-7) than COS made with CS. LMF characterised by DP~2 showed lower DPPH radical scavenging activity than HMF and NF with DP 3-7. The ability to reduce Escherichia coli increased in the given order: LMF > NF > HMF > CH.

Evaluation of biofilm formation on acrylic resins used to fabricate dental temporary restorations with the use of 3D printing technology.

BACKGROUND: Temporary implant-retained restorations are required to support function and esthetics of the masticatory system until the final restoration is completed and delivered. Acrylic resins are commonly used in prosthetic dentistry and lately they have been used in three-dimensional (3D) printing technology. Since this technology it is fairly new, the number of studies on their susceptibility to microbial adhesion is low. Restorations placed even for a short period of time may become the reservoir for microorganisms that may affect the peri-implant tissues and trigger inflammation endangering further procedures. The aim of the study was to test the biofilm formation on acrylamide resins used to fabricate temporary restorations in 3D printing technology and to assess if the post-processing impacts microbial adhesion. METHODS: Disk-shaped samples were manufactured using the 3D printing technique from three commercially available UV-curable resins consisting of acrylate and methacrylate oligomers with various time and inhibitors of polymerization (NextDent MFH bleach, NextDent 3D Plus, MazicD Temp). The tested samples were raw, polished and glazed. The ability to create biofilm by oral streptococci (S. mutans, S. sanguinis, S. oralis, S. mitis) was tested, as well as species with higher pathogenic potential: Staphylococcus aureus, Staphylococcus epidermidis and Candida albicans. The roughness of the materials was measured by an atomic force microscope. Biofilm formation was assessed after 72 h of incubation by crystal violet staining with absorbance measurement, quantification of viable microorganisms, and imaging with a scanning electron microscope (SEM). RESULTS: Each tested species formed the biofilm on the samples of all three resins. Post-production processing resulted in reduced roughness parameters and biofilm abundance. Polishing and glazing reduced roughness parameters significantly in the NextDent resin group, while glazing alone caused significant surface smoothing in Mazic Temp. A thin layer of microbial biofilm covered glazed resin surfaces with a small number of microorganisms for all tested strains except S. oralis and S. epidermidis, while raw and polished surfaces were covered with a dense biofilm, rich in microorganisms. CONCLUSIONS: UV-curing acrylic resins used for fabricating temporary restorations in the 3D technology are the interim solution, but are susceptible to adhesion and biofilm formation by oral streptococci, staphylococci and Candida. Post-processing and particularly glazing process significantly reduce bacterial biofilm formation and the risk of failure of final restoration.

Biophysico-Chemical Properties of Alginate Oligomers Obtained by Acid and Oxidation Depolymerization.

The aim of the study was to obtain alginate oligosaccharides by using two degradation methods of sodium alginate (SA): with hydrochloric acid (G-guluronate, M-mannuronate and G + M fractions) and hydrogen peroxide (HAS-hydrolyzed SA), in order to assess and compare their biological activity and physico-chemical properties, with an attempt to produce gels from the obtained hydrolysates. The efficiency of each method was determined in order to select the fastest and most efficient process. The ferric ion reducing antioxidant power (FRAP), the ability to scavenge DPPH free radicals, rheological properties, Fourier Transformed Spectroscopy (FTIR) and the microbiological test against Escherichia coli and Staphylococcus aureus were performed. In order to check the functional properties of the obtained oligosaccharides, the texture profile analysis was assessed. The hydrolysis yield of acid SA depolymerization was 28.1% and from hydrogen peroxide SA, depolymerization was 87%. The FTIR analysis confirmed the degradation process by both tested methods in the fingerprint region. The highest ferric reducing antioxidant power was noted for HSA (34.7 µg), and the highest hydroxyl radical scavenging activity was obtained by G fraction (346 µg/Trolox ml). The complete growth inhibition (OD = 0) of alginate hydrolysates was 1%. All tested samples presented pseudoplastic behavior, only HSA presented the ability to form gel.

The Effect of Using Micro-Clustered Water as a Polymer Medium.

The aim of the study was to investigate the changes within the physicochemical properties of gelatin, carrageenan, and sodium alginate hydrosols prepared on the basis of micro-clustered (MC) water. The rheological parameters, contact angle and antioxidant activity of hydrosols were investigated. Moreover, the pH, oxidation-reduction potential (ORP) and electrical conductivity (EC) were measured. The hydrosols with MC water were characterized by a lower pH, decreased viscosity, a lower contact angle, and only slightly lower antioxidant activity than control samples. The results showed that hydrosol's properties are significantly changed by MC water, which can lead to enhancement of its applicability but requires further investigation.

Impact of Processing on the Protein Quality of Pinto Bean (Phaseolus vulgaris) and Buckwheat (Fagopyrum esculentum Moench) Flours and Blends, As Determined by in Vitro and in Vivo Methodologies.

Blending of protein sources can increase protein quality by compensating for limiting amino acids present in individual sources, whereas processing grain flours by extrusion or baking can also alter protein quality. To determine the effect of baking and extrusion on the protein quality of blended flours from buckwheat and pinto beans, a rodent bioassay was performed and compared to an in vitro method of protein quality determination. Overall, extruded products had higher protein efficiency ratio values, increased digestibility, and greater protein digestibility corrected amino acid score (PDCAAS) values than baked products, with the extruded buckwheat/pinto blend having the greatest PDCAAS value of the experimental diets investigated. A correlation was found between both digestibility and PDCAAS values generated from in vitro and in vivo methods. The use of in vitro digestibility analysis should be investigated as a potential replacement for the current rodent assay for nutrient content claim purposes.

Polysaccharide-Based Edible Coatings Containing Cellulase for Improved Preservation of Meat Quality during Storage.

The objectives of this study were to optimize the composition of edible food coatings and to extend the shelf-life of pork meat. Initially, nine meat samples were coated with solutions containing chitosan and hydroxypropyl methylcellulose at various cellulase concentrations: 0%, 0.05%, and 0.1%, stored for 0, 7, and 14 days. Uncoated meat served as the controls. The samples were tested for pH, water activity (aw), total number of microorganisms (TNM), psychrotrophs (P), number of yeast and molds (NYM), colour, and thiobarbituric acid-reactive substances (TBARS). The pH and aw values varied from 5.42 to 5.54 and 0.919 to 0.926, respectively. The reductions in the TNM, P, and NYM after 14 days of storage were approximately 2.71 log cycles, 1.46 log cycles, and 0.78 log cycles, respectively. The enzyme addition improved the stability of the red colour. Significant reduction in TBARS was noted with the inclusion of cellulase in the coating material. Overall, this study provides a promising alternative method for the preservation of pork meat in industry.

Effect of Film-Forming Alginate/Chitosan Polyelectrolyte Complex on the Storage Quality of Pork.

Meat is one of the most challenging food products in the context of maintaining quality and safety. The aim of this work was to improve the quality of raw/cooked meat by coating it with sodium alginate (A), chitosan (C), and sodium alginate-chitosan polyelectrolyte complex (PEC) hydrosols. Antioxidant properties of A, C, and PEC hydrosols were determined. Subsequently, total antioxidant capacity (TAC), sensory quality of raw/cooked pork coated with experimental hydrosols, and antimicrobial efficiency of those hydrosols on the surface microbiota were analysed. Application analyses of hydrosol were performed during 0, 7, and 14 days of refrigerated storage in MAP (modified atmosphere packaging). Ferric reducing antioxidant power (FRAP) and (2,2-diphenyll-picrylhydrazyl (DPPH) analysis confirmed the antioxidant properties of A, C, and PEC. Sample C (1.0%) was characterized by the highest DPPH value (174.67 µM Trolox/mL) of all variants. PEC samples consisted of A 0.3%/C 1.0% and A 0.6%/C 1.0% were characterized by the greatest FRAP value (~7.21 µM Fe2+/mL) of all variants. TAC losses caused by thermal treatment of meat were reduced by 45% by coating meat with experimental hydrosols. Application of PEC on the meat surface resulted in reducing the total number of micro-organisms, psychrotrophs, and lactic acid bacteria by about 61%, and yeast and molds by about 45% compared to control after a two-week storage.

The Effects of Using Sodium Alginate Hydrosols Treated with Direct Electric Current as Coatings for Sausages.

We investigated the effect of sodium alginate hydrosols (1%) with 0.2% of NaCl treated with direct electric current (DC) used as a coating on microbial (Total Viable Counts, Psychrotrophic bacteria, yeast and molds, Lactic acid bacteria, Enterobacteriaceae), physiochemical (pH, lipid oxidation, antioxidant activity, weight loss, color) and sensory properties of skinned pork sausages or with artificial casing stored at 4 °C for 28 days. Moreover, the cytotoxicity analysis of sodium alginate hydrogels was performed. The results have shown that application of experimental coatings on the sausage surface resulted in reducing all tested groups of microorganisms compared to control after a 4-week storage. The cytotoxicity analysis revealed that proliferation of RAW 264.7 and L929 is not inhibited by the samples treated with 200 mA. Ferric reducing antioxidant power (FRAP) and free radical scavenging activity (DPPH) analyses showed that there are no significant differences in antioxidant properties between control samples and those covered with sodium alginate. After 28 days of storage, the highest value of thiobarbituric acid-reactive substances (TBARS) was noticed for variants treated with 400 mA (1.07 mg malondialdehyde/kg), while it was only slightly lower for the control sample (0.95 mg MDA/kg). The obtained results suggest that sodium alginate treated with DC may be used as a coating for food preservation because of its antimicrobial activity and lack of undesirable impact on the quality factors of sausages.

Cross-linked alginate/chitosan polyelectrolytes as carrier of active compound and beef color stabilizer.

The aim of this work was to develop polyelectrolyte material suitable for active beef steaks coatings, by complexation of chitosan (CH) and sodium alginate (ALG) in a broad range of alginate/chitosan ratios (R). The rheological analysis confirmed significant effect of polymers ratios on the physical properties of sodium alginate-chitosan (ACH) hydrosols. The shear thinning non-Newtonian nature, thixotropic behavior and gel-like structure of solutions were displayed. Obtained complexes possess DPPH radical scavenging activity corresponding to 5.33, 17.06 and 41.41µMTrolox/ml for 0ppm, 500ppm and 1000pmm of sodium erythorbate dose, respectively. Application of ACH hydrosols enriched with 1000ppm of sodium erythorbate enhanced redness and color stability of beef steaks during storage (∆E after 2-weeks of storage=1.44±0.08) compared to uncovered beef (∆E after 2-weeks of storage=3.53±0.13). The limited solubility in range of 0%-54.56% as well as favorable wetting properties (contact angle between 45°-66°) of polyelectrolyte ACH films were obtained.

Antimicrobial and Antioxidant Activity of Chitosan/Hydroxypropyl Methylcellulose Film-Forming Hydrosols Hydrolyzed by Cellulase.

The aim of this study was to evaluate the impact of cellulase (C) on the biological activity of chitosan/hydroxypropyl methylcellulose (CH/HPMC) film-forming hydrosols. The hydrolytic activity of cellulase in two concentrations (0.05% and 0.1%) was verified by determination of the progress of polysaccharide hydrolysis, based on viscosity measurement and reducing sugar-ends assay. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect, the ferric reducing antioxidant power (FRAP), and microbial reduction of Pseudomonas fluorescens, Yersinia enterocolitica, Bacillus cereus, and Staphylococcus aureus were studied. During the first 3 h of reaction, relative reducing sugar concentration increased progressively, and viscosity decreased rapidly. With increasing amount of enzyme from 0.05% to 0.1%, the reducing sugar concentration increased, and the viscosity decreased significantly. The scavenging effect of film-forming solutions was improved from 7.6% at time 0 and without enzyme to 52.1% for 0.1% cellulase after 20 h of reaction. A significant effect of cellulase addition and reaction time on antioxidant power of the tested film-forming solutions was also reported. Film-forming hydrosols with cellulase exhibited a bacteriostatic effect on all tested bacteria, causing a total reduction.

Potential Biomedical Application of Enzymatically Treated Alginate/Chitosan Hydrosols in Sponges-Biocompatible Scaffolds Inducing Chondrogenic Differentiation of Human Adipose Derived Multipotent Stromal Cells.

Current regenerative strategies used for cartilage repair rely on biomaterial functionality as a scaffold for cells that may have potential in chondrogenic differentiation. The purpose of the research was to investigate the biocompatibility of enzymatically treated alginate/chitosan hydrosol sponges and their suitability to support chondrogenic differentiation of human adipose derived multipotent stromal cells (hASCs). The alginate/chitosan and enzyme/alginate/chitosan sponges were formed from hydrosols with various proportions and were used as a biomaterial in this study. Sponges were tested for porosity and wettability. The porosity of each sponge was higher than 80%. An equal dose of alginate and chitosan in the composition of sponges improved their swelling ability. It was found that equal concentrations of alginate and chitosan in hydrosols sponges assure high biocompatibility properties that may be further improved by enzymatic treatment. Importantly, the high biocompatibility of these biomaterials turned out to be crucial in the context of hydrosols' pro-chondrogenic function. After exposure to the chondrogenic conditions, the hASCs in N/A/C and L/A/C sponges formed well developed nodules and revealed increased expression of collagen type II, aggrecan and decreased expression of collagen type I. Moreover, in these cultures, the reactive oxygen species level was lowered while superoxide dismutase activity increased. Based on the obtained results, we conclude that N/A/C and L/A/C sponges may have prospective application as hASCs carriers for cartilage repair.

Study of Enzymatically Treated Alginate/Chitosan Hydrosols in Sponges Formation Process.

The aim of the study was to produce 3D sponges based on enzymatically modified lysozyme selected polysaccharides and assess their physicochemical properties. The alginate/chitosan sponges were formed from polymers hydrosols in different proportions at a final concentration of 1% polysaccharides. Hydrosols were modified by lysozyme addition of 1000 U. Hydrosols without or with enzyme were analyzed for their reducing sugar content, rheological properties and ability to scavenge free radicals. Sponges formed from hydrosols were tested for solubility and compressive properties. Only chitosan was hydrolyzed by lysozyme. The morphology of sponges was investigated by scanning electron microscopy (SEM). It was proven that the antioxidant properties of hydrosols are dependent on the concentration of chitosan. It was also shown that the addition of lysozyme negatively affected the free radical scavenging ability of single hydrosols of alginate and chitosan, and their mixtures. The Ostwald de Waele as well as Herschel⁻Bulkley models of rheological properties fitted the experimental data well (R² is between 0.947 and 1.000). Increase in textural features values of sponges was observed. Sponges with pure alginate and pure chitosan were almost completely soluble. The enzyme addition significantly changed the characteristics of the cross-section structure of sponges, and made the surface smoother.

Study on Alginate⁻Chitosan Complex Formed with Different Polymers Ratio.

Biomaterials based on polyelectrolyte complexation are an innovative concept of coatings and packaging production to be applied in a wide range of food products. The aim of this study was to obtain and characterize a sodium alginate⁻chitosan complex material with variable degree of polyion interactions by complexation of oppositely charged polysaccharides. In order to characterize polyelectrolyte complexes, theromogravimetric analysis (TGA), dynamic mechanical thermal analysis (DMTA), nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FT-IR), matrix-assisted laser desorption/ionization technique with time of flight analyzer (MALDI-TOF), and scanning electron microscopy (SEM) were performed. TGA analysis showed that thermal decomposition temperature depends on the polymer ratio (R) and thermal resistance of samples was improved by increasing chitosan dosage. Accordingly to DMTA results, polyelectrolyte complexation led to obtain more flexible and resistant to mechanical deformation materials. Comparative analysis of the FTIR spectra of single polyelectrolytes, chitosan and alginate, and their mixtures indicated the formation of the polyelectrolyte complex without addition of reinforcing substances. MALDI-TOF analysis confirms the creation of polyelectrolyte aggregates (~197 Da) in samples with R ≥ 0.8; and their chemical stability and safety were proven by NMR analysis. The higher R the greater the number of polyanion⁻polycation aggregates seen in SEM as film morphology roughness.

The use of chitosan, lysozyme, and the nano-silver as antimicrobial ingredients of edible protective hydrosols applied into the surface of meat.

The aim of this study was to design and produce biologically active edible hydrosols, which, when applied to the surface of food products, will protect them from oxidative changes, spoilage and growth of microorganisms. Verification of testing hypothesis and the degree of aim realization were performed by assessing a DPPH radical scavenging activity and microbial reduction of experimental hydrosols on the basis of hydroxypropylmethylcellulose (HPMC), chitosan (CH), lysozyme (L) and nanocolloidal silver (NAg). Antimicrobial activity of different concentrations of CH, L and NAg hydrosols against Gram (+) bacteria: Bacilllus cereus and Micrococcus flavus and Gram (-) bacteria: Escherichia coli and Pseudomonas fluorescens, which exist more often in food, were analyzed using serial dilution test. Total number of microorganisms was determined on meat sample covered by tested sols. Hydrosols containing chitosan and other bioactive substances caused death of each tested microorganism. Lack of chitosan in hydrosols is reflected in a slight inhibition of M. flavus, E. coli and P. fluorescens. Simultaneous influence of CH, L and NAg addition and storage time on total number of bacteria in meat samples with hydrosols was showed. The addition of lysozyme to sols composition significantly increases antioxidant activity.