Effect of Callus Cell Immobilization on the Textural and Rheological Properties, Loading, and Releasing of Grape Seed Extract from Pectin Hydrogels.

The purpose of the present study was to prepare pectin hydrogels with immobilized Lemna minor callus cells and to identify the effect of cell immobilization on the textural, rheological, and swelling properties; loading; and releasing of grape seed extract (GSE) from the hydrogels. Hardness, adhesiveness, elasticity, the strength of linkage, and complex viscosity decreased with increasing cell content in the hydrogels based on pectin with a degree of methyl esterification (DM) of 5.7% (TVC) and during incubation in gastrointestinal fluids. An increase in the rheological properties and fragility of pectin/callus hydrogels based on pectin with a DM of 33.0% (CP) was observed at a cell content of 0.4 g/mL. TVC-based pectin/callus beads increased their swelling in gastrointestinal fluids as cell content increased. TVC-based beads released GSE very slowly into simulated gastric and intestinal fluids, indicating controlled release. The GSE release rate in colonic fluid decreased with increasing cell content, which was associated with the accumulation of GSE in cells. CP-based beads released GSE completely in the intestinal fluid due to weak textural characteristics and rapid degradation within 10 min. Pectin/callus hydrogels have the ability to preserve GSE for a long time and may have great potential for the development of proanthocyanidin delivery systems due to their novel beneficial physicochemical and textural properties.

Effect of Homogenized Callus Tissue on the Rheological and Mechanical Properties of 3D-Printed Food.

The aim of the study was to develop ink enriched with a high content of lupine callus tissue (CT) suitable for 3D printing. Printable ink obtained using mashed potatoes (20 g/100 mL) and a 3% agar solution was used as the parent CT-free ink (CT0). Viscosity increased from 9.6 to 75.4 kPa·s during the cooling of the CT0 ink from 50 to 20 °C, while the viscosity of the ink with 80 g/100 mL of CT (CT80) increased from 0.9 to 5.6 kPa·s under the same conditions. The inclusion of CT was shown to decrease the hardness of 3D-printed food gel from 0.32 ± 0.03 to 0.21 ± 0.03 N. The storage modulus G' value was 7.9 times lower in CT80 samples than in CT0 samples. The values of fracture stress for CT80 and CT0 inks were 1621 ± 711 and 13,241 ± 2329 Pa, respectively. The loss tangent and the limiting strain did not differ in CT0 and CT80, although the value of the fracture strain was 1.6 times higher in the latter. Thus, the present study demonstrates that CT may be added to printing ink in order to enhance food with plant cell material and enable the 3D printing of specially shaped foods.

Effect of Chitosan on Rheological, Mechanical, and Adhesive Properties of Pectin-Calcium Gel.

In the present study, chitosan was included in the pectin ionotropic gel to improve its mechanical and bioadhesive properties. Pectin-chitosan gels P-Ch0, P-Ch1, P-Ch2, and P-Ch3 of chitosan weight fractions of 0.00, 0.25, 0.50, and 0.75 were prepared and characterized by dynamic rheological tests, penetration tests, and serosal adhesion ex vivo assays. The storage modulus (G') and loss modulus (G″) values, gel hardness, and elasticity of P-Ch1 were significantly higher than those of P-Ch0 gel. However, a further increase in the content of chitosan in the gel significantly reduced these parameters. The inclusion of chitosan into the pectin gel led to a decrease in weight and an increase in hardness during incubation in Hanks' solution at pH 5.0, 7.4, and 8.0. The adhesion of P-Ch1 and P-Ch2 to rat intestinal serosa ex vivo was 1.3 and 1.7 times stronger, whereas that of P-Ch3 was similar to that of a P-Ch0 gel. Pre-incubation in Hanks' solution at pH 5.0 and 7.4 reduced the adhesivity of gels; however, the adhesivity of P-Ch1 and P-Ch2 exceeded that of P-Ch0 and P-Ch3. Thus, serosal adhesion combined with higher mechanical stability in a wide pH range appeared to be advantages of the inclusion of chitosan into pectin gel.

Effect of Hogweed Pectin on Rheological, Mechanical, and Sensory Properties of Apple Pectin Hydrogel.

This study aims to develop hydrogels from apple pectin (AP) and hogweed pectin (HP) in multiple ratios (4:0; 3:1; 2:2; 1:3; and 0:4) using ionotropic gelling with calcium gluconate. Rheological and textural analyses, electromyography, a sensory analysis, and the digestibility of the hydrogels were determined. Increasing the HP content in the mixed hydrogel increased its strength. The Young's modulus and tangent after flow point values were higher for mixed hydrogels than for pure AP and HP hydrogels, suggesting a synergistic effect. The HP hydrogel increased the chewing duration, number of chews, and masticatory muscle activity. Pectin hydrogels received the same likeness scores and differed only in regard to perceived hardness and brittleness. The galacturonic acid was found predominantly in the incubation medium after the digestion of the pure AP hydrogel in simulated intestinal (SIF) and colonic (SCF) fluids. Galacturonic acid was slightly released from HP-containing hydrogels during chewing and treatment with simulated gastric fluid (SGF) and SIF, as well as in significant amounts during SCF treatment. Thus, new food hydrogels with new rheological, textural, and sensory properties can be obtained from a mixture of two low-methyl-esterified pectins (LMPs) with different structures.

Enrichment of 3D-Printed k-Carrageenan Food Gel with Callus Tissue of Narrow-Leaved Lupin Lupinus angustifolius.

The aim of the study is to develop and evaluate the printability of k-carrageenan inks enriched with callus tissue of lupin (L. angustifolius) and to determine the effect of two lupin calluses (LA14 and LA16) on the texture and digestibility of 3D-printed gel. The results demonstrated that the enriched ink was successfully 3D printed at concentrations of 33 and 50 g/100 mL of LA14 callus and 33 g/100 mL of LA16 callus. The feasibility of 3D printing is extremely reduced at higher concentrations of callus material in the ink. The hardness, cohesiveness, and gumminess of the 3D-printed gel with LA16 callus were weakened compared to the gel with LA14 callus. The results of rheological measurements showed that an increase in the content of LA16 callus interfered with the formation of a k-carrageenan gel network, while LA14 callus strengthened the k-carrageenan gel with increasing concentration. Gel samples at different concentrations of LA14 and LA16 calluses formed a spongy network structure, but the number of pores decreased, and their size increased, when the volume fraction occupied by LA14 and LA16 calluses increased. Simple polysaccharides, galacturonic acid residues, and phenolic compounds (PCs) were released from A-FP gels after sequential in vivo oral and in vitro gastrointestinal digestion. PCs were released predominantly in the simulated intestinal and colonic fluids. Thus, incorporating lupin callus into the hydrocolloid ink for food 3D printing can be a promising approach to developing a gelling material with new mechanical, rheological, and functional properties.

Polysaccharides of Salsola passerina: Extraction, Structural Characterization and Antioxidant Activity.

The above-ground part of the Salsola passerine was found to contain ~13% (w/w) of polysaccharides extractable with water and aqueous solutions of ammonium oxalate and sodium carbonate. The fractions extracted with aqueous sodium carbonate solutions had the highest yield. The polysaccharides of majority fractions are characterized by similar monosaccharide composition; namely, galacturonic acid and arabinose residues are the principal components of their carbohydrate chains. The present study focused on the determination of antioxidant activity of the extracted polysaccharide fractions and elucidation of the structure of polysaccharides using nuclear magnetic resonance (NMR) spectroscopy. Homogalacturonan (HG), consisting of 1,4-linked residues of α-D-galactopyranosyluronic acid (GalpA), rhamnogalacturonan-I (RG-I), which contains a diglycosyl repeating unit with a strictly alternating sequence of 1,4-linked D-GalpA and 1,2-linked L-rhamnopyranose (Rhap) residues in the backbone, and arabinan, were identified as the structural units of the obtained polysaccharides. HMBC spectra showed that arabinan consisted of alternating regions formed by 3,5-substituted and 1,5-linked arabinofuranose residues, but there was no alternation of these residues in the arabinan structure. Polysaccharide fractions scavenged the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical at 0.2-1.8 mg/mL. The correlation analysis showed that the DPPH scavenging activity of polysaccharide fractions was associated with the content of phenolic compounds (PCs).

Enrichment of Agar Gel with Antioxidant Pectin from Fireweed: Mechanical and Rheological Properties, Simulated Digestibility, and Oral Processing.

The aims of the study were to evaluate the influence of pectin isolated from fireweed (FP) on the mechanical and rheological properties of agar (A) gel, to investigate the release of phenolic compounds (PCs) and pectin from A-FP gels at simulated digestion in vitro, and to evaluate the oral processing and sensory properties of A-FP gels. The hardness of A-FP gels decreased gradually with the increase in the concentration of FP added (0.1, 0.4, and 1.6%). The hardness of A-FP1.6 gel was 41% lower than A gel. Rheological tests found A gel was a strong physical gel (storage modulus (G') >>loss modulus (G″)), and the addition of FP up to 1.6% did not significantly change its G'. The G″ value decreased in A-FP gels compared to A gel. The release of galacturonic acid (GalA) was 3.4 ± 0.5, 0.5 ± 0.2, 2.4 ± 1.0, and 2.2 ± 0.7 mg/mL after digestion of A-FP1.6 gel in the oral in vivo phase (OP) and subsequent incubation in simulated gastric (SGF), intestinal (SIF), and colonic (SCF) fluids in vitro. The incubation medium after OP, SGF, and SIF digestion of A-FP1.6 contained 24−64 µg GAE/mL of PCs, while SCF contained 144 µg GAE/mL, supposing a predominant release of antioxidant activity from the gel in the colon. Chewing to readiness for swallowing A-FP gel required less time and fewer chews with less activity of the masseter and temporalis muscles. A-FP1.6 gel had a lower likeness score for taste and consistency and a similar score for appearance and aroma when compared with A gel. Thus, A-FP gels were weakened compared to A gel and required less time and muscle activity for oral processing. A-FP gel had antioxidant activity due to the PCs associated with pectin, while A gel had no antioxidant activity.

Swelling, Protein Adsorption, and Biocompatibility In Vitro of Gel Beads Prepared from Pectin of Hogweed Heracleum sosnówskyi Manden in Comparison with Gel Beads from Apple Pectin.

The study aims to develop gel beads with improved functional properties and biocompatibility from hogweed (HS) pectin. HS4 and AP4 gel beads were prepared from the HS pectin and apple pectin (AP) using gelling with calcium ions. HS4 and AP4 gel beads swelled in PBS in dependence on pH. The swelling degree of HS4 and AP4 gel beads was 191 and 136%, respectively, in PBS at pH 7.4. The hardness of HS4 and AP4 gel beads reduced 8.2 and 60 times, respectively, compared with the initial value after 24 h incubation. Both pectin gel beads swelled less in Hanks' solution than in PBS and swelled less in Hanks' solution containing peritoneal macrophages than in cell-free Hanks' solution. Serum protein adsorption by HS4 and AP4 gel beads was 118 ± 44 and 196 ± 68 µg/cm2 after 24 h of incubation. Both pectin gel beads demonstrated low rates of hemolysis and complement activation. However, HS4 gel beads inhibited the LPS-stimulated secretion of TNF-α and the expression of TLR4 and NF-κB by macrophages, whereas AP4 gel beads stimulated the inflammatory response of macrophages. HS4 gel beads adsorbed 1.3 times more LPS and adhered to 1.6 times more macrophages than AP4 gel beads. Thus, HS pectin gel has advantages over AP gel concerning swelling behavior, protein adsorption, and biocompatibility.

Characterization and Biocompatibility Properties In Vitro of Gel Beads Based on the Pectin and κ-Carrageenan.

This study aimed to investigate the influence of kappa (κ)-carrageenan on the initial stages of the foreign body response against pectin gel. Pectin-carrageenan (P-Car) gel beads were prepared from the apple pectin and κ-carrageenan using gelling with calcium ions. The inclusion of 0.5% κ-carrageenan (Car0.5) in the 1.5 (P1.5) and 2% pectin (P2) gel formulations decreased the gel strength by 2.5 times. Car0.5 was found to increase the swelling of P2 gel beads in the cell culture medium. P2 gel beads adsorbed 30-42 mg/g of bovine serum albumin (BSA) depending on pH. P2-Car0.2, P2-Car0.5, and P1.5-Car0.5 beads reduced BSA adsorption by 3.1, 5.2, and 4.0 times compared to P2 beads, respectively, at pH 7. The P1.5-Car0.5 beads activated complement and induced the haemolysis less than gel beads of pure pectin. Moreover, P1.5-Car0.5 gel beads allowed less adhesion of mouse peritoneal macrophages, TNF-α production, and NF-κB activation than the pure pectin gel beads. There were no differences in TLR4 and ICAM-1 levels in macrophages treated with P and P-Car gel beads. P2-Car0.5 hydrogel demonstrated lower adhesion to serous membrane than P2 hydrogel. Thus, the data obtained indicate that the inclusion of κ-carrageenan in the apple pectin gel improves its biocompatibility.

Isolation, Chemical Characterization and Antioxidant Activity of Pectic Polysaccharides of Fireweed (Epilobium angustifolium L.).

The aim of this study was to isolate pectins with antioxidant activity from the leaves of Epilobium angustifolium L. Two pectins, EA-4.0 and EA-0.8, with galacturonic acid contents of 88 and 91% were isolated from the leaves of E. angustifolium L. by the treatment of plant raw materials with aqueous hydrochloric acid at pH 4.0 and 0.8, respectively. EA-4.0 and EA-0.8 were found to scavenge the DPPH radical in a concentration-dependent manner at 17-133 µg/mL, whereas commercial apple pectin scavenged at 0.5-2 mg/mL. The antioxidant activity of EA-4.0 was the highest and exceeded the activity of EA-0.8 and a commercial apple pectin by 2 and 39 times (IC50-0.050, 0.109 and 1.961 mg/mL), respectively. Pectins EA-4.0 and EA-0.8 were found to possess superoxide radical scavenging activity, with IC50s equal to 0.27 and 0.97 mg/mL, respectively. Correlation analysis of the composition and activity of 32 polysaccharide fractions obtained by enzyme hydrolysis and anionic exchange chromatography revealed that the antioxidant capacity of fireweed pectins is mainly due to phenolics and is partially associated with xylogalacturonan chains. The data obtained demonstrate that pectic polysaccharides appeared to be bioactive components of fireweed leaves with high antioxidant activity, which depend on pH at their extraction.

Pectin-glycerol gel beads: Preparation, characterization and swelling behaviour.

Low methyl-esterified pectin (AU701) was found to form gel beads with glycerol. Wet AU701-glycerol gel beads exhibited similar diameter and hardness compared to the AU701-Ca gel beads, prepared by ionotropic gelation with Ca2+ and used for comparison. The morphology of dry pectin gel beads determined by scanning electron microscopy revealed that the beads exhibited rough and grooved surface. The AU701-glycerol gel beads absorbed more grams of water than AU701-Ca gel beads (12.2 g vs 3.9 g per 1 g of the beads). Rheological properties and hardness of the AU701-glycerol gel beads improved with the increase of the pectin/glycerol ratio. Swelling behavior of the AU701-glycerol gel beads was determined after sequential incubation in simulated gastric (SGF) and intestinal (SIF) fluids. The AU701-glycerol gel beads swelled in SGF to a greater extent and revealed higher stability in SIF than the gel beads cross-linked by Ca2+.

Pectin gelling in acidic gastric condition increases rheological properties of gastric digesta and reduces glycaemic response in mice.

The rheological characteristics and transit time of gastric digesta and the postprandial glycaemic response in mice orally administered with water (control) or pectin solutions supplemented (AP-Ca) or not supplemented (AP) with CaCO3 were elucidated. AP and AP-Ca increased viscosity, storage and loss moduluses (G' and G'') of mice gastric digesta. The gelling capacity of AP-Ca in acidic gastric conditions appeared to provide a greater enhancement of gastric digesta viscosity compared with AP. The postprandial blood glucose concentration was lower in mice orally administered with AP or AP-Ca compared with control mice. The transit time of gastric digesta and the blood glucose concentration were affected in mice orally administered with AP during the early postprandial period. The effect of AP-Ca on the gastric digesta rheology and transit time was stronger than that of AP. Both of the pectin solutions failed to reduce food intake in mice.

Pectin-silica gels as matrices for controlled drug release in gastrointestinal tract.

The synthesis of pectin-silica gels for controlled drug release in gastrointestinal tract (GIT) using low methoxyl (LM) and high methoxy (HM) pectins and tetraethoxysilane (TEOS) as precursor is described. The FTIR spectra of the pectin-silica gels show intense absorption bands at 1246cm-1 and 802cm-1 corresponding to the vibrations COSi bonds, which absent in the FTIR spectra of the native pectins that indicate the formation covalent bond between silica and pectin macromolecules in the pectin-silica gels. Pectin-TEOS, pectin-Ca-TEOS and pectin-TEOS-Ca beads with mesalazine are synthesized by different combinations of sol-gel method using TEOS and ionotropic gelation method using calcium chloride. The best resistant of pectin-TEOS and pectin-Ca-TEOS beads during incubation in simulated gastric fluid for 2h and subsequently in simulated intestinal fluids for 18h is indicated. Pectin-TEOS beads are characterized by higher encapsulation efficiency (to 28%) than pectin-Ca-TEOS beads (to 16%). The drug release of pectin-silica beads in simulated GIT occurs gradually up to 80% and is directly dependent on the hardness of the beads. The surface morphology of beads is shown. The use of pectin-silica beads is promising with regard to the development of controlled release of drug formulations.

In vitro gastrointestinal-resistant pectin hydrogel particles for ß-glucuronidase adsorption.

Pectin hydrogel particles (PHPs) were prepared by ionotropic gelation of low methylesterified pectin of Tanacetum vulgare L. with calcium ions. Wet PHPs prepared from TVF exhibited a smaller diameter and the lower weight as well as exhibited the best textural properties in terms of hardness and elasticity compared to the PHPs prepared from commercial low methylesterified pectin (CU701) used for comparison. Upon air drying, PHPs prepared from CU701 became small and dense microspheres whereas the dry PHPs prepared from TVF exhibited a drop-like shape. The morphology of dry PHPs determined by scanning electron microscopy revealed that the surface of the TVF beads exhibited fibred structures, whereas the PHPs prepared from CU701 exhibited a smooth surface. The characterization of surface roughness using atomic force microscopy indicated less roughness profile of the PHPs prepared from TVF than CU701. PHPs prepared from TVF were found to possess in vitro resistance to successive incubations in simulated gastric (SGF), intestinal (SIF), and colonic fluid (SCF) at 37 °C for 2, 4 and 18 h, respectively. The PHPs prepared from CU701 swelled in SGF and then lost their spherical shape and were fully disintegrated after 4 h of incubation in SIF. The PHPs from TVF, which were subjected to treatment with SGF, SIF and SCF, were found to adsorb microbial ß-glucuronidase (ßG) in vitro. The data obtained offered the prospect for the development of the PHPs from TVF as sorbents of colonic ßG for the inhibition of re-absorption of estrogens.

Impact of cabbage pectin-protein complex on microbial ß-glucuronidase activity.

We previously demonstrated that pectin-protein complex (PPC) isolated from white cabbage adsorbs the ß-glucuronidase (ßG) enzyme of E. coli. Concurrently, we discovered a significant increase in ßG activity in the presence of PPC. The aim of this study is to identify the structural components of PPC that are responsible for ßG adsorption and activation. PPC was isolated from white cabbage using a saline solution containing hydrochloric acid (pH 1.5) at 37 °C for 4 h. PPC proteins were precipitated by aqueous 10% (m/v) trichloroacetic acid to yield the pectin-protein fractions PPC1 and PPC2. PPC was digested using 1,4-α-d-galacturonase, yielding the PPC6 fraction. Partial acid hydrolysis of PPC revealed the galacturonan fraction, PPC3, to be the core of the macromolecule. The purified PPC4 and PPC5 fractions were isolated from PPC by ion-exchange chromatography on DEAE-cellulose. ßG activity and its adsorption in the PPC fractions were studied in vitro. Crystalline cellulose was used as a control. This study found that the PPC3 fraction (the galacturonan core) does not adsorb ßG and does not affect its activity. The adsorption of ßG in the PPC samples is inversely proportional to the degree of methyl esterification of its carbohydrate component. The PPC4 and PPC5 fractions adsorb the highest proportion of ßG (51.2% and 54%, respectively). The stimulation of ßG enzyme activity is directly proportional to the protein content of the PPC sample. The PPC and PPC1 samples have the greatest ability to increase ßG activity (57.6% and 52.1%, respectively).

Interaction of microbial ß-glucuronidase with vegetable pectins.

The purpose of this study was to investigate the in vitro effects of vegetable carbohydrates on the activity of microbial ß-glucuronidase (ßG) and the adsorption of the enzyme on carbohydrates. This study used pectin-protein complexes (PPCs) with molecular weights of 300 kDa isolated under conditions simulating a gastric environment from cabbage (HCl-PPCC and HCl+pepsin-PPCCP) and sweet pepper (PPCP and PPCPP). As a sample for comparison, microcrystalline cellulose was used. The activity of ßG from Escherichia coli was determined spectrophotometrically by the formation of the colored product from the breakdown of phenolphthalein-ß-D-glucuronide. Adsorption of ßG on biopolymers was studied by the retention of the enzyme on the membrane of a concentrator with a pore diameter of 300 kDa and by native PAGE. PPCCP and PPCC were established to increase the activity of ßG by 50 and 100%, respectively. Cellulose had a weak effect, whereas pepper PPC had no effect. All studied carbohydrates adsorb on ßG. The maximum ßG adsorption (15%) was observed with PPCC, whereas PPCCP absorbed 5% of the enzyme. Pepper PPCs and cellulose adsorbed up to 10% of the enzyme. There was a positive correlation between the increase of ßG activity in the presence of carbohydrates and enzyme adsorption on the polymers (r=0.80; P<0.01). The activity of the enzyme in the gel after electrophoresis of the PPCC+ßG mixture was inversely proportional to the concentration of PPCC in the mixture. A model explaining the effects of cabbage PPCs on the excretion of estrogens is proposed.