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Echo-planar magnetic resonance imaging of Gaviscon alginate rafts in-vivo.

Liquid Gaviscon and Gaviscon Advance are established reflux suppressant formulations. This study describes the use of echo-planar magnetic resonance imaging (EPI) to visualise non-invasively intragastric alginate rafts of Liquid Gaviscon and Gaviscon Advance in healthy subjects. Secondly, the feasibility of using relaxation rate (T2(-1)) measurements to monitor changes in the physicochemical properties of the rafts in-vivo is evaluated. Six subjects ingested 500 mL of a liquid meal and received a single dose of 20 mL Liquid Gaviscon or 10 mL Gaviscon Advance on 2 separate visits each and were imaged every 15 min. An alginate raft was observed in the stomach for all subjects and both treatments. The raft was observed to consist of a few large fragments on the majority of the scans for both products. At t = 60 min a raft was still present in all cases. Three-dimensional volume reconstructions showed, for the first time, the spatial distribution of the rafts within the gastric lumen. The T2(-1) data showed potential for assessment of dynamic changes in the physicochemical properties of the alginate rafts in-vivo. We conclude that EPI shows great potential in assessing alginate rafts formation in-vivo.

Adult↗

Alginate-pectin-poly-L-lysine particulate as a potential controlled release formulation.

Drug delivery particulates were prepared using alginate, polylysine and pectin. Theophylline, chlorothiazide and indomethacin were used as the model drugs for in-vitro assessments, and mannitol was the model for assessing paracellular drug absorption across Caco-2 cell monolayers. Alginate and pectin served as the core polymers and polylysine helped to strengthen the particulates. Use of pectin specially helped in forming a more robust particulate that was more resistant in acidic pH and modulated the release profiles of the encapsulated model drugs in the alkaline pH. Alginate and pectin were also found to enhance the paracellular absorption of mannitol across Caco-2 cell monolayers by about three times. The release rate could be described as a first-order or square-root time process depending on the drug load. Use of alginate-polylysine-pectin particulates is expected to combine the advantages of bioadhesion, absorption enhancement, and sustained release. This particulate system may have potential use as a carrier for drugs that are poorly absorbed after oral administration.

Adhesiveness↗

PEC films prepared from Chitosan-Alginate coacervates.

Chitosan-alginate polyelectrolyte complex (PEC) have been prepared in situ in beads and microspheres. This study examines the preparation of suitable chitosan-alginate coacervates for casting into homogeneous PEC films for potential applications in packaging, controlled release systems and wound dressings. Coacervation between chitosan and alginate was rapid, but the rate may be controlled with the addition of water miscible organic solvents. Compared with ethanol and PEG200, acetone was the more promising solvent moderator. Suspensions of fine, uniformly dispersed coacervates were produced by a dropwise addition of 0.25% w/v chitosan solution (solvent: 1: 1 v/v of 2% acetic acid and acetone) into 0.25% w/v sodium alginate solution in water under rapid agitation. The PEC films were transparent and flexible. They exhibited high permeability to water vapor, but resisted complete dissolution in 0.1 M HCI, distilled water and pH 7.4 phosphate buffer solution. Microscopic heterogeneity in the films could be reduced by immersion in aqueous media, but this was accompanied by modifications in the thickness, permeability and mechanical property of the films.

Alginates↗

Purification of alginate oligosaccharides with root growth-promoting activity toward lettuce.

Sodium alginate was degraded by alginate lyase from Corynebacterium sp., and the product was purified by an ultrafiltration (UF) membrane module. The UF treatment was carried out at a transmembrane pressure of 0.15 MPa and a flow velocity of 0.6 m/s in the cross-flow mode, and non-degraded alginate was almost completely removed. The alginate oligosaccharide obtained was a mixture of di- to octasaccharides and had promoting activity toward lettuce root elongation (about 2-fold compared with the control) in the concentration range of 200-3000 microg/ml. The effect of the degree of polymerization on this activity was examined by using each oligosaccharide fractionated by gel chromatography. The tri-, tetra-, penta- and hexasaccharides were each found to have root growth-promoting activity in a lettuce bioassay.

Alginates↗

Purification and characterization of bifunctional alginate lyase from Alteromonas sp. strain no. 272 and its action on saturated oligomeric substrates.

A marine bacterium (strain No. 272) isolated from sea mud in Omura Bay produced an alginate lyase and was classified as an Alteromonas species. The enzyme was purified from the culture medium of the bacterium by DEAE-Cellulofine, Sephadex G-100 gel chromatography to an electrophoretically homogeneous state in the presence and absence of SDS. The molecular mass of the enzyme was 23 and 33.9 kDa on Sephadex G-100 column chromatography and SDS-polyacrylamide gel electrophoresis, respectively, with an isoelectric point of 3.8. The predominant secondary structure of the enzyme was found to be most likely beta-structure by circular dichroism. The enzyme was most active at pH 7.5-8.0 and stable around pH 5-11. The enzyme was more labile in Tris-HCI buffer (pH 7.0) to heat treatment, than in phosphate buffer (pH 7.0). No of metal ions significantly affected the enzyme activity. The enzyme acted on sodium alginate in an endo-type manner and on two components of alginate, poly-alpha1,4-L-guluronate and poly-beta1,4-D-mannuronate, as judged by routine ultraviolet assay (235 nm) and circular dichroic spectral changes of the substrates. However, the coexisting poly-alpha1,4-L-guluronate and poly-beta1,4-D-mannuronate apparently interacted with the enzyme in a competitive manner. Although the enzyme depolymerized alginate in an endo-type, it did not act on trimeric guluronate and mannuronate, but on the tetramers or more. The kinetic analyses showed that kcat/Km for each oligomer was larger for the guluronate oligomers than for the mannuronate ones, and that the subsite structure of the enzyme most likely consisted of six binding sites from the intrinsic reaction rate constant (kint) and intrinsic substrate binding constant (Kint).

Alginates↗

Advanced wound care materials: developing an alginate fibre containing branan ferulate.

OBJECTIVE: This laboratory-based study set out to establish whether branan ferulate--a polysaccharide compound available in gels such as as Sterigel (SSL International, UK)--could be successfully added to the fibre of an alginate dressing to provide a superior wound-care dressing. METHOD: A wet extruder (Howden Engineering Services, UK) was used to produce the fibres. Researchers examined the effect of spinning-bath calcium chloride concentration (used as the coagulant in the fibre-spinning process) on the tensile and absorption properties of alginate fibres containing 25% w/w branan ferulate. Sodium alginate and branan ferulate were separately dissolved in de-ionised water and then stirred together to make up a 5% dope solution. The dope was then forced through a spinneret and into the calcium chloride bath, where fibres were precipitated and then carried over the first set of rollers. The fibres were washed in the next bath and drawn at different rates to enhance their tensile properties. At the 'winding-up' stage the filaments were either passed over a contact heater and into a furnace (dry pick-up) or transferred into acetone baths of increasing concentrations of 50, 80 and 100% aqueous acetate to remove the water (wet pick-up). The moisture content of standard alginate fibres and those with branan ferulate were measured and compared. Breaking load, tenacity and breaking extension, and the quantity of branan ferulate in the fibres were also measured. The process was repeated four times, using different strengths of calcium chloride. Using various draw ratios, the effects of liquid uptake were measured using water, saline and a solution called A (to mimic human blood and exudate-type fluids). RESULTS: Out of the four calcium chloride concentrations used, 1% concentration appeared to give the highest and most conclusive results for fibre-breaking load, tenacity and breaking extension. CONCLUSION: A 1% w/v concentration of calcium chloride provides the optimum conditions for achieving suitably strong fibres with adequate absorption capacity, while allowing the least branan ferulate loss during wash and post-production treatments. In a laboratory setting the product showed promise. Research is now needed in a clinical setting and for comparison with existing wound-care products.

Alginates↗

Improvement of in vivo stability of alginate-polylysine capsules.

Dextran (MW 70,000) and rat islets were encapsulated in alginate-polylysine membrane. The capsules were further coated with tolylene 2,4-diisocyanate. Both of positive and negative charges were detected on the surface of alginate-polylysine capsules, which were absent after coating the capsules with tolylene 2,4-diisocyanate. In the peritoneal cavity, alginate-polylysine capsules were entrapped by the peritoneum, and 89% of dextran was leaked out of the capsules. Alginate-polylysine capsules coated with tolylene 2,4-diisocyanate were present freely in the peritoneal cavity and no leakage of dextran from the capsules were detected. Insulin release from the encapsulated islets was almost the same in both of tolylene 2,4-diisocyanate-coated and uncoated capsules. This study demonstrates that in vivo stability of capsules is dependent on the charge on the surface and can be improved by coating with tolylene 2,4-diisocyanate.

Alginates↗

Calcium-alginate gel bead cross-linked with gelatin as microcarrier for anchorage-dependent cell culture.

Valuable products obtainedfrom the cultivation of anchorage-dependent mammalian cells require large-scale processes to obtain commercially useful quantities. It is generally accepted that suspension culture is the ideal mode of operation. Because anchorage-dependent cells need surfaces to be able to attach and spread, the incorporation of microcarriers to suspension culture is indispensable. Since the dextran-based microcarrier wasfirst introduced, many different types of microcarriers have been developed and commercialized. In this study, alginate-based microcarriers were made in the following order: (i) calcium-alginate gel beads prepared by dropping a blend of sodium alginate and propylene glycol alginate (PGA) into calcium chloride solution, (ii) the PGA section of gel beads cross-linked with gelatin in alkaline solution (i.e., via the transacylation reaction between the ester group of PGA and amino group of gelatin), and (iii) gelatin membrane around the beads further cross-linked by glutaraldehyde. The glutaraldehyde-treated gelatintransacylated PGA/alginate microcarrier showed superior features in high stability under phosphate-containing solution, density close to that of culture medium, and transparency. Moreover, the Chinese hamster ovary CHO-KI and amphotropic retrovirus producer PA317 cells cultivated on the newly synthesized microcarriers exhibited similar growth kinetics of these two types of cell lines cultured on commercial polystyrene microcarriers. However, cell morphology was easily monitored on the transparent microcarriers made in this study.

Alginates↗

Collagen-coated Ba(2+)-alginate microcarriers for the culture of anchorage-dependent mammalian cells.

Several types of microcarriers suitable for large-scale cultivation of mammalian cells are commercially available. However, many of these carriers have disadvantages, e.g., the need for enzymatic digestion for cell harvesting, size limitations and insufficient biocompatibility. These limitations have been overcome by the development of collagen-coated Ba(2+)-alginate microcarriers. Ba(2+)-alginate microspheres, made with the air-jet droplet generator technique, were collagen-coated by incubation in a 0.5% collagen solution, with subsequent gelling of the collagen layer around the alginate microspheres. Human chang liver (CCL-13) and mouse fibroblast (L929) cell lines were cultivated in stationary, unstirred cultures as model systems. After a lag phase of nearly 24 h, the cells grew rapidly on these microcarriers and reached confluence after 3 days. The microcarrier cultures were stable for an additional 4-9 days and longer. Cells were harvested either by trypsinization or by dissolution of the alginate matrix using 5 mM EDTA. The main advantages of this new microcarrier system are that the preparation procedure is easy and can be accomplished on demand with standard laboratory equipment.

Alginates↗

The release of fluoride from two products of alginate impression materials.

The purpose of this study was to determine the fluoride content of two products of alginate and the possible fluoride transfer to the teeth, saliva and blood. The total fluoride content of Zelgan normal-set and Kerr alginate fastset powder was assayed by direct diffusion and diffusion of the ash. The soluble fluoride leaching out in water over 24 hour was also determined. The results show that the fluoride contents of Zelgan and Kerr alginate powders are about 1.9% and 1.5% fluoride, respectively. Of the fluoride present in Zelgan and Kerr approximately 6.5% and 5.8%, respectively, leached out in 400 ml deionized water. The fluoride uptake was estimated in two adjacent enamel layers each approximately 7 micrometers thick, using 10 teeth exposed for 5 minutes and 18 h to the alginate gel (Zelgan). The results of acid etch microsamplings indicate a significant increase in the fluoride concentration of the first enamel layer after both 5 min and 18 h exposure. Fluoride uptake within the second enamel layer was insignificant, however. Fluoride transfer to the oral saliva and to the blood was evident after impression taking.

Alginates↗

Coating alginate beads with cross-linked biopolymers: a novel method based on a transacylation reaction.

Stable membranes were formed around alginate beads using a transacylation reaction between polysaccharidic esters, namely propylene glycol alginate (PGA) or pectin, and various proteins (human serum albumin (HSA), ovalbumin, bovine hemoglobin, lactoserum proteins). In a standard procedure, two reagents (PGA or pectin+protein) were added to a Na-alginate solution: beads were formed by dropwise addition into a calcium solution. Then the transacylation reaction was started by alkalinization of the bead suspension. A membrane was formed around the beads, made of a protein directly bound to a polysaccharide through amide linkages. The thickness of the membranes and the lysis time in trypsin were increased by raising the amount of NaOH used for the transacylation step. In a modified procedure, coated beads were obtained, incorporating PGA in the initial Na-alginate solution, and HSA in the transacylation bath. Activated charcoal was encapsulated in HSA-PGA beads, giving particles with adsorption properties towards creatinine. Assays were performed using PGA associated with alkaline phosphatase as the membrane-forming protein. Stable beads were obtained having a relative activity of 39.3%, as compared with free enzyme.

Acylation↗

Effect of alginate and calcium on secretion of IL-1 by immobilized Swiss peritoneal macrophages.

Murine peritoneal macrophages were immobilized in calcium alginate gel macrobeads, using 1.5% Na-alginate and 50 mM CaCl2. Secretion of IL-1 by immobilized macrophages increased during time and reached 10 fold than IL-1 quantities secreted by adherent macrophages. Calcium and alginate individually enhance production of IL-1 by macrophages and act in synergy when macrophages are immobilized in calcium alginate matrix.

Alginates↗

In vitro slow release profile of endothelial cell growth factor immobilized within calcium alginate microbeads.

Although a variety of angiogenic growth factors have been isolated, its appropriate in vivo delivery remains problematic due to nonspecific, uncontrolled delivery by conventional methods. We have investigated calcium alginate microbeads as a vehicle for the controlled slow-release of endothelial cell growth factor (ECGF). Three different microbead compositions, dependent on ECGF amount and alginate percentage were studied. Microbeads were incubated in a 1.5% calcium chloride solution and release of ECGF into solution was measured spectrophotometrically at specific timepoints. Our results show release rate and amount released after the first 2 hours are dependent on initial quick delivery of ECGF in the first 2 hours after which a sustained controlled release occurred for 4-5 days. Beyond this point, release at a slower rate was noted for at least approximately 2 weeks. Calcium alginate microbeads demonstrated a controlled and predictable rate of release and that the amount of ECGF delivered can be varied by varying the initial concentration of ECGF in the microbeads. Based on these observations we conclude that calcium alginate microbeads are a convenient and practical vehicle for sustained ECGF delivery.

Alginates↗

Poor fermentability of "mekabu" (sporophyll of Undaria pinnatifida) alginic acid in batch culture using pig cecal bacteria.

Sodium alginate, mannuronic acid-rich and guluronic acid-rich fractions were prepared from "Mekabu" (sporophyll of Undaria pinnatifida). The production of short-chain fatty acids such as acetic, propionic and n-butyric acids from these fractions in mini-scale batch culture using pig cecal bacteria was studied, and the gas released from the culture was monitored. The volume of released gas corrected for blank value decreased in the order: glucose (a reference substrate) > guluronic acid-rich fraction > sodium alginate = mannuronic acid-rich fraction. The amounts of short-chain fatty acids produced from three fractions of alginic acid were smaller than that of glucose. These results suggested that alginic acid was poorly fermentable for hindgut bacteria and that its contribution to the host energy pool via microbial metabolism is small.

Acetates↗

Comparison of phenotypic characterization between "alginate bead" and "pellet" culture systems as chondrogenic differentiation models for human mesenchymal stem cells.

Chondrogenesis involves the recruitment of mesenchymal cells to differentiate into chondroblasts, and also the cells must synthesize a cartilage-specific extracellular matrix. There were two representative culture systems that promoted the chondrogenic differentiation of human mesenchymal stem cells. These systems were adaptations of the "pellet" culture system, which was originally described as a method for preventing the phenotypic modulation of chondrocytes, and the "alginate bead" culture system, which was used to maintain encapsulated cells at their differentiated phenotype over time, and also it was used to maintain the cells' proteoglycan synthesis at a rate similar to that of primary chondrocytes. We performed test on the differences of phenotypic characterization with the two methods of differentiating human mesenchymal stem cells into chondrocytes. The typical gene for articular cartilage, collagen type II, was more strongly expressed in the "alginate bead" system than in the "pellet" culture system, in addition, specific gene for hypertrophic cartilage, collagen type X, was more rapidly expressed in the "pellet" system than in "alginate bead" culture system. Therefore, the "alginate bead" culture system is a more phenotypical, practical and appropriate system to differentiate human mesenchymal stem cells into articular chondrocytes than the "pellet" culture system.

Adult↗

Effects of disinfecting alginate impressions on the scratch hardness of stone models.

This study investigated the effects of disinfecting alginate impressions on the scratch depth of resultant stone models. Eleven brands of alginate impression material and two disinfectants, 1% sodium hypochlorite and 2% glutaraldehyde, were used. Impressions were immersed in disinfectant solutions or stored in sealed bags after spraying with disinfectants, and then poured with a type V dental stone. The scratch depth of the stone model obtained from disinfected impression was measured. The storage of alginate impressions after spraying with disinfectants did not increase the scratch depth of resultant stone models. However, the effect of immersion in disinfectants on scratch depth varied with the brand of the alginate impression material.

Alginates↗

Conjugative plasmid transfer between Pseudomonas strains within alginate bead microcosms: effect of the internal gel structure.

Because microorganisms frequently live in an immobilized state in natural habitats, a cell-confined system was used to study bacterial conjugation. Two Pseudomonas putida strains were introduced together within calcium alginate gels. Different alginate beads were designed by varying the polysaccharide and the gelation solution concentrations. Microscopic examinations showed that 2% gels were quite homogeneous, but that 1.5% and 1% gels were rather heterogeneous. In these two last cases, shaft-shaped macrostructures were present. They were colonized during the culture by great densities of highly motile bacteria. Gene transfers due to conjugation were investigated in such alginate gel bead microcosms, in batch and continuous cultures. High-initial transfer frequencies were detected whatever the gel, but no conjugation events seemed to occur with further growth in the beads. Transfer frequency values were roughly similar in the different tested systems. Alginate gels used as artificial microcosms may be valuable to study the effect of cell microenvironment on genetic transfers in complex systems.

Alginates↗

[The effect of alginate purity on microencapsule overgrowth and function of microencapsulated hepatocyte in peritoneal transplantation].

OBJECTIVE: To study the effect of alginate purity on microencapsule overgrowth, histology and function of microencapsulated hepatocyte (MH). METHODS: Alginate was purified by filtration and chloroform/butanol extraction. Using purified and crude alginate to make two kinds of MHs, we transplanted each kind of MHs into the peritoneal cavities of rats. After 2 weeks, 1, 2 and 6 months, MHs were washed out to study overgrowth of MH, histology, and expression of enzyme and albumin in hepatocytes. Two kinds of MHs (the purified and the crude) were transplanted to FHF rats separately to study their differences in liver function support. RESULTS: The amount of retrieved MHs was larger in the purified than in the crude (P < 0.001). In the purified group, most hepatocytes had normal morphology, positive enzymatic histochemistry and albumin immuno-histochemistry stain in one month of transplantation but were damaged in the crude group only 2 weeks later. Microencapsules were smooth and no overgrowth was observed during 6 months in the purified but all overgrowth one month later after transplantation in the crude. The purified MHs had excellent liver function support in the FHF comparing with the crude and the control. CONCLUSION: The purified alginate may significantly reduce the overgrowth of MHs and improve the function of hepatocytes.

Alginates↗