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STUDIES ON THE INHIBITION OF INTESTINAL ABSORPTION OF RADIOACTIVE STRONTIUM. 3. THE EFFECT OF ADMINISTRATION OF SODIUM ALGINATE IN FOOD AND IN DRINKING WATER.

A method is reported which permits selective suppression of absorption of radioactive strontium from ingested food material, permitting calcium to be available to the body. Studies were carried out by measuring bone uptake of Sr(89) and Ca(45) when various amounts of sodium alginate were fed with the diet. Long-term studies were made in which two different levels of radioactivity were used, to determine the pattern of Sr(89) deposition with continuous intake of binding agent. It was found that administration of sodium alginate as a jelly overcomes the problem of constipation and effectively reduces Sr(89) uptake, up to 83%. This fact represents a significant finding with respect to the use of the compound in human subjects. Addition of sodium alginate to drinking water is effective with low levels of Sr(89) intake.This naturally occurring water-soluble macromolecular substance possesses several advantages in use for the suppression of absorption of radioactive strontium when compared with synthetic ion exchange resins: there is no disturbance of electrolyte balance; efficiency is not reduced by treatment over a prolonged period of time; and finally, the product is palatable.

Alginates↗

Dielectric properties of alginate beads and bound water relaxation studied by electrorotation.

The electrical and dielectric properties of Ba2+ and Ca2+ cross-linked alginate hydrogel beads were studied by means of single-particle electrorotation. The use of microstructured electrodes allowed the measurements to be performed over a wide range of medium conductivity from about 5 mS/m to 1 S/m. Within a conductivity range, the beads exhibited measurable electrorotation response at frequencies above 0.2 MHz with two well-resolved co- and antifield peaks. With increasing medium conductivity, both peaks shifted toward higher frequency and their magnitudes decreased greatly. The results were analyzed using various dielectric models that consider the beads as homogeneous spheres with conductive loss and allow the complex rotational behavior of beads to be explained in terms of conductivity and permittivity of the hydrogel. The rotation spectra could be fitted very accurately by assuming (a) a linear relationship between the internal hydrogel conductivity and the medium conductivity, and (b) a broad internal dispersion of the hydrogel centered between 20 and 40 MHz. We attribute this dispersion to the relaxation of water bound to the polysaccharide matrix of the beads. The dielectric characterization of alginate hydrogels is of enormous interest for biotechnology and medicine, where alginate beads are widely used for immobilization of cells and enzymes, for drug delivery, and as microcarriers for cell cultivation.

Alginates↗

The immobilization of microbial cells, subcellular organelles, and enzymes in calcium alginate gels. Reprinted from Biotechnology and Bioengineering, Vol. XIX, No. 3, Pages 387-397 (1977).

Saccharomyces cerevisiae cells, Kluyveromyces marxianus cells, inulase, glucose oxidase, chloroplasts, and mitochondria were immobilized in calcium alginate gels. Ethanol production from glucose solutions by an immobilized preparation of S. cerevisiae was demonstrated over a total of twenty-three days, and the half-life of such a preparation was shown to be about ten days. Immobilized K. marxianus, inulase, and glucose oxidase preparations were used to demonstrate the porosity and retraining properties of calcium alginate gels. Calcium alginate-immobilized chloroplasts were shown to perform the Hill reaction. Some experiments with immobilized mitochondria are reported.

Alginates↗

Cell encapsulation with alginate and alpha-phenoxycinnamylidene-acetylated poly(allylamine).

In general, microcapsules prepared from alginate and polycations lack mechanical strength because the interaction between alginate and polycations is ionic instead of covalent, which represents a much stronger bond. To increase the mechanical strength of the capsule, we prepared photosensitive microcapsules that could form covalent bonds between polymers in the capsular membrane by light irradiation. Two types of photosensitive poly(allylamine), with 5% and 10% of amino groups modified by alpha-phenoxycinnamylidene acetylchloride, were synthesized. Both photopolymers exhibited an absorption maximum at 325 nm and were capable of crosslinking upon light exposure. These photosensitive polymers were used for the preparation of microcapsules. The capsules formed from this photosensitive poly(allylamine) and alginate were strengthened significantly by light irradiation. Only 28% of the microcapsules prepared from the 5%-modified photopolymer fractured after 48 h of shaking at 150 rpm. This fracture percentage is much lower when compared with the 60% of capsules fractured when prepared from the untreated poly(allylamine). By using poly(allylamine) at 10% modification, the mechanical strength was improved only slightly, with 26% of capsules fractured. Analysis of the permeability test indicated that the photo-crosslinked capsular membrane was freely permeable to cytochrome c and myoglobin, but less permeable to serum albumin. The encapsulation method was used to entrap and culture IW32 mouse leukemia cells. The cells proliferated to a density of about 1.1 x 10(7) cells/mL in the capsules after 7 days of cultivation. Concurrently, the concentration of erythropoietin in the microcapsules increased to 800 mU/mL. This new encapsulation technique has great potential in the application of a bioindustrial cell-culturing process.

Alginates↗

Sodium alginate sponges with or without sodium hyaluronate: in vitro engineering of cartilage.

Studies are underway to design biosystems containing embedded chondrocytes to fill osteochondral defects and to produce a tissue close to native cartilage. In the present report, a new alginate three-dimensional support for chondrocyte culture is described. A sodium alginate solution, with or without hyaluronic acid (HA), was freeze-dried to obtain large-porosity sponges. This formulation was compared with a hydrogel of the same composition. In the sponge formulation, macroscopic and microscopic studies demonstrated the formation of a macroporous network (average pore size, 174 microm) associated with a microporous one (average pore size, 250 nm). Histological and biochemical studies showed that, when loaded with HA, the sponge provides an adapted environment for proteoglycan and collagen synthesis by chondrocytes. Cytoskeleton organization was studied by three-dimensional fluorescence microscopy (CellScan EPR). Chondrocytes exhibit a marked spherical shape with a nonoriented and sparse actin microfilament network. Type II collagen was detected in both types of sponges (with or without HA) using immunohistochemistry. In conclusion, the sponge formulation affords new perspectives with respect to the in vitro production of "artificial" cartilage. Furthermore, the presence of hyaluronate within the alginate sponge mimics a functional environment, suitable for the production by embedded chondrocytes of an extracellular matrix.

Alginates↗

Fruit-alginate interactions in novel restructured products.

Novel, healthy, value-added restructured fruit products meet consumer demand for an improved diet containing increasing amounts of fruit. As primary ingredients, fruit purees promise to provide new outlets for visually imperfect fruit or fruit that is too small for the fresh or canned markets. Generally these new product forms require a texturizing agent such as alginate to control the functional properties of the final restructured fruit products. Traditional alginate and pectin gel systems are reviewed in this manuscript as are mixed gel systems. Recent research results describing the production and properties of novel restructured products containing high-guluronic alginate and peach puree without any additional calcium or sugar source are reviewed. Effects of fruit/alginate interactions on gel formation conditions and texture profile results are evaluated.

Alginates↗

Alginate-chitosan complex coacervation for cell encapsulation: effect on mechanical properties and on long-term viability.

The use of chitosan in complexation with alginate appears to be a promising strategy for cell microencapsulation, due to the biocompatibility of both polymers and the high mechanical properties attributed by the use of chitosan. The present work focuses on the optimization and characterization of the alginate-chitosan system to achieve long-term cell encapsulation. Microcapsules were prepared from four types of chitosan using one- and two-stage encapsulation procedures. The effect of reaction time and pH on long-term cell viability and mechanical properties of the microcapsules was evaluated. Using the single-stage encapsulation procedure led to increase of at least fourfold in viability compared with the two-stage procedure. Among the four types of chitosan, the use of high molecular weight (MW) chitosan glutamate and low MW chitosan chloride provided high viability levels as well as good mechanical properties, i.e., more than 93% intact capsules. The high viability levels were found to be independent of the reaction conditions when using high MW chitosan. However, when using low MW chitosan, better viability levels (195%) were obtained when using a pH of 6 and a reaction time of 30 min. An alginate-chitosan cell encapsulation system was devised to achieve high cell viability levels as well as to improve mechanical properties, thus holding great potential for future clinical application.

Alginates↗

Arsenite oxidation in batch reactors with alginate-immobilized ULPAs1 strain.

Arsenic is one of the major groundwater contaminants worldwide. It was previously demonstrated that the beta-proteobacterium Cenibacterium arsenoxidans has an efficient As[III] oxidation ability. The present study was conducted to evaluate the performance of alginate-immobilized ULPAs1 in the oxidation of As[III] to As[V] in batch reactors. A two-level full factorial experimental design was applied to investigate the influence of main parameters involved in the oxidation process, i.e., pH (7-8), temperature (4 degrees C-25 degrees C), kind of nutrient media (2%-20% sauerkraut brine), and arsenic concentration (10-100 mg/L). One hundred milligram per liter of As[III] was fully oxidized by calcium-alginate immobilized cells in 1 h. It was found that the temperature as well as the kind of nutrient media used were significant parameters at a 95% confidence interval whereas only temperature was a significant parameter at a 99% confidence interval. The immobilization of the As[III] oxidizing strain in alginate beads offers a promising way to implement new treatment processes in the remediation of arsenic contaminated waters.

Alginates↗

Encapsulation of plant growth-promoting bacteria in alginate beads enriched with humic acid.

The key to achieving successful, reproducible results following the introduction of beneficial microbes into soil relies on the survival rate of the inoculated bacteria in a heterogeneous soil environment and hence an improved encapsulation method was developed. Owing to the constraints associated with the inoculum formulation, in this study, encapsulation of a plant growth promoting bacteria (PGPB) isolate Bacillus subtilis CC-pg104 was attempted with alginate by enriching the bead microenvironment with humic acid. High viability of the encapsulated bacteria was observed with minimum cell loss upon storage for 5 months. Steady and constant cell release from the bead was observed for 1 week at different pH. Encapsulated cells remained active as evidenced by their ability to solubilize calcium phosphate in vitro. Successful plant growth promotion of lettuce by the encapsulated bacteria under gnotobiotic and sterile environment was also achieved. Feasibility of this improved encapsulation technique is mainly due to the dual benefits of humic acid to microbe and plant and its chemical properties allowing an easy mixing with alginate without interfering in the formation of the alginate gel beads by cross-linking with Ca2+ ions. Thus, the encapsulation method described in this study can be effectively used to protect the PGPB inoculum from adverse conditions of the soil for their successful establishment in the rhizosphere.

Alginates↗

Enhancing oxygen tension and cellular function in alginate cell encapsulation devices through the use of perfluorocarbons.

Encapsulation devices are often hindered by the inability to achieve sufficient oxygen levels for sustaining long-term cell survival both in vivo and in vitro. We have investigated the use of synthetic oxygen carriers in alginate gels to improve metabolic activity and viability of HepG2 cells over time. Perfluorocarbons (PFCs), specifically perfluorotributylamine (PFTBA) and perfluorooctylbromide (PFOB), were emulsified with alginate and used to encapsulate HepG2 cells in a spherical geometry. Cellular state was assessed using the MTT assay and Live/Dead stain as well as through analysis of both lactate and lactate dehydrogenase (LDH) levels which are indirect indicators of oxygen availability. Addition of 1% surfactant resulted in stable emulsions with evenly dispersed PFC droplets of the order of 1-2 microm in diameter, with no influence on cell viability. Both PFCs evaluated were effective in increasing cellular metabolic activity over alginate-only gels. The presence of 10% PFOB significantly increased cellular growth rate by 10% and reduced both intracellular LDH and extracellular lactate levels by 20-40%, improving glucose utilization efficiency. The characteristic drop in cellular metabolic activity upon encapsulation was eliminated with addition of 10% PFC and viability was better maintained throughout the bead, with a significant decrease in necrotic core size. Results were consistent under a physiologically relevant 5% oxygen environment. The incorporation of PFC synthetic oxygen carriers into encapsulation matrices has been successfully applied to improve cell function and viability with implication for a variety of tissue engineering applications.

Alginates↗

Alginate microparticles as novel carrier for oral insulin delivery.

Alginate microparticles produced by emulsification/internal gelation were investigated as a promising carrier for insulin delivery. The procedure involves the dispersion of alginate solution containing insulin protein, into a water immiscible phase. Gelation is triggered in situ by instantaneous release of ionic calcium from carbonate complex via gentle pH adjustment. Particle size is controlled through the emulsification parameters, yielding insulin-loaded microparticles. Particle recovery was compared using several washing protocols. Recovery strategies are proposed and the influence on particle mean size, morphology, recovery yield (RY), encapsulation efficiency, insulin release profile, and structural integrity of released insulin were evaluated. Spherical micron-sized particles loaded with insulin were produced. The recovery process was optimized, improving yield, and ensuring removal of residual oil from the particle surface. The optimum recovery strategy consisted in successive washing with a mixture of acetone/hexane/isopropanol coupled with centrifugation. This strategy led to small spherical particles with an encapsulation efficiency of 80% and a RY around 70%. In vitro release studies showed that alginate itself was not able to suppress insulin release in acidic media; however, this strategy preserves the secondary structure of insulin. Particles had a mean size lower than the critical diameter necessary to be orally absorbed through the intestinal mucosa followed by their passage to systemic circulation and thus can be considered as a promising technology for insulin delivery.

Administration, Oral↗

The immobilization of microbial cells, subcellular organelles, and enzymes in calcium alginate gels.

Saccharomyces cerevisiae cells, Kluyveromyces marxianus cells, inulase, glucose oxidase, chloroplasts, and mitochondria were immobilized in calcium alginate gels. Ethanol production from glucose solutions by an immobilized preparation of S. cerevisiae was deomonstrated over a total of twenty-three days, and the half-life of such a preparation was shown to be about ten days. Immobilized K. marxianus, inulase, and glucose oxidase preparations were used to demonstrate the porosity and retraining properties of calcium alginate gels. Calcium alginate-immobilized chloroplasts were shown to perform the Hill reaction. Some experiments with immobilized mitochondria are reported.

Alginates↗

[Rheological properties of a calcium alginate-potato starch hydrolysate-water system].

The rheological properties of liquid solutions and gels of sodium alginate, calcium gluconate and gelling maltodextrin were investigated and compared with the behaviour of pastes and gels of non-degraded potato starch. The dependence of the rheological properties upon the calcium gluconate concentration is extreme. A marked increase in the viscosity of the liquid solutions and maximal values for the hardness and the breaking strength of the gels are obtained at a calcium gluconate/sodium alginate ratio of 0.3. As to breaking strength, elasticity and hardness, the maltodextrin-based gels are many times inferior to the potato starch-based gels. The breaking strength, hardness and elasticity of maltodextrin gels are increased by the addition of sodium alginate and calcium gluconate, which increases their effectiveness when used in foods.

Alginates↗

Preparation, characterization, and in vitro release of gentamicin from coralline hydroxyapatite-alginate composite microspheres.

In this work, composite microspheres were prepared from bioactive ceramics such as coralline hydroxyapatite [Ca(10)(PO(4))(6)(OH)(2)] granules, a biodegradable polymer, sodium alginate, and an antibiotic, gentamicin. Previously, we have shown a gentamicin release from coralline hydroxyapatite granules-chitosan composite microspheres. In the present investigation, we attempted to prepare composite microspheres containing coralline hydroxyapatite granules and sodium alginate by the dispersion polymerization technique with gentamicin incorporated by absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. Fourier transform infrared spectra clearly indicated the presence of per-acid of sodium alginate, phosphate, and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs showed that the composite microspheres were spherical in shape and porous in nature. The particle size of composite microspheres was analyzed, and the average size was found to be 15 microns. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns.

Alginates↗

Production of microspheres based on hydrophobically associating alginate derivatives by dispersion/gelation in aqueous sodium chloride solutions.

A new "all aqueous" procedure for the preparation of stable polysaccharide microparticles was developed. The method consists of dispersing a water solution of an amphiphilic alginate derivative (in the current work, alginate substituted with low amounts of dodecyl chains) first fluidified under mechanical stress, into an NaCl solution. The procedure exploits the ability of amphiphilic associative derivatives to form strong hydrogels in the presence of nonchaotropic salts and their shear-thinning/thixotropic properties. Depending on the experimental conditions, the size of the microparticles can be varied from 10 microm to several hundred micrometers. Their mechanical properties can eventually be reinforced by addition of low concentrations of calcium chloride. The resulting microparticles exhibit a better stability than that of plain Ca(2+)-alginate particles, as they are not disrupted when nongelling cations or calcium-sequestering agents are added to the solution. In addition, the particles can be easily redispersed after being centrifuged or freeze-dried.

Alginates↗

Covalent immobilization of aminoacylase to alginate for L-phenylalanine production.

Aminoacylase I (EC. 3.5.1.14) was immobilized by covalent crosslinking to alginate molecules with 1-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide HCl followed by calcium alginate bead formation for the production of L-phenylalanine from the racemic mixtures of N-acetyl-DL-phenylalanine. Different concentrations of the coupling reagent were tested and the coupling process was optimized. The immobilized and the partially purified aminoacylase were characterized in terms of the activity, operational stability, thermal stability, pH and temperature optima and kinetic constants, Km and Vmax. The activity of the enzyme covalently immobilized in calcium alginate beads was enhanced by about 75% compared to that of free enzyme. The beads showed stable activity under operational conditions, they lost about 40% of their activity after four reaction cycles. The immobilized aminoacylase was more stable over a broader pH range. Thus this simple method provides irreversible immobilization of aminoacylase to give a biocatalyst with good operational stability and enhanced activity.

Alginates↗

Enhanced proliferation and differentiation of human articular chondrocytes when seeded at low cell densities in alginate in vitro.

Dedifferentiated human articular chondrocytes exhibited a wide variation in their capacity to proliferate and redifferentiate in an alginate suspension culture system. The greatest extent of proliferation and redifferentiation was seen to be dependent on the formation of clonal populations of chondrocytes and correlated inversely with the initial cell seeding density. Redifferentiating chondrocytes seeded at low density (1 x 10(4) cells/ml alginate) compared with chondrocytes that were seeded at high density (1 x 10(6) cells/ml alginate) showed a nearly 3-fold higher median increase in cell number. a 19-fold greater level of type-II collagen mRNA expression, a 4-fold greater level of aggrecan mRNA expression, and a 6-fold greater level of sulfated glycosaminoglycan deposition at 4 weeks of culture. Matrix molecules from low-density cultures were assembled into chondrocyte-encapsulated, spherical extracellular matrices that were readily visualized in sections from 12-week cultures stained with antibodies against types I and II collagen and aggrecan. Ultrastructural analysis of 12-week low-density cultures confirmed the presence of thin collagen fibrils throughout the matrix.

Adult↗

Characterization of riboflavin-photosensitized changes in aqueous solutions of alginate by dynamic light scattering.

The effect of irradiation, in the wavelength range of 310-800 nm, on aqueous solutions (pH = 7.4) of alginate in the presence of the photosensitizer riboflavin (RF) has been investigated with the aid of dynamic light scattering (DLS). Under aerobic conditions light irradiation of RF causes scission of the polymer chains which affects the polymer dynamics. The time correlation data obtained from DLS experiments showed at all conditions the existence of two relaxation modes: one single exponential at short times, followed by a stretched exponential at longer times. The slow relaxation time revealed, over the whole considered concentration range, lower values for the alginate/RF system, whereas no effect of photochemical degradation was observed for the fast relaxation time in the semidilute regime. The results suggest that the photochemically induced fragmentation of alginate affects the slow relaxation mode, associated with disengagement relaxation of individual chains or cluster relaxation, in a similar way as the zero-shear viscosity. These findings provide detailed insight into the dynamics of the polymer matrix, and this knowledge can be useful in the context of controlled-release delivery of drugs. The chemical units of alginate (M = mannuronic acid and G = guluronic acid).

Alginates↗