Xylitol production from wood hydrolyzates by entrapped Debaryomyces hansenii and Candida guilliermondii cells

Debaryomyces hansenii cells were entrapped in Ca-alginate beads and used for producing xylitol from wood hydrolyzates. Batch experiments showed that bioconversion was severely hindered when Ca-alginate beads were hardened with Al³⁺ solutions. As an alternative to Al³⁺ hardening, the improvements in both mechanical stability of bioparticles and fermenting ability of the immobilized system derived from using increased concentrations of sodium alginate were assessed. The best results were obtained using a 4% (w/v) Na-alginate solution in the gelification step. This concentration was selected to perform continuous fermentations in a packed-bed reactor using raw or charcoal-treated hydrolyzates (15.5 g of xylose/L) with two different yeasts: Candida guilliermondii and Debaryomyces hansenii. With a final cell concentration of about 50 g of cells/L (0.075 g of cells/g of beads), the volumetric productivities reached with these yeasts in media made from charcoal-treated hydrolyzates were 0.58 and 0.91 g/L·h, respectively.     

Preparation of calcium alginate nanoparticles using water-in-oil (W/O) nanoemulsions

A procedure for the preparation of calcium alginate nanoparticles in the aqueous phase of water-in-oil (W/O) nanoemulsions was developed. The emulsions were produced from mixtures of the nonionic surfactant tetraethylene glycol monododecyl ether (C(12)E(4)), decane, and aqueous solutions of up to 2 wt % sodium alginate by means of the phase inversion temperature (PIT) emulsification method. This method allows the preparation of finely dispersed emulsions without a large input of mechanical energy. With alginate concentrations of 1-2 wt % in the aqueous phase, emulsions showed good stability against Ostwald ripening and narrow, monomodal distributions of droplets with radii <100 nm. Gelation of the alginate was induced by the addition of aqueous CaCl(2) to the emulsions under stirring, and particles formed were collected using a simple procedure based on extraction of the surfactant on addition of excess oil. The final particles were characterized using cryo-transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS). They were found to be essentially spherical with a homogeneous interior, and their size was similar to that of the initial emulsion droplets. The herein presented "low-energy" method for preparation of biocompatible nanoparticles has the potential to be used in various applications, e.g., for the encapsulation of sensitive biomacromolecules.     

Drug release of pH/temperature-responsive calcium alginate/poly(N-isopropylacrylamide) semi-IPN beads

A series of semi-interpenetrating, polymer network (semi-IPN), hydrogel beads, composed of calcium alginate (Ca-alginate) and poly(N-isopropylacrylamide) (PNIPAAM), were prepared for a pH/temperature-sensitive drug delivery study. The equilibrium swelling showed the independent pH- and thermo- responsive nature of the developed materials. At pH=2.1, the release amount of indomethacin incorporated into these beads was about 10% within 400 min, while this value approached to 95% at pH=7.4. The release rate of the drug was higher at 37 degrees C than that at 25 degrees C and increased slightly with increasing PNIPAAM content. These results suggest that the Ca-alginate/PNIPAAM beads have the potential to be used as an effective pH/temperature sustainable delivery system of bioactive agents. [GRAPHS: SEE TEXT] A summary of the temperature- and pH-dependence on the release of the drug over a period of 450 min. The effect of the temperature on the swelling of the beads is shown in the inset.     

Speciation dependent studies on chromium absorption using calcium alginate and iron doped calcium alginate biopolymer

Attempt has been made to develop methodologies for preconcentration of chromium in the biodegradable polymer beads. The uptake behaviors for chromium have been studied with Ca-alginate (CA) and Fe-doped calcium alginate (Fe-CA) beads. The work also aims to study the differential attitude of CA and Fe-CA towards Cr(III) and Cr(VI) so that, depending on the oxidation state of chromium effluent, environmentally sustainable methodologies can be prescribed for removal of chromium. Radiotracer ⁵¹Cr has been chosen as precursor of stable chromium throughout the experiment. It was found that Fe-CA beads are suitable for removal of Cr(III) and Cr(VI) while CA beads can be used for the speciation and separation of Cr(III) and Cr(VI) at pH 5.     

Calcium-Alginate Nanoparticles Formed by Reverse Microemulsion as Gene Carriers

Natural biopolymers are widely used in the field of drug and gene delivery. In this study, alginate nanoparticles were prepared using water-in-oil microemulsion as a template followed by calcium crosslinking of guluronic acid units of alginate polymer. After collected by ultracentrifugation, alginate nanoparticles were analyzed by electron microscopy to obtain the size and morphology which were varied with the ratio of water, oil, and surfactant used. To examine the potency of Ca-alginate nanoparticles as carriers for gene delivery, GFP-encoding plasmids were encapsulated in these nanoparticles to investigate the degree of endocytosis by NIH 3T3 cells and ensuing transfection rate. Our results showed that Ca-alginate nanoparticles with an average size around 80 nm in diameter are very efficient gene carriers, in comparison with plasmid DNA condensed by polyethyleneimine (PEI).     

Preparation of filled temperature-sensitive poly(N-isopropylacrylamide) gel beads

Temperature-sensitive filled poly(N-isopropylacrylamide) (PNIPAAm) gel beads with diameters in the range of millimeters were prepared using the alginate technique. The polymerization and cross-linking reaction of NIPAAm in the presence of inorganic filling particles was performed in spherical networks of Ca-alginate forming interpenetrating networks (IPN). Thermo-sensitive gel beads could be obtained by washing these IPN with EDTA solution. The PNIPAAm gel beads were analyzed by optical methods to observe there swollen diameter in dependence on the temperature. The diameters of the swollen gel beads were in the range of 0.1 - 2 mm. The influence of the monomer to cross-linker ratio (MCR) and the filling materials (ferrofluid, BaTiO₃, TiO₂, and Ni,) were studied. The phase transition temperature (T_pt) was only weakly influenced by the MCR and the filling material remaining at around 34°C.     

Preparation and property of layer-by-layer alginate hydrogel beads based on multi-phase emulsion technique

Layer-by-layer (LbL) alginate beads, which were prepared by multi-phase emulsion technique, had been fabricated via the ionic crosslinking between calcium ion (Ca²⁺) and the carboxylic group of alginate. The prepared beads were spherical, smooth-surfaced and non-aggregated. The SEM analysis displayed the LbL structure of the beads clearly. It had been demonstrated that the size of the beads was controllable and had a correlation with concentration of sodium alginate (SA) and CaCl₂, ratio of water phase and oil phase (W/O), stirring speed, pH value of the water phase, viscosity of SA as well as the temperature for solidification. Stability studies showed that the beads degraded slowly in simulated gastric fluid and simulated intestinal fluid but degraded sharply when they were moved to simulated colonic fluid. Cytotoxicity study by MTT assay indicated that the prepared beads are slightly toxic. It is hoped that this kind of novel beads could be used in pharmaceutical area and cell culture area.     

Gas-Phase Silicon Alkoxide Reactivity vs. Na-Alginate Droplets for Conjugation of Alginate and Sol-Gel Technologies

Solutions with Na-alginate concentrations ranging from 0.5 to 2.5% w/v are processed to prepare Ca-alginate beads using a nozzle ejector under constant He-flow. Beads were spherical in shape and their size distributions were determined; in all samples the average diameter fell in the 120–140 μm interval. Volumetric yields were found to be linearly dependent of the original Na-alginate load whereas the bead diameters were almost constant, according to a constant hindrance of Ca-alginate macromolecular units in the final Ca-alginate gel. The rheology of Na-alginate solutions was studied, with determination of intrinsic viscosity; experimental evidence of microsphere formation, even at the lowest Na-alginate concentrations, indicated that ejection processing changes the rheological parameters controlling bead formation in ordinary dropping processing. Gaseous silicon alkoxides – Si(OEt)₄ and MeSi(OEt)₃ – carried by a He flow were deposited on Na-alginate droplets during ejection. The process was studied by continuous mass spectrometry analysis before and after Na-alginate ejection during the 5-min treatment; in all cases results indicated a deposition yield of 58%. Traces of alcohol in the mass spectrometry analysis of the out-flow gas excluded instantaneous formation of sol-gel silica on the Na-alginate droplets during their residence in the gas phase. For various Na-alginate concentrations, ethanol released by silica gel formation is constant as well as the amount of deposited SiO₂; a siliceous layer ranging from 0.08 to 0.17 μm thick on the surface of the Ca-alginate beads was calculated.     

Oxidative stability and effect of stress factors on flaxseed oil-in-water emulsions stabilized by sodium caseinate–sodium alginate–chitosan interfacial membrane

The susceptibility of heart healthy ω-3 fatty acids to lipid oxidation has hindered its incorporation into healthful foods and beverages. In this study, plant-based flaxseed oil rich in ω-3 fatty acids were dispersed into primary, secondary and tertiary emulsion system. A primary emulsion containing sodium caseinate-stabilized cationic droplets was prepared by homogenizing flaxseed oil as oil phase and sodium caseinate solution as the aqueous phase in an ultrasonicator. A secondary emulsion comprising of sodium caseinate–sodium alginate anionic droplets were produced by diluting appropriate primary emulsion with alginate solution. Further, a tertiary emulsion composed of sodium caseinate–sodium alginate–chitosan-coated cationic droplets was produced by diluting secondary emulsion with chitosan solution. The resistance of primary, secondary and tertiary emulsions with the same lipid concentration to destabilization by thermal treatment (30–90 °C for 30 min), sodium chloride addition (≤70 mM NaCl) and oxidative degradation (hydroperoxide concentration and TBARS) was determined. The results showed that secondary emulsions could resist variation in environmental stresses of salt and heat as well as protect the oil phase from decomposition better than primary and tertiary emulsions. Interfacial engineering could be used to design emulsion system with desirable characteristics.     

Extended release of Gliclazide from highly stabilized calcium alginate/poly(acrylamide) beads for diabetes management

In the present work, stability of calcium alginate beads has been remarkably improved by a novel strategy which consists of in situ formation of poly (acrylamide) within the calcium ions cross-linked sodium alginate beads. The resulting beads have been found to be stable for more than 48 h, in the physiological fluid (PF) of pH 7.4, while the plain alginate beads disintegrated within a couple of hours. The release of the anti-diabetic drug Gliclazide (Glz) from the beads was investigated under physiological conditions. The enhanced stability and prolonged release was also confirmed by an in vivo study on Albino Wistar rats.     

Double emulsion droplets as microreactors for synthesis of magnetic macroporous polymer beads

An easy method is presented to fabricate monodisperse magnetic macroporous polymer beads（MMPBs）. Waterin-oil high internal phase emulsion（HIPE） is prepared by emulsifying aqueous iron ions solution in an oil phase containing monomers. The HIPE is introduced into a simple microfluidic device to fabricate monodisperse（water-in-oil）-in-water double emulsion droplets. The droplets serve as microreactors to synthesize Fe3O4 nanoparticles and are on-line polymerized to form MMPBs. The prepared MMPBs display uniform size, interconnected porous structure, superparamagnetic behavior and uniform distribution of Fe3O4 in polymer matrix. The MMPBs are characterized by scanning electron microscopy（SEM）, Fourier transform infrared spectroscopy（FTIR）, X-ray diffraction（XRD）, transmission electron microscopy（TEM）, vibrating sample magnetometry（VSM）. We believe that this method is a universal technique in preparing macroporous nanocomposite beads.     

Biopolymer microparticle and nanoparticle formation within a microfluidic device

This paper reports a novel microfluidic method for the production of cross-linked alginate microparticles and nanoparticles. We describe a continuous process relying on both thermodynamic and hydrodynamic factors to form microdroplets. A rapid cross-linking reaction thereafter allows solidification of the polymer droplets either within the microfluidic device or "off-chip" to form alginate micro- and nanoparticles. Monodisperse droplets are generated by extruding an aqueous alginate solution using an axisymmetric flow-focusing design. As they flow downstream in the channel, due to water and the continuous phase being partially miscible, the water diffuses very slowly out of the polymeric droplets into the transport fluid, which causes the shrinkage of the drops and the condensation of the polymer phase. The resulting size of the solid particles depends on the polymer concentration and the ensuing balance between the kinetics of the cross-linking reaction and the volume loss due to solvent diffusion. This work details both a single-step microfluidic technique for the formation of alginate microparticles of sizes ranging from 1 to 50 microm via near-equilibrium solvent diffusion within a microfluidic device and thereafter a two-step method, which was shown to generate biopolymer nanoparticles of sizes ranging from 10 to 300 nm. These novel methodologies are extremely flexible and can be extended to the preparation of micro- and nanoparticles from a wide range of single or mixed synthetic and biologically derived polymers.     

Early stages of crystallization of calcium carbonate revealed in picoliter droplets

In this work, we studied the heterogeneous nucleation and growth of CaCO(3) within regular arrays of picoliter droplets created on patterned self-assembled monolayers (SAMs). The SAMs provide well-defined substrates that offer control over CaCO(3) nucleation, and we used these impurity-free droplet arrays to study crystal growth in spatially and chemically controlled, finite-reservoir environments. The results demonstrate a number of remarkable features of precipitation within these confined volumes. CaCO(3) crystallization proceeds significantly more slowly in the droplets than in the bulk, allowing the mechanism of crystallization, which progresses via amorphous calcium carbonate, to be easily observed. In addition, the precipitation reaction terminates at an earlier stage than in the bulk solution, revealing intermediate growth forms. Confinement can therefore be used as a straightforward method for studying the mechanisms of crystallization on a substrate without the requirement for specialized analytical techniques. The results are also of significance to biomineralization processes, where crystallization typically occurs in confinement and in association with organic matrices, and it is envisaged that the method is applicable to many crystallizing systems.     

Multiple phase inversion of emulsions stabilized by in situ surface activation of CaCO3 nanoparticles via adsorption of fatty acids

The in situ surface activation of raw CaCO(3) nanoparticles by interaction with a series of sodium carboxylates of chain length between 6 and 12 as well as sodium 2-ethylhexylsulfosuccinate (AOT) was studied, and the impact of this on the stabilization and phase inversion of toluene-water emulsions was assessed. By using complementary experiments including measurement of particle zeta potentials, adsorption isotherms of amphiphile, and relevant contact angles, the mechanism of this activation was revealed. The results show that hydrophilic CaCO(3) nanoparticles can be surface activated by interaction with sodium carboxylates and AOT even if they are not surface-active themselves. Both the electrostatic interaction between the positive charges on particle surfaces and the negative charges of anionic amphiphile headgroups and the chain-chain interactions of the amphiphile result in monolayer adsorption of the amphiphile at the particle-water interface. This transforms the particles from hydrophilic to partially hydrophobic such that they become surface-active and stabilize oil-in-water O/W(1) emulsions and induce O/W(1) → water-in-oil W/O phase inversion, depending on the chain length of the carboxylate molecules. At high amphiphile concentration, bilayer or hemimicelle adsorption may occur at the particle-water surface, rendering particles hydrophilic again and causing their desorption from the oil-water interface. A second phase inversion, W/O → O/W(2), may occur depending on the surface activity of the amphiphile. CaCO(3) nanoparticles can therefore be made good stabilizers of both O/W and W/O emulsions once surface activated by mixing with traces of suitable anionic amphiphile.     

pH-dependent release from ethylcellulose microparticles containing alginate and calcium carbonate

Ethylcellulose microparticles containing alginate and calcium carbonate nanoparticles were prepared by spray drying water-in-oil emulsion. Alginate solution (3%) in distilled water was used as an aqueous phase, ethylcellulose solution (5%) in dichloromethane as an oil phase, and sorbitan sesquioleate as an emulsifier. The nanoparticles of calcium carbonate were dispersed into the emulsion. By spray-drying the emulsion, ethylcellulose microparticles containing alginate and calcium carbonate were obtained. When the ratios of alginate to calcium carbonate were 4:1 and 2:1, the pH dependency of the release was marked and the degree of release was suppressed in acidic conditions. When the ratio increased to 1:2, the degree of release increased while the pH-dependent release profiles were maintained. Cavities created by the dissolution of calcium carbonate could account for the increased release.     

Formulation characterization and in vitro evaluation of acacia gum–calcium alginate beads for oral drug delivery systems

In the present work, new matrix bead formulations based on linear and branched polysaccharides have been developed using an ionic gelation technique, and their potential use as oral drug carriers has been evaluated. Using calcium chloride as a cross‐linking agent and sodium diclofenac (SD), as a model drug, acacia gum–calcium alginate matrix beads were formulated. The response surface methodology based on 3² factorial design was used as a statistical method to evaluate and optimize the effects of the biopolymers‐blend ratio and the concentration of calcium chloride on the particle size (mm), density (g/cm³), drug encapsulation efficiency (%), and the cumulative drug release after 8 hours (R_8h,%). The optimized beads with the highest drug encapsulation efficiency were examined for a drug‐excipients compatibility by powder X‐ray diffraction, differential scanning calorimetry, thermo‐gravimetric analysis, and Fourier transform‐infrared spectroscopy analyses. The swelling and degradation of the matrix beads were found to be influenced by the pH of medium. Higher degrees of swelling were observed in intestinal pH than in stomach pH. Accordingly, the drug release study showed that the amount of SD released from the acacia gum–calcium alginate beads was higher in intestinal pH than in stomach pH. Therefore, the in vitro drug release from the SD‐loaded beads appears to follow the controlled‐release (Hixson‐Crowell) pattern involving a case‐2 transport mechanism operated by swelling and relaxation of the polymeric blend matrix.     

Biosorption of toxic,heavy, no-carrier-added radionuclides by calcium alginate beads

Summary Studies on adsorption behavior of heavy radionuclides, present altogether in no-carrier-added state, e.g., ^197-200Pb, ^197-200Tl and ¹⁹⁷Hg, have been carried out with calcium alginate beads. High lead (100%) and moderate thallium removal (~65%) was achieved in pH range 2-7. 100% mercury removal was also achieved at pH 2 and 4. Effort has been made to recover all three radionuclides adsorbed in the calcium alginate beads using various chemicals, such as HCl, thiourea, sodium acetate, sodium oxalate and sodium nitrite. It was found that 0.1M HCl and 0.1M thiourea could remove at pH 1 80-90% of adsorbed Pb. Tl recovery was possible by all chemicals mentioned above. Hg was also recovered by all chemicals except HCl.     

Preparation of monodisperse crosslinked polymelamine microcapsules by phase separation method

Monodisperse polymelamine microcapsules were prepared by phase separation method. Control of microcapsule diameter was investigated using the uniform-sized oil-in-water emulsion droplets as the capsule core. The monodisperse emulsion droplets were prepared using the Shirasu porous glass (SPG) membrane emulsification technique. The effects of the diameter of the oil droplet and concentration of sodium dodecyl sulfate (SDS), which is a typical emulsifier in SPG membrane emulsification, on microencapsulation were investigated. The microcapsules were aggregated when oil droplets with small size were microencapsulated at high SDS concentration. To reduce the SDS concentration, the creamed emulsion was used. The monodisperse polymelamine microcapsules were successfully prepared by using the creamed emulsion. The microcapsule diameter was almost similar to the diameter of the encapsulated oil droplet. The coefficient of variation values was about 10% for all microcapsules prepared in this study. Control of microcapsule diameter was achieved in the range of 5–60 μm.     

Gold and silver uptake and nanoprecipitation on calcium alginate beads

Calcium alginate beads were investigated for their biosorption performance in the removal of gold and silver from aqueous solutions. It was found that uptake capacities were significantly affected by the solution pH, with optimum pH values of 2 and 4 for gold and silver, respectively. Kinetic and isotherm experiments were carried out at the optimum pH. The maximum uptake capacities were 290 mg/g for Au and 52 mg/g for Ag. FTIR analysis indicated that both carboxylic and hydroxylic functional groups in alginate beads are involved in the metal binding and later reduction of gold (+3) and silver (+1) to gold (0) and silver (0). SEM and X-ray diffraction confirmed the formation of gold and silver nanoparticles.     

Shape-controlled production of biodegradable calcium alginate gel microparticles using a novel microfluidic device

In this paper we describe a novel method of manufacturing shape-controlled calcium alginate gel microparticles in a microfluidic device. Both manufacturing shape-controlled microparticles and synthesizing hydrogel microparticles could be performed simultaneously in the microfluidic device. The novel microfluidic device comprised of two individual flow-focusing channels and a synthesizing channel was successfully applied as a continuous microfluidic reactor to synthesize gel microparticles with size and shape control. By passive control based on the microchannel geometric confinement and liquid-phase flow rates, we succeeded in producing monodisperse sodium alginate microparticles with diverse shapes (such as plugs, disks, microspheres, rods, and threads) in the flow-focusing channels of the microfluidic device. The shape and size of the sodium alginate microparticles could be tuned by adjusting the flow rates of the various streams. Further stages of the chemical reaction could be initiated by mixing sodium alginate microparticles and calcium chloride (CaCl2) solution in the synthesizing channel. The shapes of the sodium alginate microparticles could be permanently preserved by the synthesis of calcium alginate gel microparticles. The preparation conditions of size- and shape-controlled calcium alginate microparticles and influence factors were studied.