Synthesis and characterization of composite magnetic microspheres of artemisia seed gum and chitosan

In this study, composite magnetic microspheres of artemisia seed gum and chitosan were prepared in a well‐shaped spherical form with a size range of 230–460 μm by the suspension crosslinking technique for use in the application of magnetic carrier technology. The magnetic material used in the preparation of the composite microspheres was prepared by precipitation from FeCl₃ and FeSO₄ solution in basic medium. The morphological, magnetic properties, and the functional groups of the microspheres were characterized by different techniques (i.e., SEM, magnetometry, and FTIR). The results demonstrated that the stirring rate of the suspension and the Fe₃O₄/chitosan ratio are the most effective parameters for the average of the size distributions and the magnetic quality of the microspheres, while the amount of artemisia seed gum and Tween‐80 have no significant effect on these properties. The best magnetic quality of the composite magnetic microspheres is around 4.08 emu/g microspheres at 10 KG magnetic field intensity. The thermal stability of the composite magnetic microspheres was measured by using DSC methods. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 3045–3049, 2007     

Application of tris(hydroxymethyl)phosphine as a coupling agent for β‐galactosidase immobilized on chitosan to produce galactooligosaccharides

β‐Galactosidase was immobilized on chitosan using tris(hydroxymethyl)phosphine (THP) as a coupling agent to produce galactooligosaccharides (GOS) from lactose. Both the THP‐immobilized and the free enzymes were maximally achieved at pH 5.0 and the optimal temperature was 55 °C. The residual activities for the THP‐immobilized enzyme and the free enzyme were 75 and 25%, respectively, after being incubated in 0.1 mol dm^?3 sodium acetate buffer (pH 5.0) at 55 °C for 13 days. The formation of GOS was catalyzed by free and THP‐immobilized β‐galactosidase from lactose. The yield of GOS produced by the free enzyme from the lactose solution (36%, w/v) at 55 °C was 43% on a dry weight basis, which was similar to the 41% GOS yield produced by the THP‐immobilized enzyme system. Copyright © 2005 Society of Chemical Industry     

Chitosan and chitosan/β‐cyclodextrin microspheres as sustained‐release drug carriers

The main aim of this study was to compare two microspheres, chitosan (CTS) and CTS/β‐cyclodextrin (β‐CD), made by spray‐drying, as pulmonary sustained drug‐delivery carriers. Theophylline (TH) was used as a model drug. The characteristics of the microspheres and in vitro release were studied. The yield of CTS/β‐CD microspheres was 46.1%, which was higher than that of the CTS microspheres (36.5%). The drug loads of the CTS and CTS/β‐CD microspheres were 22.7 and 21.1%, respectively, whereas the encapsulation efficiencies were 90.7 and 91.4%, respectively. The distribution of 50% [(diameter) d (0.5)] of the CTS microspheres was below 6.49 μm and that of the CTS/β‐CD microspheres was below 4.90 μm. Scanning electron microscopy showed that both microspheres yielded a spherical shape with smooth or wrinkled surfaces. Fourier transform infrared spectroscopy demonstrated that the carbonyl group of TH formed hydrogen bonds with the amide group of CTS and the hydroxyl group of β‐CD. The swelling ability of the two microspheres was more than three times their weight, and their humidity rates attained equilibrium within 24 h. The ciliary beat movement times of CTS and CTS/β‐CD microspheres were 493.00 and 512.33 min, respectively, which indicated that the two microspheres effectively reduced the ciliotoxicity and possessed better adaptability. In vitro release of TH from CTS/β‐CD microspheres was slower than that from CTS microspheres at pH 6.8 and provided a sustained release of 72.0% within 12 h. The results suggest that CTS/β‐CD microspheres are a promising carrier for sustained release for pulmonary delivery. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 1183–1190, 2007     

Preparation of guar gum bonded with β‐cyclodextrin microspheres and the absorption on basic fuchsine

β‐Cyclodextrin microspheres (β‐CDMs) were prepared and then bonded to guar gum (GG). The structures of the precursor material and product were characterized by X‐ray diffraction and Fourier transform infrared spectroscopy. Phenolphthalein and spectrophotometry were used to determine the content of β‐cyclodextrin (β‐CD) in products. By changing environmental conditions and types of adsorbents, the adsorption capacities of polymers on basic fuchsine were studied. The results indicate that the absorption capacity was decreased accordingly when the pH value was reduced. The increase of temperature had no obvious influence on improving the absorption capacity from 40 to 60°C. When the mass ratio of chloropropyl hydroxyl GG to β‐CDMs to sodium carbonate was 0.6 : 0.8 : 1.0, with the reaction time of 2 h, the best adsorption capacity of 24 mg/g was achieved; this was in accordance with the results of the measurement of the content of β‐CD in the products. The higher content of β‐CDM was beneficial for improving the absorption capacity. By comparing its properties with the absorption capacity of traditional flocculating agents polyacrylamide and polaluminum chloride, the best absorption results for GG bonded with β‐CDMs were obtained. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Microbial production,immobilization and applications of β‐D‐galactosidase

β‐D ‐Galactosidase (β‐D ‐galactoside galactohydrolase, E.C. 3.2.1.23), most commonly known as lactase, is one of the most important enzymes used in food processing, which catalyses the hydrolysis of lactose to its constituent monosaccharides, glucose and galactose. The enzyme has been isolated and purified from a wide range of microorganisms but most commonly used β‐D ‐galactosidases are derived from yeasts and fungal sources. The major difference between yeast and fungal enzyme is the optimum pH for lactose hydrolysis. The application of β‐D ‐galactosidase for lactose hydrolysis in milk and whey offers nutritional, technological and environmental applications to human life. In this review, the main emphasis has been given to elaborate the various techniques used in recent times for the production, purification, immobilization and applications of β‐D ‐galactosidase. Copyright © 2006 Society of Chemical Industry     

Production of β‐galactosidase from recombinant Saccharomyces cerevisiae grown on lactose

Improved productivity and costs reduction in fermentation processes may be attained by using flocculating cell cultures. The production of extracellular heterologous β‐galactosidase by recombinant flocculating Saccharomyces cerevisiae cells, expressing the lacA gene (coding for β‐galactosidase) of Aspergillus niger under the ADHI promotor and terminator in a bioreactor was studied. The effects of lactose concentration and yeast extract concentration on β‐galactosidase production in a semi‐synthetic medium were analysed. The extracellular β‐galactosidase activity increased linearly with increasing initial lactose concentrations (5–150 g dm^?3). β‐Galactosidase production also increased with increased yeast extract concentration. During the entire fermentation, no accumulation of the hydrolysed sugars, glucose and galactose, was observed. The catabolic repression of the recombinant strain when cultured in a medium containing equal amounts of glucose and galactose was confirmed. In complete anaerobiosis, the fermentation of lactose resulted in a very slow fermentation pattern with lower levels of β‐galactosidase activity. The bioreactor operation together with optimisation of culture conditions (lactose and yeast extract concentration) led to a 21‐fold increase in the extracellular β‐galactosidase activity produced when compared with preliminary Erlenmeyer fermentations. Copyright © 2004 Society of Chemical Industry     

Guided tissue regeneration with use of β‐TCP/chitosan composite membrane

Guided tissue regeneration (GTR) membranes with bioabsorbable characteristics have been employed, in recent years, for periodontal procedures to deflect the growth of gingival tissues away from root surface. They provide an isolated space over regions with defective tissues and allow the relatively slow growing periodontal ligament fibroblasts to be repopulated over the root surface. In this study, we have employed chitosan and tricalcium phosphate (β‐TCP) as viable membrane materials and evaluated their roles in GTR applications. Three types of β‐TCP/chitosan membranes, weight ratio of β‐TCP/chitosan 65 : 35, 33 : 67, and 10 : 90, were prepared for three categories: the mechanical strength to create an effective space; the rapid rate to reach hydrolytic equilibrium in phosphate buffer solution; and the ease of clinical manipulations. Consequently, standardized, transosseous, and critical‐sized (cavity of 8 mm) skull defects were made in adult rabbits, and the defective regions were covered with the specifically prepared chitosan membranes. After 4 weeks of recovering, varying degrees of bone healing were observed beneath the β‐TCP/chitosan membranes in comparison to the control group. The β‐TCP/chitosan membranes covered regions showed a clear boundary space between connective tissues and bony tissues. Over all, good cell‐occlusion and beneficial osteogensis effects by these bioabsorbable materials toward the wound recovery were indicated. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Synthesis of a new chitosan immobilized with β‐cyclodextrins and its adsorption properties for bilirubin

A new chitosan derivative of chitosan immobilized with β‐cyclodextrins (CTC) has been synthesized by the reaction of chitosan and allyl‐substituted β‐cyclodextrins (ASC). Its structure has been proved by Fourier transform infrared spectral analysis, X‐ray powder diffraction analysis, and chemical analysis. Its adsorption of bilirubin has been also studied. By the experiment, it has been found that this new product has good adsorption ability on bilirubin. At initial 2 h, the adsorption increased largely. The adsorption mechanism was also dealt with. It predicted that this new chitosan derivative could have application in removing the bilirubin. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 99: 1264–1268, 2006     

Application of chitosan‐entrapped β‐galactosidase in a packed‐bed reactor system

The model enzyme β‐galactosidase was entrapped in chitosan gel beads and tested for hydrolytic activity and its potential for application in a packed‐bed reactor. The chitosan beads had an enzyme entrapment efficiency of 59% and retained 56% of the enzyme activity of the free enzyme. The Michaelis constant (K_m) was 0.0086 and 0.011 μmol/mL for the free and immobilized enzymes, respectively. The maximum velocity of the reaction (V_max) was 285.7 and 55.25 μmol mL^?1 min^?1 for the free and immobilized enzymes, respectively. In pH stability tests, the immobilized enzyme exhibited a greater range of pH stability and shifted to include a more acidic pH optimum, compared to that of the free enzyme. A 2.54 × 16.51‐cm tubular reactor was constructed to hold 300 mL of chitosan‐immobilized enzyme. A full‐factorial test design was implemented to test the effect of substrate flow (20 and 100 mL/min), concentration (0.0015 and 0.003M), and repeated use of the test bed on efficiency of the system. Parameters were analyzed using repeated‐measures analysis of variance. Flow (p < 0.05) and concentration (p < 0.05) significantly affected substrate conversion, as did the interaction progressing from Run 1 to Run 2 on a bed (p < 0.05). Reactor stability tests indicated that the packed‐bed reactor continued to convert substrate for more than 12 h with a minimal reduction in conversion efficiency. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 1294–1299, 2004     

Gentamicin loaded β‐tricalcium phosphate/gelatin composite microspheres as biodegradable bone fillers

In this study, novel composite bone fillers with microspherical shape, biodegradable property, and antibacterial effect were designed and prepared. Various fillers with different β‐tricalcium phosphate (β‐TCP)/gelatin (G)/glutaraldeyde (GA) compositions were loaded with a model antibiotic, gentamicin. The effect of composition and preparation conditions on the release of gentamicin was investigated in in vitro conditions. Complete release were observed in 12 h for pure β‐TCP powder, and this period was extended up to 96 h as the gelatin content increased in the microspheres. Morphological and chemical structures of the microspheres, before and after the release studies, were investigated by scanning electron microscopy and Fourier transform infrared, respectively. Antibacterial activities were examined against Escherichia coli by using disc diffusion method and promising results were obtained. It is proposed that these novel β‐TCP/G/GA microspheres can be applied locally to prevent and/or eliminate infection that might occur around a defected region of hard tissue and supports the healing process. POLYM. COMPOS., 2012. © 2012 Society of Plastics Engineers     

Preparation and characterization of β‐cyclodextrin‐linked chitosan microparticles

Hydrophobically modified chitosan containing β‐cyclodextrin (CD) units was synthesized by using tosylated β‐CD. The final product was characterized by Fourier transform infrared (FTIR) spectroscopy, elemental analysis and TGA, and rheometry. The polymer bearing β‐CD moieties was used to obtain crosslinked microparticles by spray‐drying which could then be used in a controlled release system for drugs. FTIR confirmed the formation of an amide linkage between cyclodextrin and chitosan. As fluorescence spectroscopy demonstrated, hydrophobic microenvironments were formed by chitosan bearing cyclodextrin in solution at lower concentrations than for chitosan. Rheometry and FTIR showed the crosslinking of the new polymer using genipin, a molecule of natural origin. Microspheres (MS) obtained by spray‐drying showed narrow size distribution when β‐CD was grafted onto chitosan and ξ‐potential of MS was slightly lower although it remained positive. In conclusion, β‐CD linked chitosan polymer can be considered as a very promising controlled drug delivery system for drugs. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Immobilization of β‐galactosidase from Escherichia coli onto modified natural silk fibers

A polymeric support based on the natural silk fibers was prepared and characterized for covalent immobilization of β‐galactosidase from Escherichia coli. The silk fibers were grafted using polyacrylonitrile in presence of benzophenone as a photo‐initiator. The grafted fibers were then activated by treatment with hydrazine hydrate followed by glyoxal cross‐linker. FTIR spectra, scanning electron microscope (SEM) in addition to the staining test derived from the Coomassie protein assay were utilized for investigation of the modification and immobilization steps. Also, the activity of both free and immobilized β‐galactosidase was evaluated as a function of the various important parameters like grafting percentage, pH, and temperature. In addition, the kinetic parameters K_m and v_max for both free and immobilized enzyme were anticipated using Michaelis–Menten equation. The results in this study indicated that the prepared modified woven silk fibers could be used effectively as a polymeric support for immobilization of β‐galactosidase. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 2923–2931, 2013     

α‐Amylase immobilized by Fe3O4/poly(styrene‐co‐maleic anhydride) magnetic composite microspheres: Preparation and characterization

Fe₃O₄/poly(styrene‐co‐maleic anhydride) core–shell composite microspheres, suitable for binding enzymes, were prepared using magnetite particles as seeds by copolymerization of styrene and maleic anhydride. The magnetite particles were encapsulated by polyethylene glycol, which improved the affinity between the magnetite particles and the monomers, thus showing that the size of the microspheres, the amount of the surface anhydrides, and the magnetite content in the composite are highly dependent on magnetite particles, comonomer ratio, and dispersion medium used in the polymerization. The composite microspheres, having 0.08–0.8 μm diameter and containing 100–800 μg magnetite/g microspheres and 0–18 mmol surface‐anhydride groups/g microsphere, were obtained. Free α‐amylase was immobilized on the microspheres containing reactive surface‐anhydride groups by covalent binding. The effects of immobilization on the properties of the immobilized α‐amylase [magnetic immobilized enzyme (MIE)] were studied. The activity of MIE and protein binding capacity reached 113,800 U and 544.3 mg/g dry microspheres, respectively. The activity recovery was 47.2%. The MIE had higher optimum temperature and pH compared with those of free α‐amylase and showed excellent thermal, storage, pH, and operational stability. Furthermore, it can be easily separated in a magnetic field and reused repeatedly. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 95: 328–335, 2005     

Synthesis and characterization of grafting β‐cyclodextrin with chitosan

Two new adsorbents [β‐cyclodextrin–chitosan (β‐CD–CTS) and β‐cyclodextrin‐6–chitosan (β‐CD‐6‐CTS)] were synthesized by the reaction of β‐cyclodextrin (β‐CD) with epoxy‐activated chitosan (CTS) and the sulfonation of the C‐6 hydroxyl group of β‐cyclodextrin with CTS, respectively. Their structures were confirmed by IR spectral analysis and X‐ray diffraction analysis, and their apparent amount of grafting was determined by ultraviolet spectroscopy. The adsorption properties of β‐CD‐CTS and β‐CD‐6‐CTS for p‐dihydroxybenzene were studied. The experimental results showed that the two new adsorbents exerted adsorption on the carefully chosen target. The highest saturated capacity of p‐dihydroxybenzene of β‐CD‐CTS and β‐CD‐6‐CTS were 51.68 and 46.41 mg/g, respectively. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 860–864, 2004     

Heparin‐immobilized poly(2‐hydroxyethylmethacrylate)‐based microspheres

Poly(2‐hydroxyethylmethacrylate) (PHEMA)‐based microspheres (150–200 µm in diameter) were produced by a modified suspension polymerization of different type of comonomers—namely, acrylic acid, dimethylaminoethyl‐methacrylate, and methylmethacrylate. These microspheres were activated with cyanogen bromide (CNBr) at pH 11.5, and heparin molecules were then immobilized through covalent bonds. The amount of immobilized heparin was controlled by changing the initial concentration of CNBr and heparin. The increase in the initial concentrations of both CNBr and heparin caused an increase in the amount of heparin immobilized onto microspheres for all polymer surfaces. The maximum heparin immobilization was observed on the PHEMA homopolymer microspheres (180 mg/g). The plain and heparin‐immobilized microspheres were contacted with blood in in vitro systems and in ex vivo animal experiments. Loss of the blood cells and clotting times were followed. Anticoagulant effect of the immobilized heparin was clearly observed with blood coagulation experiments. Loss of cells in the blood contacting with heparin‐immobilized microspheres was significantly lower than those observed with the plain microspheres. Bovine serum albumin adsorption onto the microspheres containing heparin on their surfaces was also studied. High albumin adsorption values (up to 127 mg/g) were observed in which the heparin‐immobilized PHEMA microspheres were used. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 74: 655–662, 1999     

Poly(ε‐caprolactone) composite scaffolds loaded with gentamicin‐containing β‐tricalcium phosphate/gelatin microspheres for bone tissue engineering applications

In this study, novel poly(ε‐caprolactone) (PCL) composite scaffolds were prepared for bone tissue engineering applications, where gentamicin‐loaded β‐tricalcium phosphate (β‐TCP)/gelatin microspheres were added to PCL. The effects of the amount of β‐TCP/gelatin microspheres added to the PCL scaffold on various properties, such as the gentamicin release rate, biodegradability, morphology, mechanical strength, and pore size distribution, were investigated. A higher amount of filler caused a reduction in the mechanical properties and an increase in the pore size and led to a faster release of gentamicin. Human osteosarcoma cells (Saos‐2) were seeded on the prepared composite scaffolds, and the viability of cells having alkaline phosphatase (ALP) activity was observed for all of the scaffolds after 3 weeks of incubation. Cell proliferation and differentiation enhanced the mechanical strength of the scaffolds. Promising results were obtained for the development of bone cells on the prepared biocompatible, biodegradable, and antimicrobial composite scaffolds. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40110.     

Adsorption and drug release based on β‐cyclodextrin‐grafted hydroxyapatite composite

In this study, β‐cyclodextrin (β‐CD) was covalently grafted on hydroxyapatite (HA) using a coupling agent to improve the drug loading capacity and prolong the drug release. The binding of β‐CD on the HA surface was confirmed by Fourier transformation infrared spectroscopy, thermal gravimetric analysis, and X‐ray powder diffraction. The adsorption capacity of ofloxacin on β‐CD‐grafted hydroxyapatite (β‐CD‐g‐HA) composite was found to be 30 mg g^?1 at 37°C and 24 h. The adsorption process is spontaneous, given the negative values of free energy change. Compared with the release of ofloxacin loaded on HA, the release of ofloxacin loaded on β‐CD‐g‐HA was slowed down 28% and 21% in pH 2.0 and pH 7.4 buffer media at 2 h, respectively. Biocompatibility of β‐CD‐g‐HA was assessed by MTT assay, and the result showed that it had no cytotoxicity. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Covalent immobilization of β‐galactosidase onto electrospun nanofibers of poly (AN‐co‐MMA) copolymer

Immobilization of β‐galactosidase in poly (acrylonitrile‐co‐methyl methacrylate) poly (AN‐co‐MMA) Nanofibers was studied by electrospinning, and a spacer‐arm i.e., (Polyethyleneimine (PEI)) was covalently attached by the reaction of carbonyl groups of Poly (AN‐co‐MMA) nanofibers. β‐galactosidase was then covalently immobilized through the spacer‐arm of the Poly (AN‐co‐MMA) nanofibers by using glutaraldehyde (GA) as a coupling agent. Nanofibers mode of interaction was proven by FTIR and thermal gravimetric analysis and supported by morphological changes recognized through SEM examination. Factors affecting the modification process such as PEI concentration, reaction time, and reaction temperature have been studied. Its influence on the amount of coupled PEI was consequently correlated to the changes of the catalytic activity and the retained activity of immobilized enzyme, the main parameters judging the success of the immobilization process. Evidences of Poly (AN‐co‐MMA) nanofibers modification were extracted from morphological changes recognized through SEM examination. The maximum activity (V_max) and michaelis constant (K_m) of immobilized enzyme were found to be 8.8 μmole/min mg protein and 236.7 mM, respectively. Stabilities of the immobilized β‐galactosidase were obviously improved. The optimum temperature for β‐galactosidase immobilized on the spacer‐arm attached nanofiber was 5°C higher than that of the free enzyme and was also significantly broader. The immobilized β‐galactosidase had better resistance to temperature inactivation than did the free form. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Preparation and permeation characterization of β‐zeolite‐incorporated composite membranes

In this work, several β‐zeolite‐incorporated polymer composite membranes were fabricated with the solution‐casting method. The zeolite loadings were 10, 20, and 30 wt %, respectively. Scanning electron microscopy characterization showed that the zeolite particles could be uniformly distributed in the whole polymer matrix. Gas permeation results demonstrated that after the incorporation of the same β‐zeolite, the polyimide exhibited a significant increase in gas permeability but a decrease in permselectivity, and both were quite pronounced at high zeolite loadings; this resulted from the loose structure that formed. The poly(ether sulfone) composite membranes showed obvious increases in both permeability and selectivity, and the permeability increase was considerably greater at higher zeolite loadings. The permselectivity increase could possibly be attributed to the pore sieving and preferential adsorption of β‐zeolite entities for the test gases as the heat treatment may have resulted in the formation of a defect‐free microstructure. However, breakthrough of the upper‐bound line was not achieved for these composite membranes, as reflected by Robeson plots. Our results suggest that changes in membrane performance not only are attributable to the properties and content of β‐zeolite particles but also depend on the heterogeneous microstructure created by zeolite entities. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Maximum volumetric production of β‐galactosidase by Kluyveromyces fragilis in fed‐batch culture with automatic control

The production of β‐galactosidase by Kluyveromyces fragilis was studied in different culture systems, with dissolved oxygen concentration control and using defined media. An operating strategy of fed‐batch culture with automatic control of substrate addition regulated by dissolved oxygen concentration, consisting of the replacement of variable volumes of broth by fresh medium (once the fed‐batch culture has finished), was designed. The volumetric enzyme productivity (Q_p, 13 600 UI dm^?3 h^?1) obtained was 38% higher than that reached in continuous culture of K fragilis with dissolved oxygen concentration control and far higher than that obtained by batch culture of K fragilis under the same aeration conditions. © 2002 Society of Chemical Industry