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Sample records for alginate gel matrix

  1. Calcium alginate gel as encapsulation matrix for coimmobilized enzyme systems.

    PubMed

    Blandino, A; Macías, M; Cantero, D

    2003-07-01

    Encapsulation within calcium alginate gel capsules was used to produce a coimmobilized enzyme system. Glucose oxidase (GOD) and catalase (CAT) were chosen as model enzymes. The same values of Vmax and Km app for the GOD encapsulated system and for the GOD-CAT coencapsulated system were calculated. When gel beads and capsules were compared, the same catalyst deactivation sequence for the two enzymes was observed. However, when capsules were employed as immobilization support, GOD efficiencies were higher than for the gel beads. These results were explained in terms of the structure of the capsules.

  2. Nonlinear elasticity of alginate gels

    NASA Astrophysics Data System (ADS)

    Hashemnejad, Seyed Meysam; Kundu, Santanu

    Alginate is a naturally occurring anionic polysaccharide extracted from brown algae. Because of biocompatibility, low toxicity, and simple gelation process, alginate gels are used in biomedical and food applications. Here, we report the rheological behavior of ionically crosslinked alginate gels, which are obtained by in situ gelation of alginates with calcium salts, in between two parallel plates of a rheometer. Strain stiffening behavior was captured using large amplitude oscillatory shear (LAOS) experiments. In addition, negative normal stress was observed for these gels, which has not been reported earlier for any polysaccharide networks. The magnitude of negative normal stress increases with applied strain and can exceed that of the shear stress at large strain. Rheological results fitted with a constitutive model that considers both stretching and bending of chains indicate that nonlinearity is likely related to the stretching of the chains between the crosslink junctions. The results provide an improved understanding of the deformation mechanism of ionically crosslinked alginate gel and the results will be important in developing synthetic extracellular matrix (ECM) from these materials.

  3. Calcium alginate gels as stem cell matrix-making paracrine stem cell activity available for enhanced healing after surgery.

    PubMed

    Schmitt, Andreas; Rödel, Philipp; Anamur, Cihad; Seeliger, Claudine; Imhoff, Andreas B; Herbst, Elmar; Vogt, Stephan; van Griensven, Martijn; Winter, Gerhard; Engert, Julia

    2015-01-01

    Regeneration after surgery can be improved by the administration of anabolic growth factors. However, to locally maintain these factors at the site of regeneration is problematic. The aim of this study was to develop a matrix system containing human mesenchymal stem cells (MSCs) which can be applied to the surgical site and allows the secretion of endogenous healing factors from the cells. Calcium alginate gels were prepared by a combination of internal and external gelation. The gelling behaviour, mechanical stability, surface adhesive properties and injectability of the gels were investigated. The permeability of the gels for growth factors was analysed using bovine serum albumin and lysozyme as model proteins. Human MSCs were isolated, cultivated and seeded into the alginate gels. Cell viability was determined by AlamarBlue assay and fluorescence microscopy. The release of human VEGF and bFGF from the cells was determined using an enzyme-linked immunoassay. Gels with sufficient mechanical properties were prepared which remained injectable through a syringe and solidified in a sufficient time frame after application. Surface adhesion was improved by the addition of polyethylene glycol 300,000 and hyaluronic acid. Humans MSCs remained viable for the duration of 6 weeks within the gels. Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks. This work shows that calcium alginate gels can function as immobilization matrices for human MSCs.

  4. Calcium Alginate Gels as Stem Cell Matrix – Making Paracrine Stem Cell Activity Available for Enhanced Healing after Surgery

    PubMed Central

    Schmitt, Andreas; Rödel, Philipp; Anamur, Cihad; Seeliger, Claudine; Imhoff, Andreas B.; Herbst, Elmar; Vogt, Stephan; van Griensven, Martijn; Winter, Gerhard; Engert, Julia

    2015-01-01

    Regeneration after surgery can be improved by the administration of anabolic growth factors. However, to locally maintain these factors at the site of regeneration is problematic. The aim of this study was to develop a matrix system containing human mesenchymal stem cells (MSCs) which can be applied to the surgical site and allows the secretion of endogenous healing factors from the cells. Calcium alginate gels were prepared by a combination of internal and external gelation. The gelling behaviour, mechanical stability, surface adhesive properties and injectability of the gels were investigated. The permeability of the gels for growth factors was analysed using bovine serum albumin and lysozyme as model proteins. Human MSCs were isolated, cultivated and seeded into the alginate gels. Cell viability was determined by AlamarBlue assay and fluorescence microscopy. The release of human VEGF and bFGF from the cells was determined using an enzyme-linked immunoassay. Gels with sufficient mechanical properties were prepared which remained injectable through a syringe and solidified in a sufficient time frame after application. Surface adhesion was improved by the addition of polyethylene glycol 300,000 and hyaluronic acid. Humans MSCs remained viable for the duration of 6 weeks within the gels. Human VEGF and bFGF was found in quantifiable concentrations in cell culture supernatants of gels loaded with MSCs and incubated for a period of 6 weeks. This work shows that calcium alginate gels can function as immobilization matrices for human MSCs. PMID:25793885

  5. Inter-grade and inter-batch variability of sodium alginate used in alginate-based matrix tablets.

    PubMed

    Fu, Shao; Buckner, Ira S; Block, Lawrence H

    2014-10-01

    The purpose of this study is to characterize the inter-grade and inter-batch variability of sodium alginate used in the formulation of matrix tablets. Four different grades and three batches of one grade of sodium alginate were used to prepare matrix tablets. Swelling, erosion, and drug release tests of sodium alginate matrix tablets were conducted in a USP dissolution apparatus. Substantial differences in swelling and erosion behavior of sodium alginate matrix tablets were evident among different viscosity grades. Even different batches of the same grade exhibit substantial differences in the swelling and erosion behavior of their matrix tablets. The erosion behavior of sodium alginate matrix tablets can be partly explained by their rheological properties (both apparent viscosity and viscoelasticity) in solution. Sodium alginate with higher apparent viscosity and viscoelasticity in solution show slower erosion rate and higher swelling rate. Compacts prepared from grades or batches with higher viscosity and higher viscoelasticity show slower drug release. For grades or batches with similar apparent viscosities, apparent viscosities of sodium alginate solution at low concentration alone are not sufficient to predict the functionality of sodium alginate in matrix tablets. Viscoelastic properties of sodium alginate solutions at one high concentration corresponding to the polymer gel state, may be suitable indicia of the extended release behavior of sodium alginate matrix tablets.

  6. Enzyme-catalyzed phase transition of alginate gels and gelatin-alginate interpenetrated networks.

    PubMed

    Doumèche, Bastien; Picard, Julien; Larreta-Garde, Véronique

    2007-11-01

    The enzyme-catalyzed gel-sol transition of calcium-alginate obtained by internal gelling strategy with the help of an entrapped alginate lyase is described. We show that alginate molecules and enzyme-produced oligoalginates shorten the gel time of physical gelatin gels (5% and 1.5%), probably due to local protein concentration increase. Interpenetrated networks composed of calcium-alginate and of gelatin were obtained only if elongation of gelatin helices inside a pre-existing calcium-alginate network could occur and only for low gelatin concentration (1.5%). The physical gelatin network is almost reversible inside the alginate one. Both networks can be obtained in the presence of alginate lyase, but gel-sol transition of calcium-alginate cannot be obtained in the presence of gelatin.

  7. Relevance of rheological properties of sodium alginate in solution to calcium alginate gel properties.

    PubMed

    Fu, Shao; Thacker, Ankur; Sperger, Diana M; Boni, Riccardo L; Buckner, Ira S; Velankar, Sachin; Munson, Eric J; Block, Lawrence H

    2011-06-01

    The purpose of this study is to determine whether sodium alginate solutions' rheological parameters are meaningful relative to sodium alginate's use in the formulation of calcium alginate gels. Calcium alginate gels were prepared from six different grades of sodium alginate (FMC Biopolymer), one of which was available in ten batches. Cylindrical gel samples were prepared from each of the gels and subjected to compression to fracture on an Instron Universal Testing Machine, equipped with a 1-kN load cell, at a cross-head speed of 120 mm/min. Among the grades with similar % G, (grades 1, 3, and 4), there is a significant correlation between deformation work (L(E)) and apparent viscosity (η(app)). However, the results for the partial correlation analysis for all six grades of sodium alginate show that L(E) is significantly correlated with % G, but not with the rheological properties of the sodium alginate solutions. Studies of the ten batches of one grade of sodium alginate show that η(app) of their solutions did not correlate with L(E) while tan δ was significantly, but minimally, correlated to L(E). These results suggest that other factors--polydispersity and the randomness of guluronic acid sequencing--are likely to influence the mechanical properties of the resultant gels. In summary, the rheological properties of solutions for different grades of sodium alginate are not indicative of the resultant gel properties. Inter-batch differences in the rheological behavior for one specific grade of sodium alginate were insufficient to predict the corresponding calcium alginate gel's mechanical properties.

  8. Small-angle X-ray scattering and rheological characterization of alginate gels. 3. Alginic acid gels.

    PubMed

    Draget, Kurt Ingar; Stokke, Bjørn T; Yuguchi, Yoshiaki; Urakawa, Hiroshi; Kajiwara, Kanji

    2003-01-01

    Alginic acid gels were studied by small-angle X-ray scattering and rheology to elucidate the influence of alginate chemical composition and molecular weight on the gel elasticity and molecular structure. The alginic acid gels were prepared by homogeneous pH reduction throughout the sample. Three alginates with different chemical composition and sequence, and two to three different molecular weights of each sample were examined. Three alginate samples with fractions of guluronic acid residues of 0.39 (LoG), 0.50 (InG), and 0.68 (HiG), covering the range of commercially available alginates, were employed. The excess scattering intensity I of the alginic acid gels was about 1 order of magnitude larger and exhibited a stronger curvature toward low q compared to ionically cross-linked alginate. The I(q) were decomposed into two components by assuming that the alginic acid gel is composed of aggregated multiple junctions and single chains. Time-resolved experiments showed a large increase in the average size of aggregates and their weight fraction within the first 2 h after onset of gelling, which also coincides with the most pronounced rheological changes. At equilibrium, little or no effect of molecular weight was observed, whereas at comparable molecular weights, an increased scattering intensity with increasing content of guluronic acid residues was recorded, probably because of a larger apparent molecular mass of domains. The results suggest a quasi-ordered junction zone is formed in the initial stage, followed by subsequent assembling of such zones, forming domains in the order of 50 A. The average length of the initial junction zones, being governed by the relative fraction of stabilizing G-blocks and destabilizing alternating (MG) blocks, determines the density of the final random aggregates. Hence, high-G alginates give alginic acid gels of a higher aggregate density compared to domains composed of loosely packed shorter junction zones in InG or LoG system.

  9. Ion exchange in alginate gels--dynamic behaviour revealed by electron paramagnetic resonance.

    PubMed

    Ionita, Gabriela; Ariciu, Ana Maria; Smith, David K; Chechik, Victor

    2015-12-14

    The formation of alginate gel from low molecular weight alginate and very low molecular weight alginate in the presence of divalent cations was investigated using Electron Paramagnetic Resonance (EPR) spectroscopy. The transition from sol to gel in the presence of divalent cations was monitored by the changes in the dynamics of spin labelled alginate. The immobilisation of the spin labelled alginate in the gel reflects the strength of interaction between the cation and alginate chain. Diffusion experiments showed that both the cation and alginate polyanion in the gel fibres can exchange with molecules in solution. In particular, we showed that dissolved alginate polyanions can replace alginates in the gel fibres, which can hence diffuse through the bulk of the gel. This illustrates the surprisingly highly dynamic nature of these gels and opens up the possibility of preparing multicomponent alginate gels via polyanion exchange process.

  10. A honeycomb composite of mollusca shell matrix and calcium alginate.

    PubMed

    You, Hua-jian; Li, Jin; Zhou, Chan; Liu, Bin; Zhang, Yao-guang

    2016-03-01

    A honeycomb composite is useful to carry cells for application in bone, cartilage, skin, and soft tissue regenerative therapies. To fabricate a composite, and expand the application of mollusca shells as well as improve preparing methods of calcium alginate in tissue engineering research, Anodonta woodiana shell powder was mixed with sodium alginate at varying mass ratios to obtain a gel mixture. The mixture was frozen and treated with dilute hydrochloric acid to generate a shell matrix/calcium alginate composite. Calcium carbonate served as the control. The composite was transplanted subcutaneously into rats. At 7, 14, 42, and 70 days after transplantation, frozen sections were stained with hematoxylin and eosin, followed by DAPI, β-actin, and collagen type-I immunofluorescence staining, and observed using laser confocal microscopy. The composite featured a honeycomb structure. The control and composite samples displayed significantly different mechanical properties. The water absorption rate of the composite and control group were respectively 205-496% and 417-586%. The composite (mass ratio of 5:5) showed good biological safety over a 70-day period; the subcutaneous structure of the samples was maintained and the degradation rate was lower than that of the control samples. Freezing the gel mixture afforded control over chemical reaction rates. Given these results, the composite is a promising honeycomb scaffold for tissue engineering.

  11. Understanding Alginate Gel Development for Bioclogging and Biogeophysical Experiments

    NASA Astrophysics Data System (ADS)

    Brown, I.; Atekwana, E. A.; Abdel Aal, G. Z.; Atekwana, E. A.; Sarkisova, S.; Patrauchan, M.

    2012-12-01

    Bioremediation strategies to mitigate the transport of heavy metals and radionuclides in subsurface sediments have largely targeted to increase the mobility and/or solubility of these compounds by the stimulation of biogeochemical activity of the metal- and sulfate-reducing bacteria. The latter secrete and/or release out diverse biochemical molecule including, first of all, organic acids and biopolymers such as alginic acid, proteins and DNA. Alginate gel is one of the major components determining the structure of biofilm which causes clogging in porous media. Biopolymers composing biofilm having, at least, two main functions: to be a scaffold for a microbial biofilm, and to regulate the exchange of metabolites and ions between an environment and bacterial cells. Additionally, the accumulation of biopolymers and a matured biofilm within porous media was shown to contribute to a detectable biogeophysical signal, spectral induced polarization (SIP), in particular. Our objective is to understand the role of different biofilm components on the SIP response as the latter has been proposed as a non-invasive tool to monitor biofilm development and rate of clogging in the subsurface. Understanding the process of alginate gel development may aid in the understanding of the fate and transport of mineralized heavy metals and radionuclides in contaminated soils. Here we describe the reciprocal relationship between environmental chemistry and alginate gel development. Commercial (Sigma) alginic acid (AA) was used as a substratum for the preparation of a model gel. AA was solubilized by adjusting solutions with pH up to 4 with 0.1 NaOH. Both Ca(OH)2 or CaCl2 were used to initiate the gelation of alginate. pH, fluid conductivity, soluble Ca2+ concentration, and a yield of gelated alginate were monitored in both liquid and porous media after the interaction of calcium compounds with alginate. This study confirms the critical role of Ca2+ for alginate gelation, biofilm development

  12. Kefiran-alginate gel microspheres for oral delivery of ciprofloxacin.

    PubMed

    Blandón, Lina M; Islan, German A; Castro, Guillermo R; Noseda, Miguel D; Thomaz-Soccol, Vanete; Soccol, Carlos R

    2016-09-01

    Ciprofloxacin is a broad-spectrum antibiotic associated with gastric and intestinal side effects after extended oral administration. Alginate is a biopolymer commonly employed in gel synthesis by ionotropic gelation, but unstable in the presence of biological metal-chelating compounds and/or under dried conditions. Kefiran is a microbial biopolymer able to form gels with the advantage of displaying antimicrobial activity. In the present study, kefiran-alginate gel microspheres were developed to encapsulate ciprofloxacin for antimicrobial controlled release and enhanced bactericidal effect against common pathogens. Scanning electron microscopy (SEM) analysis of the hybrid gel microspheres showed a spherical structure with a smoother surface compared to alginate gel matrices. In vitro release of ciprofloxacin from kefiran-alginate microspheres was less than 3.0% and 5.0% at pH 1.2 (stomach), and 5.0% and 25.0% at pH 7.4 (intestine) in 3 and 21h, respectively. Fourier transform infrared spectroscopy (FTIR) of ciprofloxacin-kefiran showed the displacement of typical bands of ciprofloxacin and kefiran, suggesting a cooperative interaction by hydrogen bridges between both molecules. Additionally, the thermal analysis of ciprofloxacin-kefiran showed a protective effect of the biopolymer against ciprofloxacin degradation at high temperatures. Finally, antimicrobial assays of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhymurium, and Staphylococcus aureus demonstrated the synergic effect between ciprofloxacin and kefiran against the tested microorganisms.

  13. Alginate gel particles-A review of production techniques and physical properties.

    PubMed

    Ching, Su Hung; Bansal, Nidhi; Bhandari, Bhesh

    2017-04-13

    The application of hydrocolloid gel particles is potentially useful in food, chemical, and pharmaceutical industries. Alginate gel particles are one of the more commonly used hydrocolloid gel particles due to them being biocompatible, nontoxic, biodegradable, cheap, and simple to produce. They are particularly valued for their application in encapsulation. Encapsulation in alginate gel particles confers protective benefits to cells, DNA, nutrients, and microbes. Slow release of flavors, minerals, and drugs can also be achieved by encapsulation in gel particles. The particle size and shape of the gel particles are crucial for specific applications. In this review, current methods of producing alginate gel particles will be discussed, taking into account their advantages, disadvantages, scalability, and impact on particle size. The physical properties of alginate gel particles will determine the effectiveness in different application conditions. This review will cover the current understanding of the alginate biopolymer, gelation mechanisms and factors affecting release properties, gel strength, and rheology of the alginate gel particle systems.

  14. Alginate gels with a combination of calcium and chitosan oligomer mixtures as crosslinkers.

    PubMed

    Feng, Yiming; Kopplin, Georg; Sato, Kimihiko; Draget, Kurt I; Vårum, Kjell M

    2017-01-20

    Alginates are polysaccharides that are widely used in relation to their ability to form gels. Recently we reported that alginates may also form gels with chitosan oligomers as crosslinkers (Khong, Aarstad, Skjåk-Bræk, Draget, & Vårum, 2013). The purpose of the present study was to characterize alginate gels crosslinked with calcium and chitosan oligomers. Using two different alginates of similar molecular weights but different chemical composition, i.e. guluronic acid content of 46 and 68%, we found that both alginates could form homogeneous gels with calcium and chitosan oligomers separately and without syneresis. Systematic combinations of calcium and chitosan oligomers as crosslinkers were tested, showing that up to 50% of the calcium could be substituted with chitosan oligomers without reduction in gel strength or increased syneresis for the alginate with the lowest guluronic acid content. Furthermore, the kinetics of the combined gels were different from pure calcium alginate gels.

  15. Composite alginate gels for tunable cellular microenvironment mechanics

    PubMed Central

    Khavari, Adele; Nydén, Magnus; Weitz, David A.; Ehrlicher, Allen J.

    2016-01-01

    The mechanics of the cellular microenvironment can be as critical as biochemistry in directing cell behavior. Many commonly utilized materials derived from extra-cellular-matrix create excellent scaffolds for cell growth, however, evaluating the relative mechanical and biochemical effects independently in 3D environments has been difficult in frequently used biopolymer matrices. Here we present 3D sodium alginate hydrogel microenvironments over a physiological range of stiffness (E = 1.85 to 5.29 kPa), with and without RGD binding sites or collagen fibers. We use confocal microscopy to measure the growth of multi-cellular aggregates (MCAs), of increasing metastatic potential in different elastic moduli of hydrogels, with and without binding factors. We find that the hydrogel stiffness regulates the growth and morphology of these cell clusters; MCAs grow larger and faster in the more rigid environments similar to cancerous breast tissue (E = 4–12 kPa) as compared to healthy tissue (E = 0.4–2 kpa). Adding binding factors from collagen and RGD peptides increases growth rates, and change maximum MCA sizes. These findings demonstrate the utility of these independently tunable mechanical/biochemistry gels, and that mechanical confinement in stiffer microenvironments may increase cell proliferation. PMID:27484403

  16. Composite alginate gels for tunable cellular microenvironment mechanics

    NASA Astrophysics Data System (ADS)

    Khavari, Adele; Nydén, Magnus; Weitz, David A.; Ehrlicher, Allen J.

    2016-08-01

    The mechanics of the cellular microenvironment can be as critical as biochemistry in directing cell behavior. Many commonly utilized materials derived from extra-cellular-matrix create excellent scaffolds for cell growth, however, evaluating the relative mechanical and biochemical effects independently in 3D environments has been difficult in frequently used biopolymer matrices. Here we present 3D sodium alginate hydrogel microenvironments over a physiological range of stiffness (E = 1.85 to 5.29 kPa), with and without RGD binding sites or collagen fibers. We use confocal microscopy to measure the growth of multi-cellular aggregates (MCAs), of increasing metastatic potential in different elastic moduli of hydrogels, with and without binding factors. We find that the hydrogel stiffness regulates the growth and morphology of these cell clusters; MCAs grow larger and faster in the more rigid environments similar to cancerous breast tissue (E = 4–12 kPa) as compared to healthy tissue (E = 0.4–2 kpa). Adding binding factors from collagen and RGD peptides increases growth rates, and change maximum MCA sizes. These findings demonstrate the utility of these independently tunable mechanical/biochemistry gels, and that mechanical confinement in stiffer microenvironments may increase cell proliferation.

  17. Alginate/Poly(γ-glutamic Acid) Base Biocompatible Gel for Bone Tissue Engineering

    PubMed Central

    Chan, Wing P.; Kung, Fu-Chen; Kuo, Yu-Lin; Yang, Ming-Chen; Lai, Wen-Fu Thomas

    2015-01-01

    A technique for synthesizing biocompatible hydrogels by cross-linking calcium-form poly(γ-glutamic acid), alginate sodium, and Pluronic F-127 was created, in which alginate can be cross-linked by Ca2+ from Ca–γ-PGA directly and γ-PGA molecules introduced into the alginate matrix to provide pH sensitivity and hemostasis. Mechanical properties, swelling behavior, and blood compatibility were investigated for each hydrogel compared with alginate and for γ-PGA hydrogel with the sodium form only. Adding F-127 improves mechanical properties efficiently and influences the temperature-sensitive swelling of the hydrogels but also has a minor effect on pH-sensitive swelling and promotes anticoagulation. MG-63 cells were used to test biocompatibility. Gelation occurred gradually through change in the elastic modulus as the release of calcium ions increased over time and caused ionic cross-linking, which promotes the elasticity of gel. In addition, the growth of MG-63 cells in the gel reflected nontoxicity. These results showed that this biocompatible scaffold has potential for application in bone materials. PMID:26504784

  18. Effect of alginate culture and mechanical stimulation on cartilaginous matrix synthesis of rat dedifferentiated chondrocytes.

    PubMed

    Wang, Yun; de Isla, Natalia; Huselstein, Céline; Wang, Binghua; Netter, Patrick; Stoltz, Jean-François; Muller, Sylvaine

    2008-01-01

    To investigate whether the application of alginate culture and mechanical stimulation will improve the synthesis of cartilaginous matrix in dedifferentiated chondrocytes, rat chondrocytes underwent dedifferentiation upon serial monolayer culture up to passage 6, and then were encapsulated in 2% alginate gel and subject to static culture. After 28 days culture in static, the beads were exposed to 48 h of mechanical stimulation with continuous agitation. The sGAG content in alginate bead was measured by alcian blue staining. The expression of collagen protein was detected using immunofluorescence. After 28 days culture in alginate bead, the dedifferentiated chondrocytes remained round in shape and re-synthesized the chondrocyte-specific matrix. Compared with static culture, mechanical stimulation induced statistically increases in the production of glycosaminoglycan (p< or =0.01), as well as in the synthesis of collagen type II protein (p< or =0.05). On the contrary, no positive expression of collagen type I protein was observed at the end of culture. Our results demonstrated that both of alginate culture and mechanical stimulation help to restore chondrocyte phenotype and promotes the synthesis of cartilaginous matrix.

  19. Alginate gel-coated oil-entrapped alginate-tamarind gum-magnesium stearate buoyant beads of risperidone.

    PubMed

    Bera, Hriday; Boddupalli, Shashank; Nandikonda, Sridhar; Kumar, Sanoj; Nayak, Amit Kumar

    2015-01-01

    A novel alginate gel-coated oil-entrapped calcium-alginate-tamarind gum (TG)-magnesium stearate (MS) composite floating beads was developed for intragastric risperidone delivery with a view to improving its oral bioavailability. The TG-blended alginate core beads containing olive oil and MS as low-density materials were accomplished by ionotropic gelation technique. Effects of polymer-blend ratio (sodium alginate:TG) and crosslinker (CaCl2) concentration on drug entrapment efficiency (DEE, %) and cumulative drug release after 8 h (Q8h, %) were studied to optimize the core beads by a 3(2) factorial design. The optimized beads (F-O) exhibited DEE of 75.19±0.75% and Q8h of 78.04±0.38% with minimum errors in prediction. The alginate gel-coated optimized beads displayed superior buoyancy and sustained drug release property. The drug release profiles of the drug-loaded uncoated and coated beads were best fitted in Higuchi kinetic model with Fickian and anomalous diffusion driven mechanisms, respectively. The optimized beads yielded a notable sustained drug release profile as compared to marketed immediate release preparation. The uncoated and coated Ca-alginate-TG-MS beads were also characterized by SEM, FTIR and P-XRD analyses. Thus, the newly developed alginate-gel coated oil-entrapped alginate-TG-MS composite beads are suitable for intragastric delivery of risperidone over a prolonged period of time.

  20. Comparison of some biochemical properties of artichoke polyphenol oxidase entrapped in alginate-carrageenan and alginate gels.

    PubMed

    Yagar, Hulya; Kocaturk, Selin

    2014-08-01

    Polyphenol oxidase (PPO, EC.1.14.18.1) isolated from artichoke (Cynara scolymus) was entrapped within alginate and alginate+ carrageenan beads, and the catecholase and cresolase activities of both entrapped enzymes were determined. Some properties of these immobilized enzymes such as optimum pH and temperature, kinetic parameters (Km and Vmax), thermal, and storage stability were determined and compared to each other. The highest catecholase activity was observed in alginate gel (370 U/g bead) while the highest cresolase activity was in alginate+ carrageenan gel (90 U/g bead). For catecholase and cresolase activities, optimum pHs of alginate and alginate+ carrageenan beads were determined to be 7.0 and 4.0, respectively. Optimum temperatures for catecholase activity were determined to be 40°C for both entrapped enzymes. These values for cresolase activity were 30°C and 20°C, respectively. Immobilized artichoke PPOs greatly preserved their thermal stability which exists anyway. The catalytic efficiency value (Vmax/Km) of the alginate beads is approximately high as two-and-a-half folds of that of alginate+κ-carrageenan beads for cresolase activity. These values were very close for catecholase activity. Immobilized beads saved their both activities after 30 days of storage at 4°C.

  1. Preparation of carbon nanotube-alginate nanocomposite gel for tissue engineering.

    PubMed

    Kawaguchi, Minoru; Fukushima, Tadao; Hayakawa, Toru; Nakashima, Naotoshi; Inoue, Yusuke; Takeda, Shoji; Okamura, Kazuhiko; Taniguchi, Kunihisa

    2006-12-01

    A novel scaffold material based on an alginate hydrogel which contained carbon nanotubes (CNTs) was prepared, and its mechanical property and biocompatibility evaluated. Soluble CNTs were prepared with acid treatment and dispersed in sodium alginate solution as a cross-linker. After which, the mechanical property (elastic deformation), saline sorption, histological reaction, and cell viability of the resultant nanocomposite gel (CNT-Alg gel) were evaluated. The CNT-Alg gel showed faster gelling and higher mechanical strength than the conventional alginate gel. Saline sorption amount of freeze-dried CNT-Alg gel was equal to that of the alginate gel. In terms of histological evaluation and cell viability assay, CNT-Alg gel exhibited a mild inflammatory response and non-cytotoxicity. These results thus suggested that CNT-Alg gel could be useful as a scaffold material in tissue engineering with the sidewalls of CNTs acting as active sites for chemical functionalization.

  2. Hyaluronate-alginate gel as a novel biomaterial: mechanical properties and formation mechanism.

    PubMed

    Oerther, S; Le Gall, H; Payan, E; Lapicque, F; Presle, N; Hubert, P; Dexheimer, J; Netter, P

    1999-04-20

    With the aim of producing a biomaterial for surgical applications, the alginate-hyaluronate association has been investigated to combine the gel-forming properties of alginate with the healing properties of hyaluronate. Gels were prepared by diffusion of calcium into alginate-hyaluronate mixtures, with an alginate content of 20 mg/mL. The hyaluronate source was shown to have significant effect on the aspect and the properties of the gels. The gels have viscoelastic behaviour and the transient measurements carried out in creep mode could be interpreted through a Kelvin-Voigt generalised model: experimental data led to the steady state hardness and a characteristic viscosity of the gel. Gels prepared from Na rooster comb hyaluronate with weight ratio up to 0.50 have satisfactory mechanical properties, and fully stable gels are obtained after a few days; on the contrary, use of lower molecular weight hyaluronate led to loose gels for hyaluronate contents over 0.25. Gel formation was investigated by measurements of the exchange fluxes between the calcium chloride solution and the forming gel, which allowed thorough investigations of the occuring diffusion phenomena of water, calcium ion and hyaluronate. Strong interactions of water with hyaluronate reduce significantly the rate of weight loss from the gel beads and allows higher water content in steady-state gels. Calcium content in the gel samples could be correlated to the actual alginate concentration, whatever the nature and the weight ratio of hyaluronate.

  3. Effects of alginate gel on rheological properties of hair-tail ( Trichiurus lepturus) surimi

    NASA Astrophysics Data System (ADS)

    Xue, Yong; Liu, Xin; Zhang, Lili; Lin, Dan; Xu, Jiachao; Xue, Changhu

    2011-06-01

    Effects of alginate gel at different concentrations on rheological properties of hair-tail ( Trichiurus lepturus) surimi were investigated. Alginate gel (1%-3%) exhibited enhanced effects, especially when alginate gel concentration increased. The rheological properties of mixture samples were studied by the time sweep, frequency sweep and temperature sweep. The critical strain values of the mixture samples for the onset of non-linear viscoelasticity were about 5%. The storage modulus G' of the mixture samples increased with time for 4 h. The frequency sweep showed that G' was greater than G″ for all the mixture gels with different alginate gel concentrations, and values of both n' and n″ for all samples were low (<0.2), these constants corresponding to G' and G″, and indicating the elasticity of mixture gels. The values of storage modulus G' decreased during heating process and increased with decreasing temperature.

  4. Combined rheological and ultrasonic study of alginate and pectin gels near the sol-gel transition.

    PubMed

    Audebrand, Michel; Kolb, Max; Axelos, Monique A V

    2006-10-01

    The sol-gel transition of biopolymer mixtures has been investigated by rheological and ultrasonic measurements. A scaling analysis of the data was performed for both types of measurements. A gel time was determined from rheology for the pure pectin samples, and the data could be fitted to a universal scaling form near the transition point. Its critical exponents are in good agreement with the predictions of scalar percolation theory. In addition, the ultrasonic signal of the pectin samples close to the transition was analyzed in terms of a high-frequency scaling approach for the attenuation and the velocity. For the alginate samples and the mixtures, for which the gel point cannot be determined reliably from rheology, the ultrasonic measurements were analyzed using the same scaling form as for the pectin sample, thus providing a method for estimating the gel point, even in the absence of rheological data.

  5. Microencapsulation of a synbiotic into PLGA/alginate multiparticulate gels.

    PubMed

    Cook, Michael T; Tzortzis, George; Charalampopoulos, Dimitris; Khutoryanskiy, Vitaliy V

    2014-05-15

    Probiotic bacteria have gained popularity as a defence against disorders of the bowel. However, the acid sensitivity of these cells results in a loss of viability during gastric passage and, consequently, a loss of efficacy. Probiotic treatment can be supplemented using 'prebiotics', which are carbohydrates fermented specifically by probiotic cells in the body. This combination of probiotic and prebiotic is termed a 'synbiotic'. Within this article a multiparticulate dosage form has been developed, consisting of poly(d,l-lactic-co-glycolic acid) (PLGA) microcapsules containing prebiotic Bimuno™ incorporated into an alginate-chitosan matrix containing probiotic Bifidobacterium breve. The aim of this multiparticulate was that, in vivo, the probiotic would be protected against gastric acid and the release of the prebiotic would occur in the distal colon. After microscopic investigation, this synbiotic multiparticulate was shown to control the release of the prebiotic during in vitro gastrointestinal transit, with the release of galacto-oligosaccharides (GOS) initially occurred over 6h, but with a triphasic release pattern giving further release over 288 h. Encapsulation of B. breve in multiparticulates resulted in a survival of 8.0 ± 0.3 logCFU/mL cells in acid, an improvement over alginate-chitosan microencapsulation of 1.4 logCFU/mL. This was attributed to increased hydrophobicity by the incorporation of PLGA particles.

  6. Drug delivery matrices based on scleroglucan/alginate/borax gels.

    PubMed

    Matricardi, Pietro; Onorati, Ilenia; Coviello, Tommasina; Alhaique, Franco

    2006-06-19

    The aim of this work is to obtain a new drug delivery matrix, especially designed for protein delivery, based on biodegradable and biocompatible polymers, and to describe its main physico-chemical properties. A polysaccharide based semi-interpenetrating polymer network (semi-IPN) was built up, composed by sodium alginate chains interspersed into a scleroglucan/borax hydrogel network. Tablets were obtained by compression of the resulting freeze-dried hydrogel. The different release and physico-chemical properties possessed by the two starting polymers in various aqueous media were combined in the new matrix. In this work, description is given of the in vitro ability of the matrix to deliver in a controlled manner a protein, Myoglobin, in distilled water, simulated gastric fluid and simulated intestinal fluid; the release, simulating a gastric passage, followed by an enteric delivery, was also carried out. Water uptake data, colorimetric experiments and scanning electron microscopy images are given for the characterization of this new solid dosage form; the importance of the borax presence is also discussed.

  7. Kinetic analysis of beer primary fermentation using yeast cells immobilized by ceramic support adsorption and alginate gel entrapment.

    PubMed

    Zhang, Yongming; Kennedy, John F; Knill, Charles J; Panesar, Parmjit S

    2006-01-01

    Yeast cells were immobilized by absorption onto porous ceramic support and evaluated for continuous beer primary fermentation using a bioreactor in comparison to yeast cells immobilized by entrapment in calcium alginate gel. The effects of temperature and flow rate as a function of reaction/fermentation time on fermentation rate were investigated. The fermentation reaction (in terms of loss of total soluble solids in the beer wort as a function of time) was first-order with half-lifes in the range of approximately 9-11 hours at approximately 10-12 degrees C at beer wort linear flow rates of approximately 0.8-1.6 cm/minute for ceramic support, compared with approximately 16 hours for Ca-alginate gel, the former support matrix being more efficient and demonstrating greater potential for future commercial application.

  8. Gelation and biocompatibility of injectable alginate-calcium phosphate gels for bone regeneration.

    PubMed

    Cardoso, D Alves; van den Beucken, J J J P; Both, L L H; Bender, J; Jansen, J A; Leeuwenburgh, S C G

    2014-03-01

    An emerging approach toward development of injectable, self-setting, and fully biodegradable bone substitutes involves the combination of injectable hydrogel matrices with a dispersed phase consisting of nanosized calcium phosphate particles. Here, novel injectable composites for bone regeneration have been developed based on the combination of ultrapure alginate as the matrix phase, crystalline CaP [monetite and poorly crystalline hydroxyapatite (HA)] powders as both a dispersed mineral phase and a source of calcium for cross-linking alginate, glucono-delta-lactone (GDL) as acidifier and glycerol as both plasticizer and temporary sequestrant. The composites were maximized with respect to CaP content to obtain the highest amount of osteoconductive filler. The viscoelastic and physicochemical properties of the precursor compounds and composites were analyzed using rheometry, elemental analysis (for calcium release and uptake), acidity [by measuring pH in simulated body fluid (SBF)], general biocompatibility (subcutaneous implantation in rabbits), and osteocompatibility (implantation in femoral condyle bone defect of rabbits). The gelation of the resulting composites could be controlled from seconds to tens of minutes by varying the solubility of the CaP phase (HA vs. monetite) or amount of GDL. All composites mineralized extensively in SBF for up to 11 days. In vivo, the composites also disintegrated upon implantation in subcutaneous or bone tissue, leaving behind less degradable but osteoconductive CaP particles. Although the composites need to be optimized with respect to the available amount of calcium for cross-linking of alginate, the beneficial bone response as observed in the in vivo studies render these gels promising for minimally invasive applications as bone-filling material.

  9. Gluten gel and film properties in the presence of cysteine and sodium alginate.

    PubMed

    Yuno-Ohta, Naoko; Yamada, Mariko; Inomata, Masako; Konagai, Hiromi; Kataoka, Tomomi

    2009-08-01

    Wheat flour has an ability of forming dough by mixing with water, which exhibits a rheological property required for making bread. The major protein is gluten, which is a valuable protein material for food industry. In this study, gluten protein gels and films were formed with cysteine and sodium alginate. Adding cysteine improved gel and film properties (stress relaxation behavior, bending strength). The gel containing 0.01 M cysteine had a longer relaxation time and was more rigid than the gel without cysteine. Although adding sodium alginate to the gluten suspension containing cysteine improved the water-holding ability and homogeneity of the gel network, the film from this gel was more brittle than the gluten film with cysteine alone. Microstructural observations of the gels and films with scanning electron microscopy suggested that water evaporation was more heterogeneous from the gel containing sodium alginate than from the gel with cysteine alone. Fourier transform-infrared (FT-IR) analysis during film formation suggested that the presence of cysteine encourages interaction between gluten molecules and results in intermolecular beta-sheet formation in earlier stages than in the no additive condition. FT-IR results also suggested that the combined effect of sodium alginate and cysteine on the protein secondary structure was remarkably different from that of cysteine alone. Our results suggest that addition of a suitable amount of cysteine (0.01 M) and heat treatment to 80 degrees C during gluten gel and film formation induces a homogenous network in the gel and film by regulating disulfide-sulfide interactions.

  10. Molecular imprinting in sol-gel matrix.

    PubMed

    Gupta, Radha; Kumar, Ashok

    2008-01-01

    Molecular imprinting is a newly developed methodology which provides molecular assemblies of desired structures and properties and is being increasingly used for several applications such as in separation processes, microreactors, immunoassays and antibody mimics, catalysis, artificial enzymes, biosensor recognition elements and bio- and chemo-sensors. The ambient processing conditions and versatility of the sol-gel process makes sol-gel glassy matrix suitable for molecular imprinting. The progress of sol-gel based molecular imprinted polymers (MIPs) for various applications can be seen from the growing number of publications. The main focus of the review is molecular imprinting in sol-gel matrix and applications of molecular imprinted sol-gel derived materials for the development of sensors. Combining sol-gel process with molecular imprinting enables to procure the sensors with greater sensitivity and selectivity necessary for sensing applications. The merits, problems, challenges and factors affecting molecular imprinting in sol-gel matrix have been discussed. Considerable attention has been drawn on recent developments like use of organically modified silane precursors (ORMOSILS) for the synthesis of hybrid molecular imprinted polymers (HMIPs) and applying surface sol-gel process for molecular imprinting. The development of molecular imprinted sol-gel nanotubes for biochemical separation and bio-imprinting is a new advancement and is under progress. Templated xerogels and molecularly imprinted sol-gel films provide a good platform for various sensor applications.

  11. Impact of solvent quality on the network strength and structure of alginate gels.

    PubMed

    Hermansson, Elin; Schuster, Erich; Lindgren, Lars; Altskär, Annika; Ström, Anna

    2016-06-25

    The influence of the mixture of water and alcohols on the solubility and properties of alginate and its calcium-induced gels is of interest for the food, wound care and pharmaceutical industries. The solvent quality of water with increasing amounts of ethanol (0-20%) on alginate was studied using intrinsic viscosity. The effect of ethanol addition on the rheological and mechanical properties of calcium alginate gels was determined. Small-angle X-ray scattering and transmission electron microscopy were used to study the network structure. It is shown that the addition of ethanol up to 15% (wt) increases the extension of the alginate chain, which correlates with increased moduli and stress being required to fracture the gels. The extension of the polymer chain is reduced at 20% (wt) ethanol, which is followed by reduced moduli and stress at breakage of the gels. The network structure of gels at high ethanol concentrations (24%) is characterized by thick and poorly connected network strands.

  12. Fundamental Characteristics of Bioprint on Calcium Alginate Gel

    NASA Astrophysics Data System (ADS)

    Umezu, Shinjiro; Hatta, Tatsuru; Ohmori, Hitoshi

    2013-05-01

    The goal of this study is to fabricate precision three-dimensional (3D) biodevices those are micro fluidics and artificial organs utilizing digital fabrication. Digital fabrication is fabrication method utilizing inkjet technologies. Electrostatic inkjet is one of the inkjet technologies. The electrostatic inkjet method has following two merits; those are high resolution to print and ability to eject highly viscous liquid. These characteristics are suitable to print biomaterials precisely. We are now applying for bioprint. In this paper, the electrostatic inkjet method is applied for fabrication of 3D biodevices that has cave like blood vessel. When aqueous solution of sodium alginate is printed to aqueous solution of calcium chloride, calcium alginate is produced. 3D biodevices are fabricated in case that calcium alginate is piled.

  13. Granular gel support-enabled extrusion of three-dimensional alginate and cellular structures.

    PubMed

    Jin, Yifei; Compaan, Ashley; Bhattacharjee, Tapomoy; Huang, Yong

    2016-06-03

    Freeform fabrication of soft structures has been of great interest in recent years. In particular, it is viewed as a critical step toward the grand vision of organ printing--the on-demand design and fabrication of three-dimensional (3D) human organ constructs for implantation and regenerative medicine. The objective of this study is to develop a novel granular gel support material-enabled, two-step gelation-based 'printing-then-gelation' approach to fabricate 3D alginate structures using filament extrusion. Specifically, a granular Carbopol microgel bath holds the ungelled alginate structure being extruded, avoiding the instantaneous gelation of each printed layer as well as resultant surface tension-induced nozzle clogging. Since Carbopol microgels react with multivalent cations, which are needed for alginate crosslinking, gelatin is introduced as a sacrificial material to make an alginate and gelatin bioink for extrusion, which gels thermally (step-one gelation) to initially stabilize the printed structure for removal from Carbopol. Then gelatin is melted and diffused away while alginate is ionically crosslinked in a 37 °C calcium chloride bath (step-two gelation), resulting in an alginate structure. The proposed 'printing-then-gelation' approach works for alginate structure fabrication, and it is also applicable for the printing of cellular constructs and other similar homogeneous soft structures using a two-step or even multi-step approach. The main conclusions are: (1) 0.8% (w/v) Carbopol bath with a neutral pH value may be most suitable for soft structure printing; (2) it is most effective to use a 0.9% (w/v) NaCl solution to facilitate the removal of residual Carbopol; and (3) alginate structures fabricated using the proposed approach demonstrate better mechanical properties than those fabricated using the conventional 'gelation-while-printing' approach.

  14. Alginate and Chitosan Gel Nanoparticles for Efficient Protein Entrapment

    NASA Astrophysics Data System (ADS)

    Masalova, O.; Kulikouskaya, V.; Shutava, T.; Agabekov, V.

    Alginate and chitosan nanoparticles were synthesized by ionic gelation of the polymers in the presence of stabilizers (PEG 1500, PEG 6000, TWEEN 80). The stability of 210-240 nm Ca-alginate colloids is affected by nanoparticles ageing and by the presence of a stabilizer. The diameter of chitosan nanoparticles is in the range of 180 to 260 nm and depends on polymer concentration in the reaction mixture, its molecular weight, and stabilizer type. The nanoparticles efficiently entrap a model protein, bovine serum albumin, in the amount up to 0.24 mg per 1 mg of polysaccharide.

  15. 11 alpha-Hydroxylation of progesterone in biphasic media using alginate-entrapped Aspergillus ochraceus gel beads coated with polyurea.

    PubMed

    Houng, J Y; Chiang, W P; Chen, K C; Tiu, C

    1994-06-01

    A novel cell-immobilization technique was developed in this study for increasing substrate partition to the gel matrix by coating a polyurea thin layer on the surface of Ca-alginate beads. The proposed method was simple and could be performed under mild conditions. The bioconversion of progesterone to 11 alpha-hydroxyprogesterone with these polyurea-coating alginate-entrapped Aspergillus ochraceus cells was investigated using different organic solvents in biphasic media. The reaction medium of ethyl acetate could markedly enhance the bioconversion rate with the existence of a hydrophobic layer, most likely resulting from the increasing partition of substrate to gel matrix. Bioconversion with higher substrate concentration was possible using an ethyl acetate-water medium. The conversion rate increased almost linearly with increasing substrate concentration from 10 to 80 g l-1. The rate with 80 g l-1 progesterone increased up to six times greater than the rate with the immobilized cells without coating, and also exhibited a much higher rate than that reported in the literature.

  16. Boron removal from aqueous solutions using alginate gel beads in fixed-bed systems

    PubMed Central

    Demey-Cedeño, Hary; Ruiz, Montserrat; Barron-Zambrano, Jesús Alberto; Sastre, Ana Maria

    2014-01-01

    Background A column sorption study was carried out using calcium alginate gel beads as adsorbent for the removal of boron from aqueous solutions. The breakthrough curve was obtained as a function of pH, initial concentration of boron, feed flow rate, adsorbent mass and column diameter. The breakthrough capacity values and adsorption percentage of calcium alginate gel for boron were calculated. Column data obtained at different conditions were described using the Adams–Bohart model and bed-depth service time (BDST), derived from the Adams–Bohart equation to predict breakthrough curves and to determine the characteristic column parameters required for process design. Results The maximum adsorption percentage of boron on calcium alginate gel beads using an initial concentration of boron of 50 mg L−1 at pH 11 and room temperature (20±1°C) was calculated to be 55.14%. Conclusion The results indicated that calcium alginate can be used in a continuous packed-bed column for boron adsorption. The optimal conditions for boron adsorption were obtained at high pH, higher initial boron concentration, increased column depth and lower flow velocity. © 2014 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. PMID:25821332

  17. Immobilization of urease from pigeonpea (Cajanus cajan L.) in polyacrylamide gels and calcium alginate beads.

    PubMed

    Das, N; Kayastha, A M; Malhotra, O P

    1998-02-01

    Urease from pigeonpea was entrapped in polyacrylamide gel with 50% immobilization at 10% total monomer (containing 5% cross-linker) with high mechanical stability of the gel. Approximately 0.61 mg of protein could be loaded per 5 ml of gel. The immobilized enzyme had a t1/2 of approx. 200 days when stored in 0.1 M Tris/acetate buffer, pH 6.5, at 4 degrees C. The gel strips were used 4-5 times for urea assay over a period of 6 h with less than 2% loss of activity. Approximately 50% immobilization of urease in calcium alginate was observed at 3% alginate with 0.12 mg protein/ml alginate. The resultant enzyme beads showed a t1/2 of approx. 75 days when stored in 0.1 M Tris/acetate buffer, pH 6.5, at 4 degrees C. The beads were used 4-5 times for urea assay over a period of 6 h with about 40% loss of activity. In both cases, the enzyme activity was directly proportional to the amount of immobilized enzyme. There was practically no leaching of the entrapped enzyme over a period of 48 h from either of the polymers. Both the immobilized enzyme preparations were used to analyse the blood urea of some clinical samples from the University hospital. The results obtained compared favourably with those obtained by the usual method employed in the clinical pathology laboratory.

  18. Rheological characterization of a gel produced using human blood plasma and alginate mixtures.

    PubMed

    Malagón-Romero, Dionisio; Hernández, Nicolás; Cardozo, Carmen; Godoy-Silva, Rubén D

    2014-06-01

    Human blood plasma is a material used to generate tissue equivalents due to presence of fibrinogen. However, gels formed using human blood plasma has weak mechanical properties. In this study, different mixtures of sodium alginate and blood plasma were performed and evaluated. By determining ζ potential can be established the stability of the plasma-alginate mixture and by dynamic rheology can determine the most suitable parameters for the gelation of the above mixtures, when calcium chloride is used as a crosslinker. Experimental results evidence an increment in ζ potential at alginate concentrations of 0.8% and 1.6% with a resulting pseudoplastic behavior of evaluated mixtures, which described the homogenization of the mixture. On the other hand, mixtures were gelled by using aspersion of calcium chloride and characterized by dynamic rheology. Solid behavior is dominant in all range of frequency sweep test between 0.1Hz and 100Hz. Finally, the ultimate tensile strength of a gel reach 6.36938±0.24320kPa, which is enough for manual handling of the gel. Between the tasks of the gel would be used for cell entrapment, for controlled release of drugs or in the manufacture of wound dressings.

  19. Influence of both cation and alginate nature on the rheological behavior of transition metal alginate gels.

    PubMed

    Agulhon, Pierre; Robitzer, Mike; Habas, Jean-Pierre; Quignard, Françoise

    2014-11-04

    The rheological properties of several ionotropic alginate hydrogels were investigated according to the nature of the divalent cation (Mn(2+), Co(2+), Cu(2+)) and the guluronic fraction of the alginate (HG and LG for "high G-content" and "low G-content"). Six hydrogels (Mn-LG, Mn-HG, Co-LG, Co-HG, Cu-LG and Cu-HG) were synthesized and studied by spectromechanical analyses. On one hand, Cu-HG, Cu-LG and Co-HG behaved as viscoelastic solids: the elastic contribution was higher than the dissipative component in all the frequency range studied (G'>G"). No flow zone (G">G') was detected even at very low values of the shearing frequency. On the other, Mn-HG, Mn-LG and Co-LG presented a spectromechanical behavior that resembled that observed classically for entangled polymers. Indeed, at high frequency, these latter materials could be compared to a viscoelastic solid but at low frequency, the flow zone was described and the viscous character became prevalent with finite relaxation time. Very good correlations with the microscopic structurations of the network were evidenced (rubbery vs. flow zone and fibrillar vs. complex morphology respectively).

  20. Characterization of smart auto-degradative hydrogel matrix containing alginate lyase to enhance levofloxacin delivery against bacterial biofilms.

    PubMed

    Islan, German A; Dini, Cecilia; Bartel, Laura C; Bolzán, Alejandro D; Castro, Guillermo R

    2015-12-30

    The aim of the present work is the characterization of smart auto-degradable microspheres composed of calcium alginate/high methoxylated pectin containing an alginate lyase (AL) from Sphingobacterium multivorum and levofloxacin. Microspheres were prepared by ionotropic gelation containing AL in its inactive form at pH 4.0. Incubation of microspheres in Tris-HCl and PBS buffers at pH 7.40 allowed to establish the effect of ion-chelating phosphate on matrix erodability and suggested an intrinsically activation of AL by turning the pH close to neutrality. Scanning electron and optical microscopies revealed the presence of holes and surface changes in AL containing microspheres. Furthermore, texturometric parameters, DSC profiles and swelling properties were showing strong changes in microspheres properties. Encapsulation of levofloxacin into microspheres containing AL showed 70% efficiency and 35% enhancement of antimicrobial activity against Pseudomonas aeruginosa biofilm. Levofloxacin release from microspheres was not changed at acidic pH, but was modified at neutral pH in presence of AL. Advantageously, only gel matrix debris were detectable after overnight incubation, indicating an autodegradative gel process activated by the pH. Absence of matrix cytotoxicity and a reduction of the levofloxacin toxicity after encapsulation were observed in mammalian CHO-K1 cell cultures. These properties make the system a potent and versatile tool for antibiotic oral delivery targeted to intestine, enhancing the drug bioavailability to eradicate bacterial biofilm and avoiding possible intestinal obstructions.

  1. Physical characterization of alginate-Pluronic F127 gel for endoluminal NABDs delivery.

    PubMed

    Abrami, Michela; D'Agostino, Ilenia; Milcovich, Gesmi; Fiorentino, Simona; Farra, Rossella; Asaro, Fioretta; Lapasin, Romano; Grassi, Gabriele; Grassi, Mario

    2014-02-07

    Here we focus the attention on the physical characteristics of a highly biocompatible hydrogel made up of crosslinked alginate and Pluronic F127 (PF127). This is a composite polymeric blend we propose for artery endoluminal delivery of an emerging class of molecules named nucleic acid based drugs (NABDs). The physical characterization of our composite gel, i.e. mesh size distribution and PF127-alginate mutual organization after crosslinking, can significantly determine the NABDs release kinetics. Thus, to explore these aspects, different technical approaches, i.e. rheology, low/high field NMR and TEM, were used. While rheology provided information at the macroscopic and nano-level, the other three approaches gave details at the nano-level. We observe that Pluronic micelles, organizing in cubic ordered domains, generate, upon alginate crosslinking, the formation of meshes (≈ 150 nm) larger than those occurring in a Pluronic-free alginate network (≈ 25 nm). Nevertheless, smaller alginate meshes are still on and can just host un-structured Pluronic micelles and water. Accordingly, the gel structure is quite inhomogeneous, where big meshes (filled by crystalline Pluronic) co-exist with smaller meshes (hosting water and un-structured PF127 micelles). While big meshes offer a considerable hindering action on a diffusing solute, smaller ones represent a sort of free space where solute diffusion is faster. The presence of big and small meshes indicates that drug release may follow a double kinetics characterized by a fast and slow release. Notably, this behavior is considered appropriate for endoluminal drug release to the arterial wall.

  2. Development of PVA-alginate as a matrix for enzymatic decolorization of textile dye in bioreactor system

    NASA Astrophysics Data System (ADS)

    Yanto, Dede Heri Yuli; Zahara, Syifa; Laksana, Raden Permana Budi; Anita, Sita Heris; Oktaviani, Maulida; Sari, Fahriya Puspita

    2017-01-01

    An immobilization technique using polyvinyl alcohol (PVA) crosslinked with sodium alginate as a matrix has been developed for textile dyes decolorization. Textiles use dye as an addition to the aesthetic value of the product. Dyes are generally used is a textile dye where the waste will be released directly into the waters around 2-20%. Therefore, it is important to develop an enzyme immobilization method using PVA-Alginate as a matrix. Based on the results of the study showed that the PVA-Alginate beads produced high decolorization percent compared to beads which contains only Ca-alginate alone and formula matrix is optimum at PVA 6% and alginate 1.5%. Encapsulation with boric acid at 7% showed optimum decolorization and reduction for enzyme leakage during decolorization. This study suggested that immobilization of enzymes into PVA-alginate matrix might be used as a biodecolorating agent.

  3. Assessment of molecular events during in vitro re-epithelialization under honey-alginate matrix ambience.

    PubMed

    Barui, Ananya; Mandal, Naresh; Majumder, Subhadipa; Das, Raunak Kumar; Sengupta, Sanghamitra; Banerjee, Provas; Ray, Ajoy Kumar; RoyChaudhuri, Chirosree; Chatterjee, Jyotirmoy

    2013-08-01

    Re-epithelialization is one of the most important stages of cutaneous regeneration and its success requires supportive micro-ambience which may be provided with suitable bio-matrix. Biocompatibility and efficacy of such bio-matrix in re-epithelialization could be explored by multimodal analysis of structural and functional attributes of in vitro wound healing model including evaluation of prime molecular expressions of the epithelial cells during repair. Present study examines the influence of honey-alginate and alginate matrices on re-epithelialization in keratinocyte (HaCaT) population in a 2-D wound model. Cellular viability, proliferation and cell-cell adhesion status were assessed during wound closure using live/dead cell assay and by evaluating expressions of Ki67, p63 and E-cadherin along-with % change in cellular electrical impedance. Efficacy of honey-alginate matrix in comparison to only alginate one was demonstrated by a quicker reduction in wound gap, improved cellular viability, enhanced expressions of Ki67, p63 and its isoforms (TAp63, ΔNp63) as well as E-cadherin. Faster restoration of electrical attribute (% of impedance change) after wounding also indicated better impact of honey-alginate matrix in re-epithelialization.

  4. Rapid 3D Printing of Multifunctional Calcium Alginate Gel Pipes using Coaxial Jet Extruder

    NASA Astrophysics Data System (ADS)

    Rykaczewski, Konrad; Damle, Viraj

    2014-11-01

    Calcium alginate (CA) forms when solution containing sodium alginate (SA) comes in contact with a CaCl2 solution. The resulting gel is biocompatible as well as edible and is used in production of bio-scaffolds, artificial plant seeds, and edible substances. In the latter application, referred to in the culinary world as ``spherification,'' flavored liquids are mixed with the SA and dripped into CaCl2 solution to form gel encapsulated flavored ``marbles.'' Previously, crude 3D printing of CA structures has been achieved by stacking of such flavored liquid filled marbles. In turn, solid CA rods have been fabricated by properly mixing flow of the two solutions using a microfluidic device. Here we show that by using two circular cross-section coaxial nozzles to produce coaxial jets of the SA and CaCl2 solutions, liquid filled CA micro-to-mili scale gel pipes can be produced at speeds around ~ 150 mm/s. Such extrusion rate is compatible with most commercially available 3D printers, facilitating adoption of the CA pipe coaxial jet extruder. Here, the impact of inner and outer liquid properties and flow speeds on the gel pipe extrusion process is discussed. KR acknowledges startup funding from ASU.

  5. Synthesis and Characterization of Sodium Alginate Conjugate and Study of Effect of Conjugation on Drug Release from Matrix Tablet

    PubMed Central

    Satheeshababu, B. K.; Mohamed, I.

    2015-01-01

    The aim of the present research work to study the effect of conjugation of the polymer on drug release from the matrix tablets. Sodium alginate L-cysteine conjugate was achieved by covalent attachment of thiol group of L-cysteine with the primary amino group of sodium alginate through the amide bonds formed by primary amino groups of the sodium alginate and the carboxylic acid group of L-cysteine. The synthesised sodium alginate L-cysteine conjugate was characterised by determining of charring point, Fourier transmission-infrared and differential scanning calorimetric analysis. To study the effect of conjugation on drug release pattern, the matrix tablets were prepared using various proportions of sodium alginate and sodium alginate L-cysteine conjugate along with atorvastatin calcium as model drug. The wet granulation technique was adopted and prepared matrix tablets were evaluated for various physical parameters. The in vitro drug release study results suggested that tablet formulated in combination of sodium alginate and sodium alginate L-cysteine conjugate S4 showed 100% after 8 h drug release whereas formulated with only sodium alginate S0 released 40% in 8 h. PMID:26798173

  6. Hierarchical cross-linking in physical alginate gels: a rheological and dynamic light scattering investigation

    NASA Astrophysics Data System (ADS)

    Larobina, Domenico; Cipelletti, Luca

    We investigate the dynamics of alginate gels, an important class of biopolymer-based viscoelastic materials, by combining mechanical tests and non-conventional, time-resolved light scattering methods. Two relaxation modes are observed upon applying a compressive or shear stress. Dynamic light scattering and diffusive wave spectroscopy measurements reveal that these modes are associated with discontinuous rearrangement events that restructure the gel network via anomalous, non-diffusive microscopic dynamics. We show that these dynamics are due to both thermal activation and internal stress stored during gelation and propose a scenario where a hierarchy of cross-links with different life times is responsible for the observed complex behavior. Measurements at various temperatures and sample ages are presented to support this scenario.

  7. Determination of diffusion coefficients and diffusion characteristics for chlorferon and diethylthiophosphate in Ca-alginate gel beads.

    PubMed

    Ha, Jiyeon; Engler, Cady R; Lee, Seung Jae

    2008-07-01

    Diffusion characteristics of chlorferon and diethylthiophosphate (DETP) in Ca-alginate gel beads were studied to assist in designing and operating bioreactor systems. Diffusion coefficients for chlorferon and DETP in Ca-alginate gel beads determined at conditions suitable for biodegradation studies were 2.70 x 10(-11) m(2)/s and 4.28 x 10(-11) m(2)/s, respectively. Diffusivities of chlorferon and DETP were influenced by several factors, including viscosity of the bulk solution, agitation speed, and the concentrations of diffusing substrate and immobilized cells. Diffusion coefficients increased with increasing agitation speed, probably due to poor mixing at low speed and some attrition of beads at high speeds. Diffusion coefficients also increased with decreasing substrate concentration. Increased cell concentration in the gel beads caused lower diffusivity. Theoretical models to predict diffusivities as a function of cell weight fraction overestimated the effective diffusivities for both chlorferon and DETP, but linear relations between effective diffusivity and cell weight fraction were derived from experimental data. Calcium-alginate gel beads with radii of 1.65-1.70 mm used in this study were not subject to diffusional limitations: external mass transfer resistances were negligible based on Biot number calculations and effectiveness factors indicated that internal mass transfer resistance was negligible. Therefore, the degradation rates of chlorferon and DETP inside Ca-alginate gel beads were reaction-limited.

  8. 3D Cell Culture in Alginate Hydrogels

    PubMed Central

    Andersen, Therese; Auk-Emblem, Pia; Dornish, Michael

    2015-01-01

    This review compiles information regarding the use of alginate, and in particular alginate hydrogels, in culturing cells in 3D. Knowledge of alginate chemical structure and functionality are shown to be important parameters in design of alginate-based matrices for cell culture. Gel elasticity as well as hydrogel stability can be impacted by the type of alginate used, its concentration, the choice of gelation technique (ionic or covalent), and divalent cation chosen as the gel inducing ion. The use of peptide-coupled alginate can control cell–matrix interactions. Gelation of alginate with concomitant immobilization of cells can take various forms. Droplets or beads have been utilized since the 1980s for immobilizing cells. Newer matrices such as macroporous scaffolds are now entering the 3D cell culture product market. Finally, delayed gelling, injectable, alginate systems show utility in the translation of in vitro cell culture to in vivo tissue engineering applications. Alginate has a history and a future in 3D cell culture. Historically, cells were encapsulated in alginate droplets cross-linked with calcium for the development of artificial organs. Now, several commercial products based on alginate are being used as 3D cell culture systems that also demonstrate the possibility of replacing or regenerating tissue. PMID:27600217

  9. Gelatinized Copper–Capillary Alginate Gel Functions as an Injectable Tissue Scaffolding System for Stem Cell Transplants

    PubMed Central

    Willenberg, Bradley Jay; Zheng, Tong; Meng, Fan-Wei; Meneses, Juan Carlos; Rossignol, Candace; Batich, Christopher D.; Terada, Naohiro; Steindler, Dennis A.; Weiss, Michael D.

    2013-01-01

    In severe hypoxic–ischemic brain injury, cellular components such as neurons and astrocytes are injured or destroyed along with the supporting extracellular matrix. This presents a challenge to the field of regenerative medicine since the lack of extracellular matrix and supporting structures makes the transplant milieu inhospitable to the transplanted cells. A potential solution to this problem is the use of a biomaterial to provide the extracellular components needed to keep cells localized in cystic brain regions, allowing the cells to form connections and repair lost brain tissue. Ideally, this biomaterial would be combined with stem cells, which have been proven to have therapeutic potentials, and could be delivered via an injection. To study this approach, we derived a hydrogel biomaterial tissue scaffold from oligomeric gelatin and copper–capillary alginate gel (GCCAG). We then demonstrated that our multipotent astrocytic stem cells (MASCs) could be maintained in GCCAG scaffolds for up to 2 weeks in vitro and that the cells retained their multipotency. We next performed a pilot transplant study in which GCCAG was mixed with MASCs and injected into the brain of a neonatal rat pup. After a week in vivo, our results showed that: the GCCAG biomaterial did not cause a significant reactive gliosis; viable cells were retained within the injected scaffolds; and some delivered cells migrated into the surrounding brain tissue. Therefore, GCCAG tissue scaffolds are a promising, novel injectable system for transplantation of stem cells to the brain. PMID:20699061

  10. Small-angle X-ray scattering and rheological characterization of alginate gels. 2. Time-resolved studies on ionotropic gels

    NASA Astrophysics Data System (ADS)

    Yuguchi, Y.; Urakawa, H.; Kajiwara, K.; Draget, K. I.; Stokke, B. T.

    2000-10-01

    Gelation was observed by time-resolved small-angle X-ray scattering and rheology on 10 mg/ml Ca-alginate gels prepared by in situ release of Ca 2+ from CaEGTA or CaCO 3 with total Ca 2+ concentration in the range 10-20 mM. This was carried out for alginates having a fraction of α- L-GulA (G) of FG=0.39 and 0.68, respectively, obtained by the selection of alginates isolated from two different brown algae, Ascophyllum nodosum and Laminaria hyperborea stipe. Correlation between the rheological data and SAXS data shows that a large fraction of the lateral association precedes the formation of a continuous network through the sample cell. Following the initial association of chain segments in junction zones, the analysis using two-component broken rod model indicates the formation of larger bundles, and that the relative weight of these bundles increases with increasing time. The molecular model for the bundles is proposed by associating 2-16 units (G-blocks) composed of 14 (1→4) linked residues of α- L-GulA in parallel according to the available crystallographic data. The storage modulus increases as the bundles composed of associated alginate chains grow during the gel formation. The gel elasticity is mainly sustained by single chains in the alginate sample with a low fraction of α- L-GulA. The alginates with a high fraction of α- L-GulA associate into thicker bundles which join to form a network. Here the gel elasticity seems to be due to the flexible joints between bundles, since the fraction of single chains is extremely low.

  11. Biodegradation of crystal violet using Burkholderia vietnamiensis C09V immobilized on PVA-sodium alginate-kaolin gel beads.

    PubMed

    Cheng, Ying; Lin, HongYan; Chen, Zuliang; Megharaj, Mallavarapu; Naidu, Ravi

    2012-09-01

    The strain, Burkholderia vietnamiensis C09V was immobilized on PVA-alginate-kaolin gel beads as a biomaterial to improve the degradation of crystal violet from aqueous solution. The results show that 98.6% (30 mg L(-1)) crystal violet was removed from aqueous solution using immobilized cells on PVA-alginate-kaolin gel beads, while 94.0% crystal violet was removed by free cells after degradation at the pH 5 and 30°C for 30 h. Kinetics studies show that the pseudo-second-order kinetics well described the adsorption of crystal violet on the PVA-alginate-kaolin beads. Biodegradation of crystal violet on immobilized cells was fitted well by first-order reaction kinetics, indicating that CV was adsorbed onto kaolin and followed their degradation by immobilized cells onto the the PVA-alginate-kaolin beads. Characterization with SEM shows that cells attached well to the surface of PVA-alginate-kaolin beads, leading to improved crystal violet transfer from aqueous solution to immobilized cells. In addition, UV-vis show that the absorption peak at 588 nm was reduced by the degraded N-bond linkages, as well as the formation of degrading products were observed by Fourier transform infrared (FTIR). These results suggest that crystal violet was biodegraded to N,N-dimethylaminophenol and Michler's Ketone prior to these intermediates being further degraded.

  12. Xanthan-alginate composite gel beads: molecular interaction and in vitro characterization.

    PubMed

    Pongjanyakul, Thaned; Puttipipatkhachorn, Satit

    2007-02-22

    Xanthan gum (XG), a trisaccharide branched polymer, was applied to reinforce calcium alginate beads in this study. Composite beads consisting of XG and sodium alginate (SA) were prepared using ionotropic gelation method. Diclofenac calcium-alginate (DCA) beads incorporated with different amounts of XG were produced as well. Molecular interaction between SA and XG in the composite beads and the XG-DCA beads was investigated using FTIR spectroscopy. Physical properties of the XG-DCA beads such as entrapment efficiency of diclofenac sodium (DS), thermal property, water uptake, swelling and DS release in various media were examined. XG could form intermolecular hydrogen bonding with SA in the composite beads with or without DS. Differential scanning calorimetric study indicated that XG did not affect thermal property of the DCA beads. The DS entrapment efficiency of the DCA beads increased with increasing amount of XG added. The XG-DCA beads showed higher water uptake and swelling in pH 6.8 phosphate buffer and distilled water than the DCA beads. A longer lag time and a higher DS release rate of the XG-DCA beads in pH 6.8 phosphate buffer were found. In contrast, the 0.3%XG-DCA beads could retard the drug release in distilled water because interaction between XG and SA gave higher tortuosity of the bead matrix. However, higher content of XG in the DCA beads increased the release rate of DS. This can be attributed to erosion of small aggregates of XG on the surface of the DCA beads. This finding suggested that XG could modulate physicochemical properties and drug release of the DCA beads, which based on the existence of molecular interaction between XG and SA.

  13. The Bactericidal Effect of Dendritic Copper Microparticles, Contained in an Alginate Matrix, on Escherichia coli

    PubMed Central

    Thomas, Simon F.; Rooks, Paul; Rudin, Fabian; Atkinson, Sov; Goddard, Paul; Bransgrove, Rachel; Mason, Paul T.; Allen, Michael J.

    2014-01-01

    Although the bactericidal effect of copper has been known for centuries, there is a current resurgence of interest in the use of this element as an antimicrobial agent. During this study the use of dendritic copper microparticles embedded in an alginate matrix as a rapid method for the deactivation of Escherichia coli ATCC 11775 was investigated. The copper/alginate produced a decrease in the minimum inhibitory concentration from free copper powder dispersed in the media from 0.25 to 0.065 mg/ml. Beads loaded with 4% Cu deactivated 99.97% of bacteria after 90 minutes, compared to a 44.2% reduction in viability in the equivalent free copper powder treatment. There was no observed loss in the efficacy of this method with increasing bacterial loading up to 106 cells/ml, however only 88.2% of E. coli were deactivated after 90 minutes at a loading of 108 cells/ml. The efficacy of this method was highly dependent on the oxygen content of the media, with a 4.01% increase in viable bacteria observed under anoxic conditions compared to a >99% reduction in bacterial viability in oxygen tensions above 50% of saturation. Scanning electron micrographs (SEM) of the beads indicated that the dendritic copper particles sit as discrete clusters within a layered alginate matrix, and that the external surface of the beads has a scale-like appearance with dendritic copper particles extruding. E. coli cells visualised using SEM indicated a loss of cellular integrity upon Cu bead treatment with obvious visible blebbing. This study indicates the use of microscale dendritic particles of Cu embedded in an alginate matrix to effectively deactivate E. coli cells and opens the possibility of their application within effective water treatment processes, especially in high particulate waste streams where conventional methods, such as UV treatment or chlorination, are ineffective or inappropriate. PMID:24831035

  14. Method To immobilize the aphid-pathogenic fungus erynia neoaphidis in an alginate matrix for biocontrol

    PubMed

    Shah; Aebi; Tuor

    1998-11-01

    Erynia neoaphidis is an important fungal pathogen of aphid pests worldwide. There have been few reported attempts to formulate this natural agent for use in biocontrol. In the current study, factors involved in the immobilization of E. neoaphidis hyphae in an alginate matrix were investigated. Hyphae of two isolates cultured in liquid medium were 220 to 620 &mgr;m in length and 7 to 19 &mgr;m in diameter with a 74 to 83% cytoplasmic content. The optimal concentration of low-viscosity sodium alginate for production of conidia from entrapped hyphae was 1.5% (wt/vol), and 0.1 and 0.25 M calcium chloride were equally suitable for use as the gelling solution. Alginate beads were rinsed with 10% sucrose after gelling. However, beads should not be left for longer than 40 min in 0.1 M calcium chloride or 10% sucrose to prevent a 10% loss in conidial production. A 40% (vol/vol) concentration of fungal biomass produced significantly more conidia than either 20% or the standard concentration of 10%. This effect persisted even after beads were dried overnight in a laminar flow hood and stored at 4 degreesC for 4 days. Conidia from freshly produced alginate beads caused 27 to 32% infection in Pea aphids as determined by standardized laboratory bioassays. This finding was not significantly different from infections in aphids inoculated with fresh mycelial mats or plugs from Petri dish cultures. In conclusion, algination appears to be a promising technique for utilizing E. neoaphidis in the biocontrol of aphid pests.

  15. Non-Invasive Evaluation of Alginate/Poly-L-lysine/Alginate Microcapsules by Magnetic Resonance Microscopy

    PubMed Central

    Constantinidis, Ioannis; Grant, Samuel C.; Celper, Susanne; Gauffin-Holmberg, Isabel; Agering, Kristina; Oca-Cossio, Jose A.; Bui, Jonathan D.; Flint, Jeremy; Hamaty, Christine; Simpson, Nicholas E.; Blackband, Stephen J.

    2007-01-01

    In this report, we present data to demonstrate the utility of 1H MR microscopy to noninvasively examine alginate/poly-L-lysine/alginate (APA) microcapsules. Specifically, high-resolution images were used to visualize and quantify the poly-L-lysine (PLL) layer, and monitor temporal changes in the alginate gel microstructure during a month long in vitro culture. The thickness of the alginate/PLL layer was quantified to be 40.6±6.2 μm regardless of the alginate composition used to generate the beads or the time of alginate/PLL interaction (2, 6, or 20 minutes). However, there was a notable difference in the contrast of the PLL layer that depended upon the guluronic content of the alginate and the alginate/PLL interaction time. The T2 relaxation time and the apparent diffusion coefficient (ADC) of the alginate matrix were measured periodically throughout the month long culture period. Alginate beads generated with a high guluronic content alginate demonstrated a temporal decrease in T2 over the duration of the experiment, while ADC was unaffected. This decrease in T2 is attributed to a reorganization of the alginate microstructure due to periodic media exchanges that mimicked a regular feeding regiment for cultured cells. In beads coated with a PLL layer, this temporal decrease in T2 was less pronounced suggesting that the PLL layer helped maintain the integrity of the initial alginate microstructure. Conversely, alginate beads generated with a high mannuronic content alginate (with or without a PLL layer) did not display temporal changes in either T2 or ADC. This observation suggests that the microstructure of high mannuronic content alginate beads is less susceptible to culture conditions. PMID:17239948

  16. Effect of ultrasound on the diffusion properties of casein entrapped in alginate-chitosan gel.

    PubMed

    Huang, Zhenghua; Cao, Yanping; Xu, Duoxia; Wang, Chao; Zhang, Dandan

    2015-09-01

    The effects of ultrasound-assisted and pre-ultrasound treatment on the diffusion properties of casein imbedded by alginate-chitosan gel were investigated. The fluorescence spectrophotometry for determining the fluorescence intensity of casein was established to calculate the diffusion coefficient (De). Scanning electron microscope (SEM) was used to observe the microstructure of gel beads. The results showed that two different kinds of ultrasonic treatments had obvious distinctions on the casein diffusion. As the frequency increased, the value of De decreased from 28.56 × 10(-4)m(2)s(-1) (28 kHz) to 2.57 × 10(-4)m(2)s(-1) (135 kHz) during the ultrasound-assisted process. While, the minimum De of 8.6 × 10(-4)m(2)s(-1) was achieved at the frequency of 50 kHz for the pre-ultrasound treatment. The impact of power on the diffusion showed that De increased with the increase of ultrasound power until it reached the highest value 28.56 × 10(-4)m(2)s(-1) (0.45 W/cm(2)) in the ultrasound-assisted process. It would reach the maximum value (16 × 10(-4)m(2)s(-1)) when the power was 0.25 W/cm(2) in the pretreatment ultrasound process. SEM analysis exhibited that the gel structural changes (area ratio) were in accordance with De through different ultrasonic treatment. This was mainly due to the mechanical action and cavitation of the ultrasonic treatment. This study is important to explain the diffusion properties of large molecules and explore the mechanism of enzyme immobilization treated by ultrasound.

  17. Alginate hydrogel enriched with enamel matrix derivative to target osteogenic cell differentiation in TiO2 scaffolds.

    PubMed

    Pullisaar, Helen; Verket, Anders; Szoke, Krisztina; Tiainen, Hanna; Haugen, Håvard J; Brinchmann, Jan E; Reseland, Janne E; Østrup, Esben

    2015-01-01

    The purpose of bone tissue engineering is to employ scaffolds, cells, and growth factors to facilitate healing of bone defects. The aim of this study was to assess the viability and osteogenic differentiation of primary human osteoblasts and adipose tissue-derived mesenchymal stem cells from various donors on titanium dioxide (TiO2) scaffolds coated with an alginate hydrogel enriched with enamel matrix derivative. Cells were harvested for quantitative reverse transcription polymerase chain reaction on days 14 and 21, and medium was collected on days 2, 14, and 21 for protein analyses. Neither coating with alginate hydrogel nor alginate hydrogel enriched with enamel matrix derivative induced a cytotoxic response. Enamel matrix derivative-enriched alginate hydrogel significantly increased the expression of osteoblast markers COL1A1, TNFRSF11B, and BGLAP and secretion of osteopontin in human osteoblasts, whereas osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells seemed unaffected by enamel matrix derivative. The alginate hydrogel coating procedure may have potential for local delivery of enamel matrix derivative and other stimulatory factors for use in bone tissue engineering.

  18. Alginate hydrogel enriched with enamel matrix derivative to target osteogenic cell differentiation in TiO2 scaffolds

    PubMed Central

    Pullisaar, Helen; Verket, Anders; Szoke, Krisztina; Tiainen, Hanna; Haugen, Håvard J; Brinchmann, Jan E; Reseland, Janne E

    2015-01-01

    The purpose of bone tissue engineering is to employ scaffolds, cells, and growth factors to facilitate healing of bone defects. The aim of this study was to assess the viability and osteogenic differentiation of primary human osteoblasts and adipose tissue–derived mesenchymal stem cells from various donors on titanium dioxide (TiO2) scaffolds coated with an alginate hydrogel enriched with enamel matrix derivative. Cells were harvested for quantitative reverse transcription polymerase chain reaction on days 14 and 21, and medium was collected on days 2, 14, and 21 for protein analyses. Neither coating with alginate hydrogel nor alginate hydrogel enriched with enamel matrix derivative induced a cytotoxic response. Enamel matrix derivative–enriched alginate hydrogel significantly increased the expression of osteoblast markers COL1A1, TNFRSF11B, and BGLAP and secretion of osteopontin in human osteoblasts, whereas osteogenic differentiation of human adipose tissue–derived mesenchymal stem cells seemed unaffected by enamel matrix derivative. The alginate hydrogel coating procedure may have potential for local delivery of enamel matrix derivative and other stimulatory factors for use in bone tissue engineering. PMID:26090086

  19. Injectable alginate/hydroxyapatite gel scaffold combined with gelatin microspheres for drug delivery and bone tissue engineering.

    PubMed

    Yan, Jingxuan; Miao, Yuting; Tan, Huaping; Zhou, Tianle; Ling, Zhonghua; Chen, Yong; Xing, Xiaodong; Hu, Xiaohong

    2016-06-01

    Injectable and biodegradable alginate-based composite gel scaffolds doubly integrated with hydroxyapatite (HAp) and gelatin microspheres (GMs) were cross-linked via in situ release of calcium cations. As triggers of calcium cations, CaCO3 and glucono-D-lactone (GDL) were fixed as a mass ratio of 1:1 to control pH value ranging from 6.8 to 7.2 during gelation. Synchronously, tetracycline hydrochloride (TH) was encapsulated into GMs to enhance bioactivity of composite gel scaffolds. The effects of HAp and GMs on characteristics of gel scaffolds, including pH value, gelation time, mechanical properties, swelling ratio, degradation behavior and drug release, were investigated. The results showed that HAp and GMs successfully improved mechanical properties of gel scaffolds at strain from 0.1 to 0.5, which stabilized the gel network and decreased weight loss, as well as swelling ratio and gelation time. TH could be released from this composite gel scaffold into the local microenvironment in a controlled fashion by the organic/inorganic hybrid of hydrogel network. Our results demonstrate that the HAp and GMs doubly integrated alginate-based gel scaffolds, especially the one with 6% (w/v) HAp and 5% (w/v) GMs, have suitable physical performance and bioactive properties, thus provide a potential opportunity to be used for bone tissue engineering. The potential application of this gel scaffold in bone tissue engineering was confirmed by encapsulation behavior of osteoblasts. In combination with TH, the gel scaffold exhibited beneficial effects on osteoblast activity, which suggested a promising future for local treatment of pathologies involving bone loss.

  20. Gelation time, homogeneity, and rupture testing of alginate-calcium carbonate-hydrogen peroxide gels for use as wound dressings.

    PubMed

    Alexander, Brendan R; Murphy, Kathleen E; Gallagher, Joanne; Farrell, Garrett F; Taggart, Gertie

    2012-02-01

    The care of chronic wounds carries a heavy financial burden on the healthcare industry, with billons being spent annually on their treatment. This, coupled with a decreased quality of life for sufferers, has led to a real urgency in developing inexpensive wound dressings that promote wound healing. Alginate gels for application as wound dressings were formed by varying alginate (0%-6% w/v), calcium carbonate (0%-1% w/v), hydrogen peroxide (0%-3.75% v/v), and hyaluronic acid (0-1.25 mg/L) content. The aging effects on the physical properties of the gels over a 14-day period were also investigated. The results indicated that the concentration of calcium carbonate and hydrogen peroxide, as well as sample age, all had a significant effect on the rupture characteristics and gelation time of the gels. Increased calcium carbonate content caused an increase in rupture force and rupture energy values, whereas increased hydrogen peroxide content and sample age resulted in a decrease in rupture force and rupture energy measurements. Increased calcium carbonate and hydrogen peroxide content produced a decrease in the time required for gel formation. Statistical models were also produced to provide a means of estimating rupture characteristics and gelation times for gels containing other concentrations of these components.

  1. Enrichment of cancer stem cell-like cells by culture in alginate gel beads.

    PubMed

    Xu, Xiao-xi; Liu, Chang; Liu, Yang; Yang, Li; Li, Nan; Guo, Xin; Sun, Guang-wei; Ma, Xiao-jun

    2014-05-10

    Cancer stem cells (CSCs) are most likely the reason of cancer reoccurrence and metastasis. For further elucidation of the mechanism underlying the characteristics of CSCs, it is necessary to develop efficient culture systems to culture and expand CSCs. In this study, a three-dimensional (3D) culture system based on alginate gel (ALG) beads was reported to enrich CSCs. Two cell lines derived from different histologic origins were encapsulated in ALG beads respectively and the expansion of CSCs was investigated. Compared with two-dimensional (2D) culture, the proportion of cells with CSC-like phenotypes was significantly increased in ALG beads. Expression levels of CSC-related genes were greater in ALG beads than in 2D culture. The increase of CSC proportion after being cultured within ALG beads was further confirmed by enhanced tumorigenicity in vivo. Moreover, increased metastasis ability and higher anti-cancer drug resistance were also observed in 3D-cultured cells. Furthermore, we found that it was hypoxia, through the upregulation of hypoxia-inducible factors (HIFs) that occurred in ALG beads to induce the increasing of CSC proportion. Therefore, ALG bead was an efficient culture system for CSC enrichment, which might provide a useful platform for CSC research and promote the development of new anti-cancer therapies targeting CSCs.

  2. Drug release characteristics from chitosan-alginate matrix tablets based on the theory of self-assembled film.

    PubMed

    Li, Liang; Wang, Linlin; Shao, Yang; Ni, Rui; Zhang, Tingting; Mao, Shirui

    2013-06-25

    The aim of this study was to better understand the underlying drug release characteristics from chitosan-alginate matrix tablets containing different types of drugs. Theophylline, paracetamol, metformin hydrochloride and trimetazidine hydrochloride were used as model drugs exhibiting significantly different solubilities (12, 16, 346 and >1000 mg/ml at 37 °C in water). A novel concept raised was that drugs were released from chitosan-alginate matrix tablets based on the theory of a self-assembled film-controlled release system. The film was only formed on the surface of tablets in gastrointestinal environment and originated from chitosan-alginate polyelectrolyte complex, confirmed by differential scanning calorimetry characterization. The formed film could decrease the rate of polymer swelling to a degree, also greatly limit the erosion of tablets. Drugs were all released through diffusion in the hydrated matrix and polymer relaxation, irrespective of the drug solubility. The effects of polymer level and initial drug loading on release depended on drug properties. Drug release was influenced by the change of pH. In contrast, the impact of ionic strength of the release medium within the physiological range was negligible. Importantly, hydrodynamic conditions showed a key factor determining the superiority of the self-assembled film in controlling drug release compared with conventional matrix tablets. The new insight into chitosan-alginate matrix tablets can help to broaden the application of this type of dosage forms.

  3. Encapsulated human hepatocellular carcinoma cells by alginate gel beads as an in vitro metastasis model

    SciTech Connect

    Xu, Xiao-xi; Liu, Chang; Liu, Yang; Li, Nan; Guo, Xin; Wang, Shu-jun; Sun, Guang-wei; Wang, Wei; Ma, Xiao-jun

    2013-08-15

    Hepatocellular carcinoma (HCC) is the most common primary liver cancer and often forms metastases, which are the most important prognostic factors. For further elucidation of the mechanism underlying the progression and metastasis of HCC, a culture system mimicking the in vivo tumor microenvironment is needed. In this study, we investigated the metastatic ability of HCC cells cultured within alginate gel (ALG) beads. In the culture system, HCC cells formed spheroids by proliferation and maintained in nuclear abnormalities. The gene and protein expression of metastasis-related molecules was increased in ALG beads, compared with the traditional adhesion culture. Furthermore, several gene expression levels in ALG bead culture system were even closer to liver cancer tissues. More importantly, in vitro invasion assay showed that the invasion cells derived from ALG beads was 7.8-fold higher than adhesion cells. Our results indicated that the in vitro three-dimensional (3D) model based on ALG beads increased metastatic ability compared with adhesion culture, even partly mimicked the in vivo tumor tissues. Moreover, due to the controllable preparation conditions, steady characteristics and production at large-scale, the 3D ALG bead model would become an important tool used in the high-throughput screening of anti-metastasis drugs and the metastatic mechanism research. -- Highlights: •We established a 3D metastasis model mimicking the metastatic ability in vivo. •The invasion ability of cells derived from our model was increased significantly. •The model is easy to reproduce, convenient to handle, and amenable for large-scale.

  4. Sol/Gel Processing Techniques for Glass Matrix Composites.

    DTIC Science & Technology

    1987-11-01

    development of a general technique (i.e., Pyrex is less susceptible to devitrification than SiO2 or TiO2 -SiO 2 ). In addition. the properties of these sol / gel ...of a sol / gel process for SIC 2 and SiO2 - TiO2 - together with a data base for their densification - are prerequisite to the successful fabrication of...S~%ad~ 5~ ~ ~ *~~~~;:>;::L-; 1: ’*~~’~ ’S. AFWL-TN-86-59 AFWL-TN- 86-59 00 SOL / GEL PROCESSING TECHNIQUES FOR GLASS MATRIX COMPOSITES 0) C. G

  5. Molecular mechanism of lysozyme adsorption onto chemically modified alginate guar gum matrix.

    PubMed

    Brassesco, Ma Emilia; Woitovich Valetti, Nadia; Picó, Guillermo

    2017-03-01

    The equilibrium isotherms and adsorption kinetics of lysozyme (LZ) on epichlorohydrin (Epi) cross-linked alginate-guar gum (Alg-GG) matrix were studied. Adsorption kinetics followed a pseudo-first-order model while the equilibrium isotherm could be represented by the Freundlich equation. The maximal amount of LZ adsorbed onto this matrix was around 2.4mg per g of hydrated matrix at pH 7.00. The adsorption mechanism was associated to a simple diffusion process with a weak columbic interaction between LZ and the matrix. The presence of NaCl 0.3M induced a total displacement of the LZ from the matrix. Under this condition, the percentage of desorbed protein was 95%. Successive cycles of adsorption-washing-elution were performed and the results showed the reversibility of the process and the usefulness of the method for enzyme purification and separation. A last successful step was carried out for the purification of LZ from egg white as natural source. The model proved to be useful applied as a platform design in the isolation and purification of proteins.

  6. New polymer gel dosimeters consisting of less toxic monomers with radiation-crosslinked gel matrix

    NASA Astrophysics Data System (ADS)

    Hiroki, A.; Yamashita, S.; Sato, Y.; Nagasawa, N.; Taguchi, M.

    2013-06-01

    New polymer gel dosimeters consisting of less toxic methacrylate-type monomers such as 2-hydroxymethyl methacrylate (HEMA) and polyethylene glycol 400 dimethacrylate (9G) with hydroxypropyl cellulose (HPC) gel were prepared. The HPC gels were obtained by using a radiation-induced crosslinking technique to be applied in a matrix instead of a gelatin, which is conventionally used in earlier dosimeters, for the polymer gel dosimeters. The prepared polymer gel dosimeters showed cloudiness by exposing to 60Co γ-ray, in which the cloudiness increased with the dose up to 10 Gy. At the same dose, the increase in the cloudiness appeared with increasing concentration of 9G. As a result of the absorbance measurement, it was found that the dose response depended on the composition ratio between HEMA and 9G.

  7. Survivability of probiotics encapsulated in alginate gel microbeads using a novel impinging aerosols method.

    PubMed

    Sohail, Asma; Turner, Mark S; Coombes, Allan; Bostrom, Thor; Bhandari, Bhesh

    2011-01-31

    Encapsulation of probiotic bacteria in cross-linked alginate beads is of major interest for improving the survivability in harsh acid and bile environment and also in food matrices. Alginate micro beads (10-40 μm) containing the probiotics Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM were produced by a novel technique based on dual aerosols of alginate solution and CaCl(2) cross linking solution. Extruded macro beads (approximately 2mm diameter) produced by the conventional method and micro beads produced by novel aerosols technique offered comparable protection to L. rhamnosus in high acid and bile environment. Chitosan coating of micro beads resulted in a significant increase in survival time of L. rhamnosus from 40 to 120 min in acid condition and the reduction in cell numbers was confined to 0.94 log over this time. Alginate macro beads are more effective than micro beads in protecting L. acidophilus against high acid and bile. Chitosan coating of micro beads resulted in similar protection to L. acidophilus in macro beads in acid and extended the survival time from 90 to at least 120 min. Viability of this organism in micro beads was 3.5 log after 120 min. The continuous processing capability and scale-up potential of the dual aerosol technique offers potential for an efficient encapsulation of probiotics in very small alginate micro beads below sensorial detection limits while still being able to confer effective protection in acid and bile environment.

  8. Efficient Pb(II) removal using sodium alginate-carboxymethyl cellulose gel beads: Preparation, characterization, and adsorption mechanism.

    PubMed

    Ren, Huixue; Gao, Zhimin; Wu, Daoji; Jiang, Jiahui; Sun, Youmin; Luo, Congwei

    2016-02-10

    Alginate-carboxymethyl cellulose (CMC) gel beads were prepared in this study using sodium alginate (SA) and sodium CMC through blending and cross-linking. The specific surface area and aperture of the prepared SA-CMC gel beads were tested. The SA-CMC structure was characterized and analyzed via infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. Static adsorption experiment demonstrated that Pb(II) adsorption of SA-CMC exceeded 99% under the optimized conditions. In addition, experiments conducted under the same experimental conditions showed that the lead ion removal efficiency of SA-CMC was significantly higher than that of conventional adsorbents. The Pb(II) adsorption process of SA-CMC followed the Langmuir adsorption isotherm, and the dynamic adsorption model could be described through a pseudo-second-order rate equation. Pb(II) removal mechanisms of SA-CMC, including physical, chemical, and electrostatic adsorptions, were discussed based on microstructure analysis and adsorption kinetics. Chemical adsorption was the main adsorption method among these mechanisms.

  9. Oxidized alginate hydrogels as niche environments for corneal epithelial cells.

    PubMed

    Wright, Bernice; De Bank, Paul A; Luetchford, Kim A; Acosta, Fernando R; Connon, Che J

    2014-10-01

    Chemical and biochemical modification of hydrogels is one strategy to create physiological constructs that maintain cell function. The aim of this study was to apply oxidised alginate hydrogels as a basis for development of a biomimetic niche for limbal epithelial stem cells that may be applied to treating corneal dysfunction. The stem phenotype of bovine limbal epithelial cells (LEC) and the viability of corneal epithelial cells (CEC) were examined in oxidised alginate gels containing collagen IV over a 3-day culture period. Oxidation increased cell viability (P ≤ 0.05) and this improved further with addition of collagen IV (P ≤ 0.01). Oxidised gels presented larger internal pores (diameter: 0.2-0.8 µm) than unmodified gels (pore diameter: 0.05-0.1 µm) and were significantly less stiff (P ≤ 0.001), indicating that an increase in pore size and a decrease in stiffness contributed to improved cell viability. The diffusion of collagen IV from oxidised alginate gels was similar to that of unmodified gels suggesting that oxidation may not affect the retention of extracellular matrix proteins in alginate gels. These data demonstrate that oxidised alginate gels containing corneal extracellular matrix proteins can influence corneal epithelial cell function in a manner that may impact beneficially on corneal wound healing therapy.

  10. Oxidized alginate hydrogels as niche environments for corneal epithelial cells

    PubMed Central

    Wright, Bernice; De Bank, Paul A; Luetchford, Kim A; Acosta, Fernando R; Connon, Che J

    2014-01-01

    Chemical and biochemical modification of hydrogels is one strategy to create physiological constructs that maintain cell function. The aim of this study was to apply oxidised alginate hydrogels as a basis for development of a biomimetic niche for limbal epithelial stem cells that may be applied to treating corneal dysfunction. The stem phenotype of bovine limbal epithelial cells (LEC) and the viability of corneal epithelial cells (CEC) were examined in oxidised alginate gels containing collagen IV over a 3-day culture period. Oxidation increased cell viability (P ≤ 0.05) and this improved further with addition of collagen IV (P ≤ 0.01). Oxidised gels presented larger internal pores (diameter: 0.2–0.8 µm) than unmodified gels (pore diameter: 0.05–0.1 µm) and were significantly less stiff (P ≤ 0.001), indicating that an increase in pore size and a decrease in stiffness contributed to improved cell viability. The diffusion of collagen IV from oxidised alginate gels was similar to that of unmodified gels suggesting that oxidation may not affect the retention of extracellular matrix proteins in alginate gels. These data demonstrate that oxidised alginate gels containing corneal extracellular matrix proteins can influence corneal epithelial cell function in a manner that may impact beneficially on corneal wound healing therapy. © 2013 The Authors. Journal of Biomedical Materials Research Part A Published byWiley Periodicals, Inc. Part A: 102A: 3393–3400, 2014. PMID:24142706

  11. An acid/alkaline stress and the addition of amino acids induce a prolonged viability of Lactobacillus plantarum loaded into alginate gel.

    PubMed

    Bevilacqua, Antonio; Sinigaglia, Milena; Corbo, Maria Rosaria

    2010-08-15

    This study reports on the investigation on the effects of the conditions used throughout the step of biomass production on the survival of Lactobacillus plantarum loaded into alginate gels. L. plantarum was grown under different conditions (MRS or a laboratory medium-LB(2)-at acidic or alkaline pHs, with NaCl, phenols, vitamins or amino acids) and immobilized in sodium alginate; cell number was evaluated throughout the storage and death (delta(stand)) and first-reduction times (delta) were calculated. The storage of alginate gels at 4 degrees C prolonged cell viability up to 60 days (ca. 20 days for cells produced in MRS and stored at 30 degrees C); however, a similar prolongation was achieved for cells produced in LB(2) adjusted to pH 5.0 and 9.0 or added with amino acids (death time>50-60 days).

  12. Preparation and characterization of rifampicin-PLGA microspheres/sodium alginate in situ gel combination delivery system.

    PubMed

    Hu, Chunhui; Feng, Hanzhou; Zhu, Chunyan

    2012-06-15

    We prepared a complex drug delivery system consisted of rifampicin-poly(lactic-co-glycolic acid) (PLGA) microspheres in combination with sodium alginate in situ gel. The microspheres were obtained by using a solvent evaporation method, the mean diameter was 1.748 μm and the span of particle distribution was 0.78. The combination delivery system was obtained by adding microspheres to sodium alginate solution followed by physically mixing. In an in vitro study of drug release monitored for 11 days, the release of rifampicin from combination delivery system was slower than microspheres. The cumulative release percent of rifampicin from combination delivery system was 91.83 ± 1.26%, which was lower than 97.36 ± 3.41% of rifampicin released from microspheres. An in vivo fluorescence imaging study suggests that the gel adhered to lungs within 24h, and microspheres stayed in lungs at least for 504 h (21 days). In vivo drug release study indicates that the maximum local rifampicin concentration in lungs was 48.60 ± 15.67 μg mL(-1) 5h after administration. After 21 days, the local rifampicin concentration was 0.81±0.14μgmL(-1), which was above the minimum inhibitory concentration of rifampicin. The combination delivery system significantly prolonged RFP release compared to microspheres, from which RFP released could only be detected for 10 days. This approach to control the release of rifampicin using PLGA microspheres/in situ gel combination delivery system in conjunction with interventional technology is useful for improving anti-tuberculosis treatment effectiveness for patients.

  13. Structure of alginate gels: interaction of diuronate units with divalent cations from density functional calculations.

    PubMed

    Agulhon, Pierre; Markova, Velina; Robitzer, Mike; Quignard, Françoise; Mineva, Tzonka

    2012-06-11

    The complexation of (1→4) linked α-L-guluronate (G) and β-D-mannuronate (M) disaccharides with Mg(2+), Ca(2+), Sr(2+), Mn(2+), Co(2+), Cu(2+), and Zn(2+) cations have been studied with quantum chemical density functional theory (DFT)-based method. A large number of possible cation-diuronate complexes, with one and two GG or MM disaccharide units and with or without water molecules in the inner coordination shells have been considered. The computed bond distances, cation interaction energies, and molecular orbital composition analysis revealed that the complexation of the transition metal (TM) ions to the disaccharides occurs via the formation of strong coordination-covalent bonds. On the contrary, the alkaline earth cations form ionic bonds with the uronates. The unidentate binding is found to be the most favored one in the TM hydrated and water-free complexes. By removing water molecules, the bidentate chelating binding also occurs, although it is found to be energetically less favored by 1 to 1.5 eV than the unidentate one. A good correlation is obtained between the alginate affinity trend toward TM cations and the interaction energies of the TM cations in all studied complexes, which suggests that the alginate affinities are strongly related to the chemical interaction strength of TM cations-uronate complexes. The trend of the interaction energies of the alkaline earth cations in the ionic complexes is opposite to the alginate affinity order. The binding strength is thus not a limiting factor in the alginate gelation in the presence of alkaline earth cations at variance with the TM cations.

  14. A simple, quantitative method using alginate gel to determine rat colonic tumor volume in vivo.

    PubMed

    Irving, Amy A; Young, Lindsay B; Pleiman, Jennifer K; Konrath, Michael J; Marzella, Blake; Nonte, Michael; Cacciatore, Justin; Ford, Madeline R; Clipson, Linda; Amos-Landgraf, James M; Dove, William F

    2014-04-01

    Many studies of the response of colonic tumors to therapeutics use tumor multiplicity as the endpoint to determine the effectiveness of the agent. These studies can be greatly enhanced by accurate measurements of tumor volume. Here we present a quantitative method to easily and accurately determine colonic tumor volume. This approach uses a biocompatible alginate to create a negative mold of a tumor-bearing colon; this mold is then used to make positive casts of dental stone that replicate the shape of each original tumor. The weight of the dental stone cast correlates highly with the weight of the dissected tumors. After refinement of the technique, overall error in tumor volume was 16.9% ± 7.9% and includes error from both the alginate and dental stone procedures. Because this technique is limited to molding of tumors in the colon, we utilized the Apc(Pirc/+) rat, which has a propensity for developing colonic tumors that reflect the location of the majority of human intestinal tumors. We have successfully used the described method to determine tumor volumes ranging from 4 to 196 mm³. Alginate molding combined with dental stone casting is a facile method for determining tumor volume in vivo without costly equipment or knowledge of analytic software. This broadly accessible method creates the opportunity to objectively study colonic tumors over time in living animals in conjunction with other experiments and without transferring animals from the facility where they are maintained.

  15. A Simple, Quantitative Method Using Alginate Gel to Determine Rat Colonic Tumor Volume In Vivo

    PubMed Central

    Irving, Amy A; Young, Lindsay B; Pleiman, Jennifer K; Konrath, Michael J; Marzella, Blake; Nonte, Michael; Cacciatore, Justin; Ford, Madeline R; Clipson, Linda; Amos-Landgraf, James M; Dove, William F

    2014-01-01

    Many studies of the response of colonic tumors to therapeutics use tumor multiplicity as the endpoint to determine the effectiveness of the agent. These studies can be greatly enhanced by accurate measurements of tumor volume. Here we present a quantitative method to easily and accurately determine colonic tumor volume. This approach uses a biocompatible alginate to create a negative mold of a tumor-bearing colon; this mold is then used to make positive casts of dental stone that replicate the shape of each original tumor. The weight of the dental stone cast correlates highly with the weight of the dissected tumors. After refinement of the technique, overall error in tumor volume was 16.9% ± 7.9% and includes error from both the alginate and dental stone procedures. Because this technique is limited to molding of tumors in the colon, we utilized the ApcPirc/+ rat, which has a propensity for developing colonic tumors that reflect the location of the majority of human intestinal tumors. We have successfully used the described method to determine tumor volumes ranging from 4 to 196 mm3. Alginate molding combined with dental stone casting is a facile method for determining tumor volume in vivo without costly equipment or knowledge of analytic software. This broadly accessible method creates the opportunity to objectively study colonic tumors over time in living animals in conjunction with other experiments and without transferring animals from the facility where they are maintained. PMID:24674588

  16. Chondrocyte extracellular matrix synthesis and turnover are influenced by static compression in a new alginate disk culture system.

    PubMed

    Ragan, P M; Chin, V I; Hung, H H; Masuda, K; Thonar, E J; Arner, E C; Grodzinsky, A J; Sandy, J D

    2000-11-15

    The goal of this study was to examine the effects of mechanical compression on chondrocyte biosynthesis of extracellular matrix (ECM) components during culture in a new alginate disk culture system. Specifically, we have examined chondrocyte biosynthesis rates, and the structure of aggrecan core protein species present in the cell-associated matrix (CM), in the further removed matrix (FRM) and in the surrounding culture medium. In this alginate disk culture system, chondrocytes can be subjected to mechanical deformations similar to those experienced in vivo. Our results show that over an 8-week culture period, chondrocytes synthesize a functional ECM and can respond to mechanical forces similarly to chondrocytes maintained in native cartilage. In the alginate disk system, static compression was shown to decrease and dynamic compression to increase synthesis of aggrecan of bovine chondrocytes. Western blot analysis of the core proteins of aggrecan molecules identified a number of different species that were present in different relative amounts in the CM, FRM, and medium. Over 21 days of culture, the predominant form of aggrecan found in the ECM was a full-length link-stabilized species. In addition, our data show that the application of 40 h of static compression caused an increase in the proportion of newly synthesized aggrecan molecules released into the medium. However, this was not accompanied by a significant change in the size and composition of aggrecan and aggrecan fragments in the different compartments, suggesting that mechanical compression did not alter the catabolic pathways. Together, these data show that chondrocyte function is maintained in an alginate disk culture system and that this culture system is a useful model to examine chondrocyte ECM assembly and some aspects of catabolism normally found in vivo.

  17. Sustained Delivery of Bioactive GDNF from Collagen and Alginate-Based Cell-Encapsulating Gel Promoted Photoreceptor Survival in an Inherited Retinal Degeneration Model

    PubMed Central

    Chan, Barbara P.; Lo, Amy C. Y.

    2016-01-01

    Encapsulated-cell therapy (ECT) is an attractive approach for continuously delivering freshly synthesized therapeutics to treat sight-threatening posterior eye diseases, circumventing repeated invasive intravitreal injections and improving local drug availability clinically. Composite collagen-alginate (CAC) scaffold contains an interpenetrating network that integrates the physical and biological merits of its constituents, including biocompatibility, mild gelling properties and availability. However, CAC ECT properties and performance in the eye are not well-understood. Previously, we reported a cultured 3D CAC system that supported the growth of GDNF-secreting HEK293 cells with sustainable GDNF delivery. Here, the system was further developed into an intravitreally injectable gel with 1x104 or 2x105 cells encapsulated in 2mg/ml type I collagen and 1% alginate. Gels with lower alginate concentration yielded higher initial cell viability but faster spheroid formation while increasing initial cell density encouraged cell growth. Continuous GDNF delivery was detected in culture and in healthy rat eyes for at least 14 days. The gels were well-tolerated with no host tissue attachment and contained living cell colonies. Most importantly, gel-implanted in dystrophic Royal College of Surgeons rat eyes for 28 days retained photoreceptors while those containing higher initial cell number yielded better photoreceptor survival. CAC ECT gels offers flexible system design and is a potential treatment option for posterior eye diseases. PMID:27441692

  18. Sustained release of a water-soluble drug from alginate matrix tablets prepared by wet granulation method.

    PubMed

    Mandal, Sanchita; Basu, Sanat Kumar; Sa, Biswanath

    2009-01-01

    Alginate matrix tablet of diltiazem hydrochloride (DTZ), a water-soluble drug, was prepared using sodium alginate (SAL) and calcium gluconate (CG) by the conventional wet granulation method for sustained release of the drug. The effect of formulation variables like SAL/CG ratio, drug load, microenvironmental pH modulator, and processing variable like compression force on the extent of drug release was examined. The tablets prepared with 1:2 w/w ratio of SAL/CG produced the most sustained release of the drug extending up to 13.5 h. Above and below this ratio, the drug release was faster. The drug load and the hardness of the tablets produced minimal variation in drug release. The addition of alkaline or acidic microenvironmental modulators did not extend the release; instead, these excipients produced somewhat faster release of diltiazem. This study revealed that proper selection of SAL/CG ratio is important to produce alginate matrix tablet by wet granulation method for sustained release of DTZ.

  19. Efficacy of alginate-based reflux suppressant and magnesium-aluminium antacid gel for treatment of heartburn in pregnancy: a randomized double-blind controlled trial

    PubMed Central

    Meteerattanapipat, Pontip; Phupong, Vorapong

    2017-01-01

    The aim of this study was to compare the therapeutic efficacy of alginate-based reflux suppressant and magnesium-aluminium antacid gel for treatment of heartburn in pregnancy. A double-blinded, randomized, controlled trial was conducted. One hundred pregnant women at less than 36 weeks gestation with heartburn at least twice per week were randomized to either alginate-based reflux suppressant or to magnesium-aluminium antacid gel. Details of heartburn were recorded before beginning the treatment and the second week of study. Primary outcome measure was the improvement of heartburn frequency after treatment and secondary outcome were the improvement of heartburn intensity, quality of life, maternal satisfaction, maternal side effects, pregnancy and neonatal outcomes. There was no difference between treatment and control groups in improvement of heartburn frequency (80% vs 88%, p = 0.275), 50% reduction of frequency of heartburn (56% vs 52%, p = 0.688), improvement of heartburn intensity (92% vs 92%, p = 1.000) and 50% reduction of heartburn intensity (68% vs 80% cases, p = 0.075). There were also no significant differences in quality of life, maternal satisfaction, maternal side effects, pregnancy and neonatal outcomes. Alginate-based reflux suppressant was not different from magnesium-aluminium antacid gel in the treatment of heartburn in pregnancy. PMID:28317885

  20. Fed batch bioconversion of 2-propanol by a solvent tolerant strain of Alcaligenes faecalis entrapped in Ca-alginate gel.

    PubMed

    Mohammad, Balsam T; Bustard, Mark T

    2008-07-01

    A gram-negative, rod-shaped, aerobe, capable of converting 2-propanol (isopropanol, IPA) to acetone was isolated from an oil/sump, and identified by 16 S rDNA analysis as Alcaligenes faecalis. Investigations showed this strain to be extremely solvent-tolerant and it was subsequently named ST1. In this study, A. faecalis ST1 cells were immobilized by entrapment in Ca-alginate beads (3 mm in diameter), and used in the bioconversion of high concentration IPA. The biodegradation rates and the corresponding microbial growth inside the beads were measured at four different IPA concentration ranges from 2 to 15 g l(-1). The maximum cell concentration obtained was 9.59 g dry cell weight (DCW) l(-1) medium which equated to 66 g DCW l(-1) gel, at an initial IPA concentration of 15 g l(-1) after 216 h of incubation. A maximum biodegradation rate of 0.067 g IPA g cells(-1) h(-1) was achieved for 5 g l(-1) IPA where an increase in IPA concentration to 38 g l(-1) caused reduction in bead integrity. A modified growth medium was developed which allowed repeated use of the beads for more than 42 days without any loss of integrity and continued bioconversion activity.

  1. Comparison of a dimethicone/antacid (Asilone gel) with an alginate/antacid (Gaviscon liquid) in the management of reflux oesophagitis.

    PubMed

    Smart, H L; Atkinson, M

    1990-09-01

    Fifty-three patients with symptomatic reflux oesophagitis were entered into a single centre randomized study comparing the effects of a dimethicone/antacid (Asilone Gel) and an alginate/antacid (Gaviscon liquid) on symptoms and endoscopic changes over an 8-week period. Both treatments significantly improved heartburn, acid regurgitation and flatulence. Dimethicone/antacid but not alginate/antacid, produced a significant improvement in oesophagitis, oesophageal ulceration and histological grade of inflammation over the 8-week treatment period so that 14 patients treated with dimethicone/antacid and 10 with alginate/antacid had normal endoscopic oesophageal appearances at the end of the study. The difference in improvement between the two patient groups did not reach significance however, except for dimethicone/antacid improving histological changes (P less than 0.05). These findings suggest that dimethicone/antacid and alginate/antacid are equally effective in treating symptomatic reflux oesophagitis although dimethicone/antacid may have an advantage in improving oesophageal histological appearances.

  2. Chondrocyte calcium signaling in response to fluid flow is regulated by matrix adhesion in 3-D alginate scaffolds.

    PubMed

    Degala, Satish; Zipfel, Warren R; Bonassar, Lawrence J

    2011-01-01

    The interaction between chondrocytes and their surrounding extracellular matrix plays an important role in regulating cartilage metabolism in response to environmental cues. This study characterized the role of cell adhesion on the calcium signaling response of chondrocytes to fluid flow. Bovine chondrocytes were suspended in alginate hydrogels functionalized with RGD at concentrations of 0-400μM. The hydrogels were perfused and the calcium signaling response of the cells was measured over a range of fluid velocities from 0 to 68μm/s. Attachment to RGD-alginate doubled the sensitivity of chondrocytes to flows in the range of 8-13μm/s, but at higher fluid velocities, the contribution of cell adhesion to the observed calcium signaling response was no longer apparent. The enhanced sensitivity to flow was dependent on the density of RGD-ligand present in the scaffolds. The RGD-enhanced sensitivity to flow was completely inhibited by the addition of soluble RGD which acted as a competitive inhibitor. The results of this study indicate a role for matrix adhesion in regulating chondrocyte response to fluid flow through a calcium dependent mechanism.

  3. Poly(vinyl alcohol) and alginate cross-linked matrix with immobilized Prussian blue and ion exchange resin for cesium removal from waters.

    PubMed

    Lai, Yu-Chen; Chang, Yin-Ru; Chen, Man-Li; Lo, Yu-Kuo; Lai, Juin-Yih; Lee, Duu-Jong

    2016-08-01

    Cesium (Cs) removal from contaminated water bodies is an emerging issue after the disaster at the Fukushima Daiichi Nuclear Power Plant. The Prussian blue (PB) is an effective Cs adsorbent but will release hexacyanoferrate fragments from the adsorbent matrix during adsorption. Alginate is an affordable biopolymer for PB particles immobilization. This study synthesized poly(vinyl alcohol) (PVA) and alginate cross-linked matrix for immobilization of PB nano-sized particles and a surface-modified styrene-ethyl styrene divinyl benzene resin and tested their swelling stability and Cs adsorption performance in fresh water and in seawater. The PVA-alginate granules have high structural stability in both fresh water and seawater, with the Cs adsorption capability higher for the former than the latter. The adopted resin effectively remove released PB fragments from the tested granules. The transport and reaction parameters for the granules and for the sand filter bed were estimated.

  4. A surfactant-based, regularly arrayed nanostructure gel matrix for migration of small molecules.

    PubMed

    Kato, Masaru; Suwanai, Yusuke; Shimojima, Atsushi; Santa, Tomofumi

    2012-11-01

    The preparation of nanometer-scale pores, or nanopores, has become easy because of the progress in nanotechnology. Surfactants are promising materials for the preparation of nanostructures containing nanopores, because surfactants form many different phase structures, including cubic, micellar, and lamellar structures. We prepared a gel matrix with a cubic structure from a commercially available surfactant, polyoxyethylene(50) lauryl ether (C12EO50, Adekatol LA-50). This gel matrix had regularly arrayed nanopores between the packed spherical micelles. We used the gel to separate biomolecules by means of slab gel electrophoresis. The gel was applicable to migration of amino acids and peptides; however, larger molecules, such as proteins and single-walled carbon nanotubes, did not migrate through the gel. We concluded that the pore size was too small for the penetration of large molecules, and that only small molecules could penetrate the gel matrix. The migration mechanism of small molecules was similar to that observed in conventional gel electrophoresis. We concluded that the gel matrix prepared from surfactant is a promising matrix for migration and purification of small molecules. We also expect that the gel can be used as a nanoscale filter to trap large molecules, allowing only small molecules to pass.

  5. Studies of matrix vesicle-induced mineralization in a gelatin gel

    NASA Technical Reports Server (NTRS)

    Boskey, A. L.; Boyan, B. D.; Doty, S. B.; Feliciano, A.; Greer, K.; Weiland, D.; Swain, L. D.; Schwartz, Z.

    1992-01-01

    Matrix vesicles isolated from fourth-passage cultures of chondrocytes were tested for their ability to induce hydroxyapatite formation in a gelatin gel in order to gain insight into the function of matrix vesicles in in situ mineralization. These matrix vesicles did not appear to be hydroxyapatite nucleators per se since the extent of mineral accumulation in the gel diffusion system was not altered by the presence of matrix vesicles alone, and in the vesicle containing gels, mineral crystals were formed whether associated with vesicles or not. In gels with these matrix vesicles and beta-glycerophosphate, despite the presence of alkaline phosphatase activity, there was no increase in mineral deposition. This suggested that in the gel system these culture-derived vesicles did not increase local phosphate concentrations. However, when known inhibitors of mineral crystal formation and growth (proteoglycan aggregates [4 mg/ml], or ATP [1 mM], or both proteoglycan and ATP) were included in the gel, more mineral was deposited in gels with the vesicles than in comparable gels without vesicles, indicating that enzymes within these vesicles were functioning to remove the inhibition. These data support the suggestion that one function of the extracellular matrix vesicles is to transport enzymes for matrix modification.

  6. Methods of Manufacturing Bioactive Gels from Extracellular Matrix Material

    NASA Technical Reports Server (NTRS)

    Kentner, Kimberly A. (Inventor); Stuart, Katherine A. (Inventor); Janis, Abram D. (Inventor)

    2015-01-01

    The present invention is directed to methods of manufacturing bioactive gels from ECM material, i.e., gels which retain bioactivity, and can serve as scaffolds for preclinical and clinical tissue engineering and regenerative medicine approaches to tissue reconstruction. The manufacturing methods take advantage of a new recognition that bioactive gels from ECM material can be created by digesting particularized ECM material in an alkaline environment and neutralizing to provide bioactive gels.

  7. Methods of Manufacturing Bioactive Gels from Extracellular Matrix Material

    NASA Technical Reports Server (NTRS)

    Kentner, Kimberly A. (Inventor); Stuart, Katherine A. (Inventor); Janis, Abram D. (Inventor)

    2014-01-01

    The present invention is directed to methods of manufacturing bioactive gels from ECM material, i.e., gels which retain bioactivity, and can serve as scaffolds for preclinical and clinical tissue engineering and regenerative medicine approaches to tissue reconstruction. The manufacturing methods take advantage of a new recognition that bioactive gels from ECM material can be created by digesting particularized ECM material in an alkaline environment and neutralizing to provide bioactive gels.

  8. Methods of Manufacturing Bioactive Gels from Extracellular Matrix Material

    NASA Technical Reports Server (NTRS)

    Kentner, Kimberly A. (Inventor); Stuart, Katherine A. (Inventor); Janis, Abram D. (Inventor)

    2016-01-01

    The present invention is directed to methods of manufacturing bioactive gels from ECM material, i.e., gels which retain bioactivity, and can serve as scaffolds for preclinical and clinical tissue engineering and regenerative medicine approaches to tissue reconstruction. The manufacturing methods take advantage of a new recognition that bioactive gels from ECM material can be created by digesting particularized ECM material in an alkaline environment and neutralizing to provide bioactive gels.

  9. Characterization of structure, physico-chemical properties and diffusion behavior of Ca-Alginate gel beads prepared by different gelation methods.

    PubMed

    Puguan, John Marc C; Yu, Xiaohong; Kim, Hern

    2014-10-15

    Ca-Alginate beads were prepared with either external or internal calcium sources by dripping technique. It was found that beads synthesized with internal calcium source had a looser structure and bigger pore size than those produced with external calcium source. Consequently, a faster diffusion rate of Vitamin B12 (VB12) within the beads with an internal calcium source was observed. Furthermore, the concentration of calcium ion, ionic strength and pH of the external gel beads formation solution were investigated. Results showed that (a) the concentration of the calcium ion was found to be the determining factor in the gel formation phenomenon; (b) the weight and volume losses are in effect due to water removal; (c) NaCl acts as a competitor with calcium and a screen in the electrostatic repulsion; and (d) the pH controls the gel formation process by regulating the dissociation of alginate and the complexation of the calcium cations. These results are keys to understanding the behavior and performance of beads in their utilization medium.

  10. Physico-chemical properties of the encapsulation matrix and germination of carrot somatic embryos.

    PubMed

    Timbert, R; Barbotin, J N; Kersulec, A; Bazinet, C; Thomas, D

    1995-06-20

    Carrot somatic embryos were encapsulated in alginate gel beads. To improve the quality of a "synthetic seed" coating, the rheology and dehydration properties of different matrices were tested. By increasing alginate and CaCl(2) concentrations, additional mineral elements were shown to increase resistance to rupture, and to depress the germination of somatic embryos. A polysaccharide addition was found to slow the alginate matrix dehydration; alginate-gellan gum and alginate-kaolin matrices could preserve the viability of somatic embryos at low relative humidities (30% to 35% germinations at 50% relative humidity) to a greater extent than other matrices.

  11. Alginate/cashew gum floating bead as a matrix for larvicide release.

    PubMed

    Paula, Haroldo C B; de Oliveira, Erick F; Abreu, Flávia O M S; de Paula, Regina C M

    2012-08-01

    A polymeric floating system composed of Alginate (ALG) and Cashew gum (CG), loaded with an essential oil (Lippia sidoides-Ls) was prepared by ionotropic gelation, characterized regarding its physical-chemistry properties and evaluated on its potential as a controlled release system. The influence of process parameters on the buoyancy, loading, swelling and in vitro and in vivo release kinetics, was investigated. Results showed that beads produced with carbonate and Ls at high level contents exhibit good floatability (up to 5 days) and loading capacity (15.2-23.8%). In vitro release data showed a Fickian diffusion profile and in vivo experiments showed that ALG-CG floating system presented a superior and prolonged larvicide effect, in comparison with non-floating ones, presenting larvae mortality values of 85% and 33%, respectively, after 48 h. These results indicate that ALG-CG floating beads loaded with Ls presented enhanced oil entrapment efficiency, excellent floating ability, and suitable larvicide release pattern.

  12. Thermally reversible gels in electrophoresis. I - Matrix characterization

    NASA Technical Reports Server (NTRS)

    Righetti, Pier Giorgio; Snyder, Robert S.

    1988-01-01

    Two series of thermally reversible hydrogen-bonded gels have been characterized: (5 pct) PVA-(4 pct) PEG and (5 pct) PVA-(0.04 pct) borate gels. They both have extremely low melting points (16-17 C) and could be of potential interest for recovery of proteins after preparative electrophoresis. The PVA-borate gels can be exploited in the pH range 7-11 by progressively increasing the borate content in the pH interval 8 to 7 and concomitantly decreasing the borate levels in the pH zone 8 to 11. It is hypothesized that the low melting point of these gels is due to the fact that they are sparingly and sparsely hydrogen bonded along the PVA chain: on the average, 1 OH group out of 3 or 4 OH groups in the PVA polymer should be engaged in H-bond formation.

  13. An additive manufacturing-based PCL-alginate-chondrocyte bioprinted scaffold for cartilage tissue engineering.

    PubMed

    Kundu, Joydip; Shim, Jin-Hyung; Jang, Jinah; Kim, Sung-Won; Cho, Dong-Woo

    2015-11-01

    Regenerative medicine is targeted to improve, restore or replace damaged tissues or organs using a combination of cells, materials and growth factors. Both tissue engineering and developmental biology currently deal with the process of tissue self-assembly and extracellular matrix (ECM) deposition. In this investigation, additive manufacturing (AM) with a multihead deposition system (MHDS) was used to fabricate three-dimensional (3D) cell-printed scaffolds using layer-by-layer (LBL) deposition of polycaprolactone (PCL) and chondrocyte cell-encapsulated alginate hydrogel. Appropriate cell dispensing conditions and optimum alginate concentrations for maintaining cell viability were determined. In vitro cell-based biochemical assays were performed to determine glycosaminoglycans (GAGs), DNA and total collagen contents from different PCL-alginate gel constructs. PCL-alginate gels containing transforming growth factor-β (TGFβ) showed higher ECM formation. The 3D cell-printed scaffolds of PCL-alginate gel were implanted in the dorsal subcutaneous spaces of female nude mice. Histochemical [Alcian blue and haematoxylin and eosin (H&E) staining] and immunohistochemical (type II collagen) analyses of the retrieved implants after 4 weeks revealed enhanced cartilage tissue and type II collagen fibril formation in the PCL-alginate gel (+TGFβ) hybrid scaffold. In conclusion, we present an innovative cell-printed scaffold for cartilage regeneration fabricated by an advanced bioprinting technology.

  14. In vitro evaluation of a fibrin gel antibiotic delivery system containing mesenchymal stem cells and vancomycin alginate beads for treating bone infections and facilitating bone formation.

    PubMed

    Hou, Tianyong; Xu, Jianzhong; Li, Qiang; Feng, Jianghua; Zen, Ling

    2008-07-01

    Bone infection and defects are two major problems that occur in the course of treating posttraumatic open bone fractures and osteomyelitis for which local antibiotic delivery is efficacious. Further, hemostasis is an essential treatment after removal of infected bones. Herein we report a new antibiotics delivery system made of vancomycin alginate beads embedded in a fibrin gel (Vanco-AB-FG) to treat bone infections, with the addition of bone marrow-derived mesenchymal stem cells (BMMSCs) seeded in the fibrin gel to promote bone formation. The proliferation of BMMSCs was measured under different conditions of three-dimensional (3D) gel or monolayer, with or without Vanco-AB; cells were labeled by enhanced green fluorescence protein, and their morphology and distribution were observed. The alkaline phosphatase (ALP) activity, real-time RT-PCR, and von Kossa staining were used for determining the osteogenic differentiation of BMMSCs. The concentrations of vancomycin resulting from the antibiotic delivery were determined; the antibiotic activity was evaluated by an assay with standard Staphylococcus aureus (ATCC 25923) as a biological target. The results showed that for Vanco-AB-FG, vancomycin concentrations remained above the breakpoint sensitivity for 22 days. The 3D culture within the gel and the addition of Vanco-AB affected the cell behavior. The morphology of BMMSCs within the 3D gel was different from that in monolayer. The proliferation of the cells within the 3D gel was lower than that in monolayer in early stage, but in later stage the number of BMMSCs in Vanco-AB-FG was similar to that in monolayer. The ALP activity was higher in the 3D gel, and the addition of Vanco-AB slightly increased ALP activity. The osteogenic gene expression levels of ALP, osteopontin, and alpha1 chain of collagen I were higher in the 3D gel than those in monolayer, and additional Vanco-AB could also increase their expression. The von Kossa staining showed that the deposition of

  15. Alginate encapsulation of Begonia microshoots for short-term storage and distribution

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Synthetic seeds were formed from in vitro grown Begonia cultivars (Sweetheart Mix and BabyWing White) shoot tips using 3% sodium alginate in Murashige and Skoog medium (MS) salt solution as the gel matrix and 100 mM calcium chloride for complexation. Synthetic seed formation was achieved by releasin...

  16. Influence of hydrophobic modification in alginate-based hydrogels for biomedical applications

    NASA Astrophysics Data System (ADS)

    Choudhary, Soumitra

    Alginate has been exploited commercially for decades in foods, textiles, paper, pharmaceutical industries, and also as a detoxifier for removing heavy metals. Alginate is also popular in cell encapsulation because of its relatively mild gelation protocol and simple chemistry with which biological active entities can be immobilized. Surface modification of alginate gels has been explored to induce desired cell interactions with the gel matrix. These modifications alter the bulk properties, which strongly determine on how cells feel and response to the three-dimensional microenvironment. However, there is a need to develop strategies to engineer functionalities into bulk alginate hydrogels that not only preserve their inherent qualities but are also less toxic. In this thesis, our main focus was to optimize the mechanical properties of alginate-based hydrogels, and by doing so control the performance of the biomaterials. In the first scheme, we used alginate and hydrophobically modified ethyl hydroxy ethyl cellulose as components in interpenetrating polymer network (IPN) gels. The second network was used to control gelation time and rheological properties. We believe these experiments also may provide insight into the mechanical and structural properties of more complex biopolymer gels and naturally-occurring IPNs. Next, we worked on incorporating a hydrophobic moiety directly into the alginate chain, resulting in materials for extended release of hydrophobic drugs. We successfully synthesized hydrophobically modified alginate (HMA) by attaching octylamine groups onto the alginate backbone by standard carbodiimide based amide coupling reaction. Solubility of several model hydrophobic drugs in dilute HMA solutions was found to be increased by more than an order of magnitude. HMA hydrogels, prepared by crosslinking the alginate chains with calcium ions, were found to exhibit excellent mechanical properties (modulus ˜100 kPa) with release extended upto 5 days. Ability

  17. Scaling law and microstructure of alginate hydrogel.

    PubMed

    Liu, Sijun; Li, Huijun; Tang, Bijun; Bi, Shuguang; Li, Lin

    2016-01-01

    The gelation of alginate in aqueous solution was studied as a function of Ca(2+) concentration. At each given concentration of alginate, a critical gel concentration [Formula: see text] , was successfully determined for the first time using the Winter-Chambon criterion. The critical gel concentration [Formula: see text] was found to increase linearly with alginate concentration. At the same time, the critical relaxation exponent n decreased and the critical gel strength Sg increased linearly with alginate concentration. An improved egg-box model was proposed to describe the change in gel junction and gel network. In the stable gel state, the plateau modulus Ge of alginate gel depended on Ca(2+) concentration according to a power-law scaling, Ge=kɛ(1.5), where ɛ is the relative distance of a gelling variable (Ca(2+) concentration in this case) from the gel point ( [Formula: see text] ). The FESEM images verified the microstructure of alginate gel in which alginate chains associated into fibrils in the presence of Ca(2+) ions. The fibrillar diameter and network density increased with increasing Ca(2+) ion concentration while alginate concentration had a weak influence on fibrillar diameter.

  18. Impact of phase separation of soy protein isolate/sodium alginate co-blending mixtures on gelation dynamics and gels properties.

    PubMed

    Pan, Hongyang; Xu, Xueming; Tian, Yaoqi; Jiao, Aiquan; Jiang, Bo; Chen, Jie; Jin, Zhengyu

    2015-07-10

    The influence of sodium alginate (SA) on soy protein isolate (SPI)-based co-blending system gelling properties was studied under thermodynamic compatibility and incompatibility conditions using a direct addition (SPI/SA) or co-drying (SPI/SA-CO) process. For an SPI/SA (30:1) or SPI/SA-CO (30:1) system, the addition of too little SA did not significantly modify the SPI, and the gelation temperature (Tgel) and storage modulus (G') were similar to an SPI solution alone. For SPI/SA (20:1) and SPI/SA-CO (10:1), the Tgel and G' were between the values for solutions of SPI or SA alone; however, SPI/SA-CO (20:1) and SPI/SA-CO (10:1) gels could nearly double the equilibrium value of G' (Geq'), thus improving the barrier and mechanical properties of the final formed films. The cryo-transmission electron microscope morphology of the SPI/SA-CO (20:1) and SPI/SA-CO (10:1) systems after heating was of the core-shell type in which the core comprised SPI gel.

  19. Nitric Oxide Sensors Obtained Through the Entrapment of Iron Complexes in Sol-Gel Matrix

    DTIC Science & Technology

    2002-04-05

    iron complexes in sol-gel matrix Juliana C. Biazzotto, Jodo F. Borin, Roberto Mendonga Faria’ and Carlos F.O Graeff Departamento de Fisica e...Matemdtica-FFCLRP-USP, Av. Bandeirantes 3900, 14040-901 Ribeirdo Preto, Brazil 1-Instituto de Fisica de S~o Carlos-USP, C.P. 369, 13560-970 Sdo Carlos, Brazil

  20. Structural transition in the humic matrix of soil gels and its effect on the soil properties

    NASA Astrophysics Data System (ADS)

    Fedotov, G. N.; Shoba, S. A.; Pozdnyakov, A. I.; Puzanova, A. E.

    2014-09-01

    The analysis of drying-wetting cycles in soils has shown that the existence of the humic matrix of soil gels and, hence, the soil structure is ensured by hydrophilic bonds in dry soils and hydrophobic bonds in wet soils. This suggests that the structural transition from one mechanism controlling the stability of the soil gels and the existence of the soil structure to another mechanism occurs in the humic matrix of soil gels in a specific range of water content. The experimental results have confirmed the effect of the structural transition on the water stability of the soil structure, the pHwater, the hydrophilicity of the soil particle surface, and the structural-mechanical properties of the soils.

  1. Investigation of optical properties of anthocyanin doped into sol-gel based matrix

    NASA Astrophysics Data System (ADS)

    Hashim, Hasrina; Abdul Aziz, Nik Mohd Azmi Nik; Isnin, Aishah

    2012-06-01

    Anthocyanin dye was extracted from petal of Hibiscus rosasinensis (Bunga Raya) and doped into sol-gel based matrix to investigate an effect of pH change on its optical properties. Sol-gel matrix based on Vinyl triethoxysilene (VTES) as a precursor was prepared through Sol-gel process at pH 7. The sol was doped with 0.1% of Anthocyanin and the same amount of dye was also dissolved in ethanol as a comparative sample. Hydrochloric Acid, HCl and Tetramethylammonium Hydroxide, TMAH were used to change the pH value by adding them at various concentrations into each sample. The emission spectra and chemical structures of the samples were measured by Spectrofluorometer and Fourier Transform Infrared (FTIR) respectively. When excited at 410 nm, two emission peaks at about 492 and 574 nm were observed for Anthocyanin in acidic environment both in ethanol and VTES sol. In base environment however, only Anthocyanin dissolved in ethanol produced emission peak with a single peak at about 539 nm. The sensitivity of Anthocyanin dye toward pH changes in VTES open a possibility to use it as sensing element in which sol-gel based matrix are known to have higher mechanical strength and thermal stability.

  2. Biosorption of cadmium, lead and copper with calcium alginate xerogels and immobilized Fucus vesiculosus.

    PubMed

    Mata, Y N; Blázquez, M L; Ballester, A; González, F; Muñoz, J A

    2009-04-30

    This paper determines the effect of immobilized brown alga Fucus vesiculosus in the biosorption of heavy metals with alginate xerogels. Immobilization increased the kinetic uptakes and intraparticle diffusion rates of the three metals. The Langmuir maximum biosorption capacity increased twofold for cadmium, 10 times for lead, and decreased by half for copper. According to this model, the affinity of the metals for the biomass was as follows: Cu>Pb>Cd without alga and Pb>Cu>Cd with alga. FITR confirmed that carboxyl groups were the main groups involved in the metal uptake. Calcium in the gels was displaced by heavy metals from solution according to the "egg-box" model. The restructured gel matrix became more uniform and organized as shown by scanning electron microscopy (SEM) characterization. F. vesiculosus immobilized in alginate xerogels constitutes an excellent biosorbent for cadmium, lead and copper, sometimes surpassing the biosorption performance of alginate alone and even the free alga.

  3. A new agarose matrix for single-strand conformation polymorphism (SSCP), heteroduplex (HTX), and gel shift analyses

    SciTech Connect

    Dumais, M.M.; White, H.W.; Rashid, M.R.

    1994-09-01

    Detection of mutation, by SSCP or heteroduplex analysis, is important in medical genetics and oncology. Analysis of DNA binding proteins is a powerful tool in molecular biology research. Traditionally, these methods are performed using nondenaturing gel electrophoresis on poly-acrylamide or polyacrylamide-type matrices. Here we report the development of a new agarose gel matrix that can be used for all three methods. SSCP analyses were performed using the prototype agarose gel matrix for wild-type, polymorphic, and mutant samples from c-Kras exon 12, p53 exons 8 and 9, and HOX2B. We performed SSCP analyses using both isotopic and nonisotopic methods. We also analyzed the samples by deliberate HTX formation and subsequent gel analysis. Using the prototype agarose matrix, we detected single and multiple DNA sequence variants in 150-350 bp fragments with an efficiency comparable to polyacrylamide gels run under similar conditions. For SSCP and HTX assays, we achieved optimal resolution in gels run in vertical formats. However, some HTX samples could be resolved in horizontal gel systems. In addition, based on our studies, we have developed a useful battery of controls and standards for quality control of SSCP and HTX assays. We analyzed several different DNA/protein complexes (SP1, AP2, and octamer binding protein) using the prototype agarose matrix. We obtained good resolution in both vertical and horizontal gel formats. The horizontal gel system is generally superior for this application, due to its ease of use and slightly better resolution. This new prototype gel matrix offers an alternative for researchers performing analyses that previously could only be done on polyacrylamide-type gel matrices. For some applications, this new matrix offers the ease of horizontal gel casting. For all applications, this matrix offers the safety of a nontoxic system and the reproducibility of a thermally gelling system.

  4. Alginate: properties and biomedical applications

    PubMed Central

    Lee, Kuen Yong; Mooney, David J.

    2011-01-01

    Alginate is a biomaterial that has found numerous applications in biomedical science and engineering due to its favorable properties, including biocompatibility and ease of gelation. Alginate hydrogels have been particularly attractive in wound healing, drug delivery, and tissue engineering applications to date, as these gels retain structural similarity to the extracellular matrices in tissues and can be manipulated to play several critical roles. This review will provide a comprehensive overview of general properties of alginate and its hydrogels, their biomedical applications, and suggest new perspectives for future studies with these polymers. PMID:22125349

  5. Immobilization and characterization of the transmembrane ion channel peptide gramicidin in a sol-gel matrix

    NASA Astrophysics Data System (ADS)

    Esquembre, Rocío; Poveda, José Antonio; Mallavia, Ricardo; Mateo, C. Reyes

    2007-05-01

    Immobilization of ion channels requires of a methodology able to retain the physical properties of the lipid bilayer where their activity is performed. However, most of lipid membrane immobilization methods have been observed to alter the structural properties of the bilayers. Use of sol-gel routes seems to be an interesting alternative, although unstable liposomes were obtained when conventional sol-gel methodology was employed for immobilizing. Recently, we have suggested that use of alcohol-free sol-gel routes combined with negatively charged lipids could minimize effects exerted by host matrix on liposome structure, increasing its stability. Here we confirm this assumption by analysing the physical properties of a series of zwitterionic and anionic liposomes entrapped in a sol-gel matrix and we develop a methodology able to retain the physical properties of the lipid bilayer. This methodology has been successfully used to immobilize the transmembrane ion channel peptide gramicidin. Gramicidin was reconstituted in anionic liposomes and its immobilization was confirmed from changes observed in the photophysical properties of the tryptophan residues. Ion channel activity was determined using the fluorescent dye pyrene-1,3,6,8-tetrasulphonic acid (PTSA) and long term stability of the immobilized system was checked from steady-state fluorescence anisotropy measurements.

  6. RGD-peptide modified alginate by a chemoenzymatic strategy for tissue engineering applications.

    PubMed

    Sandvig, Ioanna; Karstensen, Kristin; Rokstad, Anne Mari; Aachmann, Finn Lillelund; Formo, Kjetil; Sandvig, Axel; Skjåk-Bræk, Gudmund; Strand, Berit Løkensgard

    2015-03-01

    One of the main challenges in tissue engineering and regenerative medicine is the ability to maintain optimal cell function and survival post-transplantation. Biomaterials such as alginates are commonly used for immunoisolation, while they may also provide structural support to the cell transplants by mimicking the extracellular matrix. In this study, arginine-glycine-aspartate (RGD)-peptide-coupled alginates of tailored composition were produced by adopting a unique chemoenzymatic strategy for substituting the nongelling mannuronic acid on the alginate. Alginates with and without RGD were produced with high and low content of G. Using carbodiimide chemistry 0.1-0.2% of the sugar units were substituted by peptide. Furthermore, the characterization by (1)H-nuclear magnetic resonance (NMR) revealed by-products from the coupling reaction that partly could be removed by coal filtration. Olfactory ensheathing cells (OECs) and myoblasts were grown in two-dimensional (2D) and 3D cultures of RGD-peptide modified or unmodified alginates obtained by the chemoenzymatically strategy and compared to native alginate. Both OECs and myoblasts adhered to the RGD-peptide modified alginates in 2D cultures, forming bipolar protrusions. OEC encapsulation resulted in cell survival for up to 9 days, thus demonstrating the potential for short-term 3D culture. Myoblasts showed long-term survival in 3D cultures, that is, up to 41 days post encapsulation. The RGD modifications did not result in marked changes in cell viability in 3D cultures. We demonstrate herein a unique technique for tailoring peptide substituted alginates with a precise and flexible composition, conserving the gel forming properties relevant for the use of alginate in tissue engineering.

  7. Silica sol-gel matrix doped with Photolon molecules for sensing and medical therapy purposes.

    PubMed

    Podbielska, Halina; Ulatowska-Jarza, Agnieszka; Müller, Gerhard; Holowacz, Iwona; Bauer, Joanna; Bindig, Uwe

    2007-11-01

    Photolon is one of the new photosensitisers that has found application in photodynamic therapy (PDT). Its chemical structure has a partially reduced porphyrin moiety and its molecular structure is comparable to chlorin e(6), which can be isolated after hydrolysis of the 5-membered exocyclic beta-ketoester moiety of pheophorbide a. For this study, a Photolon doped sol-gel matrix was produced in the form of coatings deposited on silica fibers cores. The material was produced from sols prepared from the silicate precursor TEOS mixed with ethyl alcohol. The sol-gel films were prepared with factor R=20, where R denotes the solvent-to-precursor molar ratio. Hydrochloric acid was added as a catalyst in the correct proportion to ensure acid hydrolysis (pH approximately 2). The mixture was stirred at room temperature for 4h using a magnetic stirrer (speed 400 rpm). The coated fibers were examined in different environments, liquid and gaseous, at different pH values and with various zinc cation concentrations. The chemical reactions were studied by means of spectroscopic methods, whereby the fluorescence response was studied. It was demonstrated that Photolon immobilized in a sol-gel matrix is accessible for the environment and shows visible response to the external changes. Furthermore, it was observed that these reactions are reversible. These biomaterials are also examined as carriers for PDT. It was also proved that a toxic effect is observed an environment with microorganisms, meaning that doped coatings have photodynamic activity.

  8. [Comparative evaluation of physical-mechanical properties and surface morphology of the samples of base self cured acrylic resin "Redont-kolir" polymerized in the silicone and alginate matrixes].

    PubMed

    2014-01-01

    Determination of advantages of using silicone or alginate impression material as a matrix is decisive for quality of immediate and transitional dentures manufactured by the direct method using self-cured acrylic resins. The aim of this study was a comparative evaluation of physical-mechanical properties and surface morphology of the samples of base self-cured acrylic resin "Redont-kolir" polymerized in the silicone and alginate matrix. The samples were polymerized in the C-silicone - "Zeta plus-putty" ("Zhermack", Italy) and alginate -"Ypeen" ("Spofa Dental", Czech Republic) matrixes under different regimes: 1) in the pneumopolymerizer "Averon" at an air pressure of 3 atm., a temperature of 450C for 15 minutes, and 2) polymerization in water at 450C for 15 minutes. We determined the following physical and mechanical properties: bending load, toughness, bending stress at break, hardness by Heppler, conical point of fluidity and water absorption. Electron microscopy studies of the samples have been conducted on electronic raster microscope JSM-840 ("Jeol", Japan). As a result of studies, it was found that the optimum regime of polymerization for acrylate "Redont-kolir" is in the pneumopolymerizer "Averon" at an air pressure of 3 atm., a temperature of 450 C for 15 minutes. By the results of studying the surface morphology of the samples we can draw a conclusion that the use of an alginate impression material as matrix allows to obtain a qualitatively better surface of denture. But taking into account the technological properties of the alginate impression materials, namely an expressed shrinkage, their use for this purpose must be limited by the time during which the impression matrix remain stable in size, which is specified by manufacturer's recommendations.

  9. Glucose microbiosensor based on alumina sol-gel matrix/electropolymerized composite membrane.

    PubMed

    Chen, Xiaohong; Hu, Yibai; Wilson, George S

    2002-12-01

    A procedure is described that provides co-immobilization of enzyme and bovine serum albumin (BSA) within an alumina sol-gel matrix and a polyphenol layer permselective for endogenous electroactive species. BSA has first been employed for the immobilization of glucose oxidase (GOx) on a Pt electrode in a sol-gel to produce a uniform, thin and compact film with enhanced enzyme activity. Electropolymerization of phenol was then employed to form an anti-interference and protective polyphenol film within the enzyme layer. In addition, a stability-reinforcing membrane derived from (3-aminopropyl)-trimethoxysilane was constructed by electrochemically-assisted crosslinking. This hybrid film outside the enzyme layer contributed both to the improved stability and to permselectivity. The resulting glucose sensor was characterized by a short response time (<10 s), high sensitivity (10.4 nA/mM mm(2)), low interference from endogenous electroactive species, and a working lifetime of at least 60 days.

  10. Alginate-modifying enzymes: biological roles and biotechnological uses

    PubMed Central

    Ertesvåg, Helga

    2015-01-01

    Alginate denotes a group of industrially important 1-4-linked biopolymers composed of the C-5-epimers β-D-mannuronic acid (M) and α-L-guluronic acid (G). The polysaccharide is manufactured from brown algae where it constitutes the main structural cell wall polymer. The physical properties of a given alginate molecule, e.g., gel-strength, water-binding capacity, viscosity and biocompatibility, are determined by polymer length, the relative amount and distribution of G residues and the acetyl content, all of which are controlled by alginate modifying enzymes. Alginate has also been isolated from some bacteria belonging to the genera Pseudomonas and Azotobacter, and bacterially synthesized alginate may be O-acetylated at O-2 and/or O-3. Initially, alginate is synthesized as polymannuronic acid, and some M residues are subsequently epimerized to G residues. In bacteria a mannuronan C-5-epimerase (AlgG) and an alginate acetylase (AlgX) are integral parts of the protein complex necessary for alginate polymerization and export. All alginate-producing bacteria use periplasmic alginate lyases to remove alginate molecules aberrantly released to the periplasm. Alginate lyases are also produced by organisms that utilize alginate as carbon source. Most alginate-producing organisms encode more than one mannuronan C-5 epimerase, each introducing its specific pattern of G residues. Acetylation protects against further epimerization and from most alginate lyases. An enzyme from Pseudomonas syringae with alginate deacetylase activity has been reported. Functional and structural studies reveal that alginate lyases and epimerases have related enzyme mechanisms and catalytic sites. Alginate lyases are now utilized as tools for alginate characterization. Secreted epimerases have been shown to function well in vitro, and have been engineered further in order to obtain enzymes that can provide alginates with new and desired properties for use in medical and pharmaceutical applications

  11. Encapsulation of ammonium molybdophosphate and zirconium phosphate in alginate matrix for the sorption of rubidium(I).

    PubMed

    Krys, Pawel; Testa, Flaviano; Trochimczuk, Andrzej; Pin, Christian; Taulemesse, Jean-Marie; Vincent, Thierry; Guibal, Eric

    2013-11-01

    Ammonium molybdophosphate and Phozir (alone or in combination) have been encapsulated in alginate beads for the synthesis of rubidium sorbents. SEM and SEM-EDX analyses confirm the homogeneity of the sorbents in terms of composition and metal binding. AMP sorbent is less sensitive to pH than Phozir, and optimum pH is close to pH 3 for the binding of Rb(I). The Langmuir equation fitted well sorption isotherms and the maximum sorption capacities were in the range 0.65-0.74 mmol Rb g(-1). The resistance to intraparticle diffusion contributes to control uptake kinetics (effect of particle size) though the presence of solid inorganic particles reduces the impact of drying alginate capsules (preventing the collapse of the porous structure during the drying step). Breakthrough curves demonstrate the potential of these sorbents for the dynamic sorption of Rb(I) while using ammonium chloride (combined to nitric acid) allows recovering Rb(I) from loaded sorbents.

  12. Adsorption isotherms, kinetics and thermodynamic studies towards understanding the interaction between cross-linked alginate-guar gum matrix and chymotrypsin.

    PubMed

    Woitovich Valetti, Nadia; Picó, Guillermo

    2016-02-15

    The adsorption kinetics of chymotrypsin, a pancreatic serine protease, onto an alginate-gum guar matrix cross-linked with epichlorohydrin has been performed using a batch-adsorption technique. The effect of various experimental parameters such as pH, salt presence, contact time and temperature were investigated. The pseudo-first-order and pseudo-second-order kinetic models were used to describe the kinetic data which shows that the adsorption of the enzyme followed the pseudo-second-order rate expression. The Langmuir, Freundlich and Hill adsorption isotherm models were applied to describe the equilibrium isotherms, and the isotherm constants were determined. It was found that Hill model was more suitable for our data because the isotherm data showed a sigmoidal behavior with the free enzyme concentration increasing in equilibrium. At 8°C and at pH 5.0, 1g hydrate matrix adsorbed about 7mg of chymotrypsin. In the desorption process 80% of the biological activity of chymotrypsin was recovered under the condition of 50mM phosphate buffer, pH 7.00-500mM NaCl. When successive cycles of adsorption/washing/desorption were performed, it was observed that the matrix remained functional until the fourth cycle of repeated batch enzyme adsorption. These results are important in terms of diminishing of cost and waste generation.

  13. Encapsulation and culture of mammalian cells including corneal cells in alginate hydrogels.

    PubMed

    Hunt, Nicola C; Grover, Liam M

    2013-01-01

    The potential of cell therapy for the regeneration of diseased and damaged tissues is now widely -recognized. As a consequence there is a demand for the development of novel systems that can deliver cells to a particular location, maintaining viability, and then degrade at a predictable rate to release the cells into the surrounding tissues. Hydrogels have attracted much attention in this area, as the hydrogel structure provides an environment that is akin to that of the extracellular matrix. One widely investigated hydrogel is alginate, which has been used for cell encapsulation for more than 30 years. Alginate gels have the potential to be used as 3D cell culture systems and as prosthetic materials, both are applied to regeneration of the cornea. Here, we describe an alginate-based process that has been used for encapsulation of mammalian cells including corneal cells, with high levels of viability, and which allows subsequent retrieval of cell cultures for further characterization.

  14. Extracellular Matrix Fibronectin Stimulates the Self-Assembly of Microtissues on Native Collagen Gels

    PubMed Central

    Sevilla, Carlos A.; Dalecki, Diane

    2010-01-01

    Fibronectin is an adhesive glycoprotein that is polymerized into extracellular matrices via a tightly regulated, cell-dependent process. Here, we demonstrate that fibronectin matrix polymerization induces the self-assembly of multicellular structures in vitro, termed tissue bodies. Fibronectin-null mouse embryonic fibroblasts adherent to compliant gels of polymerized type I collagen failed to spread or proliferate. In contrast, addition of fibronectin to collagen-adherent fibronectin-null mouse embryonic fibroblasts resulted in a dose-dependent increase in cell number, and induced the formation of three-dimensional (3D) multicellular structures that remained adherent and well-spread on the native collagen substrate. An extensive fibrillar fibronectin matrix formed throughout the microtissue. Blocking fibronectin matrix polymerization inhibited both cell proliferation and microtissue formation, demonstrating the importance of fibronectin fibrillogenesis in triggering cellular self-organization. Cell proliferation, tissue body formation, and tissue body shape were dependent on both fibronectin and collagen concentrations, suggesting that the relative proportion of collagen and fibronectin fibrils polymerized into the extracellular matrix influences the extent of cell proliferation and the final shape of microtissues. These data demonstrate a novel role for cell-mediated fibronectin fibrillogenesis in the formation and vertical assembly of microtissues, and provide a novel approach for engineering complex tissue architecture. PMID:20673131

  15. The endogenous fluorescence of fibroblast in collagen gels as indicator of stiffness of the extracellular matrix

    NASA Astrophysics Data System (ADS)

    Padilla-Martinez, J. P.; Ortega-Martinez, A.; Franco, W.

    2016-03-01

    The stiffness or rigidity of the extracellular matrix (ECM) regulates cell response. Established mechanical tests to measure stiffness, such as indentation and tensile tests, are invasive and destructive to the sample. Endogenous or native molecules to cells and ECM components, like tryptophan and cross-links of collagen, display fluorescence upon irradiation with ultraviolet light. Most likely, the concentration of these endogenous fluorophores changes as the stiffness of the ECM changes. In this work we investigate the endogenous fluorescence of collagen gels containing fibroblasts as a non-invasive non-destructive method to measure stiffness of the ECM. Human fibroblast cells were cultured in three-dimensional gels of type I collagen (50,000 cells/ml). This construct is a simple model of tissue contraction. During contraction, changes in the excitation-emission matrix (a fluorescence map in the 240-520/290-530 nm range) of constructs were measured with a spectrofluoremeter, and changes in stiffness were measured with a standard indentation test over 16 days. Results show that a progressive increase in fluorescence of the 290/340 nm excitation-emission pair correlates with a progressive increase in stiffness (r=0.9, α=0.5). The fluorescence of this excitation-emission pair is ascribed to tryptophan and variations in the fluorescence of this pair correlate with cellular proliferation. In this tissue model, the endogenous functional fluorescence of proliferating fibroblast cells is a biomechanical marker of stiffness of the ECM.

  16. Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation.

    PubMed

    Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

    2014-03-04

    Two mathematical models were developed for studying the effect of main fermentation temperature (TMF), immobilized cell mass (MIC) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model.

  17. Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation

    PubMed Central

    Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

    2014-01-01

    Two mathematical models were developed for studying the effect of main fermentation temperature (T MF), immobilized cell mass (M IC) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model. PMID:26019512

  18. Engineering alginate for intervertebral disc repair.

    PubMed

    Bron, Johannes L; Vonk, Lucienne A; Smit, Theodoor H; Koenderink, Gijsje H

    2011-10-01

    Alginate is frequently studied as a scaffold for intervertebral disc (IVD) repair, since it closely mimics mechanical and cell-adhesive properties of the nucleus pulposus (NP) of the IVD. The aim of this study was to assess the relation between alginate concentration and scaffold stiffness and find preparation conditions where the viscoelastic behaviour mimics that of the NP. In addition, we measured the effect of variations in scaffold stiffness on the expression of extracellular matrix molecules specific to the NP (proteoglycans and collagen) by native NP cells. We prepared sample discs of different concentrations of alginate (1%-6%) by two different methods, diffusion and in situ gelation. The stiffness increased with increasing alginate concentration, while the loss tangent (dissipative behaviour) remained constant. The diffusion samples were ten-fold stiffer than samples prepared by in situ gelation. Sample discs prepared from 2% alginate by diffusion closely matched the stiffness and loss tangent of the NP. The stiffness of all samples declined upon prolonged incubation in medium, especially for samples prepared by diffusion. The biosynthetic phenotype of native cells isolated from NPs was preserved in alginate matrices up to 4 weeks of culturing. Gene expression levels of extracellular matrix components were insensitive to alginate concentration and corresponding matrix stiffness, likely due to the poor adhesiveness of the cells to alginate. In conclusion, alginate can mimic the viscoelastic properties of the NP and preserve the biosynthetic phenotype of NP cells but certain limitations like long-term stability still have to be addressed.

  19. Human platelet lysate gel provides a novel three dimensional-matrix for enhanced culture expansion of mesenchymal stromal cells.

    PubMed

    Walenda, Gudrun; Hemeda, Hatim; Schneider, Rebekka K; Merkel, Rudolf; Hoffmann, Bernd; Wagner, Wolfgang

    2012-12-01

    Cell culture in regenerative medicine needs to facilitate efficient expansion according to good manufacturing practice requirements. Human platelet lysate (HPL) can be used as a substitute for fetal calf serum without the risk of xenogeneic immune reactions or transmission of bovine pathogens. Heparin needs to be added as anticoagulant before addition of HPL to culture medium; otherwise, HPL-medium forms a gel within 1 h. Here, we demonstrated that such HPL-gels provide a suitable 3D-matrix for cell culture that-apart from heparin-consists of the same components as the over-layered culture medium. Mesenchymal stromal cells (MSCs) grew in several layers at the interface between HPL-gel and HPL-medium without contact with any artificial biomaterials. Notably, proliferation of MSCs was much higher on HPL-gel compared with tissue culture plastic. Further, the frequency of initial fibroblastoid colony forming units (CFU-f) increased on HPL-gel. The viscous consistency of HPL-gel enabled passaging with a convenient harvesting and reseeding procedure by pipetting cells together with their HPL-matrix-this method does not require washing steps and can easily be automated. The immunophenotype and in vitro differentiation potential toward adipogenic, osteogenic, and chondrogenic lineage were not affected by culture-isolation on HPL-gel. Taken together, HPL-gel has many advantages over conventional plastic surfaces: it facilitates enhanced CFU-f outgrowth, increased proliferation rates, higher cell densities, and nonenzymatic passaging procedures for culture expansion of MSCs.

  20. Encapsulation of fluorescence vegetable extracts within a templated sol-gel matrix

    NASA Astrophysics Data System (ADS)

    Lacatusu, Ioana; Badea, Nicoleta; Nita, Rodica; Murariu, Alina; Miculescu, Florin; Iosub, Ion; Meghea, Aurelia

    2010-04-01

    The sol-gel encapsulation of labile substances with specific properties and recognition functions within robust polymer matrices remains a challenging task, despite the considerable research that has been focused on this field. Numerous studies have been reported in the field of sol-gel processes regarding different physical and chemical packing of sensitive biomolecules encapsulated in silica matrix. In this paper the classical sol-gel synthesis has been used under mild conditions in order to minimize denaturizing effects on encapsulated active vegetable extracts from flavones class. The silica templated matrix was obtained by using two types of surfactants with different alkyl chain (didodecyldimethyl-ammonium bromide and trioctadecylmetilammonium bromide) as structure-directing agents for the silicon oxide framework. An organic precursor of silicic acid (triethoxymethylsilane) has been used and it was processed by competitive hydrolysis and polycondensation reactions under controlled directions assured by the presence of oriented template. Silica materials thus obtained are used for encapsulation of two flavonoid samples containing as active principles two sources: rutin and a vegetable extract from Begonia plant. The synthesis of encapsulated nanocompounds has been achieved taking into consideration the specific interaction between the colloidal gel precursors and molecular structures of selected biomolecules. The main objective was to improve the encapsulation conditions for specific biomolecules, searching for the highest stability and functionality without loosing the quality of the flavonoid properties, particularly optical properties like fluorescence. The structural properties of the encapsulated samples have been studied by FT-IR and UV-VIS spectroscopy, thermal analysis and SEM/EDX analysis. The fluorescence experiments showed that, in the case of all four encapsulated samples, the fluorescence spectra manifest a significant increase in intensity

  1. Biopolymer gel matrix as acellular scaffold for enhanced dermal tissue regeneration.

    PubMed

    Judith, Rangasamy; Nithya, Mariappan; Rose, Chellan; Mandal, Asit Baran

    2012-07-01

    Biological grafts have drawbacks such as donor scarcity, disease transmission, tissue infection, while the scaffolds of either collagen or chitosan fabrics fail to become part of the tissue at the wound site, though they favor the formation of connective tissue matrix. This study developed a novel composite consisting of the combination of atelocollagen and chitosan in order to provide a biodegradable molecular matrix in gel form as a biomimetic surface for cell attachment, to promote the wound healing in excision wounds. We found that the topical application of biopolymer composite on the wound promoted cell proliferation, migration and collagen deposition overtime. The enhanced cellular activity in the collagen-chitosan treated wound tissue was also assed by increased levels of Platelet derived growth factor (PDGF) and Nerve growth factor (NGF) associated with elevated levels of antioxidants and decreased level of lipid peroxidation. The acellular matrix-like topical application material is designed to guide the eventual re-establishment of an anatomically normal skin. The results of this study demonstrate the feasibility of multi-cell regeneration on a molecular system that mimics tissue engineering in vivo.

  2. Effects of Matrix Metalloproteinases on the Performance of Platelet Fibrin Gel Spiked With Cardiac Stem Cells in Heart Repair

    PubMed Central

    Shen, Deliang; Tang, Junnan; Hensley, Michael Taylor; Li, Taosheng; Caranasos, Thomas George; Zhang, Tianxia

    2016-01-01

    Stem cells and biomaterials have been studied for therapeutic cardiac repair. Previous studies have shown the beneficial effects of platelet fibrin gel and cardiac stem cells when cotransplanted into rodent hearts with myocardial infarction (MI). We hypothesized that matrix metalloproteinases (MMPs) play an important role in such protection. Thus, the present study is designed to elucidate the effects of MMP inhibition on the therapeutic benefits of intramyocardial injection of platelet fibrin gel spiked with cardiac stem cells (cell-gel) in a rat model of acute MI. In vitro, broad-spectrum MMP inhibitor GM6001 undermines cell spreading and cardiomyocyte contraction. In a syngeneic rat model of myocardial infarction, MMP inhibition blunted the recruitment of endogenous cardiovascular cells into the injected biomaterials, therefore hindering de novo angiogenesis and cardiomyogenesis. Echocardiography and histology 3 weeks after treatment revealed that metalloproteinase inhibition diminished the functional and structural benefits of cell-gel in treating MI. Reduction of host angiogenesis, cardiomyocyte cycling, and MMP-2 activities was evident in animals treated with GM6001. Our findings suggest that MMPs play a critical role in the therapeutic benefits of platelet fibrin gel spiked with cardiac stem cells for treating MI. Significance In this study, the effects of matrix metalloproteinase inhibition on the performance of platelet gel spiked with cardiac stem cells (cell-gel) for heart regeneration are explored. The results demonstrate that matrix metalloproteinases are required for cell-gel to exert its benefits in cardiac repair. Inhibition of matrix metalloproteinases reduces cell engraftment, host angiogenesis, and recruitment of endogenous cardiovascular cells in rats with heart attack. PMID:27112177

  3. Spectrofluorimetric assessment of hydrochlorothiazide using optical sensor nano-composite terbium ion doped in sol-gel matrix.

    PubMed

    Youssef, A O

    2012-05-01

    A new, simple, sensitive and selective spectrofluorimetric method for the determination of Hydrochlorothiazide was developed in acetonitrile at pH 6.2. The Hydrochlorothiazide can remarkably enhance the luminescence intensity of the Tb(3+) ion doped in sol-gel matrix at λ(ex) = 370 nm. The intensity of the emission band of Tb(3+) ion doped in sol-gel matrix was increased due to the energy transfer from the triplet excited state of Hydrochlorothiazide to ((5)D(4)) excited energy state of Tb(3) ion. The enhancement of the emission band of Tb(3+) ion doped in sol-gel matrix at ((5)D(4)→(7)F(5)) 545 nm was directly proportion to the concentration of Hydrochlorothiazide with a dynamic ranges of 5.0 × 10(-10)-5.0 × 10(-6) mol L(-1) and detection limit of 2.2 × 10(-11) mol L(-1).

  4. Development of functionalized multi-walled carbon-nanotube-based alginate hydrogels for enabling biomimetic technologies

    NASA Astrophysics Data System (ADS)

    Joddar, Binata; Garcia, Eduardo; Casas, Atzimba; Stewart, Calvin M.

    2016-08-01

    Alginate is a hydrogel commonly used for cell culture by ionically crosslinking in the presence of divalent Ca2+ ions. However these alginate gels are mechanically unstable, not permitting their use as scaffolds to engineer robust biological bone, breast, cardiac or tumor tissues. This issue can be addressed via encapsulation of multi-walled carbon nanotubes (MWCNT) serving as a reinforcing phase while being dispersed in a continuous phase of alginate. We hypothesized that adding functionalized MWCNT to alginate, would yield composite gels with distinctively different mechanical, physical and biological characteristics in comparison to alginate alone. Resultant MWCNT-alginate gels were porous, and showed significantly less degradation after 14 days compared to alginate alone. In vitro cell-studies showed enhanced HeLa cell adhesion and proliferation on the MWCNT-alginate compared to alginate. The extent of cell proliferation was greater when cultured atop 1 and 3 mg/ml MWCNT-alginate; although all MWCNT-alginates lead to enhanced cell cluster formation compared to alginate alone. Among all the MWCNT-alginates, the 1 mg/ml gels showed significantly greater stiffness compared to all other cases. These results provide an important basis for the development of the MWCNT-alginates as novel substrates for cell culture applications, cell therapy and tissue engineering.

  5. Development of functionalized multi-walled carbon-nanotube-based alginate hydrogels for enabling biomimetic technologies

    PubMed Central

    Joddar, Binata; Garcia, Eduardo; Casas, Atzimba; Stewart, Calvin M.

    2016-01-01

    Alginate is a hydrogel commonly used for cell culture by ionically crosslinking in the presence of divalent Ca2+ ions. However these alginate gels are mechanically unstable, not permitting their use as scaffolds to engineer robust biological bone, breast, cardiac or tumor tissues. This issue can be addressed via encapsulation of multi-walled carbon nanotubes (MWCNT) serving as a reinforcing phase while being dispersed in a continuous phase of alginate. We hypothesized that adding functionalized MWCNT to alginate, would yield composite gels with distinctively different mechanical, physical and biological characteristics in comparison to alginate alone. Resultant MWCNT-alginate gels were porous, and showed significantly less degradation after 14 days compared to alginate alone. In vitro cell-studies showed enhanced HeLa cell adhesion and proliferation on the MWCNT-alginate compared to alginate. The extent of cell proliferation was greater when cultured atop 1 and 3 mg/ml MWCNT-alginate; although all MWCNT-alginates lead to enhanced cell cluster formation compared to alginate alone. Among all the MWCNT-alginates, the 1 mg/ml gels showed significantly greater stiffness compared to all other cases. These results provide an important basis for the development of the MWCNT-alginates as novel substrates for cell culture applications, cell therapy and tissue engineering. PMID:27578567

  6. Zirconium carbonitride pellets by internal sol gel and spark plasma sintering as inert matrix fuel material

    NASA Astrophysics Data System (ADS)

    Hedberg, Marcus; Cologna, Marco; Cambriani, Andrea; Somers, Joseph; Ekberg, Christian

    2016-10-01

    Inert matrix fuel is a fuel type where the fissile material is blended with a solid diluent material. In this work zirconium carbonitride microspheres have been produced by internal sol gel technique, followed by carbothermal reduction. Material nitride purities in the produced materials ranged from Zr(N0.45C0.55) to Zr(N0.74C0.26) as determined by X-ray diffraction and application of Vegard's law. The zirconium carbonitride microspheres have been pelletized by spark plasma sintering (SPS) and by conventional cold pressing and sintering. In all SPS experiments cohesive pellets were formed. Maximum final density reached by SPS at 1700 °C was 87% theoretical density (TD) compared to 53% TD in conventional sintering at 1700 °C. Pore sizes in all the produced pellets were in the μm scale and no density gradients could be observed by computer tomography.

  7. Photoconductivity and stabilization of dopamine embedded in sol-gel TiO2 matrix

    NASA Astrophysics Data System (ADS)

    Prado-Prone, Gina; Valverde-Aguilar, Guadalupe; García-Macedo, Jorge; Vergara-Aragón, P.

    2012-09-01

    This work reports the synthesis of amorphous TiO2 matrix by sol-gel method at atmospheric conditions. DA was encapsulated in a TiO2 matrix to reduce its chemical instability. To TiO2/DA sample was added the 15C5 to diminish the oxidation process. The stabilization process was followed by absorption spectra, colour change and infrared spectroscopy. Oxidation processes of the DA were identified by the presence of DA-quinone and DA-chrome. The TiO2/DA complex retarded the oxidation process for 30 days, while the TiO2/DA/15C5 complex this period was extended for 47 days. Photoconductivity studies were performed on both kinds of samples to analyze their charge transports. The experimental data were fitted with straight lines at darkness and under illumination at 320 nm, 400 nm, and 515 nm. This indicates an ohmic behavior. Transport parameters were calculated. The conductive effect is stronger under darkness than under illumination at 320 nm because the oxidation process in the darkness is less intense than under illumination.

  8. Optimizing the calcium content of a copolymer acrylamide gel matrix for dark-grown seedlings

    NASA Technical Reports Server (NTRS)

    Myers, P. N.; Mitchell, C. A.

    1998-01-01

    A copolymer acrylamide acrylate gel was investigated as the sole root matrix for dark-grown seedlings of soybean (Glycine max Merr. 'Century 84'). Increasing Ca2+ in the hydrating solution of the hydrogel from 1 to 10 mM decreased its water-holding capacity from 97 to 46 mL g-1, yet water potential of the medium remained high, sufficient for normal plant growth at all Ca2+ concentrations tested. Elongation rate of dark-grown soybean seedlings over a 54-hour period was 0.9, 1.5, and 1.8 mm h-1 with 1.0, 2.5, or 5.0 mM Ca2+, respectively, but did not increase with further increases in Ca2+ concentration. Further study revealed that Na+ was released from the hydrogel medium and was taken up by the seedlings as Ca2+ increased in the medium. In dry hypocotyl tissue, sodium content correlated negatively with calcium content. Despite the presence of Na+ in the hydrogel, seedling growth was normal when adequate Ca2+ was added in the hydrating solution. Acrylamide hydrogels hold good potential as a sole growth matrix for short-term experiments with dark-grown seedlings without irrigation.

  9. Interpenetrated Si-HPMC/alginate hydrogels as a potential scaffold for human tissue regeneration.

    PubMed

    Viguier, Alexia; Boyer, Cecile; Chassenieux, Christophe; Benyahia, Lazhar; Guicheux, Jérôme; Weiss, Pierre; Rethore, Gildas; Nicolai, Taco

    2016-05-01

    Interpenetrated gels of biocompatible polysaccharides alginate and silanized hydroxypropyl methyl cellulose (Si-HPMC) have been studied in order to assess their potential as scaffolds for the regeneration of human tissues. Si-HPMC networks were formed by reduction of the pH to neutral and alginate networks were formed by progressive in situ release of Ca(2+). Linear and non-linear mechanical properties of the mixed gels at different polymer and calcium concentrations were compared with those of the corresponding single gels. The alginate/Si-HPMC gels were found to be stiffer than pure Si-HPMC gels, but weaker and more deformable than pure alginate gels. No significant difference was found for the maximum stress at rupture measured during compression for all these gels. The degrees of swelling or contraction in excess water at pH 7 as well as the release of Ca(2+) was measured as a function of time. Pure alginate gels contracted by as much as 50 % and showed syneresis, which was much reduced or even eliminated for mixed gels. The important release of Ca(2+) upon ageing for pure alginate gels was much reduced for the mixed gels. Furthermore, results of cytocompatibility assays indicated that there was no cytotoxicity of Si-HPMC/alginate hydrogels in 2D and 3D culture of human SW1353 cells. The results show that using interpenetrated Si-HPMC/alginate gels has clear advantages over the use of single gels for application in tissue regeneration.

  10. Delaying cluster growth of ionotropic induced alginate gelation by oligoguluronate.

    PubMed

    Padoł, Anna Maria; Maurstad, Gjertrud; Draget, Kurt Ingar; Stokke, Bjørn Torger

    2015-11-20

    Alginates form gels in the presence of various divalent ions, such as Ca(2+) that mediate lateral association of chain segments. Various procedures exist that introduce Ca(2+) to yield alginate hydrogels with overall homogeneous or controlled gradients in the concentration profiles. In the present study, the effect of adding oligomers of α-l-guluronic acid (oligoGs) to gelling solutions of alginate was investigated by determination of the cluster growth stimulated by in situ release of Ca(2+). Three different alginate samples varying in fraction of α-l-guluronic acid and molecular weights were employed. The cluster growth was determined for both pure alginates and alginates with two different concentrations of the oligoGs employing dynamic light scattering. The results show that addition of oligoG slows down the cluster growth, the more efficient for the alginates with higher fraction of α-l-guluronic acid, and the higher molecular weight. The efficiency in delaying and slowing the cluster growth induced by added oligoG were discussed in view of the molecular parameters of the alginates. These results show that oligoG can be added to alginate solutions to control the cluster growth and eventually also transition to the gel state. Quantitative relation between the concentration of added oligoG, type and molecular weight of the alginate, and concentration, can be employed as guidelines in tuning alginate cluster growth with specific properties.

  11. Designing Solvent Exchange-Induced In Situ Forming Gel from Aqueous Insoluble Polymers as Matrix Base for Periodontitis Treatment.

    PubMed

    Srichan, Tharatree; Phaechamud, Thawatchai

    2017-01-01

    An in situ forming gel is a dosage form which is promised for site-specific therapy such as periodontal pocket of periodontitis treatment. Ethylcellulose, bleached shellac, and Eudragit RS were applied in this study as a polymeric matrix for in situ forming gel employing N-methyl pyrrolidone (NMP) as solvent. Solutions comprising ethylcellulose, bleached shellac, and Eudragit RS in NMP were evaluated for viscosity, rheology, and rate of water penetration. Ease of administration by injection was determined as the force required to expel polymeric solutions through a needle using texture analyzer. In vitro gel formation and in vitro gel degradation were conducted after injection into phosphate buffer solution pH 6.8. Ethylcellulose, bleached shellac, and Eudragit RS could form the in situ gel, in vitro. Gel viscosity and pH value depended on percentage amount of the polymer, whereas the water diffusion at early period likely relied on types of polymer. Furthermore, the solutions containing higher polymer concentration exhibited the lower degree of degradation. All the preparations were acceptable as injectable dosage forms because the applied force was lower than 50 N. All of them inhibited Staphylococcus aureus, Escherichia coli, Candida albicans, Streptococcus mutans, and Porphyrommonas gingivalis growth owing to antimicrobial activity of NMP which exhibited a potential use for periodontitis treatment. Moreover, the developed systems presented as the solvent exchange induced in situ forming gel and showed capability to be incorporated with the suitable antimicrobial active compounds for periodontitis treatment which should be further studied.

  12. 3D porous sol-gel matrix incorporated microdevice for effective large volume cell sample pretreatment.

    PubMed

    Lee, Chan Joo; Jung, Jae Hwan; Seo, Tae Seok

    2012-06-05

    In this study, we demonstrated an effective sample pretreatment microdevice that could perform the capture, purification, and release of pathogenic bacteria with a large-volume sample and at a high speed and high-capture yield. We integrated a sol-gel matrix into the microdevice which forms three-dimensional (3D) micropores for the cell solution to pass through and provides a large surface area for the immobilization of antibodies to capture the target Staphylococcus aureus (S. aureus) cells. The antibody was linked to the surface of the sol-gel via a photocleavable linker, allowing the cell-captured antibody moiety to be released by UV irradiation. In addition to the optimization of the antibody immobilization and UV cleavage processes, the cell-capture efficiency was maximized by controlling the sample flow rate with a pumping scheme (3 steps, 5 steps: 3 steps with one flutter step, 7 steps: 3 steps with two flutter steps) and the pumping time (100, 200, and 300 ms). A quantitative capture analysis was performed by targeting a specific gene site of protein A of S. aureus in real-time PCR (RT-PCR). While the 3-step process with an actuation time of 100 ms showed the fastest flow rate (1 mL sample processing time in 10 min), the pumping scheme with the 7-step process and the 300 ms actuation time revealed the highest cell-capture efficiency. A limit of detection study with the 7-step and the 300 ms pumping scheme demonstrated that 100 cells per 100 μL were detected with a 70% yield, and even a single cell could be analyzed via on-chip sample preparation. Thus, our novel sol-gel based microdevice was proven more cost-effective, simple, and efficient in terms of its sample pretreatment ability compared to the use of a conventional 2D flat microdevice. This proposed sample pretreatment device can be further incorporated to an analytical functional unit to realize a micrototal analysis system (μTAS) with sample-in-answer-out capability in the fields of biomedical

  13. Efficient functionalization of alginate biomaterials.

    PubMed

    Dalheim, Marianne Ø; Vanacker, Julie; Najmi, Maryam A; Aachmann, Finn L; Strand, Berit L; Christensen, Bjørn E

    2016-02-01

    Peptide coupled alginates obtained by chemical functionalization of alginates are commonly used as scaffold materials for cells in regenerative medicine and tissue engineering. We here present an alternative to the commonly used carbodiimide chemistry, using partial periodate oxidation followed by reductive amination. High and precise degrees of substitution were obtained with high reproducibility, and without formation of by-products. A protocol was established using l-Tyrosine methyl ester as a model compound and the non-toxic pic-BH3 as the reducing agent. DOSY was used to indirectly verify covalent binding and the structure of the product was further elucidated using NMR spectroscopy. The coupling efficiency was to some extent dependent on alginate composition, being most efficient on mannuronan. Three different bioactive peptide sequences (GRGDYP, GRGDSP and KHIFSDDSSE) were coupled to 8% periodate oxidized alginate resulting in degrees of substitution between 3.9 and 6.9%. Cell adhesion studies of mouse myoblasts (C2C12) and human dental stem cells (RP89) to gels containing various amounts of GRGDSP coupled alginate demonstrated the bioactivity of the material where RP89 cells needed higher peptide concentrations to adhere.

  14. Enhancement in dose sensitivity of polymer gel dosimeters composed of radiation-crosslinked gel matrix and less toxic monomers

    NASA Astrophysics Data System (ADS)

    Hiroki, A.; Yamashita, S.; Taguchi, M.

    2015-01-01

    Polymer gel dosimeters based on radiation-crosslinked hydroxypropyl cellulose gel were prepared, which comprised 2-hydroxyethyl methacrylate (HEMA) and polyethylene glycol #400 dimethacrylate (9G) as less toxic monomers and tetrakis (hydroxymethyl) phosphonium chloride (THPC) as an antioxidant. The dosimeters exposed to 60Co γ-rays became cloudy at only 1 Gy. The irradiated dosimeters were optically analyzed by using a UV- vis spectrophotometer to evaluate dose response. Absorbance of the dosimeters linearly increased in the dose range from 0 to 10 Gy, in which dose sensitivity increased with increasing 9G concentration. The dose sensitivity of the dosimeters with 2 wt% HEMA and 3 wt% 9G was also enhanced by increment in THPC.

  15. Alginate hydrogel-mediated crystallization of calcium carbonate

    SciTech Connect

    Ma, Yufei; Feng, Qingling

    2011-05-15

    We documented a specific method for combining calcium ions and alginate molecules slowly and continuously in the mineralization system for the purpose of understanding the mediating function of alginate on the crystallization of calcium carbonate. The alginate was involved in the nucleation and the growth process of CaCO{sub 3}. The crystal size, morphology and roughness of crystal surface were significantly influenced by the type of the alginate, which could be accounted for by the length of the G blocks in alginate. A combination of Fourier transform infrared spectroscopy and thermogravimetric analysis showed that there were the chemical interactions between the alginate and the mineral phase. This strategic approach revealed the biologically controlled CaCO{sub 3} mineralization within calcium alginate hydrogels via the selective nucleation and the confined crystallization of CaCO{sub 3}. The results presented here could contribute to the understanding of the mineralization process in hydrogel systems. -- Graphical abstract: Schematic illustration of the growth of calcite aggregates with different morphologies obtained from (a) Low G alginate gels and (b) High G alginate gels. Display Omitted highlights: > We use a specific method for combining calcium ions and alginate molecules slowly and continuously in the mineralization system to understand the mediating function of alginate on the crystallization of CaCO{sub 3} crystals. > The crystal size, morphology and crystal surface roughness are influenced by the length of G blocks in alginate. There are chemical interactions between the alginate and the mineral phase. > We propose a potential mechanism of CaCO{sub 3} crystallization within High G and Low G calcium alginate hydrogel.

  16. Concentric gel system to study the biophysical role of matrix microenvironment on 3D cell migration.

    PubMed

    Kurniawan, Nicholas Agung; Chaudhuri, Parthiv Kant; Lim, Chwee Teck

    2015-04-03

    The ability of cells to migrate is crucial in a wide variety of cell functions throughout life from embryonic development and wound healing to tumor and cancer metastasis. Despite intense research efforts, the basic biochemical and biophysical principles of cell migration are still not fully understood, especially in the physiologically relevant three-dimensional (3D) microenvironments. Here, we describe an in vitro assay designed to allow quantitative examination of 3D cell migration behaviors. The method exploits the cell's mechanosensing ability and propensity to migrate into previously unoccupied extracellular matrix (ECM). We use the invasion of highly invasive breast cancer cells, MDA-MB-231, in collagen gels as a model system. The spread of cell population and the migration dynamics of individual cells over weeks of culture can be monitored using live-cell imaging and analyzed to extract spatiotemporally-resolved data. Furthermore, the method is easily adaptable for diverse extracellular matrices, thus offering a simple yet powerful way to investigate the role of biophysical factors in the microenvironment on cell migration.

  17. Concentric Gel System to Study the Biophysical Role of Matrix Microenvironment on 3D Cell Migration

    PubMed Central

    Kurniawan, Nicholas Agung; Chaudhuri, Parthiv Kant; Lim, Chwee Teck

    2015-01-01

    The ability of cells to migrate is crucial in a wide variety of cell functions throughout life from embryonic development and wound healing to tumor and cancer metastasis. Despite intense research efforts, the basic biochemical and biophysical principles of cell migration are still not fully understood, especially in the physiologically relevant three-dimensional (3D) microenvironments. Here, we describe an in vitro assay designed to allow quantitative examination of 3D cell migration behaviors. The method exploits the cell’s mechanosensing ability and propensity to migrate into previously unoccupied extracellular matrix (ECM). We use the invasion of highly invasive breast cancer cells, MDA-MB-231, in collagen gels as a model system. The spread of cell population and the migration dynamics of individual cells over weeks of culture can be monitored using live-cell imaging and analyzed to extract spatiotemporally-resolved data. Furthermore, the method is easily adaptable for diverse extracellular matrices, thus offering a simple yet powerful way to investigate the role of biophysical factors in the microenvironment on cell migration. PMID:25867104

  18. The effect of an autologous cellular gel-matrix integrated implant system on wound healing

    PubMed Central

    2010-01-01

    Background This manuscript reports the production and preclinical studies to examine the tolerance and efficacy of an autologous cellular gel-matrix integrated implant system (IIS) aimed to treat full-thickness skin lesions. Methods The best concentration of fibrinogen and thrombin was experimentally determined by employing 28 formula ratios of thrombin and fibrinogen and checking clot formation and apparent stability. IIS was formed by integrating skin cells by means of the in situ gelification of fibrin into a porous crosslinked scaffold composed of chitosan, gelatin and hyaluronic acid. The in vitro cell proliferation within the IIS was examined by the MTT assay and PCNA expression. An experimental rabbit model consisting of six circular lesions was utilized to test each of the components of the IIS. Then, the IIS was utilized in an animal model to cover a 35% body surface full thickness lesion. Results The preclinical assays in rabbits demonstrated that the IIS was well tolerated and also that IIS-treated rabbit with lesions of 35% of their body surface, exhibited a better survival rate (p = 0,06). Conclusion IIS should be further studied as a new wound dressing which shows promising properties, being the most remarkable its good biological tolerance and cell growth promotion properties. PMID:20565787

  19. Design and Fabrication of Anatomical Bioreactor Systems Containing Alginate Scaffolds for Cartilage Tissue Engineering

    PubMed Central

    Gharravi, Anneh Mohammad; Orazizadeh, Mahmoud; Ansari-Asl, Karim; Banoni, Salem; Izadi, Sina; Hashemitabar, Mahmoud

    2012-01-01

    The aim of the present study was to develop a tissue-engineering approach through alginate gel molding to mimic cartilage tissue in a three-dimensional culture system. The perfusion biomimetic bioreactor was designed to mimic natural joint. The shear stresses exerting on the bioreactor chamber were calculated by Computational Fluid Dynamic (CFD). Several alginate/bovine chondrocyte constructs were prepared, and were cultured in the bioreactor. Histochemical and immunohistochemical staining methods for the presence of glycosaminoglycan(GAG), overall matrix production and type II collagen protein were performed, respectively. The dynamic mechanical device applied a linear mechanical displacement of 2 mm to 10 mm. The CFD modeling indicated peak velocity and maximum wall shear stress were 1.706×10−3 m/s and 0.02407 dyne/cm 2, respectively. Histochemical and immunohistochemical analysis revealed evidence of cartilage-like tissue with lacunas similar to those of natural cartilage and the production of sulfated GAG of matrix by the chondrons, metachromatic territorial matrix-surrounded cells and accumulation of type II collagen around the cells. The present study indicated that when chondrocytes were seeded in alginate hydrogel and cultured in biomimetic cell culture system, cells survived well and secreted newly synthesized matrix led to improvement of chondrogenesis. PMID:23408660

  20. Matrix molecularly imprinted mesoporous sol-gel sorbent for efficient solid-phase extraction of chloramphenicol from milk.

    PubMed

    Samanidou, Victoria; Kehagia, Maria; Kabir, Abuzar; Furton, Kenneth G

    2016-03-31

    Highly selective and efficient chloramphenicol imprinted sol-gel silica based inorganic polymeric sorbent (sol-gel MIP) was synthesized via matrix imprinting approach for the extraction of chloramphenicol in milk. Chloramphenicol was used as the template molecule, 3-aminopropyltriethoxysilane (3-APTES) and triethoxyphenylsilane (TEPS) as the functional precursors, tetramethyl orthosilicate (TMOS) as the cross-linker, isopropanol as the solvent/porogen, and HCl as the sol-gel catalyst. Non-imprinted sol-gel polymer (sol-gel NIP) was synthesized under identical conditions in absence of template molecules for comparison purpose. Both synthesized materials were characterized by Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FT-IR) and nitrogen adsorption porosimetry, which unambiguously confirmed their significant structural and morphological differences. The synthesized MIP and NIP materials were evaluated as sorbents for molecularly imprinted solid phase extraction (MISPE) of chloramphenicol in milk. The effect of critical extraction parameters (flow rate, elution solvent, sample and eluent volume, selectivity coefficient, retention capacity) was studied in terms of retention and desorption of chloramphenicol. Competition and cross reactivity tests have proved that sol-gel MIP sorbent possesses significantly higher specific retention and enrichment capacity for chloramphenicol compared to its non-imprinted analogue. The maximum imprinting factor (IF) was found as 9.7, whereas the highest adsorption capacity of chloramphenicol by sol-gel MIP was 23 mg/g. The sol-gel MIP was found to be adequately selective towards chloramphenicol to provide the necessary minimum required performance limit (MRPL) of 0.3 μg/kg set forth by European Commission after analysis by LC-MS even without requiring time consuming solvent evaporation and sample reconstitution step, often considered as an integral part in solid phase extraction work-flow. Intra and

  1. Structure and dynamics of spin-labeled insulin entrapped in a silica matrix by the sol-gel method.

    PubMed

    Vanea, E; Gruian, C; Rickert, C; Steinhoff, H-J; Simon, V

    2013-08-12

    The structure and conformational dynamics of insulin entrapped into a silica matrix was monitored during the sol to maturated-gel transition by electron paramagnetic resonance (EPR) spectroscopy. Insulin was successfully spin-labeled with iodoacetamide and the bifunctional nitroxide reagent HO-1944. Room temperature continuous wave (cw) EPR spectra of insulin were recorded to assess the mobility of the attached spin labels. Insulin conformation and its distribution within the silica matrix were studied using double electron-electron resonance (DEER) and low-temperature cw-EPR. A porous oxide matrix seems to form around insulin molecules with pore diameters in the order of a few nanometers. Secondary structure of the encapsulated insulin investigated by Fourier transform infrared spectroscopy proved a high structural integrity of insulin even in the dried silica matrix. The results show that silica encapsulation can be used as a powerful tool to effectively isolate and functionally preserve biomolecules during preparation, storage, and release.

  2. Biocompatibility of mannuronic acid-rich alginates.

    PubMed

    Klöck, G; Pfeffermann, A; Ryser, C; Gröhn, P; Kuttler, B; Hahn, H J; Zimmermann, U

    1997-05-01

    Highly purified algin preparations free of adverse contaminants with endotoxins and other mitogens recently became available by a new purification process (Klöck et al., Appl. Microbiol. Biotechnol., 1994, 40, 638-643). An advantage of this purification protocol is that it can be applied to alginates with various ratios of mannuronic acid to guluronic acid. High mannuronic acid alginate capsules are of particular practical interest for cell transplantation and for biohybrid organs, because mannuronate-rich alginates are usually less viscous, allowing one to make gels with a higher alginate content. This will increase their stability and reduce the diffusion permeability and could therefore protect immobilized cells more efficiently against the host immune system. Here we report the biocompatibility of purified, mannuronic acid-rich alginate (68% mannuronate residues) in a series of in vitro, as well as in vivo, assays. In contrast to raw alginate extracts, the purified product showed no mitogenic activity towards murine lymphocytes in vitro. Its endotoxin content was reduced to the level of the solvent. Animal studies with these new, purified algin formulations revealed the absence of a mitogen-induced foreign body reaction, even when the purified material (after cross-linking with Ba2+ ions) is implanted into animal models with elevated macrophage activity (diabetes-prone BB/OK rat). Thus, alginate capsules with high mannuronic acid content become available for applications such as implantation. In addition to the utilization as implantable cell reactors in therapy and biotechnology, these purified algins have broad application potential as ocular fillings, tissue replacements, microencapsulated growth factors and/or interleukins or slow-release dosage forms of antibodies, surface coatings of sensors and other invasive medical devices, and in encapsulation of genetically engineered cells for gene therapy.

  3. On-bead expression of recombinant proteins in an agarose gel matrix coated on a glass slide.

    PubMed

    Lee, Kyung-Ho; Lee, Ka-Young; Byun, Ju-Young; Kim, Byung-Gee; Kim, Dong-Myung

    2012-05-07

    A system for expression and in situ display of recombinant proteins on a microbead surface is described. Biotinylated PCR products were immobilized on microbead surfaces, which were then embedded in a gel matrix and supplied with translation machinery and substrates. Upon the incubation of the gel matrix, target proteins encoded on the bead-immobilized DNA were expressed and captured on the same bead, thus allowing bead-mediated linkage of DNA and encoded proteins. The new method combines the simplicity and convenience of solid-phase separation of genetic information with the benefits of cell-free protein synthesis, such as instant translation of genetic information, unrestricted substrate accessibility and flexible assay configuration design.

  4. Degradation potential of protocatechuate 3,4-dioxygenase from crude extract of Stenotrophomonas maltophilia strain KB2 immobilized in calcium alginate hydrogels and on glyoxyl agarose.

    PubMed

    Guzik, Urszula; Hupert-Kocurek, Katarzyna; Krysiak, Marta; Wojcieszyńska, Danuta

    2014-01-01

    Microbial intradiol dioxygenases have been shown to have a great potential for bioremediation; however, their structure is sensitive to various environmental and chemical agents. Immobilization techniques allow for the improvement of enzyme properties. This is the first report on use of glyoxyl agarose and calcium alginate as matrixes for the immobilization of protocatechuate 3,4-dioxygenase. Multipoint attachment of the enzyme to the carrier caused maintenance of its initial activity during the 21 days. Immobilization of dioxygenase in calcium alginate or on glyoxyl agarose resulted in decrease in the optimum temperature by 5 °C and 10 °C, respectively. Entrapment of the enzyme in alginate gel shifted its optimum pH towards high-alkaline pH while immobilization of the enzyme on glyoxyl agarose did not influence pH profile of the enzyme. Protocatechuate 3,4-dioygenase immobilized in calcium alginate showed increased activity towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate. Slightly lower activity of the enzyme was observed after its immobilization on glyoxyl agarose. Entrapment of the enzyme in alginate gel protected it against chelators and aliphatic alcohols while its immobilization on glyoxyl agarose enhanced enzyme resistance to inactivation by metal ions.

  5. Nonlinear behavior of ionically and covalently cross-linked alginate hydrogels

    NASA Astrophysics Data System (ADS)

    Hashemnejad, Seyedmeysam; Zabet, Mahla; Kundu, Santanu

    2015-03-01

    Gels deform differently under applied load and the deformation behavior is related to their network structures and environmental conditions, specifically, strength and density of crosslinking, polymer concentration, applied load, and temperature. Here, we investigate the mechanical behavior of both ionically and covalent cross-linked alginate hydrogel using large amplitude oscillatory shear (LAOS) and cavitation experiments. Ionically-bonded alginate gels were obtained by using divalent calcium. Alginate volume fraction and alginate to calcium ratio were varied to obtain gels with different mechanical properties. Chemical gels were synthesized using adipic acid dihdrazide (AAD) as a cross-linker. The non-linear rheological parameters are estimated from the stress responses to elucidate the strain softening behavior of these gels. Fracture initiation and propagation mechanism during shear rheology and cavitation experiments will be presented. Our results provide a better understanding on the deformation mechanism of alginate gel under large-deformation.

  6. Effect of gel structure of matrix orientation in pulsed alternating electric fields

    SciTech Connect

    Stellwagen, N.C.; Stellwagen, J.

    1993-12-31

    Four polymeric gels with different structures, LE agarose, HEEO agarose, beta-carrageenan, and polyacrylamide, were studied by transient electric birefringence to determine the importance of various structural features on the orientation of the gels in pulsed alternating electric fields. The birefrigence relaxation times observed for agarose gels in low voltage electric fields suggest that long fibers and/or domains, ranging up to tens of microns in size, are oriented by the electric field. The sign of the birefringence reverses when the direction of the electric field is reversed, suggesting that the oriented domains change their direction of orientation from parallel to perpendicular (or vice versa) when the polarity of the electric field is reversed. These anamalous orientation effects are observed with both types of agarose gels, but not with beta-carrageenan or polyacrylamide gels, suggesting that the alternating D,L galactose residues in the agarose backbone are responsible for the anomalies.

  7. Combined of ultrasound irradiation with high hydrostatic pressure (US/HHP) as a new method to improve immobilization of dextranase onto alginate gel.

    PubMed

    Bashari, Mohanad; Abbas, Shabbar; Xu, Xueming; Jin, Zhengyu

    2014-07-01

    In this research work, dextranase was immobilized onto calcium alginate beads by the combination of ultrasonic irradiation and high hydrostatic pressure (US/HHP) treatments. Effects of US/HHP treatments on loading efficiency and immobilization yield of dextranase enzyme onto calcium alginate beads were investigated. Furthermore, the activities of immobilized enzymes prepared with and without US/HHP treatments and that prepared with ultrasonic irradiation (US) and high hydrostatic pressure (HHP), as a function of pH, temperature, recyclability and enzyme kinetic parameters, were compared with that for free enzyme. The maximum loading efficiency and the immobilization yield were observed when the immobilized dextranase was prepared with US (40 W at 25 kHz for 15 min) combined with HHP (400 MPa for 15 min), under which the loading efficiency and the immobilization yield increased by 88.92% and 80.86%, respectively, compared to immobilized enzymes prepared without US/HHP treatment. On the other hand, immobilized enzyme prepared with US/HHP treatment showed Vmax, KM, catalytic and specificity constants values higher than that for the immobilized enzyme prepared with HHP treatment, indicated that, this new US/HHP method improved the catalytic kinetics activity of immobilized dextranase at all the reaction conditions studied. Compared to immobilized enzyme prepared either with US or HHP, the immobilized enzymes prepared with US/HHP method exhibited a higher: pH optimum, optimal reaction temperature, thermal stability and recyclability, and lower activation energy, which, illustrating the effectiveness of the US/HHP method. These results indicated that, the combination of US and HHP treatments could be an effective method for improving the immobilization of enzymes in polymers.

  8. Alginate composites for bone tissue engineering: a review.

    PubMed

    Venkatesan, Jayachandran; Bhatnagar, Ira; Manivasagan, Panchanathan; Kang, Kyong-Hwa; Kim, Se-Kwon

    2015-01-01

    Bone is a complex and hierarchical tissue consisting of nano hydroxyapatite and collagen as major portion. Several attempts have been made to prepare the artificial bone so as to replace the autograft and allograft treatment. Tissue engineering is a promising approach to solve the several issues and is also useful in the construction of artificial bone with materials including polymer, ceramics, metals, cells and growth factors. Composites consisting of polymer-ceramics, best mimic the natural functions of bone. Alginate, an anionic polymer owing enormous biomedical applications, is gaining importance particularly in bone tissue engineering due to its biocompatibility and gel forming properties. Several composites such as alginate-polymer (PLGA, PEG and chitosan), alginate-protein (collagen and gelatin), alginate-ceramic, alginate-bioglass, alginate-biosilica, alginate-bone morphogenetic protein-2 and RGD peptides composite have been investigated till date. These alginate composites show enhanced biochemical significance in terms of porosity, mechanical strength, cell adhesion, biocompatibility, cell proliferation, alkaline phosphatase increase, excellent mineralization and osteogenic differentiation. Hence, alginate based composite biomaterials will be promising for bone tissue regeneration. This review will provide a broad overview of alginate preparation and its applications towards bone tissue engineering.

  9. Mechanical and structural contribution of non-fibrillar matrix in uniaxial tension: a collagen-agarose co-gel model.

    PubMed

    Lake, Spencer P; Barocas, Victor H

    2011-07-01

    The mechanical role of non-fibrillar matrix and the nature of its interaction with the collagen network in soft tissues remain poorly understood, in part because of the lack of a simple experimental model system to quantify these interactions. This study's objective was to examine mechanical and structural properties of collagen-agarose co-gels, utilized as a simplified model system, to understand better the relationships between the collagen network and non-fibrillar matrix. We hypothesized that the presence of agarose would have a pronounced effect on microstructural reorganization and mechanical behavior. Samples fabricated from gel solutions containing 1.0 mg/mL collagen and 0, 0.125, or 0.25% w/v agarose were evaluated via scanning electron microscopy, incremental tensile stress-relaxation tests, and polarized light imaging. While the incorporation of agarose did not dramatically alter collagen network morphology, agarose led to concentration-dependent changes in mechanical and structural properties. Specifically, resistance of co-gels to volume change corresponded with differences in fiber reorientation and elastic/viscoelastic mechanics. Results demonstrate strong relationships between tissue properties and offer insight into behavior of tissues of varying Poisson's ratio and fiber kinematics. Results also suggest that non-fibrillar material may have significant effects on properties of artificial and native tissues even in tension, which is generally assumed to be collagen dominated.

  10. Smart designing of new hybrid materials based on brushite-alginate and monetite-alginate microspheres: bio-inspired for sequential nucleation and growth.

    PubMed

    Amer, Walid; Abdelouahdi, Karima; Ramananarivo, Hugo Ronald; Fihri, Aziz; El Achaby, Mounir; Zahouily, Mohamed; Barakat, Abdellatif; Djessas, Kamal; Clark, James; Solhy, Abderrahim

    2014-02-01

    In this report new hybrid materials based on brushite-alginate and monetite-alginate were prepared by self-assembling alginate chains and phosphate source ions via a gelation process with calcium ions. The alginate served as nanoreactor for nucleation and growth of brushite or/and monetite due to its gelling and swelling properties. The alginate gel framework, the crystalline phase and morphology of formed hybrid biomaterials were shown to be strongly dependent upon the concentration of the phosphate precursors. These materials were characterized by thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive x-ray analysis (EDX).

  11. Mechanical and microstructural properties of "wet" alginate and composite films containing various carbohydrates.

    PubMed

    Harper, B Allison; Barbut, Shai; Smith, Alexandra; Marcone, Massimo F

    2015-01-01

    Composite "wet" alginate films were manufactured from alginate-carbohydrate solutions containing 5% alginate and 0.25% pectin, carrageenan (kappa or iota), potato starch (modified or unmodified), gellan gum, or cellulose (extracted or commercial). The "wet" alginate films were used as a model to understand co-extruded alginate sausage casings that are currently being used by several sausage manufacturers. The mechanical, optical, and microstructural properties of the calcium cross-linked composite films were explored. In addition, the water holding capacity and textural profile analysis properties of the alginate-carbohydrate gels were studied. The results indicate that the mechanical properties of "wet" alginate films/casings can be modified by adding various carbohydrates to them. Alginate films with pectin, carrageenan, and modified potato starch had significantly (P < 0.05) greater elongation values than pure alginate films. The alginate-pectin films also had greater (P < 0.05) tensile strengths than the pure alginate films. Alginate films with extracted cellulose, commercial cellulose, and modified potato starch had lower (P < 0.05) puncture force, distance, and work values than the alginate control films. Transmission electron microscopy images showed a very uniform alginate network in the control films. Several large cellulose fibers were visible in the films with extracted cellulose, while the cellulose fibers in the films with commercial cellulose were difficult to distinguish. Despite these apparent differences in cellulose fiber length, the 2 cellulose films had similar puncture and tensile properties.

  12. Fabrication of high-density collagen fibril matrix gels by renaturation of triple-helix collagen from gelatin.

    PubMed

    Ohyabu, Yoshimi; Yunoki, Shunji; Hatayama, Hirosuke; Teranishi, Yoshikazu

    2013-11-01

    Collagen-based 3-D hydrogels often lack sufficient mechanical strength for tissue engineering. We developed a method for fabrication of high-density collagen fibril matrix (CFM) gels from concentrated solutions of uncleaved gelatin (UCG). Denatured random-coil UCG exhibited more rapid and efficient renaturation into collagen triple-helix than cleaved gelatin (CG) over a broad range of setting temperatures. The UCG solution formed opaque gels with high-density reconstituted collagen fibrils at 28-32 °C and transparent gels similar to CG at <25 °C. The unique gelation properties of UCG enabled the encapsulation of cultured cells in CFM of high solid volume (>5%) and elasticity (1.28 ± 0.15 kPa at 5% and 4.82 ± 0.38 kPa at 8%) with minimal cell loss. The elastic modulus of these gels was higher than that of conventional CFM containing 0.5% collagen. High-strength CFM may provide more durable hydrogels for tissue engineering and regenerative medicine.

  13. Decolorization applicability of sol–gel matrix immobilized manganese peroxidase produced from an indigenous white rot fungal strain Ganoderma lucidum

    PubMed Central

    2013-01-01

    Background An eco-friendly treatment of industrial effluents is a major environmental concern of the modern world in the face of stringent environmental legislations. By keeping in mind the extensive industrial applications of ligninolytic enzymes, this study was performed to purify, and immobilize the manganese peroxidase (MnP) produced from an indigenous strain of Ganoderma lucidum. The present study was also focused on investigating the capability of immobilized MnP for decolorization of dye containing textile effluents. Results A large magnitude of an indigenous MnP (882±13.3 U/mL) was obtained from white rot fungal strain G. lucidum in solid state bio-processing of wheat straw under optimized fermentation conditions (moisture, 50%; substrate, 5 g; pH, 5.5; temperature, 30°C; carbon source, 2% fructose; nitrogen source, 0.02% yeast extract; C: N ratio, 25:1; fungal spore suspension, 5 mL and fermentation time period, 4 days). After ammonium sulfate fractionation and Sephadex-G-100 gel filtration chromatography, MnP was 4.7-fold purified with specific activity of 892.9 U/mg. G. lucidum MnP was monomeric protein as evident by single band corresponding to 48 kDa on native and denaturing SDS-PAGE. The purified MnP (2 mg/mL) was immobilized using a sol–gel matrix of tetramethoxysilane (TMOS) and proplytrimethoxysilane (PTMS). The oxidation of MnSO4 for up to 10 uninterrupted cycles demonstrated the stability and reusability of the immobilized MnP. Shelf life profile revealed that enzyme may be stored for up to 60 days at 25°C without losing much of its activity. To explore the industrial applicability of MnP produced by G. lucidum, the immobilized MnP was tested against different textile effluents. After 4 h reaction time, the industrial effluents were decolorized to different extents (with a maximum of 99.2%). The maximally decolorized effluent was analyzed for formaldehyde and nitroamines and results showed that the toxicity parameters were below the

  14. Gel structure has an impact on pericellular and extracellular matrix deposition, which subsequently alters metabolic activities in chondrocyte-laden PEG hydrogels.

    PubMed

    Nicodemus, G D; Skaalure, S C; Bryant, S J

    2011-02-01

    While designing poly(ethylene glycol) hydrogels with high moduli suitable for in situ placement is attractive for cartilage regeneration, the impact of a tighter crosslinked structure on the organization and deposition of the matrix is not fully understood. The objectives of this study were to characterize the composition and spatial organization of new matrix as a function of gel crosslinking and study its impact on chondrocytes in terms of anabolic and catabolic gene expression and catabolic activity. Bovine articular chondrocytes were encapsulated in hydrogels with three crosslinking densities (compressive moduli 60, 320 and 590 kPa) and cultured for 25 days. Glycosaminoglycan production increased with culture time and was greatest in the gels with lowest crosslinking. Collagens II and VI, aggrecan, link protein and decorin were localized to pericellular regions in all gels, but their presence decreased with increasing gel crosslinking. Collagen II and aggrecan expression were initially up-regulated in gels with higher crosslinking, but increased similarly up to day 15. Matrix metalloproteinase (MMP)-1 and MMP-13 expression were elevated (∼25-fold) in gels with higher crosslinking throughout the study, while MMP-3 was unaffected by gel crosslinking. The presence of aggrecan and collagen degradation products confirmed MMP activity. These findings indicate that chondrocytes synthesized the major cartilage components within PEG hydrogels, however, gel structure had a significant impact on the composition and spatial organization of the new tissue and on how chondrocytes responded to their environment, particularly with respect to their catabolic expression.

  15. Apparatus for the production of gel beads containing a biocatalyst

    DOEpatents

    Scott, C.D.; Scott, T.C.; Davison, B.H.

    1998-03-19

    An apparatus is described for the large-scale and continuous production of gel beads containing a biocatalyst. The apparatus is a columnar system based on the chemical cross-linking of hydrocolloidal gels that contain and immobilize a biocatalyst, the biocatalyst being a microorganism or an enzyme. Hydrocolloidal gels, such as alginate, carrageenan, and a mixture of bone gelatin and modified alginate, provide immobilization matrices that can be used to entrap and retain the biocatalyst while allowing effective contact with substrates and release of products. Such immobilized biocatalysts are generally formulated into small spheres or beads that have high concentrations of the biocatalyst within the gel matrix. The columnar system includes a gel dispersion nozzle submerged in a heated non-interacting liquid, typically an organic liquid, that is immiscible with water to allow efficient formation of spherical gel droplets, the non-interacting liquid having a specific gravity that is less than water so that the gel droplets will fall through the liquid by the force of gravity. The heated non-interacting liquid is in direct contact with a chilled upflowing non-interacting liquid that will provide sufficient residence time for the gel droplets as they fall through the liquid so that they will be cooled below the gelling temperature and form solid spheres. The upflowing non-interacting liquid is in direct contact with an upflowing temperature-controlled aqueous solution containing the necessary chemicals for cross-linking or fixing of the gel beads to add the necessary stability. The flow rates of the two liquid streams can be varied to control the proper residence time in each liquid section to accommodate the production of gel beads of differing settling velocities. A valve is provided for continuous removal of the stabilized gel beads from the bottom of the column. 1 fig.

  16. Apparatus for the production of gel beads containing a biocatalyst

    DOEpatents

    Scott, Charles D.; Scott, Timothy C.; Davison, Brian H.

    1998-01-01

    An apparatus for the large-scale and continuous production of gel beads containing a biocatalyst. The apparatus is a columnar system based on the chemical cross-linking of hydrocolloidal gels that contain and immobilize a biocatalyst, the biocatalyst being a microorganism or an enzyme. Hydrocolloidal gels, such as alginate, carrageenan, and a mixture of bone gelatin and modified alginate, provide immobilization matrices that can be used to entrap and retain the biocatalyst while allowing effective contact with substrates and release of products. Such immobilized biocatalysts are generally formulated into small spheres or beads that have high concentrations of the biocatalyst within the gel matrix. The columnar system includes a gel dispersion nozzle submerged in a heated non-interacting liquid, typically an organic liquid, that is immiscible with water to allow efficient formation of spherical gel droplets, the non-interacting liquid having a specific gravity that is less than water so that the gel droplets will fall through the liquid by the force of gravity. The heated non-interacting liquid is in direct contact with a chilled upflowing non-interacting liquid that will provide sufficient residence time for the gel droplets as they fall through the liquid so that they will be cooled below the gelling temperature and form solid spheres. The upflowing non-interacting liquid is in direct contact with an upflowing temperature-controlled aqueous solution containing the necessary chemicals for cross-linking or fixing of the gel beads to add the necessary stability. The flow rates of the two liquid streams can be varied to control the proper residence time in each liquid section to accommodate the production of gel beads of differing settling velocities. A valve is provided for continuous removal of the stabilized gel beads from the bottom of the column.

  17. Enzymatic Hydrolysis of Alginate to Produce Oligosaccharides by a New Purified Endo-Type Alginate Lyase

    PubMed Central

    Zhu, Benwei; Chen, Meijuan; Yin, Heng; Du, Yuguang; Ning, Limin

    2016-01-01

    Enzymatic hydrolysis of sodium alginate to produce alginate oligosaccharides has drawn increasing attention due to its advantages of containing a wild reaction condition, excellent gel properties and specific products easy for purification. However, the efficient commercial enzyme tools are rarely available. A new alginate lyase with high activity (24,038 U/mg) has been purified from a newly isolated marine strain, Cellulophaga sp. NJ-1. The enzyme was most active at 50 °C and pH 8.0 and maintained stability at a broad pH range (6.0–10.0) and temperature below 40 °C. It had broad substrate specificity toward sodium alginate, heteropolymeric MG blocks (polyMG), homopolymeric M blocks (polyM) and homopolymeric G blocks (polyG), and possessed higher affinity toward polyG (15.63 mM) as well as polyMG (23.90 mM) than polyM (53.61 mM) and sodium alginate (27.21 mM). The TLC and MS spectroscopy analysis of degradation products suggested that it completely hydrolyzed sodium alginate into oligosaccharides of low degrees of polymerization (DPs). The excellent properties would make it a promising tool for full use of sodium alginate to produce oligosaccharides. PMID:27275826

  18. Immobilization of Erwinia sp. D12 Cells in Alginate-Gelatin Matrix and Conversion of Sucrose into Isomaltulose Using Response Surface Methodology

    PubMed Central

    Kawaguti, Haroldo Yukio; Carvalho, Priscila Hoffmann; Figueira, Joelise Alencar; Sato, Hélia Harumi

    2011-01-01

    Isomaltulose is a noncariogenic reducing disaccharide and also a structural isomer of sucrose and is used by the food industry as a sucrose replacement. It is obtained through enzymatic conversion of microbial sucrose isomerase. An Erwinia sp. D12 strain is capable of converting sucrose into isomaltulose. The experimental design technique was used to study the influence of immobilization parameters on converting sucrose into isomaltulose in a batch process using shaken Erlenmeyer flasks. We assessed the effect of gelatin and transglutaminase addition on increasing the reticulation of granules of Erwinia sp. D12 cells immobilized in alginate. Independent parameters, sodium alginate concentration, cell mass concentration, CaCl2 concentration, gelatin concentration, and transglutaminase concentration had all a significant effect (P < 0.05) on isomaltulose production. Erwinia sp. D12 cells immobilized in 3.0% (w/v) sodium alginate, 47.0% (w/v) cell mass, 0.3 molL−1 CaCl2, 1.7% (w/v) gelatin and 0.15% (w/v) transglutaminase presented sucrose conversion into isomaltulose, of around 50–60% in seven consecutive batches. PMID:21785708

  19. Immobilization of Erwinia sp. D12 Cells in Alginate-Gelatin Matrix and Conversion of Sucrose into Isomaltulose Using Response Surface Methodology.

    PubMed

    Kawaguti, Haroldo Yukio; Carvalho, Priscila Hoffmann; Figueira, Joelise Alencar; Sato, Hélia Harumi

    2011-01-01

    Isomaltulose is a noncariogenic reducing disaccharide and also a structural isomer of sucrose and is used by the food industry as a sucrose replacement. It is obtained through enzymatic conversion of microbial sucrose isomerase. An Erwinia sp. D12 strain is capable of converting sucrose into isomaltulose. The experimental design technique was used to study the influence of immobilization parameters on converting sucrose into isomaltulose in a batch process using shaken Erlenmeyer flasks. We assessed the effect of gelatin and transglutaminase addition on increasing the reticulation of granules of Erwinia sp. D12 cells immobilized in alginate. Independent parameters, sodium alginate concentration, cell mass concentration, CaCl(2) concentration, gelatin concentration, and transglutaminase concentration had all a significant effect (P < 0.05) on isomaltulose production. Erwinia sp. D12 cells immobilized in 3.0% (w/v) sodium alginate, 47.0% (w/v) cell mass, 0.3 molL(-1) CaCl(2), 1.7% (w/v) gelatin and 0.15% (w/v) transglutaminase presented sucrose conversion into isomaltulose, of around 50-60% in seven consecutive batches.

  20. Bioactive apatite incorporated alginate microspheres with sustained drug-delivery for bone regeneration application.

    PubMed

    Li, Haibin; Jiang, Fei; Ye, Song; Wu, Yingying; Zhu, Kaiping; Wang, Deping

    2016-05-01

    The strontium-substituted hydroxyapatite microspheres (SrHA) incorporated alginate composite microspheres (SrHA/Alginate) were prepared via adding SrHA/alginate suspension dropwise into calcium chloride solution, in which the gel beads were formed by means of crosslinking reaction. The structure, morphology and in vitro bioactivity of the composite microspheres were studied by using XRD, SEM and EDS methods. The biological behaviors were characterized and analyzed through inductively coupled plasma optical emission spectroscopy (ICP-OES), CCK-8, confocal laser microscope and ALP activity evaluations. The experimental results indicated that the synthetic SrHA/Alginate showed similar morphology to the well-known alginate microspheres (Alginate) and both of them possessed a great in vitro bioactivity. Compared with the control Alginate, the SrHA/Alginate enhanced MC3T3-E1 cell proliferation and ALP activity by releasing osteoinductive and osteogenic Sr ions. Furthermore, vancomycin was used as a model drug to investigate the drug release behaviors of the SrHA/Alginate, Alginate and SrHA. The results suggested that the SrHA/Alginate had a highest drug-loading efficiency and best controlled drug release properties. Additionally, the SrHA/Alginate was demonstrated to be pH-sensitive as well. The increase of the pH value in phosphate buffer solution (PBS) accelerated the vancomycin release. Accordingly, the multifunctional SrHA/Alginate can be applied in the field of bioactive drug carriers and bone filling materials.

  1. Microbial alginate dressings show improved binding capacity for pathophysiological factors in chronic wounds compared to commercial alginate dressings of marine origin.

    PubMed

    Fischer, Melissa; Gebhard, Florian; Hammer, Timo; Zurek, Christian; Meurer, Guido; Marquardt, Christoph; Hoefer, Dirk

    2017-01-01

    Marine alginates are well established in wound management. Compared with different modern wound dressings, marine alginates cannot prove superior effects on wound healing. Alginates from bacteria have never been studied for medical applications so far, although the microbial polymer raises expectations for improved binding of wound factors because of its unique O-acetylation. Due to its possible positive effects on wound healing, alginates from bacteria might be a superior future medical product for clinical use. To prove the binding capacity of microbial alginates to pathophysiological factors in chronic wounds, we processed microbial alginate fibres, produced from fermentation of the soil bacterium Azotobacter vinelandii ATCC 9046, into needle web dressings and compared them with commercial dressings made of marine alginate. Four dressings were assessed: Marine alginate dressings containing either ionic silver or zinc/manganese/calcium, and microbial alginate dressings with and without nanosilver. All dressings were tested in an in vitro approach for influence on chronic wound parameters such as elastase, matrix metalloproteases-2, tumour necrosis factor-α, interleukin-8, and free radical formation. Despite the alginate origin or addition of antimicrobials, all dressings were able to reduce the concentration of the proinflammatory cytokines TNF-α and IL-8. However, microbial alginate was found to bind considerable larger amounts of elastase and matrix metalloproteases-2 in contrast to the marine alginate dressings. The incorporation of zinc, silver or nanosilver into alginate fibres did not improve their binding capacity for proteases or cytokines. The addition of nanosilver slightly enhanced the antioxidant capacity of microbial alginate dressings, whereas the marine alginate dressing containing zinc/manganese/calcium was unable to inhibit the formation of free radicals. The enhanced binding affinity by microbial alginate of Azotobacter vinelandii to

  2. Synthesis rates and binding kinetics of matrix products in engineered cartilage constructs using chondrocyte-seeded agarose gels.

    PubMed

    Nims, Robert J; Cigan, Alexander D; Albro, Michael B; Hung, Clark T; Ateshian, Gerard A

    2014-06-27

    Large-sized cartilage constructs suffer from inhomogeneous extracellular matrix deposition due to insufficient nutrient availability. Computational models of nutrient consumption and tissue growth can be utilized as an efficient alternative to experimental trials to optimize the culture of large constructs; models require system-specific growth and consumption parameters. To inform models of the [bovine chondrocyte]-[agarose gel] system, total synthesis rate (matrix accumulation rate+matrix release rate) and matrix retention fractions of glycosaminoglycans (GAG), collagen, and cartilage oligomeric matrix protein (COMP) were measured either in the presence (continuous or transient) or absence of TGF-β3 supplementation. TGF-β3's influences on pyridinoline content and mechanical properties were also measured. Reversible binding kinetic parameters were characterized using computational models. Based on our recent nutrient supplementation work, we measured glucose consumption and critical glucose concentration for tissue growth to computationally simulate the culture of a human patella-sized tissue construct, reproducing the experiment of Hung et al. (2003). Transient TGF-β3 produced the highest GAG synthesis rate, highest GAG retention ratio, and the highest binding affinity; collagen synthesis was elevated in TGF-β3 supplementation groups over control, with the highest binding affinity observed in the transient supplementation group; both COMP synthesis and retention were lower than those for GAG and collagen. These results informed the modeling of GAG deposition within a large patella construct; this computational example was similar to the previous experimental results without further adjustments to modeling parameters. These results suggest that these nutrient consumption and matrix synthesis models are an attractive alternative for optimizing the culture of large-sized constructs.

  3. Cheap glass fiber mats as a matrix of gel polymer electrolytes for lithium ion batteries

    PubMed Central

    Zhu, Yusong; Wang, Faxing; Liu, Lili; Xiao, Shiyin; Yang, Yaqiong; Wu, Yuping

    2013-01-01

    Lithium ion batteries (LIBs) are going to play more important roles in electric vehicles and smart grids. The safety of the current LIBs of large capacity has been remaining a challenge due to the existence of large amounts of organic liquid electrolytes. Gel polymer electrolytes (GPEs) have been tried to replace the organic electrolyte to improve their safety. However, the application of GPEs is handicapped by their poor mechanical strength and high cost. Here, we report an economic gel-type composite membrane with high safety and good mechanical strength based on glass fiber mats, which are separator for lead-acid batteries. The gelled membrane exhibits high ionic conductivity (1.13 mS cm−1), high Li+ ion transference number (0.56) and wide electrochemical window. Its electrochemical performance is evaluated by LiFePO4 cathode with good cycling. The results show this gel-type composite membrane has great attraction to the large-capacity LIBs requiring high safety with low cost. PMID:24216756

  4. Cheap glass fiber mats as a matrix of gel polymer electrolytes for lithium ion batteries

    NASA Astrophysics Data System (ADS)

    Zhu, Yusong; Wang, Faxing; Liu, Lili; Xiao, Shiyin; Yang, Yaqiong; Wu, Yuping

    2013-11-01

    Lithium ion batteries (LIBs) are going to play more important roles in electric vehicles and smart grids. The safety of the current LIBs of large capacity has been remaining a challenge due to the existence of large amounts of organic liquid electrolytes. Gel polymer electrolytes (GPEs) have been tried to replace the organic electrolyte to improve their safety. However, the application of GPEs is handicapped by their poor mechanical strength and high cost. Here, we report an economic gel-type composite membrane with high safety and good mechanical strength based on glass fiber mats, which are separator for lead-acid batteries. The gelled membrane exhibits high ionic conductivity (1.13 mS cm-1), high Li+ ion transference number (0.56) and wide electrochemical window. Its electrochemical performance is evaluated by LiFePO4 cathode with good cycling. The results show this gel-type composite membrane has great attraction to the large-capacity LIBs requiring high safety with low cost.

  5. In situ cell-matrix mechanics in tendon fascicles and seeded collagen gels: implications for the multiscale design of biomaterials.

    PubMed

    Duncan, Neil A; Bruehlmann, Sabina B; Hunter, Christopher J; Shao, Xinxin; Kelly, Elizabeth J

    2014-01-01

    Designing biomaterials to mimic and function within the complex mechanobiological conditions of connective tissues requires a detailed understanding of the micromechanical environment of the cell. The objective of our study was to measure the in situ cell-matrix strains from applied tension in both tendon fascicles and cell-seeded type I collagen scaffolds using laser scanning confocal microscopy techniques. Tendon fascicles and collagen gels were fluorescently labelled to simultaneously visualise the extracellular matrix and cell nuclei under applied tensile strains of 5%. There were significant differences observed in the micromechanics at the cell-matrix scale suggesting that the type I collagen scaffold did not replicate the pattern of native tendon strains. In particular, although the overall in situ tensile strains in the matrix were quite similar (∼2.5%) between the tendon fascicles and the collagen scaffolds, there were significant differences at the cell-matrix boundary with visible shear across cell nuclei of >1 μm measured in native tendon which was not observed at all in the collagen scaffolds. Similarly, there was significant non-uniformity of intercellular strains with relative sliding observed between cell rows in tendon which again was not observed in the collagen scaffolds where the strain environment was much more uniform. If the native micromechanical environment is not replicated in biomaterial scaffolds, then the cells may receive incorrect or mixed mechanical signals which could affect their biosynthetic response to mechanical load in tissue engineering applications. This study highlights the importance of considering the microscale mechanics in the design of biomaterial scaffolds and the need to incorporate such features in computational models of connective tissues.

  6. Argatroban-coupled Affi-Gel matrix for the purification of thrombin from plasma.

    PubMed

    Lefkowitz, Jerry B

    2005-10-01

    Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay. None of the available purification methods easily deals with this subject. The procedure described in the present paper uses a readily available pharmaceutical agent, argatroban, to construct an affinity matrix. Argatroban has a high affinity for thrombin and its thrombin binding is reversible. Prothrombin derived from a Ba(2+) precipitate of human plasma is used as the starting material. The crude prothrombin can be bulk activated to thrombin using taipan-snake (Oxyuranus scutellatus) venom and bound to the argatroban-coupled matrix without further processing steps. The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis. This purification scheme is rapid, yielding purified thrombin within 2 days.

  7. An anomalous behavior of trypsin immobilized in alginate network.

    PubMed

    Ganachaud, Chrystelle; Bernin, Diana; Isaksson, Dan; Holmberg, Krister

    2013-05-01

    Alginate is a biopolymer used in drug formulations and for surgical purposes. In the presence of divalent cations, it forms solid gels, and such gels are of interest for immobilization of cells and enzymes. In this work, we entrapped trypsin in an alginate gel together with a known substrate, N α-benzoyl-L-arginine-4-nitroanilide hydrochloride (L-BAPNA), and in the presence or absence of D-BAPNA, which is known to be a competitive inhibitor. Interactions between alginate and the substrate as well as the enzyme were characterized with transmission electron microscopy, rheology, and nuclear magnetic resonance spectroscopy. The biocatalysis was monitored by spectrophotometry at temperatures ranging from 10 to 42 °C. It was found that at 37 and 42 °C a strong acceleration of the reaction was obtained, whereas at 10 °C and at room temperature, the presence of D-BAPNA leads to a retardation of the reaction rate. The same effect was found when the reaction was performed in a non-cross-linked alginate solution. In alginate-free buffer solution, as well as in a solution of carboxymethylcellulose, a biopolymer that resembles alginate, the normal behavior was obtained; however, with D-BAPNA acting as an inhibitor at all temperatures. A more detailed investigation of the reaction kinetics showed that at higher temperature and in the presence of alginate, the curve of initial reaction rate versus L-BAPNA concentration had a sigmoidal shape, indicating an allosteric behavior. We believe that the anomalous behavior of trypsin in the presence of alginate is due to conformational changes caused by interactions between the positively charged trypsin and the strongly negatively charged alginate.

  8. Influence of oligoguluronates on alginate gelation, kinetics, and polymer organization.

    PubMed

    Jørgensen, Tor Erik; Sletmoen, Marit; Draget, Kurt I; Stokke, Bjørn T

    2007-08-01

    Structural polysaccharides of the alginate family form gels in aqueous Ca2+-containing solutions by lateral association of chain segments. The effect of adding oligomers of alpha-l-guluronic acid (G blocks) to gelling solutions of alginate was investigated using rheology and atomic force microscopy (AFM). Ca-alginate gels were prepared by in situ release of Ca2+. The gel strength increased with increasing level of calcium saturation of the alginate and decreased with increasing amount of free G blocks. The presence of free G blocks also led to an increased gelation time. The gel point and fractal dimensionalities of the gels were determined based on the rheological characterization. Without added free G blocks the fractal dimension of the gels increased from df = 2.14 to df = 2.46 when increasing [Ca2+] from 10 to 20 mM. This increase was suggested to arise from an increased junction zone multiplicity induced by the increased concentration of calcium ions. In the presence of free G blocks (G block/alginate = 1/1) the fractal dimension increased from 2.14 to 2.29 at 10 mM Ca2+, whereas there was no significant change associated with addition of G blocks at 20 mM Ca2+. These observations indicate that free G blocks are involved in calcium-mediated bonds formed between guluronic acid sequences within the polymeric alginates. Thus, the added oligoguluronate competes with the alginate chains for the calcium ions. The gels and pregel situations close to the gel point were also studied using AFM. The AFM topographs indicated that in situations of low calcium saturation microgels a few hundred nanometers in diameter develop in solution. In situations of higher calcium saturation lateral association of a number of alginate chains are occurring, giving ordered fiber-like structures. These results show that G blocks can be used as modulators of gelation kinetics as well as local network structure formation and equilibrium properties in alginate gels.

  9. A green and environment-friendly gel polymer electrolyte with higher performances based on the natural matrix of lignin

    NASA Astrophysics Data System (ADS)

    Gong, Sheng-Dong; Huang, Yun; Cao, Hai-Jun; Lin, Yuan-Hua; Li, Yang; Tang, Shui-Hua; Wang, Ming-Shan; Li, Xing

    2016-03-01

    In order to explore one truly green and environment-friendly gel polymer electrolyte (GPE), the natural biopolymer of lignin is firstly all over the world used as matrix to prepare GPE. The electrolyte membrane based on lignin can be easily fabricated just with lignin, liquid electrolyte and distilled water. Through comprehensive investigation of obtained GPE, it is found that the liquid electrolyte uptake reaches up to 230 wt.%; before 100 °C, GPE does not lose any weight and is thermal stable; at room temperature the ion conductivity is 3.73 mS cm-1; the amazing property of lithium ion transference number is high up to 0.85; GPE expresses complete electrochemical stability before 7.5 V and favorable compatibility with lithium anode; the outstanding cell performance of C-rate and cycle capacity. All these remarkably excellent performances endow lignin with application potential in GPE used in lithium ion batteries (LIBs) with higher performances.

  10. Storage stability of low-fat sodium reduced fresh merguez sausage prepared with olive oil in konjac gel matrix.

    PubMed

    Triki, Mehdi; Herrero, Ana M; Jiménez-Colmenero, Francisco; Ruiz-Capillas, Claudia

    2013-08-01

    This paper evaluates the nutritional values and stability during refrigerated storage of fresh beef merguez sausage as affected by a reformulation process which modified the fat content both by reducing fat (replacing beef fat with konjac gel) and incorporating olive oil (replacing beef fat with olive oil stabilized in a konjac matrix) and by reducing sodium content, replacing sodium chloride with a salt mixture (containing potassium chloride, calcium chloride and magnesium chloride). A preservative (sodium metabisulphite) was also used to extend the shelf-life of the product. The fat was reduced by 32 to 80% and sodium by over 36%. The reformulation did not negatively affect the sensory evaluation. Low microbiota growth rate and biogenic amines were attributed mainly to the presence of sodium metabisulphite. This preservative could be used in the reformulation to enhance safety and/or extend the shelf-life of this type of product.

  11. An antibacterial coating based on a polymer/sol-gel hybrid matrix loaded with silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Rivero, Pedro José; Urrutia, Aitor; Goicoechea, Javier; Zamarreño, Carlos Ruiz; Arregui, Francisco Javier; Matías, Ignacio Raúl

    2011-12-01

    In this work a novel antibacterial surface composed of an organic-inorganic hybrid matrix of tetraorthosilicate and a polyelectrolyte is presented. A precursor solution of tetraethoxysilane (TEOS) and poly(acrylic acid sodium salt) (PAA) was prepared and subsequently thin films were fabricated by the dip-coating technique using glass slides as substrates. This hybrid matrix coating is further loaded with silver nanoparticles using an in situ synthesis route. The morphology and composition of the coatings have been studied using UV-VIS spectroscopy and atomic force microscopy (AFM). Energy dispersive X-ray (EDX) was also used to confirm the presence of the resulting silver nanoparticles within the thin films. Finally the coatings have been tested in bacterial cultures of genus Lactobacillus plantarum to observe their antibacterial properties. It has been experimentally demonstrated that these silver loaded organic-inorganic hybrid films have a very good antimicrobial behavior against this type of bacteria.

  12. A Thermally Conductive Composite with a Silica Gel Matrix and Carbon-Encapsulated Copper Nanoparticles as Filler

    NASA Astrophysics Data System (ADS)

    Lin, Jin; Zhang, Haiyan; Hong, Haoqun; Liu, Hui; Zhang, Xiubin

    2014-07-01

    Core-shell-structured nanocapsules with a copper core encapsulated in a carbon shell (Cu-C) were synthesized by a direct-current arc-discharge method. Morphological and microstructural characterization showed that the Cu-C consisted of a nanosized Cu core and carbon shell, with the carbon shells containing 6 to 15 ordered graphitic layers and amorphous carbon that effectively shield the metallic Cu core from oxidation. A thermally conductive composite was successfully fabricated using a silica gel matrix incorporated with Cu-C filler. The Cu-C nanoparticles were homogeneously dispersed in the silica gel. The effects of Cu-C on the thermal conductivity, electrical resistivity, and coefficient of thermal expansion (CTE) of the composite were investigated. For composites with 6.16 vol.%, 11.04 vol.%, 16.70 vol.%, and 23.34 vol.% Cu-C content, the thermal conductivity at 50°C was 0.32 W/(m K) to 0.77 W/(m K), the electrical resistivity was 1.98 × 109, 3.48 × 107, 302, and 1 Ω m, respectively, while the CTE at 200°C was 3.79 × 10-4 K-1 to 3.44 × 10-4 K-1. The results reveal that the ordered graphitic shells in the Cu-C increased both the thermal and electrical conduction, but decreased the CTE by preventing the Cu cores from expanding.

  13. Chemical tailoring of hybrid sol-gel thick coatings as hosting matrix for functional patterned microstructures.

    PubMed

    Falcaro, Paolo; Costacurta, Stefano; Malfatti, Luca; Buso, Dario; Patelli, Alessandro; Schiavuta, Piero; Piccinini, Massimo; Grenci, Gianluca; Marmiroli, Benedetta; Amenitsch, Heinz; Innocenzi, Plinio

    2011-02-01

    A phenyl-based hybrid organic - inorganic coating has been synthesized and processed by hard X-ray lithography. The overall lithography process is performed in a two-step process only (X-rays exposure and chemical etching). The patterns present high aspect ratio, sharp edges, and high homogeneity. The coating has been doped with a variety of polycyclic aromatic hydrocarbon functional molecules, such as anthracene, pentacene, and fullerene. For the first time, hard X-rays have been combined with thick hybrid functional coatings, using the sol-gel thick film directly as resist. A new technique based on a new material combined with hard X-rays is now available to fabricate optical devices. The effect due to the high-energy photon exposure has been investigated using FT-IR and Raman spectroscopy, laser scanner, optical profilometer, and confocal and electron microscope. High-quality thick hybrid fullerene-doped microstructures have been fabricated.

  14. Injectable alginate-O-carboxymethyl chitosan/nano fibrin composite hydrogels for adipose tissue engineering.

    PubMed

    Jaikumar, Dhanya; Sajesh, K M; Soumya, S; Nimal, T R; Chennazhi, K P; Nair, Shantikumar V; Jayakumar, R

    2015-03-01

    Injectable, biodegradable scaffolds are required for soft tissue reconstruction owing to its minimally invasive approach. Such a scaffold can mimic the native extracellular matrix (ECM), provide uniform distribution of cells and overcome limitations like donor site morbidity, volume loss, etc. So, here we report two classes of biocompatible and biodegradable hydrogel blend systems namely, Alginate/O-carboxymethyl chitosan (O-CMC) and Alginate/poly (vinyl alcohol) (PVA) with the inclusion of fibrin nanoparticles in each. The hydrogels were prepared by ionic cross-linking method. The developed hydrogels were compared in terms of its swelling ratio, degradation profile, compressive strength and elastic moduli. From these preliminary findings, it was concluded that Alginate/O-CMC formed a better blend for tissue engineering applications. The potential of the formed hydrogel as an injectable scaffold was revealed by the survival of adipose derived stem cells (ADSCs) on the scaffold by its adhesion, proliferation and differentiation into adipocytes. Cell differentiation studies of fibrin incorporated hydrogel scaffolds showed better differentiation was confirmed by Oil Red O staining technique. These injectable gels have potential in soft tissue regeneration.

  15. Calcium alginate matrix increases the stability and recycling capability of immobilized endo-β-1,4-xylanase from Geobacillus stearothermophilus KIBGE-IB29.

    PubMed

    Bibi, Zainab; Qader, Shah Ali Ul; Aman, Afsheen

    2015-07-01

    Exploration of microbial pool from extremely diversified ecosystem is significantly important for various industrial applications. Bacterial communities from extreme habitats including volcanic vents, hot springs, and industrial sectors are eagerly explored for the isolation of thermophiles. Geobacillus stearothermophilus KIBGE-IB29, isolated from blast furnace site of a steel processing industry, is capable of producing thermostable endo-β-1,4-xylanase. In the current study, this enzyme was immobilized within calcium alginate beads using entrapment technique. Amalgamation of sodium alginate (40.0 gL(-1)) and calcium chloride (0.4 M) was used for the formation of immobilized beads. It was observed that temperature (50 °C) and pH (7.0) optima of immobilized enzyme remained same, but enzyme-substrate reaction time increased from 5.0 to 30.0 min as compared to free enzyme. Diffusion limit of high molecular weight xylan (corncob) caused a decline in V max of immobilized enzyme from 4773 to 203.7 U min(-1), whereas K m value increased from 0.5074 to 0.5722 mg ml(-1) with reference to free enzyme. Immobilized endo-β-1,4-xylanase showed its stability even at high temperatures as compared to free enzyme and retained 18 and 9 % residual activity at 70 and 80 °C, respectively. Immobilized enzyme also exhibited sufficient recycling efficiency up to five reaction cycles which indicated that this enzyme can be a plausible candidate in paper and pulp industry.

  16. Mediator-free phenol sensor based on titania sol-gel encapsulation matrix for immobilization of tyrosinase by a vapor deposition method.

    PubMed

    Yu, Jiuhong; Liu, Songqin; Ju, Huangxian

    2003-12-30

    A novel amperometric phenol sensor was constructed by immobilizing tyrosinase in a titania sol-gel matrix. The tyrosinase entrapped sol-gel film was obtained with a vapor deposition method, which simplified the traditional sol-gel process and avoided the shrinkage and cracking of conventional sol-gel-derived glasses. This matrix provided a microenvironment for retaining the native structure and activity of the entrapped enzyme and a very low mass transport barrier to the enzyme substrates. Phenol could be oxidized by dissolving oxygen in presence of immobilized tyrosinase to form a detectable product, which was determined at -150 mV without any mediator. The phenol sensor exhibited a fast response (less than 5 s) and sensitivity as high as 103 microA/mM, which resulted from the porous structure and high enzyme loading of the sol-gel matrix. The linear range for phenol determination was from 1.2x10(-7) to 2.6x10(-4) M with a detection limit of 1.0x10(-7) M. The apparent Michaelis-Menten constant of the encapsulated tyrosinase was calculated to be (0.29+/-0.02) mM. The stability of the biosensor was also evaluated.

  17. Compatible compositions based on aqueous polyurethane dispersions and sodium alginate.

    PubMed

    Daemi, Hamed; Barikani, Mehdi; Barmar, Mohammad

    2013-01-30

    A series of aqueous polyurethane dispersions were synthesized by the reaction of polytetramethylene glycol and isophorone diisocyanate, extended with dimethylol propionic acid. Their chemical structures were characterized using FTIR, (1)H NMR, and (13)C NMR, and thermal properties were determined by DMTA. Then, a number of aqueous polyurethane dispersions-sodium alginate (PUD/SA) compositions were prepared by addition of sodium alginate solution with different concentrations into the aqueous polyurethane dispersion. Characterization of chemical structure and thermal properties of these blends were performed by FTIR, EDX and DMTA, respectively. The morphology of the alginate in polyurethane matrix was studied by SEM. The hydrophilicity of the prepared samples decreases by increasing the content of sodium alginate in blends. These observations were attributed to the increase of hydrophilicity of the blends as a consequence of addition of hydrophilic carboxylate, hydroxyl and ether functional groups of the alginate to them.

  18. In-gel expression and in situ immobilization of proteins for generation of three dimensional protein arrays in a hydrogel matrix.

    PubMed

    Byun, Ju-Young; Lee, Kyung-Ho; Lee, Ka-Young; Kim, Min-Gon; Kim, Dong-Myung

    2013-03-07

    A method has been developed for the direct conversion of DNA arrays into three dimensional protein arrays on a hydrogel matrix. An agarose gel embedded with bacterial protein synthesis machinery was used as the DNA-programmable expression gel matrix for the in situ translation of genes on a DNA array. Upon incubation of the expression gel matrix cast on a DNA array, protein synthesis took place at the interface of the two surfaces and the cell-free synthesized proteins were deposited on the gel matrix surrounding the corresponding DNA spots. Diffusional dilution of the expressed proteins was minimized by modifying the agarose with Ni-NTA moieties. This procedure resulted in the generation of localized protein spots with confined radii. The developed approach not only simplifies the procedures typically used for the preparation of protein arrays but it also provides conditions for the loading of higher amounts of proteins on the array while retaining their structural integrity and functionality over extended time periods.

  19. Three-Dimensional Nanoporous Cellulose Gels as a Flexible Reinforcement Matrix for Polymer Nanocomposites.

    PubMed

    Shi, Zhuqun; Huang, Junchao; Liu, Chuanjun; Ding, Beibei; Kuga, Shigenori; Cai, Jie; Zhang, Lina

    2015-10-21

    With the world's focus on utilization of sustainable natural resources, the conversion of wood and plant fibers into cellulose nanowhiskers/nanofibers is essential for application of cellulose in polymer nanocomposites. Here, we present a novel fabrication method of polymer nanocomposites by in-situ polymerization of monomers in three-dimensionally nanoporous cellulose gels (NCG) prepared from aqueous alkali hydroxide/urea solution. The NCG have interconnected nanofibrillar cellulose network structure, resulting in high mechanical strength and size stability. Polymerization of the monomer gave P(MMA/BMA)/NCG, P(MMA/BA)/NCG nanocomposites with a volume fraction of NCG ranging from 15% to 78%. SEM, TEM, and XRD analyses show that the NCG are finely distributed and preserved well in the nanocomposites after polymerization. DMA analysis demonstrates a significant improvement in tensile storage modulus E' above the glass transition temperature; for instance, at 95 °C, E' is increased by over 4 orders of magnitude from 0.03 MPa of the P(MMA/BMA) up to 350 MPa of nanocomposites containing 15% v/v NCG. This reinforcement effect can be explained by the percolation model. The nanocomposites also show remarkable improvement in solvent resistance (swelling ratio of 1.3-2.2 in chloroform, acetone, and toluene), thermal stability (do not melt or decompose up to 300 °C), and low coefficients of thermal expansion (in-plane CTE of 15 ppm·K(-1)). These nanocomposites will have great promising applications in flexible display, packing, biomedical implants, and many others.

  20. Ice crystal patterns in artificial gels of extracellular matrix macromolecules after quick-freezing and freeze-substitution.

    PubMed

    Allenspach, A L; Kraemer, T G

    1989-04-01

    Artificial gels, composed of collagen with or without hyaluronate (HA), a glycosaminoglycan (GAG), and chondroitin sulfate (CS), were prepared and quick-frozen for the purpose of studying the influence of composition and concentration on ice patterns. Dilute gels were spread on coverslips, plunged into a slush of 30% isopentane/70% propane (-185 degrees C), freeze-substituted, and examined by phase-contrast microscopy. Ice patterns were revealed as "ice cavities" in the gel after freeze-substitution. Ice morphology in the gels was gel-type-specific, suggesting that composition in dilute gels can influence ice pattern formation. Crystallization patterns reflecting high, intermediate, and low rates of freezing were observed in all gel types. Intermediate freezing in differentiating gel-type-specific ice patterns. Gels which included hyaluronate (HA) and chondroitin sulfate (CS) altered the ice crystal pattern commonly observed in collagen gels. Ice structure in collagen gels consisted predominantly of long, parallel crystals in the herringbone pattern. Ice crystals separated gel into thin, unbranched fibers with a primary spacing of approximately 2 microns. Ice morphology in HA gels formed a mosaic consisting of packets of ice crystals. Contiguous packets were often oriented at right angles to each other. Periodic crossbridges interconnect primary gel fibers of HA gels and interrupt the lengthwise growth of ice crystals. Smooth beads were visible on primary strands in HA gels frozen at intermediate velocities. The addition of CS to collagen gels resulted in formation of randomly oriented ice crystals in gels frozen at intermediate rates. CS has little influence on ice morphology at low freezing velocities. Primary strands in CS gels were decorated with rough-surfaced, osmiophilic aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. Preparation methods of alginate nanoparticles.

    PubMed

    Paques, Jerome P; van der Linden, Erik; van Rijn, Cees J M; Sagis, Leonard M C

    2014-07-01

    This article reviews available methods for the formation of alginate nano-aggregates, nanocapsules and nanospheres. Primarily, alginate nanoparticles are being prepared by two methods. In the "complexation method", complex formation on the interface of an oil droplet is used to form alginate nanocapsules, and complex formation in an aqueous solution is used to form alginate nano-aggregates. In a second method w/o emulsification coupled with gelation of the alginate emulsion droplet can be used to form alginate nanospheres. We review advantages and disadvantages of these methods, and give an overview of the properties of the alginate particles produced with these methods.

  2. The effect of conjugating RGD into 3D alginate hydrogels on adipogenic differentiation of human adipose-derived stromal cells.

    PubMed

    Kang, Sun-Woong; Cha, Byung-Hyun; Park, Honghyun; Park, Kwang-Sook; Lee, Kuen Yong; Lee, Soo-Hong

    2011-05-12

    The effects of RGD peptide conjugation to alginate hydrogel on the adipogenic differentiation of ASCs was investigated. After 3 d of culture, RGD-modified alginate hydrogels significantly stimulated FAK and integrin α1 gene expressions and vinculin expression in ASCs. In addition, RGD-modified alginate hydrogels significantly enhanced the adipogenic differentiation of human ASCs to exhibit higher expression levels of oil red O staining and adipogenic genes compared to those of the control group (unmodified gels). These results suggest potential applications of RGD-modified alginate gels for adipose tissue regeneration.

  3. Apparatus and method for the production of gel beads containing a biocatalyst

    DOEpatents

    Scott, Charles D.; Scott, Timothy C.; Davison, Brian H.

    1998-01-01

    An apparatus and method for the large-scale and continuous production of gel beads containing a biocatalyst. The apparatus is a columnar system based on the chemical cross-linking of hydrocolloidal gels that contain and immobilize a biocatalyst, the biocatalyst being a microorganism or an enzyme. Hydrocolloidal gels, such as alginate, carrageenan, and a mixture of bone gelatin and modified alginate, provide immobilization matrices that can be used to entrap and retain the biocatalyst while allowing effective contact with substrates and release of products. Such immobilized biocatalysts are generally formulated into small spheres or beads that have high concentrations of the biocatalyst within the gel matrix. The columnar system includes a gel dispersion nozzle submerged in a heated non-interacting liquid, typically an organic liquid, that is immiscible with water to allow efficient formation of spherical gel droplets, the non-interacting liquid having a specific gravity that is less than water so that the gel droplets will fall through the liquid by the force of gravity. The heated non-interacting liquid is in direct contact with a chilled upflowing non-interacting liquid that will provide sufficient residence time for the gel droplets as they fall through the liquid so that they will be cooled below the gelling temperature and form solid spheres. The upflowing non-interacting liquid is in direct contact with an upflowing temperature-controlled aqueous solution containing the necessary chemicals for cross-linking or fixing of the gel beads to add the necessary stability. The flow rates of the two liquid streams can be varied to control the proper residence time in each liquid section to accommodate the production of gel beads of differing settling velocities. A valve is provided for continuous removal of the stabilized gel beads from the bottom of the column.

  4. Apparatus and method for the production of gel beads containing a biocatalyst

    DOEpatents

    Scott, C.D.; Scott, T.C.; Davison, B.H.

    1998-01-27

    An apparatus and method are disclosed for the large-scale and continuous production of gel beads containing a biocatalyst. The apparatus is a columnar system based on the chemical cross-linking of hydrocolloidal gels that contain and immobilize a biocatalyst, the biocatalyst being a microorganism or an enzyme. Hydrocolloidal gels, such as alginate, carrageenan, and a mixture of bone gelatin and modified alginate, provide immobilization matrices that can be used to entrap and retain the biocatalyst while allowing effective contact with substrates and release of products. Such immobilized biocatalysts are generally formulated into small spheres or beads that have high concentrations of the biocatalyst within the gel matrix. The columnar system includes a gel dispersion nozzle submerged in a heated non-interacting liquid, typically an organic liquid, that is immiscible with water to allow efficient formation of spherical gel droplets, the non-interacting liquid having a specific gravity that is less than water so that the gel droplets will fall through the liquid by the force of gravity. The heated non-interacting liquid is in direct contact with a chilled upflowing non-interacting liquid that will provide sufficient residence time for the gel droplets as they fall through the liquid so that they will be cooled below the gelling temperature and form solid spheres. The upflowing non-interacting liquid is in direct contact with an upflowing temperature-controlled aqueous solution containing the necessary chemicals for cross-linking or fixing of the gel beads to add the necessary stability. The flow rates of the two liquid streams can be varied to control the proper residence time in each liquid section to accommodate the production of gel beads of differing settling velocities. A valve is provided for continuous removal of the stabilized gel beads from the bottom of the column. 1 fig.

  5. Online matrix removal platform for coupling gel-based separations to whole protein electrospray ionization mass spectrometry.

    PubMed

    Kim, Ki Hun; Compton, Philip D; Tran, John C; Kelleher, Neil L

    2015-05-01

    A fractionation method called gel-eluted liquid fraction entrapment electrophoresis (GELFrEE) has been used to dramatically increase the number of proteins identified in top-down proteomic workflows; however, the technique involves the use of sodium dodecyl sulfate (SDS), a surfactant that interferes with electrospray ionization. Therefore, an efficient removal of SDS is absolutely required prior to mass analysis. Traditionally, methanol/chloroform precipitation and spin columns have been used, but they lack reproducibility and are difficult to automate. Therefore, we developed an in-line matrix removal platform to enable the direct analysis of samples containing SDS and salts. Only small molecules like SDS permeate a porous membrane and are removed in a manner similar to cross-flow filtration. With this device, near-complete removal of SDS is accomplished within 5 min and proteins are subsequently mobilized into a mass spectrometer. The new platform was optimized for the analysis of GELFrEE fractions enriched for histones extracted from human HeLa cells. All four core histones and their proteoforms were detected in a single spectrum by high-resolution mass spectrometry. The new method versus protein precipitation/resuspension showed 2- to 10-fold improved signal intensities, offering a clear path forward to improve proteome coverage and the efficiency of top-down proteomics.

  6. Rheological characterization and turbidity of riboflavin-photosensitized changes in alginate/GDL systems.

    PubMed

    Baldursdóttir, Stefanía G; Kjøniksen, Anna-Lena

    2005-04-01

    Riboflavin (RF) in combination with light, in the wavelength range of 310-800 nm, is used to induce degradation of alginic acid gels. Light irradiation of alginate solutions in the presence of RF under aerobic conditions causes scission of the polymer chains. In the development process of a new drug delivery system, RF photosensitized degradation of alginic acid gels is studied by monitoring changes in the turbidity and rheological parameters of alginate/glucono-delta-lactone (GDL) systems with different concentrations of GDL. Addition of GDL induces gel formation of the samples by gradually lowering the pH-value of the system. The turbidity is measured and the cloud point determined. The turbidity starts to increase after shorter times with enhanced concentration of GDL. Enhanced viscoelasticity is detected with increasing GDL concentration in the post-gel regime, but small differences are detected at the gel point. The incipient gel is 'soft' and has an open structure independent on the GDL concentration. In the post-gel regime solid-like behavior is observed, this is more distinct for the systems with high GDL concentrations. The effect of photosensitized RF on alginate/GDL systems decreases with increasing amount of GDL in the system. The same trend is detected whether the systems are irradiated in the pre-gel or in the post-gel regime.

  7. Utilizing Core–Shell Fibrous Collagen-Alginate Hydrogel Cell Delivery System for Bone Tissue Engineering

    PubMed Central

    Perez, Roman A.; Kim, Meeju; Kim, Tae-Hyun; Kim, Joong-Hyun; Lee, Jae Ho; Park, Jeong-Hui; Knowles, Jonathan C.

    2014-01-01

    Three-dimensional matrices that encapsulate and deliver stem cells with defect-tuned formulations are promising for bone tissue engineering. In this study, we designed a novel stem cell delivery system composed of collagen and alginate as the core and shell, respectively. Mesenchymal stem cells (MSCs) were loaded into the collagen solution and then deposited directly into a fibrous structure while simultaneously sheathing with alginate using a newly designed core–shell nozzle. Alginate encapsulation was achieved by the crosslinking within an adjusted calcium-containing solution that effectively preserved the continuous fibrous structure of the inner cell-collagen part. The constructed hydrogel carriers showed a continuous fiber with a diameter of ∼700–1000 μm for the core and 200–500 μm for the shell area, which was largely dependent on the alginate concentration (2%–5%) as well as the injection rate (20–80 mL/h). The water uptake capacity of the core–shell carriers was as high as 98%, which could act as a pore channel to supply nutrients and oxygen to the cells. Degradation of the scaffolds showed a weight loss of ∼22% at 7 days and ∼43% at 14 days, suggesting a possible role as a degradable tissue-engineered construct. The MSCs encapsulated within the collagen core showed excellent viability, exhibiting significant cellular proliferation up to 21 days with levels comparable to those observed in the pure collagen gel matrix used as a control. A live/dead cell assay also confirmed similar percentages of live cells within the core–shell carrier compared to those in the pure collagen gel, suggesting the carrier was cell compatible and was effective for maintaining a cell population. Cells allowed to differentiate under osteogenic conditions expressed high levels of bone-related genes, including osteocalcin, bone sialoprotein, and osteopontin. Further, when the core–shell fibrous carriers were implanted in a rat calvarium defect, the bone

  8. Viscoelastic Behavior and Adhesion of Ionic Alginate Hydrogels

    NASA Astrophysics Data System (ADS)

    Webber, Rebecca; Shull, Kenneth

    2004-03-01

    Transient networks, polymer gels in which the physical crosslinks can be broken and recovered, have been of recent interest to the scientific community, especially due to their potential as soft, dissipative materials for biomedical applications. Alginates, naturally derived linear copolymers of mannuronic and guluronic acid residues, can form hydrogels in the presence of divalent ions. Alginate gels have been studied extensively and are useful model systems to elucidate the mechanisms behind the mechanical behavior of reversibly associating polymers. In this study, alginate hydrogels were formed by the addition of Ca ions to an aqueous solution of sodium alginate. The rheological and mechanical behavior of the hydrogels was studied using an axisymmetric probe tack apparatus with stress relaxation and cyclic movement capabilities. These hydrogels behave elastically at small strains and become viscoelastic at large strains, supporting transient network theories. During cyclic loading tests, it was found that the alginate hydrogels exhibit time-dependent adhesion. The effects of humidity, aging and ion exchange on the gel properties were also investigated.

  9. Self-crosslinked oxidized alginate/gelatin hydrogel as injectable, adhesive biomimetic scaffolds for cartilage regeneration.

    PubMed

    Balakrishnan, Biji; Joshi, Nitin; Jayakrishnan, Athipettah; Banerjee, Rinti

    2014-08-01

    Biopolymeric hydrogels that mimic the properties of extracellular matrix have great potential in promoting cellular migration and proliferation for tissue regeneration. The authors reported earlier that rapidly gelling, biodegradable, injectable hydrogels can be prepared by self-crosslinking of periodate oxidized alginate and gelatin in the presence of borax, without using any toxic crosslinking agents. The present paper investigates the suitability of this hydrogel as a minimally invasive injectable, cell-attractive and adhesive scaffold for cartilage tissue engineering for the treatment of osteoarthritis. Time and frequency sweep rheology analysis confirmed gel formation within 20s. The hydrogel integrated well with the cartilage tissue, with a burst pressure of 70±3mmHg, indicating its adhesive nature. Hydrogel induced negligible inflammatory and oxidative stress responses, a prerequisite for the management and treatment of osteoarthritis. Scanning electron microscopy images of primary murine chondrocytes encapsulated within the matrix revealed attachment of cells onto the hydrogel matrix. Chondrocytes demonstrated viability, proliferation and migration within the matrix, while maintaining their phenotype, as seen by expression of collagen type II and aggrecan, and functionality, as seen by enhanced glycosoaminoglycan (GAG) deposition with time. DNA content and GAG deposition of chondrocytes within the matrix can be tuned by incorporation of bioactive signaling molecules such as dexamethasone, chondroitin sulphate, platelet derived growth factor (PDGF-BB) and combination of these three agents. The results suggest that self-crosslinked oxidized alginate/gelatin hydrogel may be a promising injectable, cell-attracting adhesive matrix for neo-cartilage formation in the management and treatment of osteoarthritis.

  10. A combined approach of enamel matrix derivative gel and autogenous bone grafts in treatment of intrabony periodontal defects. A case report.

    PubMed

    Leung, George; Jin, Lijian

    2003-04-01

    Enamel matrix derivative (EMD) has recently been introduced as a new modality in regenerative periodontal therapy. This case report demonstrates a combined approach in topical application of EMD gel (Emdogain) and autogenous bone grafts for treatment of intrabony defects and furcation involvement defects in a patient with chronic periodontitis. The seven-month post-surgery clinical and radiographic results were presented. The combined application of EMD gel with autogenous bone grafts in intrabony osseous defects resulted in clinically significant gain of attachment on diseased root surfaces and bone fill on radiographs. Further controlled clinical studies are required to confirm the long-term effectiveness of the combination of EMD gel and autogenous bone grafts in treatment of various osseous defects in subjects with chronic periodontitis.

  11. Immobilization and characterization of 2,3-diaminonaphthalene/cyclodextrin complexes in a sol-gel matrix: a new fluorimetric sensor for nitrite.

    PubMed

    Martínez-Tomé, M J; Esquembre, R; Mallavia, R; Mateo, C R

    2009-01-01

    The aromatic diamino compound 2,3-diaminonaphthalene (DAN) has been extensively used to detect and quantify nitrite ions in biological and environmental samples. We have immobilized the DAN reagent in a porous silicate glass matrix, via previous incorporation of the dye in HP-beta-CD. Changes in fluorescence intensity were used to characterize the inclusion complexes and determine the association constant and stoichiometry of the process. Fluorescence spectrum of these complexes was also used to monitor their immobilization within the sol-gel matrix. Reactivity of the immobilized complexes was evaluated with increasing concentrations of nitrite up to 10 microM (with a detection limit around 20 nM). Results show that sol-gel immobilization does not modify the reactivity of the dye against nitrite and serves to prepare a highly sensitive ready to use fluorescence-based sensor for the specific measurement of nitrite at submicromolar concentrations with no further sample pretreatment.

  12. [Kinetic study on the in situ synthesis of nickle phthalocyanine in silica gel glass matrix by UV/Vis absorption spectra].

    PubMed

    Huang, Juan; Zheng, Chan; Feng, Miao; Zhan, Hong-Bing

    2009-01-01

    In decades, metallo-phthalocyanines (MPcs) have undergone a renaissance because of their singular and unconventional physical properties. However, for the successful application of MPcs in practical devices, it is important to disperse MPc molecules into solid state matrix to fabricate MPc doped composite with desired properties. Inorganic glass is an ideal matrix because of its transparency and high environmental stability. One attractive approach to fabricating MPc/inorganic composite is sol-gel technique. In the present paper, silica gel glass matrix was prepared by hydrolysis and poly-condensation of tetraethyloxysilane. 1,2-dicyanobenzene and analytically pure soluble nickle salt were used as the nickle phthalocyanine (NiPc) reactants and chemical synthesis technique was used to prepare NiPc doped sol-gel materials at several temperatures. During the heat treatment, four 1, 2-dicyanobenzene molecules and one nickle ion collide to form a NiPc molecule. In-situ synthesizing process of NiPc in the pores of silica gel glass matrix was traced by UV/Vis absorption spectra. Owing to the remarkable absorption band of NiPc in visible region, quantity of in-situ synthesized NiPc was calculated by the absorbance at certain wavelength of 670 nm, using composites with physically doped NiPc as a reference. The in-situ synthesized kinetics was studied in detail and found to be consistent with Avrami-Erofeev equation The reaction grades were deduced to be 4.5, 4.5, 3.7, 3.2 and 1.9 respectively at temperatures of 180 degrees C, 185 degrees C, 190 degrees C, 195 degrees C and 200 degrees C, respectively.

  13. Alginic Acid Accelerates Calcite Dissolution

    NASA Astrophysics Data System (ADS)

    Perry, T. D.; Duckworth, O. W.; McNamara, C. J.; Martin, S. T.; Mitchell, R.

    2003-12-01

    Accelerated carbonate weathering through biological activity affects both geochemical cycling and the local pH and alkalinity of terrestrial and marine waters. Microbes affect carbonate dissolution through metabolic activity, production of acidic or chelating exudates, and cation binding by cell walls. Dissolution occurs within microbial biofilms - communities of microorganisms attached to stone in an exopolymer matrix. We investigated the effect of alginic acid, a common biological polymer produced by bacteria and algae, on calcite dissolution using a paired atomic force microscopy/flow-through reactor apparatus. The alginic acid caused up to an order of magnitude increase in dissolution rate at 3 < pH < 12. Additionally, the polymer preferentially binds to the obtuse pit steps and increases step velocity. We propose that the polymer is actively chelating surficial cations reducing the activation energy and increasing dissolution rate. The role of biologically produced polymers in mineral weathering is important in the protection of cultural heritage materials and understanding of marine and terrestrial systems.

  14. Physical and structural characteristics of acrylated poly(ethylene glycol)-alginate conjugates.

    PubMed

    Davidovich-Pinhas, Maya; Bianco-Peled, Havazelet

    2011-07-01

    Transmucosal delivery of therapeutic agents is a non-invasive approach that utilizes human entry paths such as the nasal, buccal, rectal and vaginal routes. Mucoadhesive polymers have the ability to adhere to the mucus layer covering those surfaces and by that promote drug release, targeting and absorption. We have recently demonstrated that acrylated polymers display enhanced mucoadhesive properties due to their ability to covalently attach to mucus type glycoproteins. We have synthesized an acrylated poly(ethylene glycol)-alginate conjugate (alginate-PEGAc), a molecule which combines the gelation ability of alginate with the mucoadhesion properties arising from both the characteristics of poly(ethylene glycol) and the acrylate functionality. In the current investigation we introduce an in-depth characterization of the thermal, mechanical and structural properties of alginate-PEGAc aimed at gaining a better knowledge of its structure-function relations. The thermal stability, evaluated by thermal gravimetric analysis and differential scanning calorimetry, was compared with that of alginate and the intermediate product thiolated alginate. Dehydration at temperatures up to 200 °C was detected for all samples, followed by distinctive decomposition steps arising from the decomposition of the polymer backbone and side-chains. The nanostructure of the solutions and gels was evaluated from small angle X-ray scattering patterns, to which the "broken rod linked by flexible chain" model was fitted, and from rheology measurements. The maxima arising from electrostatic repulsion between the highly charged alginate chains was diminished for both modified alginate samples, suggesting that modification led to electrostatic screening. Alginate, thiolated alginate and alginate-PEGAc cross-linked with calcium ions demonstrated similar scattering patterns. However, different scattering intensities, gel strengths, and gelation kinetics were observed, suggesting a decrease in the

  15. Surfactant and metal ion effects on the mechanical properties of alginate hydrogels.

    PubMed

    Kaygusuz, Hakan; Evingür, Gülşen Akın; Pekcan, Önder; von Klitzing, Regine; Erim, F Bedia

    2016-11-01

    This paper addresses the controlled variation of the mechanical properties of alginate gel beads by changing the alginate concentration or by adding different surfactants or cross-linking cations. Alginate beads containing nonionic Brij 35 or anionic sodium dodecyl sulfate (SDS) surfactants were prepared with two different types of cations (Ca(2+), Ba(2+)) as crosslinkers. Compression measurements were performed to investigate the effect of the surfactant and cation types and their concentrations on the Young's modulus of alginate beads. The Young's modulus was determined by using Hertz theory. For all types of alginate gel beads the Young's modulus showed an increasing value for increasing alginate contents. Addition of the anionic surfactant SDS increases the Young's modulus of the alginate beads while the addition of non-ionic surfactant Brij 35 leads to a decrease in Young's modulus. This opposite behavior is related to the contrary effect of both surfactants on the charge of the alginate beads. When Ba(2+) ions were used as crosslinker cation, the Young's modulus of the beads with the surfactant SDS was found to be approximately two times higher than the modulus of beads with the surfactant Brij 35. An ion specific effect was found for the crosslinking ability of divalent cations.

  16. Laponite as a rheology modifier of alginate solutions: Physical gelation and aging evolution.

    PubMed

    Dávila, José Luis; d'Ávila, Marcos Akira

    2017-02-10

    The rheological behavior of alginate and Laponite/alginate solutions was studied. It was observed that the Cross viscosity model successfully describes the steady-state shear behavior of this polysaccharide. The scaling behavior analyzed for the entangled regime is in good agreement with polyelectrolyte solutions (Ge∼cp(3/2)), with interactions generated between the alginate and the charged surfaces of the Laponite platelets. Therefore, the effect of Laponite as a rheology modifier is influenced by the alginate concentration. Higher alginate concentrations hindered the formation of the house of cards microstructure. Frequency sweep tests were performed to analyze the transition from solid-like to liquid-like behavior in a solid-like dominated domain. Soft physical gels were obtained at low alginate concentrations. The gel point was determined (1.65wt.% of alginate and 2wt.% of Laponite) through the Kramers-Krönig damping factor, and time sweep tests revealed the evolution of the storage (G') and loss modulus (G″) as functions of the waiting time (tw). The growing elasticity revealed that Laponite/alginate solutions undergo aging.

  17. Ca(ii) and Ce(iii) homogeneous alginate hydrogels from the parent alginic acid precursor: a structural study.

    PubMed

    Sonego, Juan Manuel; Santagapita, Patricio R; Perullini, Mercedes; Jobbágy, Matías

    2016-06-14

    Alginate hydrogels are suitable for the encapsulation of biomolecules and microorganisms for the building of bioactive materials. Several alternatives to the conventional alginate formulation are being studied for a broad range of biotechnological applications; among them the crosslinking of alginate by lanthanide cations, Ln(iii), envisages expanded biomedical applications. The performance of these functional materials is highly related to the microstructure of the alginate matrix, which in turn is affected by the conditions of synthesis. In particular, when a diffusing gradient of the crosslinking cation is involved, microstructure inhomogeneities are expected at the macroscopic level. Here we discuss the subtle differences in the microstructure, as assessed by SAXS (Small Angle X-ray Scattering), established in the direction of the gradient of diffusion of Ca(ii) or Ce(iii).

  18. Preparation, characterisation and viability of encapsulated Trichoderma harzianum UPM40 in alginate-montmorillonite clay.

    PubMed

    Adzmi, Fariz; Meon, Sariah; Musa, Mohamed Hanafi; Yusuf, Nor Azah

    2012-01-01

    Microencapsulation is a process by which tiny parcels of an active ingredient are packaged within a second material for the purpose of shielding the active ingredient from the surrounding environment. This study aims to determine the ability of the microencapsulation technique to improve the viability of Trichoderma harzianum UPM40 originally isolated from healthy groundnut roots as effective biological control agents (BCAs). Alginate was used as the carrier for controlled release, and montmorillonite clay (MMT) served as the filler. The encapsulated Ca-alginate-MMT beads were characterised using Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). The FTIR results showed the interaction between the functional groups of alginate and MMT in the Ca-alginate-MMT beads. Peaks at 1595, 1420 and 1020 cm(-1) characterised alginate, and peaks at 1028 and 453 cm(-1) characterised MMT; both sets of peaks appeared in the Ca-alginate-MMT FTIR spectrum. The TGA analysis showed an improvement in the thermal stability of the Ca-alginate-MMT beads compared with the alginate beads alone. SEM analysis revealed a homogeneous distribution of the MMT particles throughout the alginate matrix. T. harzianum UPM40 was successfully encapsulated in the Ca-alginate-MMT beads. Storage analysis of the encapsulated T. harzianum UPM40 showed that the low storage temperature of 5°C resulted in significantly (p < 0.05) better storage compared with room temperature (30°C).

  19. Development of zirconia nanoparticles-decorated calcium alginate hydrogel fibers for extraction of organophosphorous pesticides from water and juice samples: Facile synthesis and application with elimination of matrix effects.

    PubMed

    Zare, Maryam; Ramezani, Zahra; Rahbar, Nadereh

    2016-11-18

    In this research, novel zirconia nanoparticles-decorated calcium alginate hydrogel fibers (ZNCAHF) were synthesized through a simple, green procedure. ZNCAHF were used as an adsorbent in the micro-solid-phase extraction (MSPE) of methyl parathion (MP), fenitrothion (FT) and malathion (MT) as model pesticides prior to gas chromatography-mass spectroscopic detection (GC-MS). The composition and morphology of the prepared fiber were characterized by Fourier transform-infrared spectroscopy (FTIR), field emission scanning electron microscopy (FESEM), energy dispersive X-ray diffraction (EDX), and differential scanning calorimetry (DSC). Various parameters affecting fabrication of the fiber (weight ratio of components) and relative extraction recovery (pH, amount of adsorbent, extraction time, salt addition, and desorption conditions) were investigated and optimized. Under optimized conditions, the calibration curves were obtained in the concentration range of 0.01-500ngmL(-1) with regression coefficients between 0.9997 and 0.9999. The limits of detection (LOD) (S/N=3) and limits of quantification (LOQ) (S/N=10) of the method ranged from 0.001 to 0.004ngmL(-1) and 0.003 to 0.012ngmL(-1), respectively. The intra-day and inter-day relative standard deviations (RSDs) were 2.2-5.9% and 3.2-7.8%, respectively. The applicability of the fabricated adsorbent was investigated by extraction of selected organophosphorous pesticides (OPPs) from real samples of juice and water. The obtained relative recoveries were in the range of 90.6-105.4%, demonstrating elimination of matrix effects which can be attributed to the remarkable affinity of OPPs toward ZNCAHF.

  20. Layer-by-Layer Self-Assembling Gold Nanorods and Glucose Oxidase onto Carbon Nanotubes Functionalized Sol-Gel Matrix for an Amperometric Glucose Biosensor

    PubMed Central

    Wu, Baoyan; Hou, Shihua; Miao, Zhiying; Zhang, Cong; Ji, Yanhong

    2015-01-01

    A novel amperometric glucose biosensor was fabricated by layer-by-layer self-assembly of gold nanorods (AuNRs) and glucose oxidase (GOD) onto single-walled carbon nanotubes (SWCNTs)-functionalized three-dimensional sol-gel matrix. A thiolated aqueous silica sol containing SWCNTs was first assembled on the surface of a cleaned Au electrode, and then the alternate self-assembly of AuNRs and GOD were repeated to assemble multilayer films of AuNRs-GOD onto SWCNTs-functionalized silica gel for optimizing the biosensor. Among the resulting glucose biosensors, the four layers of AuNRs-GOD-modified electrode showed the best performance. The sol-SWCNTs-(AuNRs-GOD)4/Au biosensor exhibited a good linear range of 0.01–8 mM glucose, high sensitivity of 1.08 μA/mM, and fast amperometric response within 4 s. The good performance of the proposed glucose biosensor could be mainly attributed to the advantages of the three-dimensional sol-gel matrix and stereo self-assembly films, and the natural features of one-dimensional nanostructure SWCNTs and AuNRs. This study may provide a new facile way to fabricate the enzyme-based biosensor with high performance.

  1. Layer-by-Layer Self-Assembling Gold Nanorods and Glucose Oxidase onto Carbon Nanotubes Functionalized Sol-Gel Matrix for an Amperometric Glucose Biosensor.

    PubMed

    Wu, Baoyan; Hou, Shihua; Miao, Zhiying; Zhang, Cong; Ji, Yanhong

    2015-09-18

    A novel amperometric glucose biosensor was fabricated by layer-by-layer self-assembly of gold nanorods (AuNRs) and glucose oxidase (GOD) onto single-walled carbon nanotubes (SWCNTs)-functionalized three-dimensional sol-gel matrix. A thiolated aqueous silica sol containing SWCNTs was first assembled on the surface of a cleaned Au electrode, and then the alternate self-assembly of AuNRs and GOD were repeated to assemble multilayer films of AuNRs-GOD onto SWCNTs-functionalized silica gel for optimizing the biosensor. Among the resulting glucose biosensors, the four layers of AuNRs-GOD-modified electrode showed the best performance. The sol-SWCNTs-(AuNRs- GOD)₄/Au biosensor exhibited a good linear range of 0.01-8 mM glucose, high sensitivity of 1.08 μA/mM, and fast amperometric response within 4 s. The good performance of the proposed glucose biosensor could be mainly attributed to the advantages of the three-dimensional sol-gel matrix and stereo self-assembly films, and the natural features of one-dimensional nanostructure SWCNTs and AuNRs. This study may provide a new facile way to fabricate the enzyme-based biosensor with high performance.

  2. Alginate-chitosan coacervation in production of artificial seeds.

    PubMed

    Tay, L F; Khoh, L K; Loh, C S; Khor, E

    1993-08-05

    Survival of secondary embryoids of winter oilseed rape (Brassica napus ssp. oleifera cv. Primor) has been used as an assay for the development of artificial seeds involving complex coacervation of alginate (polyanion) with chitosan (polycation). Germination frequency of 100% was achieved for encapsulated embryoids when alginate formed the inner matrix and chitosan the outer layer. When the matrix makeup was reversed, there was no germination of embryoids. The artificial seeds produced were hardened in dilute alkaline solutions of NaOH and Ca(OH)(2). An optimum setting time could be selected based on a quantitative measurement of resistance of hardened capsules to compression and the germination frequency of the encapsulated embryoids.

  3. Gel bead composition for metal adsorption

    SciTech Connect

    Scott, C.D.; Woodward, C.A.; Byers, C.H.

    1990-12-18

    This patent describes a gel bead consisting essentially of a sufficient amount of water and propylene glycol alginate to allow for bead formation and a sufficient amount of bone gelatin to allow for metal absorption and chemically crosslinked in an alkaline medium to form a stable structure. A gel bead contained therein a biological absorbent capable of removing metals from solution.

  4. Highly efficient up-conversion and bright white light in RE co-doped KYF4 nanocrystals in sol-gel silica matrix

    NASA Astrophysics Data System (ADS)

    Méndez-Ramos, J.; Yanes, A. C.; Santana-Alonso, A.; del-Castillo, J.

    2013-01-01

    Transparent nano-glass-ceramics comprising Yb3+, Er3+ and Tm3+ co-doped KYF4 nanocrystals have been developed from sol-gel method. A structural analysis by means of X-ray diffraction confirmed the precipitation of cubic KYF4 nanocrystals into a silica matrix. Visible luminescence has been analyzed as function of treatment temperature of precursor sol-gel glasses. Highly efficient up-conversion emissions have been obtained under 980 nm excitation and studied by varying the doping level, processing temperature and pump power. Color tuneability has been quantified in terms of CIE diagram and in particular, a white-balanced overall emission has been achieved for a certain doping level and thermal treatment.

  5. 21 CFR 184.1187 - Calcium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium alginate. 184.1187 Section 184.1187 Food... Specific Substances Affirmed as GRAS § 184.1187 Calcium alginate. (a) Calcium alginate (CAS Reg. No. 9005.... Calcium alginate is prepared by the neutralization of purified alginic acid with appropriate pH...

  6. 21 CFR 184.1610 - Potassium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Potassium alginate. 184.1610 Section 184.1610 Food... GRAS § 184.1610 Potassium alginate. (a) Potassium alginate (CAS Reg. No. 9005-36-1) is the potassium salt of alginic acid, a natural polyuronide constituent of certain brown algae. Potassium alginate...

  7. 21 CFR 184.1724 - Sodium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium alginate. 184.1724 Section 184.1724 Food and... Substances Affirmed as GRAS § 184.1724 Sodium alginate. (a) Sodium alginate (CAS Reg. No. 9005-38-3) is the sodium salt of alginic acid, a natural polyuronide constituent of certain brown algae. Sodium alginate...

  8. 21 CFR 184.1724 - Sodium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Sodium alginate. 184.1724 Section 184.1724 Food... GRAS § 184.1724 Sodium alginate. (a) Sodium alginate (CAS Reg. No. 9005-38-3) is the sodium salt of alginic acid, a natural polyuronide constituent of certain brown algae. Sodium alginate is prepared by...

  9. Alginate encapsulation of Begonia microshoots for short-term storage and distribution.

    PubMed

    Sakhanokho, Hamidou F; Pounders, Cecil T; Blythe, Eugene K

    2013-01-01

    Synthetic seeds were formed from shoot tips of two in vitro grown Begonia cultivars using 3% sodium alginate in Murashige and Skoog medium (MS) salt solution as the gel matrix and 100 mM calcium chloride for complexation. Synthetic seed formation was achieved by releasing the sodium alginate/explant combination into 100 mM calcium chloride (CaCl₂ ·H₂O) solution for 30 or 45 min. Both control and encapsulated shoots were transferred into sterile Petri dishes and stored at 4°C or 22°C for 0, 2, 4, 6, or 8 weeks. Conversion of synthetic seeds into plantlets for both storage environments was assessed in MS medium or peat-based substrate. No significant difference was found between the 30 and 45 min CaCl₂ ·H₂O treatments or the two cultivars. Encapsulation of explants improved survival rate over time irrespective of the medium type or storage environment. Survival rates of 88, 53, 28, and 11% for encapsulated microshoots versus 73, 13, 0, and 0% for control explants were achieved in microshoots stored for 2, 4, 6, and 8 weeks, respectively. The best results were obtained when synthetic seeds were stored at 4°C and germinated on MS medium. Regenerated plantlets were successfully established in potting soil.

  10. Alginate Encapsulation of Begonia Microshoots for Short-Term Storage and Distribution

    PubMed Central

    Sakhanokho, Hamidou F.; Pounders, Cecil T.; Blythe, Eugene K.

    2013-01-01

    Synthetic seeds were formed from shoot tips of two in vitro grown Begonia cultivars using 3% sodium alginate in Murashige and Skoog medium (MS) salt solution as the gel matrix and 100 mM calcium chloride for complexation. Synthetic seed formation was achieved by releasing the sodium alginate/explant combination into 100 mM calcium chloride (CaCl2·H2O) solution for 30 or 45 min. Both control and encapsulated shoots were transferred into sterile Petri dishes and stored at 4°C or 22°C for 0, 2, 4, 6, or 8 weeks. Conversion of synthetic seeds into plantlets for both storage environments was assessed in MS medium or peat-based substrate. No significant difference was found between the 30 and 45 min CaCl2·H2O treatments or the two cultivars. Encapsulation of explants improved survival rate over time irrespective of the medium type or storage environment. Survival rates of 88, 53, 28, and 11% for encapsulated microshoots versus 73, 13, 0, and 0% for control explants were achieved in microshoots stored for 2, 4, 6, and 8 weeks, respectively. The best results were obtained when synthetic seeds were stored at 4°C and germinated on MS medium. Regenerated plantlets were successfully established in potting soil. PMID:24396296

  11. Enzymatic cellulose hydrolysis: enzyme reusability and visualization of β-glucosidase immobilized in calcium alginate.

    PubMed

    Tsai, Chien-Tai; Meyer, Anne S

    2014-11-25

    The high cellulase enzyme dosages required for hydrolysis of cellulose is a major cost challenge in lignocellulosic ethanol production. One method to decrease the enzyme dosage and increase biocatalytic productivity is to re-use β-glucosidase (BG) via immobilization. In the present research, glutaraldehyde cross-linked BG was entrapped in calcium alginate gel particles. More than 60% of the enzyme activity could be recovered under optimized conditions, and glutaraldehyde cross-linking decreased leakage of BG from the calcium alginate particles. The immobilized BG aggregates were visualized by confocal laser scanning microscopy (CLSM). The CLSM images, which we believe are the first to be published, corroborate that more BG aggregates were entrapped in the matrix when the enzymes were cross-linked by glutaraldehyde as opposed to when they are not cross-linked. The particles with the immobilized BG were recycled for cellulase catalyzed hydrolysis of Avicel. No significant loss in BG activity was observed for up to 20 rounds of reaction recycle steps of the BG particles of 48 h each, verifying a significant stabilization of the BG by immobilization. Similar high glucose yields were obtained by one round of enzymatic hydrolysis of hydrothermally pretreated barley straw during a 72 h reaction with immobilized BG and free BG.

  12. Gelling process of sodium alginate with bivalent ions rich microsphere: Nature of bivalent ions

    NASA Astrophysics Data System (ADS)

    Mauri, Marco; Vicini, Silvia; Castellano, Maila

    2016-05-01

    In the paper we present a new approach for obtaining a controlled gelling process of sodium alginate, based on the quantity of bivalent ions rich alginate micro-beads added as crosslinkers. Typically, calcium ions are used in gelation of alginate solutions. In this study we present different gelling systems realized with alginate microspheres, made by electrospinning methodology, enriched with different bivalent ions (Ca2+, Ba2+ and Mg2+). The microspheres were characterized under the point of view of the morphology by OM and as the ions content. Realized gels were characterized in light of the amount of the ions added to the alginate solution, and in light of the different dimensions of the micro-beads, using rheological measurements to assess the variation in the storage modulus (G'), loss modulus (G″) and complex viscosity (η*).

  13. Microencapsulation of bioactives in cross-linked alginate matrices by spray drying.

    PubMed

    Santa-Maria, Monica; Scher, Herbert; Jeoh, Tina

    2012-01-01

    Microencapsulation of biomolecules, cells and chemicals is widely used in the food and pharmaceutical industries to improve stability, delivery and to control the release of encapsulated moieties. Among encapsulation matrices, alginate is preferred due to its low cost, biodegradability and biocompatibility. Current methods for producing stable alginate gels involve dropping alginate suspensions into divalent cation solutions. This procedure is difficult to scale-up and produces undesirably large alginate beads. In our novel encapsulation method, alginate gelation occurs during spray drying upon volatilisation of a base and rapid release of otherwise unavailable calcium ions. The resulting particles, with median particle sizes in the range 15-120 µm, are insoluble in solution. Cellulase and hemicellulase activities encapsulated by this method were not compromised during spray drying and remained stable over prolonged storage. The procedure described here offers a one-step alternative to other encapsulation methods that are costly and difficult to scale-up.

  14. Alginate: A Versatile Biomaterial to Encapsulate Isolated Ovarian Follicles.

    PubMed

    Vanacker, Julie; Amorim, Christiani A

    2017-02-28

    In vitro culture of ovarian follicles isolated or enclosed in ovarian tissue fragments and grafting of isolated ovarian follicles represent a potential alternative to restore fertility in cancer patients who cannot undergo cryopreservation of embryos or oocytes or transplantation of frozen-thawed ovarian tissue. In this regard, respecting the three-dimensional (3D) architecture of isolated follicles is crucial to maintaining their proper follicular physiology. To this end, alginate hydrogel has been widely investigated using follicles from numerous animal species, yielding promising results. The goal of this review is therefore to provide an overview of alginate applications utilizing the biomaterial as a scaffold for 3D encapsulation of isolated ovarian follicles. Different methods of isolated follicle encapsulation in alginate are discussed in this review, as its use of 3D alginate culture systems as a tool for in vitro follicle analysis. Possible improvements of this matrix, namely modification with arginine-glycine-aspartic acid peptide or combination with fibrin, are also summarized. Encouraging results have been obtained in different animal models, and particularly with isolated follicles encapsulated in alginate matrices and grafted to mice. This summary is designed to guide the reader towards development of next-generation alginate scaffolds, with enhanced properties for follicle encapsulation.

  15. Removable colored coatings based on calcium alginate hydrogels.

    PubMed

    Kobaslija, Muris; McQuade, D Tyler

    2006-08-01

    This article describes the creation of a nontoxic, biodegradable coating using calcium alginate and FD&C approved dyes. The coating is robust but is rapidly removed upon treatment with disodium ethylenediamine tetraacetate (EDTA). Dye leaching from calcium alginate films was studied, and it was determined that the efficiency of dye retention is proportional to the degree of cross-linking. Degradation rates were studied on calcium alginate beads serving as a model for a coating. We determined that degradation rates depend on the gel's cross-linking and on the amount of EDTA used. Bead size also influenced the degradation rates; smaller beads degraded faster than larger beads. We show that the coating can be used as an easily removable and environmentally friendly logotype on an artificial turf surface. Applications of these coatings can be extended to food, cosmetic, medicinal, and textile uses and to wherever nontoxic, easily removable colored coating is desired.

  16. Preparation of calcium alginate microgel beads in an electrodispersion reactor using an internal source of calcium carbonate nanoparticles.

    PubMed

    Zhao, Yinyan; Carvajal, M Teresa; Won, You-Yeon; Harris, Michael T

    2007-12-04

    An electrodispersion reactor has been used to prepare calcium alginate (Ca-alginate) microgel beads in this study. In the electrodispersion reactor, pulsed electric fields are utilized to atomize aqueous mixtures of sodium alginate and CaCO3 nanoparticles (dispersed phase) from a nozzle into an immiscible, insulating second liquid (continuous phase) containing a soluble organic acid. This technique combines the features of the electrohydrodynamic force driven emulsion processes and externally triggered gelations in microreactors (the droplets) ultimately to yield soft gel beads. The average particle size of the Ca-alginate gels generated by this method changed from 412 +/- 90 to 10 +/- 3 microm as the applied peak voltage was increased. A diagram depicting structural information for the Ca-alginate was constructed as a function of the concentrations of sodium alginate and CaCO3 nanoparticles. From this diagram, a critical concentration of sodium alginate required for sol-gel transformation was observed. The characteristic highly porous structure of Ca-alginate particles made by this technique appears suitable for microencapsulation applications. Finally, time scale analysis was performed for the electrodispersion processes that include reactions in the microreactor droplets to provide guidelines for the future employment of this technique. This electrodispersion reactor can be used potentially in the formation of many reaction-based microencapsulation systems.

  17. One-step preparation of CdS nanocrystals supported on thiolated silica-gel matrix and evaluation of photocatalytic performance.

    PubMed

    Andrade, George R S; Nascimento, Cristiane C; Neves, Erick C; Barbosa, Cintya D'Angeles Espirito Santo; Costa, Luiz P; Barreto, Ledjane S; Gimenez, Iara F

    2012-02-15

    Here we report the use of a thiol-functionalized silica-gel to prepare supported CdS nanocrystals by a facile one-step procedure. Upon changing the relative proportion of the matrix we obtained nanocrystals with different average sizes and size distributions, as evidenced by spectroscopic measurements as well as TEM images. Photoluminescence spectra also indicated that the main effect of the matrix is related to the size control since the spectral profiles were found to be strongly dependent on the excitation wavelength. The performance of the material in the photocatalytic degradation of two commercial dyes (methylene blue and rhodamine 6G) has been tested under sunlight radiation, showing promising results. Almost complete decolorization has been observed after 80 min of exposure, with no adsorption on the silica surface.

  18. Thermosensitive and mucoadhesive in situ gel based on poloxamer as new carrier for rectal administration of nimesulide.

    PubMed

    Yuan, Yuan; Cui, Ying; Zhang, Li; Zhu, Hui-Ping; Guo, Yi-Sha; Zhong, Bo; Hu, Xia; Zhang, Ling; Wang, Xiao-Hui; Chen, Li

    2012-07-01

    Poloxamer 407 has excellent thermo-sensitive gelling properties. Nevertheless, these gels possess inadequate poor bioadhesiveness and high permeability to water, which limited its' application as a thermoresponsive matrix. The main aim of the present investigation was to develop thermosensitive and mucoadhesive rectal in situ gel of nimesulide (NM) by using mucoadhesive polymers such as sodium alginate (Alg-Na) and HPMC. These gels were prepared by addition of mucoadhesive polymers (0.5%) to the formulations of thermosensitive gelling solution containing poloxamer 407 (18%) and nimesulide (2.0%). Polyethylene glycol (PEG) was used to modify gelation temperature and drug release properties. The gelation temperature and drug release rate of the prepared in situ gels were evaluated. Gelation temperature was significantly increased with incorporation of nimesulide (2.0%) in the poloxamer solution, while the addition of the mucoadhesive polymers played a reverse role on gelation temperature. The addition of PEG polymers increased the gelation temperature and the drug release rate. Among the formulations examined, the poloxamer 407/nimesulide/sodium alginate/PEG 4000 (18/2.0/0.5/1.2%) exhibited the appropriate gelation temperature, acceptable drug release rate and rectal retention at the administration site. Furthermore, the micrographic results showed that in situ gel, given at the dose of 20mg/kg, was safe for no mucosa irritation. In addition, it resulted in significantly higher initial serum concentrations, C(max) and AUC of NM compared to the solid suppository.

  19. Enzyme immobilization in novel alginate-chitosan core-shell microcapsules.

    PubMed

    Taqieddin, Ehab; Amiji, Mansoor

    2004-05-01

    Alginate-chitosan core-shell microcapsules were prepared in order to develop a biocompatible matrix for enzyme immobilization, where the protein is retained either in a liquid or solid core and the shell allows permeability control over substrates and products. The permeability coefficients of different molecular weight compounds (vitamin B2, vitamin B12, and myoglobin) were determined through sodium tripolyphosphate (Na-TPP)-crosslinked chitosan membrane. The microcapsule core was formed by crosslinking sodium alginate with either calcium or barium ions. The crosslinked alginate core was uniformly coated with a chitosan layer and crosslinked with Na-TPP. In the case of calcium alginate, the phosphate ions of Na-TPP were able to extract the calcium ions from alginate and liquefy the core. A model enzyme, beta-galactosidase, was immobilized in the alginate core and the catalytic activity was measured with o-nitrophenyl-beta-D-galactopyranoside (ONPG). Change in the activity of free and immobilized enzyme was determined at three different temperatures. Na-TPP crosslinked chitosan membranes were found to be permeable to solutes of up to 17,000Da molecular weight. The enzyme loading efficiency was higher in the barium alginate core (100%) as compared to the calcium alginate core (60%). The rate of ONPG conversion to o-nitrophenol was faster in the case of calcium alginate-chitosan microcapsules as compared to barium alginate-chitosan microcapsules. Barium alginate-chitosan microcapsules, however, did improve the stability of the enzyme at 37 degrees C relative to calcium alginate-chitosan microcapsules or free enzyme. This study illustrates a new method of enzyme immobilization for biotechnology applications using liquid or solid core and shell microcapsule technology.

  20. Alginate as a cell culture substrate for growth and differentiation of human retinal pigment epithelial cells.

    PubMed

    Heidari, Razeih; Soheili, Zahra-Soheila; Samiei, Shahram; Ahmadieh, Hamid; Davari, Maliheh; Nazemroaya, Fatemeh; Bagheri, Abouzar; Deezagi, Abdolkhalegh

    2015-03-01

    The purpose of this study was to evaluate retinal pigment epithelium (RPE) cells' behavior in alginate beads that establish 3D environment for cellular growth and mimic extracellular matrix versus the conventional 2D monolayer culture. RPE cells were encapsulated in alginate beads by dripping alginate cell suspension into CaCl2 solution. Beads were suspended in three different media including Dulbecco's modified Eagle's medium (DMEM)/F12 alone, DMEM/F12 supplemented with 10 % fetal bovine serum (FBS), and DMEM/F12 supplemented with 30 % human amniotic fluid (HAF). RPE cells were cultivated on polystyrene under the same conditions as controls. Cell phenotype, cell proliferation, cell death, and MTT assay, immunocytochemistry, and real-time RT-PCR were performed to evaluate the effect of alginate on RPE cells characteristics and integrity. RPE cells can survive and proliferate in alginate matrixes. Immunocytochemistry analysis exhibited Nestin, RPE65, and cytokeratin expressions in a reasonable number of cultured cells in alginate beads. Real-time PCR data demonstrated high levels of Nestin, CHX10, RPE65, and tyrosinase gene expressions in RPE cells immobilized in alginate when compared to 2D monolayer culture systems. The results suggest that alginate can be used as a reliable scaffold for maintenance of RPE cells' integrity and in vitro propagation of human retinal progenitor cells for cell replacement therapies in retinal diseases.

  1. Alginate Nanoparticles as a Promising Adjuvant and Vaccine Delivery System

    PubMed Central

    Sarei, F.; Dounighi, N. Mohammadpour; Zolfagharian, H.; Khaki, P.; Bidhendi, S. Moradi

    2013-01-01

    During last decades, diphtheria has remained as a serious disease that still outbreaks and can occur worldwide. Recently, new vaccine delivery systems have been developed by using the biodegradable and biocompatible polymers such as alginate. Alginate nanoparticles as a carrier with adjuvant and prolong release properties that enhance the immunogenicity of vaccines. In this study diphtheria toxoid loaded nanoparticles were prepared by ionic gelation technique and characterized with respect to size, zeta potential, morphology, encapsulation efficiency, release profile, and immunogenicity. Appropriate parameters (calcium chloride and sodium alginate concentration, homogenization rate and homogenization time) redounded to the formation of suitable nanoparticles with a mean diameter of 70±0.5 nm. The loading studies of the nanoparticles resulted in high loading capacities (>90%) and subsequent release studies showed prolong profile. The stability and antigenicity of toxoid were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and ouchterlony test and proved that the encapsulation process did not affect the antigenic integrity and activity. Guinea pigs immunized with the diphtheria toxoid-loaded alginate nanoparticles showed highest humoral immune response than conventional vaccine. It is concluded that, with regard to the desirable properties of nanoparticles and high immunogenicity, alginate nanoparticles could be considered as a new promising vaccine delivery and adjuvant system. PMID:24302799

  2. Synthesis of an imprinted hybrid organic-inorganic polymeric sol-gel matrix toward the specific binding and isotherm kinetics investigation of creatinine.

    PubMed

    Chang, Yong-Sheun; Ko, Ting-Hsien; Hsu, Ting-Jung; Syu, Mei-Jywan

    2009-03-15

    Hybrid organic-inorganic polymeric sol-gel materials imprinted with creatinine template molecules were synthesized for the specific binding of creatinine. Creatinine is a metabolite from creatine and is the final product from kidney metabolism. Therefore, creatinine can be an important index to estimate the function of the kidney. It was then chosen as the target molecule in this work. To achieve the specific binding toward creatinine, molecular imprinting was used to create a polymeric matrix for the regarding purpose. Sol-gel was further added to create a rigid network structure for the absorption of creatinine. An inorganic precursor, tetraethoxysilane (TEOS), was mixed with an organic functional monomer, 2-acrylamido-2-methylpropane-sulfonic acid (AMPS), and the creatinine template to form a hybrid organic-inorganic imprinted polymer. The chemical functionality was achieved as well as a confined matrix via the polymerization and the hydrolysis-condensation of the sol-gel. The imprinting effect from the hybrid materials against the corresponding nonimprinted was investigated. BET (Brunauer-Emmett-Teller) analysis was carried out for the imprinted and the nonimprinted materials. The specificity of the hybrid materials was further examined by capping the surface silanol groups with chloro-trimethylsilane (CTMS) and 1,1,1,3,3,3-hexamethyldisilazane (HMDS), respectively. The capping effect was compared and discussed from the binding results. Selectivity of the materials toward creatinine was obtained using mixture solutions in the presence of creatinine and its analogues. Reutilization and storage stability of the hybrid organic-inorganic imprinted material were also studied. Additionally, the affinity distribution of the hybrid imprinted materials derived from the allosteric model was also analyzed from the adsorption isotherm data.

  3. Jellyfish collagen and alginate: Combined marine materials for superior chondrogenesis of hMSC.

    PubMed

    Pustlauk, W; Paul, B; Gelinsky, M; Bernhardt, A

    2016-07-01

    Marine, hybrid constructs of porous scaffolds from fibrillized jellyfish collagen and alginate hydrogel are mimicking both of the main tissue components of cartilage, thus being a promising approach for chondrogenic differentiation of human mesenchymal stem cells (hMSC). Investigating their potential for articular cartilage repair, the present study examined scaffolds being either infiltrated with an alginate-cell-suspension (ACS) or seeded with hMSC and embedded in alginate after cell adhesion (EAS). Hybrid constructs with 2×10(5) and 4.5×10(5)hMSC/scaffold were compared to hMSC encapsulated in pure alginate discs, both chondrogenically stimulated for 21days. Typical round, chondrocyte-like morphology was observed in pure alginate gels and ACS scaffolds, while cells in EAS were elongated and tightly attached to the collagen pores. Col 2 gene expression was comparable in all scaffold types examined. However, the Col 2/Col 1 ratio was higher for pure alginate discs and ACS scaffolds compared to EAS. In contrast, cells in EAS scaffolds displayed higher gene expression of Sox 9, Col 11 and ACAN compared to ACS and pure alginate. Secretion of sulfated glycosaminoglycans (sGAG) was comparable for ACS and EAS scaffolds. In conclusion hybrid constructs of jellyfish collagen and alginate support hMSC chondrogenic differentiation and provide more stable and constructs compared to pure hydrogels.

  4. Synthesis of "click" alginate hydrogel capsules and comparison of their stability, water swelling, and diffusion properties with that of Ca(+2) crosslinked alginate capsules.

    PubMed

    Breger, Joyce C; Fisher, Benjamin; Samy, Raghu; Pollack, Steven; Wang, Nam Sun; Isayeva, Irada

    2015-07-01

    Ionically crosslinked alginate hydrogels have been extensively explored for encapsulation and immunoisolation of living cells/tissues to develop implantable cell therapies, such as islet encapsulation for bioartificial pancreas. Chemical instability of these hydrogels during long-term implantation hinders the development of viable cell therapy. The exchange between divalent crosslinking ions (e.g., Ca(+2) ) with monovalent ions from physiological environment causes alginate hydrogels to degrade, resulting in exposure of the donor tissue to the host's immune system and graft failure. The goal of this study was to improve stability of alginate hydrogels by utilizing covalent "click" crosslinking while preserving other biomedically viable hydrogel properties. Alginate was first functionalized to contain either pendant alkyne or azide functionalities, and subsequently reacted via "click" chemistry to form "click" gel capsules. Alginate functionalization was confirmed by NMR and gel permeation chromatography. When compared with Ca(+2) capsules, "click" capsules exhibited superior stability in ionic media, while showing higher permeability to small size diffusants and similar molecular weight cut-off and water swelling. Physicochemical properties of "click" alginate hydrogels demonstrate their potential utility for therapeutic cell encapsulation and other biomedical applications.

  5. Alginate-polymethacrylate hybrid hydrogels with double ionic and covalent network for tissue engineering

    NASA Astrophysics Data System (ADS)

    Schizzi, I.; Utzeri, R.; Castellano, M.; Stagnaro, P.

    2016-05-01

    Hydrogels based on alginates are very promising candidates to realize scaffolds for tissue engineering. Indeed, alginate hydrogels are able to mimic the extracellular matrix (ECM) thus promoting in vitro and/or in vivo cell growth; moreover, their capability of giving rise to highly porous structures can specifically favor the osteochondral tissue regeneration. However, mechanical properties of polymeric hydrogels are often inadequate to endow the final constructs with the required characteristics of elasticity and toughness. Here alginate/polymethacrylate hybrid hydrogels, with a suitable porous structure and characterized by a double network, ionic (from alginate) and covalent (from polymethacrylate) were designed and realized. The mechanical performance of these hybrid materials resulted, as expected, improved due to the double interconnected network, where the alginate portion provides the appropriate micro-environment mimicking the ECM, whereas the polymethacrylate portion acts as a reinforce.

  6. Alginate gelation-induced cell death during laser-assisted cell printing.

    PubMed

    Gudapati, Hemanth; Yan, Jingyuan; Huang, Yong; Chrisey, Douglas B

    2014-09-01

    Modified laser-induced forward transfer has emerged as a promising bioprinting technique. Depending on the operating conditions and cell properties, laser cell printing may cause cell injury and even death, which should be carefully elucidated for it to be a viable technology. This study has investigated the effects of alginate gelation, gelation time, alginate concentration, and laser fluence on the post-transfer cell viability of NIH 3T3 fibroblasts. Sodium alginate and calcium chloride are used as the gel precursor and gel reactant solution to form cell-laden alginate microspheres. It is found that the effects of gelation depend on the duration of gelation. Two-minute gelation is observed to increase the cell viability after 24 h incubation, mainly due to the protective cushion effect of the forming gel membrane during droplet landing. Despite the cushion effect from 10 min gelation, it is observed that the cell viability decreases after 24 h incubation because of the forming thick gel membrane that reduces nutrient and oxygen diffusion from the culture medium. In addition, the cell viability after 24 h incubation decreases as the laser fluence or alginate concentration increases.

  7. Protection of probiotic bacteria in a synbiotic matrix following aerobic storage at 4 °C.

    PubMed

    Chaluvadi, S; Hotchkiss, A T; Call, J E; Luchansky, J B; Phillips, J G; Liu, Ls; Yam, K L

    2012-09-01

    The survival of single strains of Bifidobacterium breve, Bifidobacterium longum, Lactobacillus acidophilus, and Lactobacillus reuteri was investigated in synbiotics that included 10 mg/ml of fructo-oligosaccharides, inulin and pectic-oligosaccharides in an alginate matrix under refrigerated (4 °C) aerobic storage conditions. When the matrices were cross-linked with calcium (45 mM), 102-103 cfu/ml of L. acidophilus and L. reuteri, and 0-103 cfu/ml of B. breve and B. longum survived refrigerated aerobic storage for 28 days. Following refrigerated storage, acetic (3-9 mM), butyric (0-2 mM), propionic (5-16 mM) and lactic acids (1-48 mM) were produced during the growth of probiotics in BHI broth at 37 °C, suggesting their metabolic activity after storage was stressed. When calcium cross-linking was not used in synbiotics, the matrix remained more gel-like after inoculation when compared to the calcium cross-linked matrix. At least 107 cfu/ml of probiotic bacteria survived after 21 days of storage within these gel-like alginate matrices. Significantly higher levels of B. breve, L. acidophilus and L. reuteri were obtained from the synbiotic matrices supplemented with fructo-oligosaccharides, inulin and pectic-oligosaccharides compared to alginate alone. B. longum survival was the same (~7 logs) in all gel-like synbiotic matrices. These results show that synbiotics protected probiotic bacteria and extended their shelf-life under refrigerated aerobic conditions. Synbiotics represent a viable delivery vehicle for health-promoting bacteria.

  8. 21 CFR 184.1610 - Potassium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Potassium alginate. 184.1610 Section 184.1610 Food... Specific Substances Affirmed as GRAS § 184.1610 Potassium alginate. (a) Potassium alginate (CAS Reg. No. 9005-36-1) is the potassium salt of alginic acid, a natural polyuronide constituent of certain...

  9. 21 CFR 184.1610 - Potassium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium alginate. 184.1610 Section 184.1610 Food... Specific Substances Affirmed as GRAS § 184.1610 Potassium alginate. (a) Potassium alginate (CAS Reg. No. 9005-36-1) is the potassium salt of alginic acid, a natural polyuronide constituent of certain...

  10. 21 CFR 184.1610 - Potassium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Potassium alginate. 184.1610 Section 184.1610 Food... Specific Substances Affirmed as GRAS § 184.1610 Potassium alginate. (a) Potassium alginate (CAS Reg. No. 9005-36-1) is the potassium salt of alginic acid, a natural polyuronide constituent of certain...

  11. 21 CFR 184.1610 - Potassium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Potassium alginate. 184.1610 Section 184.1610 Food... Specific Substances Affirmed as GRAS § 184.1610 Potassium alginate. (a) Potassium alginate (CAS Reg. No. 9005-36-1) is the potassium salt of alginic acid, a natural polyuronide constituent of certain...

  12. 21 CFR 184.1724 - Sodium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Sodium alginate. 184.1724 Section 184.1724 Food... Specific Substances Affirmed as GRAS § 184.1724 Sodium alginate. (a) Sodium alginate (CAS Reg. No. 9005-38-3) is the sodium salt of alginic acid, a natural polyuronide constituent of certain brown...

  13. 21 CFR 184.1724 - Sodium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium alginate. 184.1724 Section 184.1724 Food... Specific Substances Affirmed as GRAS § 184.1724 Sodium alginate. (a) Sodium alginate (CAS Reg. No. 9005-38-3) is the sodium salt of alginic acid, a natural polyuronide constituent of certain brown...

  14. 21 CFR 184.1724 - Sodium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Sodium alginate. 184.1724 Section 184.1724 Food... Specific Substances Affirmed as GRAS § 184.1724 Sodium alginate. (a) Sodium alginate (CAS Reg. No. 9005-38-3) is the sodium salt of alginic acid, a natural polyuronide constituent of certain brown...

  15. A new nano-optical sensor thin film cadmium sulfide doped in sol-gel matrix for assessment of α-amylase activity in human saliva.

    PubMed

    Attia, M S; Zoulghena, H; Abdel-Mottaleb, M S A

    2014-02-21

    A novel, simple, sensitive and precise spectrofluorimetric method is developed for measuring the activity of the α-amylase enzyme in human saliva. The remarkable quenching of the luminescence intensity at 634 nm of nano CdS doped in a sol-gel matrix by various concentrations of maltose (produced from the reaction of the enzyme with the starch substrate) was successfully used as an optical sensor for the assessment of α-amylase activity. The calibration plot was achieved over the concentration range 4.8 × 10(-10) to 1.2 × 10(-5) mol L(-1) maltose with a correlation coefficient of 0.999 and a detection limit of 5.7 × 10(-11) mol L(-1). The method was used satisfactorily for assessment of the α-amylase activity in a number of human saliva samples collected from various healthy volunteers.

  16. Inhibition of vimentin or B1 integrin reverts morphology of prostate tumor cells grown in laminin-rich extracellular matrix gels and reduces tumor growth in vivo

    SciTech Connect

    Zhang, Xueping; Fournier, Marcia V; Ware, Joy L; Bissell, Mina J; Yacoub, Adly; Zehner, Zendra E

    2008-06-12

    Prostate epithelial cells grown embedded in laminin-rich extracellular matrix (lrECM) undergo morphologic changes that closely resemble their architecture in vivo. In this study, growth characteristics of three human prostate epithelial sublines derived from the same cellular lineage, but displaying different tumorigenic and metastatic properties in vivo, were assessed in three-dimensional lrECM gels. M12, a highly tumorigenic and metastatic subline, was derived from the immortalized, prostate epithelial P69 cell line by selection in athymic, nude mice and found to contain a deletion of 19p-q13.1. The stable reintroduction of an intact human chromosome 19 into M12 resulted in a poorly tumorigenic subline, designated F6. When embedded in lrECM gels, the parental, nontumorigenic P69 line produced acini with clearly defined lumena. Immunostaining with antibodies to {beta}-catenin, E-cadherin, or {alpha}6 and {beta}1 integrins showed polarization typical of glandular epithelium. In contrast, the metastatic M12 subline produced highly disorganized cells with no evidence of polarization. The F6 subline reverted to acini-like structures exhibiting basal polarity marked with integrins. Reducing either vimentin levels via small interfering RNA interference or the expression of {alpha}6 and {beta}1 integrins by the addition of blocking antibodies, reorganized the M12 subline into forming polarized acini. The loss of vimentin significantly reduced M12-Vim tumor growth when assessed by s.c. injection in athymic mice. Thus, tumorigenicity in vivo correlated with disorganized growth in three-dimensional lrECM gels. These studies suggest that the levels of vimentin and {beta}1 integrin play a key role in the homeostasis of the normal acinus in prostate and that their dysregulation may lead to tumorigenesis. [Mol Cancer Ther 2009;8(3):499-508].

  17. Decision peptide-driven: a free software tool for accurate protein quantification using gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry.

    PubMed

    Santos, Hugo M; Reboiro-Jato, Miguel; Glez-Peña, Daniel; Nunes-Miranda, J D; Fdez-Riverola, Florentino; Carvallo, R; Capelo, J L

    2010-09-15

    The decision peptide-driven tool implements a software application for assisting the user in a protocol for accurate protein quantification based on the following steps: (1) protein separation through gel electrophoresis; (2) in-gel protein digestion; (3) direct and inverse (18)O-labeling and (4) matrix assisted laser desorption ionization time of flight mass spectrometry, MALDI analysis. The DPD software compares the MALDI results of the direct and inverse (18)O-labeling experiments and quickly identifies those peptides with paralleled loses in different sets of a typical proteomic workflow. Those peptides are used for subsequent accurate protein quantification. The interpretation of the MALDI data from direct and inverse labeling experiments is time-consuming requiring a significant amount of time to do all comparisons manually. The DPD software shortens and simplifies the searching of the peptides that must be used for quantification from a week to just some minutes. To do so, it takes as input several MALDI spectra and aids the researcher in an automatic mode (i) to compare data from direct and inverse (18)O-labeling experiments, calculating the corresponding ratios to determine those peptides with paralleled losses throughout different sets of experiments; and (ii) allow to use those peptides as internal standards for subsequent accurate protein quantification using (18)O-labeling. In this work the DPD software is presented and explained with the quantification of protein carbonic anhydrase.

  18. Cold gelation of alginates induced by monovalent cations.

    PubMed

    Karakasyan, C; Legros, M; Lack, S; Brunel, F; Maingault, P; Ducouret, G; Hourdet, D

    2010-11-08

    A new reversible gelation pathway is described for alginates in aqueous media. From various samples differing by their mannuronic/guluronic content (M/G), both enthalpic and viscoelastic experiments demonstrate that alginates having a high M content are able to form thermoreversible assemblies in the presence of potassium salts. The aggregation behavior is driven by the low solubility of M-blocks at low temperature and high ionic strength. In semidilute solutions, responsive assemblies induce a strong increase of the viscosity below a critical temperature. A true physical gel is obtained in the entangled regime, although the length scale of specific interactions between M-blocks decreases with increasing density of entanglements. Cold setting takes place at low temperatures, below 0 °C for potassium concentrations lower than 0.2 mol/kg, but the aggregation process can be easily shifted to higher temperatures by increasing the salt concentration. The self-assembling process of alginates in solution of potassium salts is characterized by a sharp gelation exotherm and a broad melting endotherm with a large hysteresis of 20-30 °C between the transition temperatures. The viscoelastic properties of alginate gels in potassium salts closely depend on thermal treatment (rate of cooling, time, and temperature of storage), polymer and salt concentrations, and monomer composition as well. In the case of alginates with a high G content, a similar aggregation behavior is also evidenced at higher salt concentrations, but the extent of the self-assembling process remains too weak to develop a true gelation behavior in solution.

  19. Terminal sterilization of alginate hydrogels: efficacy and impact on mechanical properties.

    PubMed

    Stoppel, Whitney L; White, Joseph C; Horava, Sarena D; Henry, Anna C; Roberts, Susan C; Bhatia, Surita R

    2014-05-01

    Terminal, or postprocessing, sterilization of composite biomaterials is crucial for their use in wound healing and tissue-engineered devices. Recent research has focused on optimizing traditional biomaterial formulations to create better products for commercial and academic use which incorporate hydrophobic compounds or secondary gel networks. To use a hydrogel in a clinical setting, terminal sterilization is necessary to ensure patient safety. Lyophilization, gamma-irradiation, and ethylene oxide treatment all have negative consequences when applied to alginate scaffolds for clinical use. Here, we aim to find alternative terminal sterilization methods for alginate and alginate-based composite hydrogels which maintain the structure of composite alginate networks for use in biomedical applications. A thorough investigation of the effect of common sterilization methods on swollen alginate-based hydrogels has not been reported and therefore, this work examines autoclaving, ethanol washing, and ultraviolet light as sterilization techniques for alginate and alginate/Pluronic® F68 composite hydrogels. Preservation of structural integrity is evaluated using shear rheology and analysis of water retention, and efficacy of sterilization is determined via bacterial persistence within the hydrogel. Results indicate that ethanol sterilization is the best method of those investigated because ethanol washing results in minimal effects on mechanical properties and water retention and eliminates bacterial persistence. Furthermore, this study suggests that ethanol treatment is an efficacious method for terminally sterilizing interpenetrating networks or other composite hydrogel systems.

  20. The effect of omeprazole pre-treatment on rafts formed by reflux suppressant tablets containing alginate.

    PubMed

    Dettmar, P W; Little, S L; Baxter, T

    2005-01-01

    Alginate-based reflux suppressant preparations provide symptom relief by forming a physical barrier on top of the stomach contents in the form of a neutral floating gel or raft. This study investigated whether reduced acidity in the stomach brought about by omeprazole pre-treatment affected the formation and gastric residence time of alginate rafts. It was a balanced, cross-over study in 12 healthy non-patient volunteers following a single dose of two indium-111-labelled alginate tablets in the presence or absence of 3 days' pre-treatment with omeprazole. Raft formation and gastric residence, in the presence of a technetium-99m-labelled meal, were assessed by gamma scintigraphy for 3 h after alginate tablet administration. The relative raft-forming ability of alginate tablets after omeprazole compared with alginate tablets alone was 0.950 with 95% confidence intervals of 0.882 and 1.018. Pre-treatment and co-administration with omeprazole has no significant effect on the raft-forming ability of alginate tablets.

  1. Magnetic Pycnoporus sanguineus-loaded alginate composite beads for removing dye from aqueous solutions.

    PubMed

    Yang, Chih-Hui; Shih, Ming-Cheng; Chiu, Han-Chen; Huang, Keng-Shiang

    2014-06-18

    Dye pollution in wastewater is a severe environmental problem because treating water containing dyes using conventional physical, chemical, and biological treatments is difficult. A conventional process is used to adsorb dyes and filter wastewater. Magnetic filtration is an emerging technology. In this study, magnetic Pycnoporus sanguineus-loaded alginate composite beads were employed to remove a dye solution. A white rot fungus, P. sanguineus, immobilized in alginate beads were used as a biosorbent to remove the dye solution. An alginate polymer could protect P. sanguineus in acidic environments. Superparamagnetic nanomaterials, iron oxide nanoparticles, were combined with alginate gels to form magnetic alginate composites. The magnetic guidability of alginate composites and biocompatibility of iron oxide nanoparticles facilitated the magnetic filtration and separation processes. The fungus cells were immobilized in loaded alginate composites to study the influence of the initial dye concentration and pH on the biosorption capacity. The composite beads could be removed easily post-adsorption by using a magnetic filtration process. When the amount of composite beads was varied, the results of kinetic studies of malachite green adsorption by immobilized cells of P. sanguineus fitted well with the pseudo-second-order model. The results indicated that the magnetic composite beads effectively adsorbed the dye solution from wastewater and were environmentally friendly.

  2. Mesoporous Silica Gel-Based Mixed Matrix Membranes for Improving Mass Transfer in Forward Osmosis: Effect of Pore Size of Filler

    NASA Astrophysics Data System (ADS)

    Lee, Jian-Yuan; Wang, Yining; Tang, Chuyang Y.; Huo, Fengwei

    2015-11-01

    The efficiency of forward osmosis (FO) process is generally limited by the internal concentration polarization (ICP) of solutes inside its porous substrate. In this study, mesoporous silica gel (SG) with nominal pore size ranging from 4-30 nm was used as fillers to prepare SG-based mixed matrix substrates. The resulting mixed matrix membranes had significantly reduced structural parameter and enhanced membrane water permeability as a result of the improved surface porosity of the substrates. An optimal filler pore size of ~9 nm was observed. This is in direct contrast to the case of thin film nanocomposite membranes, where microporous nanoparticle fillers are loaded to the membrane rejection layer and are designed in such a way that these fillers are able to retain solutes while allowing water to permeate through them. In the current study, the mesoporous fillers are designed as channels to both water and solute molecules. FO performance was enhanced at increasing filler pore size up to 9 nm due to the lower hydraulic resistance of the fillers. Nevertheless, further increasing filler pore size to 30 nm was accompanied with reduced FO efficiency, which can be attributed to the intrusion of polymer dope into the filler pores.

  3. Effect of Rheology and Poloxamers Properties on Release of Drugs from Silicon Dioxide Gel-Filled Hard Gelatin Capsules-A Further Enhancement of Viability of Liquid Semisolid Matrix Technology.

    PubMed

    Sultana, Misbah; Butt, Mobashar Ahmad; Saeed, Tariq; Mahmood, Rizwan; Ul Hassan, Saeed; Hussain, Khalid; Raza, Syed Atif; Ahsan, Muhammad; Bukhari, Nadeem Irfan

    2016-12-08

    The liquid and semisolid matrix technology, filling liquids, semi-solids and gels in hard gelatin capsule are promising, thus, there is a need of enhanced research interest in the technology. Therefore, the present study was aimed to investigate isoniazid (freely soluble) and metronidazole (slightly soluble) gels filled in hard gelatin capsules for the effect of poloxamers of different viscosities on release of the drugs. Gel of each drug (10% w/w, particle size 180-250 μm), prepared by mixing poloxamer and 8% w/w hydrophilic silicon dioxide (Aerosil® A200), was assessed for rheology, dispersion stability and release profile. Both the drugs remained dispersed in majority of gels for more than 30 days, and dispersions were depended on gels' viscosity, which was further depended on viscosity of poloxamers. A small change in viscosity was noted in gels on storage. FTIR spectra indicated no interactions between components of the gels. The gels exhibited thixotropic and shear-thinning behaviour, which were suitable for filling in hard gelatin capsules without any leakage from the capsules. The release of both drugs from the phase-stable gels for 30 days followed first-order kinetics and was found to be correlated to drugs' solubility, poloxamers' viscosity, polyoxyethylene contents and proportion of block copolymer (poloxamers) in the gels. The findings of the present study indicated that release of drugs of different solubilities (isoniazid and metronidazole) might be modified from gels using different poloxamers and Aerosil® A200.

  4. EVALUATION OF ALGINATE MICROSPHERES WITH METRONIDAZOLE OBTAINED BY THE SPRAY DRYING TECHNIQUE.

    PubMed

    Szekalska, Marta; Winnicka, Katarzyna; Czajkowska-Kośnik, Anna; Sosnowska, Katarzyna; Amelian, Aleksandra

    2015-01-01

    In the present study, nine formulations (F1-F9) of alginate microspheres with metronidazole were prepared by the spray drying technique with using different drug:polymer ratio (1:2, 1:1, 2:1) and different sodium alginate concentration (1, 2, 3%). The obtained microspheres were characterized for size, morphology, drug loading, (potential and swelling degree. Mucoadhesive properties were examined using texture analyzer and three different models of adhesive layers--gelatin discs, mucin gel and porcine vaginal mucosa. In vitro drug release, mathematical release profile and physical state of microspheres were also evaluated. The obtained results indicate that sodium alginate is a suitable polymer for developing mucoadhesive dosage forms of metronidazole. The optimal formulation F3 (drug:polymer ratio 1:2 and 1% alginate solution) was characterized by the highest metronidazole loading and sustained drug release. The results of this study indicate promising potential of ALG microspheres as alternative dosage forms for metronidazole delivery.

  5. In situ gelation for cell immobilization and culture in alginate foam scaffolds.

    PubMed

    Andersen, Therese; Markussen, Christine; Dornish, Michael; Heier-Baardson, Helene; Melvik, Jan Egil; Alsberg, Eben; Christensen, Bjørn E

    2014-02-01

    Essential cellular functions are often lost under culture in traditional two-dimensional (2D) systems. Therefore, biologically more realistic three-dimensional (3D) cell culture systems are needed that provide mechanical and biochemical cues which may otherwise be unavailable in 2D. For the present study, an alginate-based hydrogel system was used in which cells in an alginate solution were seeded onto dried alginate foams. A uniform distribution of NIH:3T3 and NHIK 3025 cells entrapped within the foam was achieved by in situ gelation induced by calcium ions integrated in the foam. The seeding efficiency of the cells was about 100% for cells added in a seeding solution containing 0.1-1.0% alginate compared with 18% when seeded without alginate. The NHIK 3025 cells were allowed to proliferate and form multi-cellular structures inside the transparent gel that were later vital stained and evaluated by confocal microscopy. Gels were de-gelled at different time points to isolate the multi-cellular structures and to determine the spheroid growth rate. It was also demonstrated that the mechanical properties of the gel could largely be varied through selection of type and concentration of the applied alginate and by immersing the already gelled disks in solutions providing additional gel-forming ions. Cells can efficiently be incorporated into the gel, and single cells and multi-cellular structures that may be formed inside can be retrieved without influencing cell viability or contaminating the sample with enzymes. The data show that the current system may overcome some limitations of current 3D scaffolds such as cell retrieval and in situ cell staining and imaging.

  6. Initial Fiber Alignment Pattern Alters Extracellular Matrix Synthesis in Fibroblast Populated Fibrin Gel Cruciforms and Correlates with Predicted Tension

    PubMed Central

    Sander, E.A.; Barocas, V.H.; Tranquillo, R.T.

    2013-01-01

    Human dermal fibroblasts entrapped in fibrin gels cast in cross-shaped (cruciform) geometries with 1:1 and 1:0.5 ratios of arm widths were studied to assess whether tension and alignment of the cells and fibrils affected ECM deposition. The cruciforms of contrasting geometry (symmetric vs. asymmetric), which developed different fiber alignment patterns, were harvested at 2, 5, and 10 weeks of culture. Cruciforms were subjected to planar biaxial testing, polarimetric imaging, DNA and biochemical analyses, histological staining, and SEM imaging. As the cruciforms compacted and developed fiber alignment, fibrin was degraded and elastin and collagen were produced in a geometry-dependent manner. Using a continuum mechanical model that accounts for direction-dependent stress due to cell traction forces and cell contact guidance with aligned fibers that occurs in the cruciforms, the mechanical stress environment was concluded to influence collagen deposition, with deposition being greatest in the narrow arms of the asymmetric cruciform where stress was predicted to be largest. PMID:21046467

  7. Physicochemical characterization and biocompatibility of alginate-polycation microcapsules designed for islet transplantation

    NASA Astrophysics Data System (ADS)

    Tam, Susan Kimberly

    Microencapsulation represents a method for immunoprotecting transplanted therapeutic cells or tissues from graft rejection using a physical barrier. This approach is advantageous in that it eliminates the need to induce long-term immunosuppression and allows the option of transplanting non-cadaveric cell sources, such as animal cells and stem cell-derived tissues. The microcapsules that we have investigated are designed to immunoprotect islets of Langerhans (i.e. clusters of insulin-secreting cells), with the goal of treating insulin-dependent diabetes. With the aid of techniques for physicochemical analysis, this research focused on understanding which properties of the microcapsule are the most important for determining its biocompatibility. The objective of this work was to elucidate correlations between the chemical make-up, physicochemical properties, and in vivo biocompatibility of alginate-based microcapsules. Our approach was based on the hypothesis that the immune response to the microcapsules is governed by, and can therefore be controlled by, specific physicochemical properties of the microcapsule and its material components. The experimental work was divided into five phases, each associated with a specific aim : (1) To prove that immunoglobulins adsorb to the surface of alginate-polycation microcapsules, and to correlate this adsorption with the microcapsule chemistry. (2) To test interlaboratory reproducibility in making biocompatible microcapsules, and evaluate the suitability of our materials and fabrication protocols for subsequent studies. (3) To determine which physicochemical properties of alginates affect the in vivo biocompatibility of their gels. (4) To determine which physiochemical properties of alginate-polycation microcapsules are most important for determining their in vivo biocompatibility (5) To determine whether a modestly immunogenic membrane hinders or helps the ability of the microcapsule to immunoprotect islet xenografts in

  8. Maintaining dimensions and mechanical properties of ionically crosslinked alginate hydrogel scaffolds in vitro.

    PubMed

    Kuo, Catherine K; Ma, Peter X

    2008-03-15

    Ionically crosslinked alginate hydrogels are attractive scaffolds because of their biocompatibility and mild gelation reaction that allows for gentle cell incorporation. However, the instability of ionically crosslinked hydrogels in an aqueous environment is a challenge that limits their application. This report presents a novel method to control the dimensions and mechanical properties of ionically crosslinked hydrogels via control of the ionic concentration of the medium. Homogeneous calcium-alginate gels were incubated in physiological saline baths adjusted to specific calcium ion concentrations. Swelling and shrinking occurred at low and high ionic concentrations of the medium, respectively, while an "optimal" intermediate calcium ion concentration of the medium was found to maintain original size and shape of the hydrogel. This optimal calcium ion concentration was found to be a function of crosslinking density and polymer concentration of the hydrogel and chemical composition of the alginate. The effects of optimal and high calcium ion concentrations of the medium on swelling behavior, calcium content, dry weight, and mechanical properties of the immersed hydrogels were investigated. It was found that the resulting hydrogel composition and mechanical properties depended on not only the calcium concentration of the medium, but also the crosslinking density and polymer concentration of the gel. In an 8-week experiment, controlled dimensions and mechanical properties of alginate gels in an aqueous environment were demonstrated. This new technique significantly enhances the potential of alginate hydrogels for tissue engineering and other biomedical applications.

  9. Swelling and mechanical properties of alginate hydrogels with respect to promotion of neural growth.

    PubMed

    Matyash, Marina; Despang, Florian; Ikonomidou, Chrysanthy; Gelinsky, Michael

    2014-05-01

    Soft alginate hydrogels support robust neurite outgrowth, but their rapid disintegration in solutions of high ionic strength restricts them from long-term in vivo applications. Aiming to enhance the mechanical stability of soft alginate hydrogels, we investigated how changes in pH and ionic strength during gelation influence the swelling, stiffness, and disintegration of a three-dimensional (3D) alginate matrix and its ability to support neurite outgrowth. Hydrogels were generated from dry alginate layers through ionic crosslinks with Ca(2+) (≤ 10 mM) in solutions of low or high ionic strength and at pH 5.5 or 7.4. High- and low-viscosity alginates with different molecular compositions demonstrated pH and ionic strength-independent increases in hydrogel volume with decreases in Ca(2+) concentrations from 10 to 2 mM. Only soft hydrogels that were synthesized in the presence of 150 mM of NaCl (Ca-alginate NaCl) displayed long-term volume stability in buffered physiological saline, whereas analogous hydrogels generated in NaCl-free conditions (Ca-alginate) collapsed. The stiffnesses of Ca-alginate NaCl hydrogels elevated from 0.01 to 19 kPa as the Ca(2+)-concentration was raised from 2 to 10 mM; however, only Ca-alginate NaCl hydrogels with an elastic modulus ≤ 1.5 kPa that were generated with ≤ 4 mM of Ca(2+) supported robust neurite outgrowth in primary neuronal cultures. In conclusion, soft Ca-alginate NaCl hydrogels combine mechanical stability in solutions of high ionic strength with the ability to support neural growth and could be useful as 3D implants for neural regeneration in vivo.

  10. Survival of Bifidobacterium longum immobilized in calcium alginate beads in simulated gastric juices and bile salt solution.

    PubMed

    Lee, K Y; Heo, T R

    2000-02-01

    Bifidobacterium longum KCTC 3128 and HLC 3742 were independently immobilized (entrapped) in calcium alginate beads containing 2, 3, and 4% sodium alginate. When the bifidobacteria entrapped in calcium alginate beads were exposed to simulated gastric juices and a bile salt solution, the death rate of the cells in the beads decreased proportionally with an increase in both the alginate gel concentration and bead size. The initial cell numbers in the beads affected the numbers of survivors after exposure to these solutions; however, the death rates of the viable cells were not affected. Accordingly, a mathematical model was formulated which expressed the influences of several parameters (gel concentration, bead size, and initial cell numbers) on the survival of entrapped bifidobacteria after sequential exposure to simulated gastric juices followed by a bile salt solution. The model proposed in this paper may be useful for estimating the survival of bifidobacteria in beads and establishing optimal entrapment conditions.

  11. Characterization of holmium loaded alginate microspheres for multimodality imaging and therapeutic applications.

    PubMed

    Zielhuis, S W; Seppenwoolde, J H; Bakker, C J G; Jahnz, U; Zonnenberg, B A; van het Schip, A D; Hennink, W E; Nijsen, J F W

    2007-09-15

    In this paper the preparation and characterization of holmium-loaded alginate microspheres is described. The rapid development of medical imaging techniques offers new opportunities for the visualisation of (drug-loaded) microparticles. Therefore, suitable imaging agents have to be incorporated into these particles. For this reason, the element holmium was used in this study in order to utilize its unique imaging characteristics. The paramagnetic behaviour of this element allows visualisation with MRI and holmium can also be neutron-activated resulting in the emission of gamma-radiation, allowing visualisation with gamma cameras, and beta-radiation, suitable for therapeutic applications. Almost monodisperse alginate microspheres were obtained by JetCutter technology where alginate droplets of a uniform size were hardened in an aqueous holmium chloride solution. Ho(3+) binds via electrostatic interactions to the carboxylate groups of the alginate polymer and as a result alginate microspheres loaded with holmium were obtained. The microspheres had a mean size of 159 microm and a holmium loading of 1.3 +/- 0.1% (w/w) (corresponding with a holmium content based on dry alginate of 18.3 +/- 0.3% (w/w)). The binding capacity of the alginate polymer for Ho(3+) (expressed in molar amounts) is equal to that for Ca(2+), which is commonly used for the hardening of alginate. This indicates that Ho(3+) has the same binding affinity as Ca(2+). In line herewith, dynamic mechanical analyses demonstrated that alginate gels hardened with Ca(2+) or Ho(3+) had similar viscoelastic properties. The MRI relaxation properties of the microspheres were determined by a MRI phantom experiment, demonstrating a strong R(2)* effect of the particles. Alginate microspheres could also be labelled with radioactive holmium by adding holmium-166 to alginate microspheres, previously hardened with calcium (labelling efficiency 96%). The labelled microspheres had a high radiochemical stability (94% after

  12. Fabrication of individual alginate-TCP scaffolds for bone tissue engineering by means of powder printing.

    PubMed

    Castilho, Miguel; Rodrigues, Jorge; Pires, Inês; Gouveia, Barbara; Pereira, Manuel; Moseke, Claus; Groll, Jürgen; Ewald, Andrea; Vorndran, Elke

    2015-01-06

    The development of polymer-calcium phosphate composite scaffolds with tailored architectures and properties has great potential for bone regeneration. Herein, we aimed to improve the functional performance of brittle ceramic scaffolds by developing a promising biopolymer-ceramic network. For this purpose, two strategies, namely, direct printing of a powder composition consisting of a 60:40 mixture of α/β-tricalcium phosphate (TCP) powder and alginate powder or vacuum infiltration of printed TCP scaffolds with an alginate solution, were tracked. Results of structural characterization revealed that the scaffolds printed with 2.5 wt% alginate-modified TCP powders presented a uniformly distributed and interfusing alginate TCP network. Mechanical results indicated a significant increase in strength, energy to failure and reliability of powder-modified scaffolds with an alginate content in the educts of 2.5 wt% when compared to pure TCP, as well as to TCP scaffolds containing 5 wt% or 7.5 wt% in the educts, in both dry and wet states. Culture of human osteoblast cells on these scaffolds also demonstrated a great improvement of cell proliferation and cell viability. While in the case of powder-mixed alginate TCP scaffolds, isolated alginate gels were formed between the calcium phosphate crystals, the vacuum-infiltration strategy resulted in the covering of the surface and internal pores of the TCP scaffold with a thin alginate film. Furthermore, the prediction of the scaffolds' critical fracture conditions under more complex stress states by the applied Mohr fracture criterion confirmed the potential of the powder-modified scaffolds with 2.5 wt% alginate in the educts as structural biomaterial for bone tissue engineering.

  13. Three-dimensional bioprinting of complex cell laden alginate hydrogel structures.

    PubMed

    Tabriz, Atabak Ghanizadeh; Hermida, Miguel A; Leslie, Nicholas R; Shu, Wenmiao

    2015-12-21

    Different bioprinting techniques have been used to produce cell-laden alginate hydrogel structures, however these approaches have been limited to 2D or simple three-dimension (3D) structures. In this study, a new extrusion based bioprinting technique was developed to produce more complex alginate hydrogel structures. This was achieved by dividing the alginate hydrogel cross-linking process into three stages: primary calcium ion cross-linking for printability of the gel, secondary calcium cross-linking for rigidity of the alginate hydrogel immediately after printing and tertiary barium ion cross-linking for long-term stability of the alginate hydrogel in culture medium. Simple 3D structures including tubes were first printed to ensure the feasibility of the bioprinting technique and then complex 3D structures such as branched vascular structures were successfully printed. The static stiffness of the alginate hydrogel after printing was 20.18 ± 1.62 KPa which was rigid enough to sustain the integrity of the complex 3D alginate hydrogel structure during the printing. The addition of 60 mM barium chloride was found to significantly extend the stability of the cross-linked alginate hydrogel from 3 d to beyond 11 d without compromising the cellular viability. The results based on cell bioprinting suggested that viability of U87-MG cells was 93 ± 0.9% immediately after bioprinting and cell viability maintained above 88% ± 4.3% in the alginate hydrogel over the period of 11 d.

  14. Gel bead composition for metal adsorption

    SciTech Connect

    Scott, Charles D.; Woodward, Charlene A.; Byers, Charles H.

    1990-01-01

    The invention is a gel bead comprising propylene glycol alginate and bone gelatin and is capable of removing metals such as Sr and Cs from solution without adding other adsorbents. The invention could have application to the nuclear industry's waste removal activities.

  15. Gel bead composition for metal adsorption

    DOEpatents

    Scott, Charles D.; Woodward, Charlene A.; Byers, Charles H.

    1991-01-01

    The invention is a gel bead comprising propylene glycol alginate and bone gelatin and is capable of removing metals such as Sr and Cs from solution without adding other adsorbents. The invention could have application to the nuclear industry's waste removal activities.

  16. Gelling process for sodium alginate: New technical approach by using calcium rich micro-spheres.

    PubMed

    Vicini, Silvia; Castellano, Maila; Mauri, Marco; Marsano, Enrico

    2015-12-10

    Alginate based materials have become an important class of products in many fields from the pharmaceutical industry to tissue engineering, because of their ability to create stimuli responsive hydrogels. We present a new technical approach for obtaining a controlled gelling process, based on the quantities of Ca(2+) rich alginate micro-beads added as crosslinkers. The gels have been evaluated in light of the amount of Ca(2+) added to the alginate solution, and in light of the different dimensions of the micro-beads, using rheological measurements to assess the variation in the storage modulus (G'), loss modulus (G'') and complex viscosity (η(*)) as well as swelling and deswelling tests. The methodology was developed to obtain a material with specific characteristics for application in the field of conservation. The material had to be able to create a stable gel after being applied on the artwork surface and to confine the solvent action at the interface during cleaning operations.

  17. Preparation and characterization of. beta. -D-glucosidase immobilized in calcium alginate

    SciTech Connect

    Krasniak, S. R.; Smith, R. D.

    1982-01-01

    Enzymatic hydrolysis of biomass to produce glucose may become feasible if an inexpensive method to reuse the enzyme can be found. This study investigated one such method whereby ..beta..-D-glucosidase (E.C. 3.2.1.21) was immobilized in calcium alginate gel spheres, which were shown to catalyze the hydrolysis of cellobiose to glucose. There was a loss of 49% of the enzyme from the alginate slurry during gelation. After gelation, in the stable gel spheres, there was a 37% retention of the enzyme activity that was actually immobilized. The reason for the loss in activity was investigated and may be caused by inhibition of the enzyme within the sphere by the calcium cations and the alginate anions also present. Mass transfer effects were minimal in this system and were not responsible for the activity loss.

  18. Magnetic Characterization of Iron Oxide Cross Linked Hydro gels

    NASA Astrophysics Data System (ADS)

    Senaratne, U.; Powell, N.; Kroll, E.; Tsoi, G.; Naik, R.; Naik, V.; Vaishnava, P. P.; Wenger, L. E.

    2004-03-01

    Magnetic hydro gels have potential applications in drug delivery, cells sorting, sensors, and actuating technologies. Iron oxide alginate nanocomposites were synthesized following the method of Kroll et al^1 by cross linking sodium alginate with Fe^2+ and Fe^3+ in methanol: water. The ion-cross linked alginate hydro gels are oxidized in an alkaline solution. The resulting hydro gel consists of iron oxide cross linked alginate. The alginate hydro gels are inert to the reaction conditions and therefore the reaction sequence can be repeated. The multiple loadings result in an increase in the amount of iron oxide and the size of the iron oxide nanoparticles in the cross linked hydro gels. The third and sixth loaded iron oxide alginate hydro gels were dried and characterized by X-ray diffraction (XRD), Transmission Electron Microscopy (TEM), and Superconducting Quantum Interference Device (SQUID) magnetometry. The XRD patterns have characteristic features of γ- Fe_2O3 or Fe_3O4 phases. The average particle size, calculated from the XRD peaks, for third loaded iron oxide alginate was 2 nm. The zero-field-cooled and field-cooled SQUID measurements show the iron oxide nanoparticles are superparamagnetic with blocking temperature (T_B) of approximately 35 K. Above the blocking temperature, the inverse susceptibility versus temperature relationship does not follow the Curie-Weiss law, indicating strong inter-particle interactions. The M vs. H data above the blocking temperature was fitted with a modified Langevin function to obtain additional information about the iron oxide particle size. Details of the relationship between coercive field and temperature as well as the particle size distribution obtained from XRD and TEM measurements will be presented. *Research supported by NSF grant # DGE ˜980720 **Supported by NSF REU grant # EEC-0097736 ^1E. Kroll, F.M. Winnik, and R.F. Ziolo, Chem. Mater, 8, 1594 (1996).

  19. Review: efficacy of alginate supplementation in relation to appetite regulation and metabolic risk factors: evidence from animal and human studies.

    PubMed

    Georg Jensen, M; Pedersen, C; Kristensen, M; Frost, G; Astrup, A

    2013-02-01

    This review provides a critical update on human and animal studies investigating the effect of alginate supplementation on appetite regulation, glycaemic and insulinemic responses, and lipid metabolism with discussion of the evidence on potential mechanisms, efficacy and tolerability. Dependent on vehicle applied for alginate supplementation, the majority of animal and human studies suggest that alginate consumption does suppress satiety and to some extent energy intake. Only one long-term intervention trial found effects on weight loss. In addition, alginates seem to exhibit beneficial influence on postprandial glucose absorption and insulin response in animals and humans. However, alginate supplementation was only found to have cholesterol-lowering properties in animals. Several mechanisms have been suggested for the positive effect observed, which involve delayed gastric emptying, increased viscosity of digesta and slowed nutrient absorption in the small intestine upon alginate gel formation. Despite reasonable efficacy and tolerability from the acute or short-term studies, we still realize there is a critical need for development of optimal alginate types and vehicles as well as studies on further long-term investigation on alginate supplementation in humans before inferring that it could be useful in the management of obesity and the metabolic syndrome.

  20. A dual-ion imprinted polymer embedded in sol-gel matrix for the ultra trace simultaneous analysis of cadmium and copper.

    PubMed

    Bali Prasad, Bhim; Jauhari, Darshika; Verma, Archana

    2014-03-01

    In simultaneous determination of group of elements, there are inter-metallic interactions which result in a non-linear relationship between the peak current and ionic concentration for each of the element, at bare (unmodified) electrode. To resolve this problem, we have resorted, for the first time, to develop a modified pencil graphite electrode using a typical ion imprinted polymer network (dual-ion imprinted polymer embedded in sol-gel matrix (inorganic-organic hybrid nano-material)) for the simultaneous analysis of a binary mixture of Cd(II) and Cu(II) ions, without any complication of inter-metallic interactions and competitive bindings, in real samples. The adequate resolution of differential pulse anodic stripping voltammetry peaks by 725 mV (cf, 615 mV with unmodified electrode), without any cross-reactivity and the stringent detection limits as low as, 0.050 and 0.034 ng mL(-1) (S/N=3) for Cd(II) and Cu(II) ions, respectively by the proposed sensor can be considered useful for the primitive diagnosis of several chronic diseases in clinical settings.

  1. Alginate Hydrogels Coated with Chitosan for Wound Dressing

    PubMed Central

    Straccia, Maria Cristina; Gomez d’Ayala, Giovanna; Romano, Ida; Oliva, Adriana; Laurienzo, Paola

    2015-01-01

    In this work, a coating of chitosan onto alginate hydrogels was realized using the water-soluble hydrochloride form of chitosan (CH-Cl), with the dual purpose of imparting antibacterial activity and delaying the release of hydrophilic molecules from the alginate matrix. Alginate hydrogels with different calcium contents were prepared by the internal setting method and coated by immersion in a CH-Cl solution. Structural analysis by cryo-scanning electron microscopy was carried out to highlight morphological alterations due to the coating layer. Tests in vitro with human mesenchymal stromal cells (MSC) were assessed to check the absence of toxicity of CH-Cl. Swelling, stability in physiological solution and release characteristics using rhodamine B as the hydrophilic model drug were compared to those of relative uncoated hydrogels. Finally, antibacterial activity against Escherichia coli was tested. Results show that alginate hydrogels coated with chitosan hydrochloride described here can be proposed as a novel medicated dressing by associating intrinsic antimicrobial activity with improved sustained release characteristics. PMID:25969981

  2. Alginate hydrogels coated with chitosan for wound dressing.

    PubMed

    Straccia, Maria Cristina; d'Ayala, Giovanna Gomez; Romano, Ida; Oliva, Adriana; Laurienzo, Paola

    2015-05-11

    In this work, a coating of chitosan onto alginate hydrogels was realized using the water-soluble hydrochloride form of chitosan (CH-Cl), with the dual purpose of imparting antibacterial activity and delaying the release of hydrophilic molecules from the alginate matrix. Alginate hydrogels with different calcium contents were prepared by the internal setting method and coated by immersion in a CH-Cl solution. Structural analysis by cryo-scanning electron microscopy was carried out to highlight morphological alterations due to the coating layer. Tests in vitro with human mesenchymal stromal cells (MSC) were assessed to check the absence of toxicity of CH-Cl. Swelling, stability in physiological solution and release characteristics using rhodamine B as the hydrophilic model drug were compared to those of relative uncoated hydrogels. Finally, antibacterial activity against Escherichia coli was tested. Results show that alginate hydrogels coated with chitosan hydrochloride described here can be proposed as a novel medicated dressing by associating intrinsic antimicrobial activity with improved sustained release characteristics.

  3. Alginate electrodeposition onto three-dimensional porous Co-Ni films as drug delivery platforms.

    PubMed

    García-Torres, J; Gispert, C; Gómez, E; Vallés, E

    2015-01-21

    Three-dimensional porous Co-Ni films/alginate hybrid materials have been successfully prepared by electrodeposition to be used as a steerable magnetic device for drug delivery. Firstly, 3D porous Co-Ni films were prepared as substrates for the subsequent electrodeposition of the alginate biopolymer. Cyclic voltammetry, galvanostatic and potentiostatic studies were performed to establish the best conditions to obtain porous Co-Ni films. The electrochemical experiments were carried out in an electrolyte containing the metal salts and ammonium chloride at low pHs. In a second stage, the electrochemical deposition of alginate as a biocompatible polymer drug delivery carrier was performed. The characteristics of the alginate matrix were investigated in terms of electrochemical properties, morphology and drug release. The hybrid material obtained showed soft-magnetic behavior and drug release indicating its suitability to be used as a steerable magnetic drug delivery device.

  4. Alginate Lyase Exhibits Catalysis-Independent Biofilm Dispersion and Antibiotic Synergy

    PubMed Central

    Lamppa, John W.

    2013-01-01

    More than 2 decades of study support the hypothesis that alginate lyases are promising therapeutic candidates for treating mucoid Pseudomonas aeruginosa infections. In particular, the enzymes' ability to degrade alginate, a key component of mucoid biofilm matrix, has been the presumed mechanism by which they disrupt biofilms and enhance antibiotic efficacy. The systematic studies reported here show that, in an in vitro model, alginate lyase dispersion of P. aeruginosa biofilms and enzyme synergy with tobramycin are completely decoupled from catalytic activity. In fact, equivalent antibiofilm effects can be achieved with bovine serum albumin or simple amino acids. These results provide new insights into potential mechanisms of alginate lyase therapeutic activity, and they should motivate a careful reexamination of the fundamental assumptions underlying interest in enzymatic biofilm dispersion. PMID:23070175

  5. Novel copper (II) alginate hydrogels and their potential for use as anti-bacterial wound dressings.

    PubMed

    Klinkajon, Wimonwan; Supaphol, Pitt

    2014-08-01

    The incorporation of a metal ion, with antimicrobial activity, into an alginate dressing is an attractive approach to minimize infection in a wound. In this work, copper (II) cross-linked alginate hydrogels were successfully prepared using a two-step cross-linking procedure. In the first step, solid alginate films were prepared using a solvent-casting method from soft gels of alginate solutions that had been lightly cross-linked using a copper (II) (Cu(2+)) sulfate solution. In the second step, the films were further cross-linked in a corresponding Cu(2+) sulfate solution using a dipping method to further improve their dimensional stability. Alginate solution (at 2%w/v) and Cu(2+) sulfate solution (at 2%w/v) in acetate buffer at a low pH provided soft films with excellent swelling behavior. An increase in either Cu(2+) ion concentration or cross-linking time led to hydrogels with more densely-cross-linked networks that limited water absorption. The hydrogels clearly showed antibacterial activity against Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis and Streptococcus pyogenes, which was proportional to the Cu(2+) ion concentration. Blood coagulation studies showed that the tested copper (II) cross-linked alginate hydrogels had a tendency to coagulate fibrin, and possibly had an effect on pro-thrombotic coagulation and platelet activation. Conclusively, the prepared films are likely candidates as antibacterial wound dressings.

  6. Radiation synthesis of PVP/alginate hydrogel containing nanosilver as wound dressing.

    PubMed

    Singh, Rita; Singh, Durgeshwer

    2012-11-01

    Hydrogels with polyvinyl pyrrolidone (PVP) and alginate were synthesized and silver nanoparticles were incorporated in hydrogel network using gamma radiation. PVP (10 and 15 %) in combination with 0.5 and 1 % alginate was gamma irradiated at different doses of 25 and 40 kGy. Maximum gel percent was obtained with 15 % PVP in combination with 0.5 % alginate. The fluid absorption capacity for the PVP/alginate hydrogels was about 1881-2361 % at 24 h. Moisture vapour transmission rate (MVTR) of hydrogels containing nanosilver at 24 h was 278.44 g/(m(2)h). The absorption capacity and moisture permeability of the PVP/alginate-nanosilver composite hydrogel dressings show the ability of the hydrogels to prevent fluid accumulation in exudating wound. The hydrogels containing nanosilver demonstrated strong antimicrobial effect and complete inhibition of microbial growth was observed with 70 ppm nanosilver dressings. PVP/alginate hydrogels containing nanosilver with efficient fluid handling capacity and antimicrobial activity was found suitable for use as wound dressing.

  7. Effects of alginate hydrogel cross-linking density on mechanical and biological behaviors for tissue engineering.

    PubMed

    Jang, Jinah; Seol, Young-Joon; Kim, Hyeon Ji; Kundu, Joydip; Kim, Sung Won; Cho, Dong-Woo

    2014-09-01

    An effective cross-linking of alginate gel was made through reaction with calcium carbonate (CaCO3). We used human chondrocytes as a model cell to study the effects of cross-linking density. Three different pore size ranges of cross-linked alginate hydrogels were fabricated. The morphological, mechanical, and rheological properties of various alginate hydrogels were characterized and responses of biosynthesis of cells encapsulated in each gel to the variation in cross-linking density were investigated. Desired outer shape of structure was maintained when the alginate solution was cross-linked with the applied method. The properties of alginate hydrogel could be tailored through applying various concentrations of CaCO3. The rate of synthesized GAGs and collagens was significantly higher in human chondrocytes encapsulated in the smaller pore structure than that in the larger pore structure. The expression of chondrogenic markers, including collagen type II and aggrecan, was enhanced in the smaller pore structure. It was found that proper structural morphology is a critical factor to enhance the performance and tissue regeneration.

  8. 21 CFR 582.7187 - Calcium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Calcium alginate. 582.7187 Section 582.7187 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Calcium alginate. (a) Product. Calcium alginate. (b) Conditions of use. This substance is...

  9. 21 CFR 582.7133 - Ammonium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ammonium alginate. 582.7133 Section 582.7133 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Ammonium alginate. (a) Product. Ammonium alginate. (b) Conditions of use. This substance is...

  10. 21 CFR 582.7610 - Potassium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Potassium alginate. 582.7610 Section 582.7610 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Potassium alginate. (a) Product. Potassium alginate. (b) Conditions of use. This substance is...

  11. 21 CFR 582.7610 - Potassium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Potassium alginate. 582.7610 Section 582.7610 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Potassium alginate. (a) Product. Potassium alginate. (b) Conditions of use. This substance is...

  12. 21 CFR 582.7610 - Potassium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Potassium alginate. 582.7610 Section 582.7610 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Potassium alginate. (a) Product. Potassium alginate. (b) Conditions of use. This substance is...

  13. 21 CFR 582.7610 - Potassium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Potassium alginate. 582.7610 Section 582.7610 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Potassium alginate. (a) Product. Potassium alginate. (b) Conditions of use. This substance is...

  14. 21 CFR 582.7610 - Potassium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Potassium alginate. 582.7610 Section 582.7610 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Potassium alginate. (a) Product. Potassium alginate. (b) Conditions of use. This substance is...

  15. 21 CFR 582.7724 - Sodium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Sodium alginate. 582.7724 Section 582.7724 Food... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7724 Sodium alginate. (a) Product. Sodium alginate. (b) Conditions of use. This substance is generally recognized...

  16. 21 CFR 582.7724 - Sodium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium alginate. 582.7724 Section 582.7724 Food... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7724 Sodium alginate. (a) Product. Sodium alginate. (b) Conditions of use. This substance is generally recognized...

  17. 21 CFR 582.7724 - Sodium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium alginate. 582.7724 Section 582.7724 Food... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7724 Sodium alginate. (a) Product. Sodium alginate. (b) Conditions of use. This substance is generally recognized...

  18. 21 CFR 582.7724 - Sodium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium alginate. 582.7724 Section 582.7724 Food... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7724 Sodium alginate. (a) Product. Sodium alginate. (b) Conditions of use. This substance is generally recognized...

  19. 21 CFR 582.7724 - Sodium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium alginate. 582.7724 Section 582.7724 Food... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7724 Sodium alginate. (a) Product. Sodium alginate. (b) Conditions of use. This substance is generally recognized...

  20. 21 CFR 184.1011 - Alginic acid.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1011 Alginic acid. (a) Alginic acid is a colloidal,...

  1. 21 CFR 184.1011 - Alginic acid.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DIRECT FOOD....1011 Alginic acid. (a) Alginic acid is a colloidal, hydrophilic polysaccharide obtained from...

  2. 21 CFR 184.1011 - Alginic acid.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1011 Alginic acid. (a) Alginic acid is a colloidal,...

  3. 21 CFR 184.1011 - Alginic acid.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1011 Alginic acid. (a) Alginic acid is a colloidal,...

  4. 21 CFR 184.1011 - Alginic acid.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1011 Alginic acid. (a) Alginic acid is a colloidal,...

  5. Pulse Field Gel Electrophoresis

    PubMed Central

    Sharma-Kuinkel, Batu K.; Rude, Thomas H.; Fowler, Vance G.

    2015-01-01

    Pulse Field Gel Electrophoresis (PFGE) is a powerful genotyping technique used for the separation of large DNA molecules (entire genomic DNA) after digesting it with unique restriction enzymes and applying to a gel matrix under the electric field that periodically changes direction. PFGE is a variation of agarose gel electrophoresis that permits analysis of bacterial DNA fragments over an order of magnitude larger than that with conventional restriction enzyme analysis. It provides a good representation of the entire bacterial chromosome in a single gel with a highly reproducible restriction profile, providing clearly distinct and well-resolved DNA fragments. PMID:25682374

  6. Pulse Field Gel Electrophoresis.

    PubMed

    Sharma-Kuinkel, Batu K; Rude, Thomas H; Fowler, Vance G

    2016-01-01

    Pulse Field Gel Electrophoresis (PFGE) is a powerful genotyping technique used for the separation of large DNA molecules (entire genomic DNA) after digesting it with unique restriction enzymes and applying to a gel matrix under the electric field that periodically changes direction. PFGE is a variation of agarose gel electrophoresis that permits analysis of bacterial DNA fragments over an order of magnitude larger than that with conventional restriction enzyme analysis. It provides a good representation of the entire bacterial chromosome in a single gel with a highly reproducible restriction profile, providing clearly distinct and well-resolved DNA fragments.

  7. A self-assembling matrix-forming gel can be easily and safely applied to prevent delayed bleeding after endoscopic resections

    PubMed Central

    Pioche, Mathieu; Camus, Marine; Rivory, Jérôme; Leblanc, Sarah; Lienhart, Isabelle; Barret, Maximilien; Chaussade, Stanislas; Saurin, Jean-Christophe; Prat, Frederic; Ponchon, Thierry

    2016-01-01

    Background: Endoscopic resections have low morbidity and mortality. Delayed bleeding has been reported in approximately 1 – 15 % of cases, increasing with antiplatelet/anticoagulant therapy or portal hypertension. A self-assembling peptide (SAP) forming a gel could protect the mucosal defect during early healing. This retrospective trial aimed to assess the safety and efficacy of SAP in preventing delayed bleeding after endoscopic resections. Methods: Consecutive patients with endoscopic resections were enrolled in two tertiary referral centers. Patients with a high risk of bleeding (antiplatelet agents, anticoagulation drugs with heparin bridge therapy, and cirrhosis with portal hypertension) were also included. The SAP gel was applied immediately after resection to cover the whole ulcer bed. Results: In total, 56 patients were included with 65 lesions (esophagus [n = 8], stomach [n = 22], duodenum [n = 10], ampullary [n = 3], colon [n = 7], and rectum [n = 15]) in two centers. Among those 65 lesions, 29 were resected in high risk situations (9 uninterrupted aspirin therapy, 6 heparin bridge therapies, 5 cirrhosis and portal hypertension, 1 both cirrhosis and heparin bridge, 3 both cirrhosis and uninterrupted aspirin, 3 large duodenal lesions > 2 cm, and 2 early introduction of clopidogrel at day 1). The resection technique was endoscopic submucosal dissection (ESD) in 40 cases, en bloc endoscopic mucosal resection (EMR) in 16, piecemeal EMR in 6, and ampullectomy in 3. The mean lesion size was 37.9 mm (SD: 2.2 mm) with a mean area of 6.3 cm2 (SD: 3.5 cm2). No difficulty was noted during application. Four delayed overt bleedings occurred (6.2 %) (3 hematochezia, 1 hematemesis) requiring endoscopic hemostasis. The mean hemoglobin drop off was 0.6 g/dL (– 0.6 to 3.1 g/dL). No adverse events occurred. Conclusion: The use of this novel extracellular matrix scaffold may help to reduce post-endoscopic resection

  8. Investigation of cell viability and morphology in 3D bio-printed alginate constructs with tunable stiffness.

    PubMed

    Shi, Pujiang; Laude, Augustinus; Yeong, Wai Yee

    2017-04-01

    In this article, mouse fibroblast cells (L929) were seeded on 2%, 5%, and 10% alginate hydrogels, and they were also bio-printed with 2%, 5%, and 10% alginate solutions individually to form constructs. The elastic and viscous moduli of alginate solutions, their interior structure and stiffness, interactions of cells and alginate, cell viability, migration and morphology were investigated by rheometer, MTT assay, scanning electron microscope (SEM), and fluorescent microscopy. The three types of bio-printed scaffolds of distinctive stiffness were prepared, and the seeded cells showed robust viability either on the alginate hydrogel surfaces or in the 3D bio-printed constructs. Majority of the proliferated cells in the 3D bio-printed constructs weakly attached to the surrounding alginate matrix. The concentration of alginate solution and hydrogel stiffness influenced cell migration and morphology, moreover the cells formed spheroids in the bio-printed 10% alginate hydrogel construct. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1009-1018, 2017.

  9. Matrix effects on the photocatalytic oxidation of alcohols by [nBu4N]4W10O32 incorporated into sol-gel silica.

    PubMed

    Molinari, Alessandra; Bratovcic, Amra; Magnacca, Giuliana; Maldotti, Andrea

    2010-09-07

    Two heterogeneous photocatalysts have been prepared by entrapment of [nBu(4)N](4)W(10)O(32) in a silica matrix, through a sol-gel procedure: SiO(2)/W30% and SiO(2)/W10% with 30% and 10% of decatungstate, respectively. They are characterized by the presence of micropores of about 7 A and 15 A and mesopores of about 25 A. Due to different preparation procedures, SiO(2)/W10% presents a more remarkable porous network than SiO(2)/W30%. The morphological features of SiO(2)/W30% and SiO(2)/W10% differ from those of their parent material SiO(2)/W0%, indicating that incorporation of the decatungstate induces a significant modification of the porous texture of the siliceous material. These photocatalysts demonstrate good stability in the oxygen-assisted photooxidation of 1-pentanol and 3-pentanol, which have been chosen as models of primary and secondary aliphatic alcohols. In particular, photoexcitation (lambda > 290 nm, 25 degrees C, 760 torr of O(2)) leads to conversion of these two substrates to pentanal or 3-pentanone, with a mass balance of about 90%. There is a strong effect of the solid support on the reactivity of the two alcoholic substrates. In particular, oxidation of 1-pentanol with SiO(2)/W10% is about four times faster than with [nBu(4)N](4)W(10)O(32) in homogeneous solution. Preferential adsorption phenomena, due to the hydrophilic character of silica explain the photocatalytic properties of the two heterogeneous systems, because adsorption favours the contact between the photoexcited decatungstate and the primary OH group of 1-pentanol. Moreover, some kind of shape selectivity, due to the microporous structure of the investigated materials, likely contributes to control the conversion yields.

  10. Alginate Production by Plant-Pathogenic Pseudomonads

    PubMed Central

    Fett, William F.; Osman, Stanley F.; Fishman, Marshall L.; Siebles, T. S.

    1986-01-01

    Eighteen plant-pathogenic and three non-plant-pathogenic pseudomonads were tested for the ability to produce alginic acid as an exopolysaccharide in vitro. Alginate production was demonstrated for 10 of 13 fluorescent plant-pathogenic pseudomonads tested with glucose or gluconate as the carbon source, but not for all 5 nonfluorescent plant pathogens and all 3 non-plant pathogens tested. With sucrose as the carbon source, some strains produced alginate while others produced both polyfructan (levan) and alginate. Alginates ranged from <1 to 28% guluronic acid, were acetylated, and had number-average molecular weights of 11.3 × 103 to 47.1 × 103. Polyfructans and alginates were not elicitors of the soybean phytoalexin glyceollin when applied to wounded cotyledon surfaces and did not induce prolonged water soaking of soybean leaf tissues. All or most pseudomonads in rRNA-DNA homology group I may be capable of synthesizing alginate as an exopolysaccharide. PMID:16347146

  11. Photonic monitoring of chitosan nanostructured alginate microcapsules for drug release

    NASA Astrophysics Data System (ADS)

    Khajuria, Deepak Kumar; Konnur, Manish C.; Vasireddi, Ramakrishna; Roy Mahapatra, D.

    2015-02-01

    By using a novel microfluidic set-up for drug screening applications, this study examines delivery of a novel risedronate based drug formulation for treatment of osteoporosis that was developed to overcome the usual shortcomings of risedronate, such as its low bioavailability and adverse gastric effects. Risedronate nanoparticles were prepared using muco-adhesive polymers such as chitosan as matrix for improving the intestinal cellular absorption of risedronate and also using a gastric-resistant polymer such as sodium alginate for reducing the gastric inflammation of risedronate. The in-vitro characteristics of the alginate encapsulated chitosan nanoparticles are investigated, including their stability, muco-adhesiveness, and Caco-2 cell permeability. Fluorescent markers are tagged with the polymers and their morphology within the microcapsules is imaged at various stages of drug release.

  12. Bile salt-reinforced alginate-chitosan beads.

    PubMed

    Takka, Sevgi; Cali, Aybige Gürel

    2012-01-01

    A polymeric delayed release protein delivery system was investigated with albumin as the model drug. The polysaccharide chitosan was reacted with sodium alginate in the presence of calcium chloride to form beads with a polyelectrolyte. In this study, attempts were made to extend albumin release in the phosphate buffer at pH 6.8 from the alginate-chitosan beads by reinforcing the matrix with bile salts. Sodium taurocholate was able to prevent albumin release at pH 1.2, protecting the protein from the acidic environment and extending the total albumin release at pH 6.8. This effect was explained by an interaction between the permanent negatively charged sulfonic acid of sodium taurocholate with the amino groups of chitosan. Mild formulation conditions, high bovine serum albumin (BSA) entrapment efficiency, and resistance to gastrointestinal release seem to be synergic and promising factors toward the development of an oral protein delivery form.

  13. Sol-Gel Glasses

    NASA Technical Reports Server (NTRS)

    Mukherjee, S. P.

    1985-01-01

    Multicomponent homogeneous, ultrapure noncrystalline gels/gel derived glasses are promising batch materials for the containerless glass melting experiments in microgravity. Hence, ultrapure, homogeneous gel precursors could be used to: (1) investigate the effect of the container induced nucleation on the glass forming ability of marginally glass forming compositions; and (2) investigate the influence of gravity on the phase separation and coarsening behavior of gel derived glasses in the liquid-liquid immiscibility zone of the nonsilicate systems having a high density phase. The structure and crystallization behavior of gels in the SiO2-GeO2 as a function of gel chemistry and thermal treatment were investigated. As are the chemical principles involved in the distribution of a second network former in silica gel matrix being investigated. The procedures for synthesizing noncrystalline gels/gel-monoliths in the SiO2-GeO2, GeO2-PbO systems were developed. Preliminary investigations on the levitation and thermal treatment of germania silicate gel-monoliths in the Pressure Facility Acoustic Levitator were done.

  14. Thermoplastic starch plasticized with alginate-glycerol mixtures: Melt-processing evaluation and film properties.

    PubMed

    López, Olivia V; Ninago, Mario D; Lencina, M M Soledad; García, María A; Andreucetti, Noemí A; Ciolino, Andrés E; Villar, Marcelo A

    2015-08-01

    Corn starch melt-processing in the presence of a commonly used plasticizer mixture (water/glycerol) and a non-conventional alternative (alginate/glycerol) was evaluated. All assayed formulations were successfully processed by melt-mixing and injected in circular probes. It was determined that all samples presented a typical viscoelastic behavior, observing a decrease in storage and loss modulus with water and alginate concentration, which facilitated samples processability. Concerning to thermal stability, it was not affected neither for water nor alginate presence. From injected probes, flexible films were obtained by thermo-compression. Films with the highest assayed water content presented a sticky appearance, whereas those containing alginate were non-tacky. Plasticizing action of water and alginate was evidenced by the occurrence of homogeneous fracture surfaces, without the presence of unmelted starch granules. Besides, the shift of glass transition temperature to lower values also corroborated the plasticizing effect of both additives. In conclusion, obtained results demonstrated the well-plasticizing action of sodium alginate on starch matrix, turning this additive into a promissory alternative to replace water during melt-processing of thermoplastic corn-starch.

  15. Osteogenic Differentiation of Human Mesenchymal Stem Cells in Mineralized Alginate Matrices

    PubMed Central

    Westhrin, Marita; Xie, Minli; Olderøy, Magnus Ø.; Sikorski, Pawel

    2015-01-01

    Mineralized biomaterials are promising for use in bone tissue engineering. Culturing osteogenic cells in such materials will potentially generate biological bone grafts that may even further augment bone healing. Here, we studied osteogenic differentiation of human mesenchymal stem cells (MSC) in an alginate hydrogel system where the cells were co-immobilized with alkaline phosphatase (ALP) for gradual mineralization of the microenvironment. MSC were embedded in unmodified alginate beads and alginate beads mineralized with ALP to generate a polymer/hydroxyapatite scaffold mimicking the composition of bone. The initial scaffold mineralization induced further mineralization of the beads with nanosized particles, and scanning electron micrographs demonstrated presence of collagen in the mineralized and unmineralized alginate beads cultured in osteogenic medium. Cells in both types of beads sustained high viability and metabolic activity for the duration of the study (21 days) as evaluated by live/dead staining and alamar blue assay. MSC in beads induced to differentiate in osteogenic direction expressed higher mRNA levels of osteoblast-specific genes (RUNX2, COL1AI, SP7, BGLAP) than MSC in traditional cell cultures. Furthermore, cells differentiated in beads expressed both sclerostin (SOST) and dental matrix protein-1 (DMP1), markers for late osteoblasts/osteocytes. In conclusion, Both ALP-modified and unmodified alginate beads provide an environment that enhance osteogenic differentiation compared with traditional 2D culture. Also, the ALP-modified alginate beads showed profound mineralization and thus have the potential to serve as a bone substitute in tissue engineering. PMID:25769043

  16. Effects of purified alginate sponge on the regeneration of chondrocytes: in vitro and in vivo.

    PubMed

    Song, Jeong Eun; Kim, A Ram; Lee, Cheon Jung; Tripathy, Nirmalya; Yoon, Kun Ho; Lee, Dongwon; Khang, Gilson

    2015-01-01

    Regeneration science has been studied using tissue engineering techniques due to the self-renewal difficulties of damaged or degenerated cartilage. A scaffold with biodegradability and biocompatibility features plays a key role in developing cartilage tissue similar to human biological materials. Herein, we have fabricated three-dimensional sponge using purified alginate for the regeneration of chondrocytes cells and formation of cartilage. We demonstrated that the alginate purification can effectively minimize inflammatory reaction through reducing the content of mannuronic acid causing immune rejection. Cartilage regeneration research was performed using three-dimensional non-purified and purified alginate sponges synthesized by modified Korbutt method. In vitro cell viability and specific gene expression in the cartilage cells were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) after seeding chondrocytes on the as-fabricated sponges. Specific extracellular matrix (ECM) of chondrocytes, sGAG, and the content of collagen were also measured. Histological staining was carried out after purified alginate sponge seeded with chondrocytes and was implanted in subcutaneous nude mouse followed by extraction. Compared to the non-purified ones, the purified alginate sponges showed positive effects on maintaining affinities and phenotype of chondrocytes. From these results, it can be suggested that the purified alginate sponges provide a promising platform for cartilage regeneration.

  17. Facile fabrication of poly(L-lactic acid) microsphere-incorporated calcium alginate/hydroxyapatite porous scaffolds based on Pickering emulsion templates.

    PubMed

    Hu, Yang; Ma, Shanshan; Yang, Zhuohong; Zhou, Wuyi; Du, Zhengshan; Huang, Jian; Yi, Huan; Wang, Chaoyang

    2016-04-01

    In this study, we develop a facile one-pot approach to the fabrication of poly(L-lactic acid) (PLLA) microsphere-incorporated calcium alginate (ALG-Ca)/hydroxyapatite (HAp) porous scaffolds based on HAp nanoparticle-stabilized oil-in-water Pickering emulsion templates, which contain alginate in the aqueous phase and PLLA in the oil phase. The emulsion aqueous phase is solidified by in situ gelation of alginate with Ca(2+) released from HAp by decreasing pH with slow hydrolysis of D-gluconic acid δ-lactone (GDL) to produce emulsion droplet-incorporated gels, followed by freeze-drying to form porous scaffolds containing microspheres. The pore structure of porous scaffolds can be adjusted by varying the HAp or GDL concentration. The compressive tests show that the increase of HAp or GDL concentration is beneficial to improve the compressive property of porous scaffolds, while the excessive HAp can lead to the decrease in compressive property. Moreover, the swelling behavior studies display that the swelling ratios of porous scaffolds reduce with increasing HAp or GDL concentration. Furthermore, hydrophobic drug ibuprofen (IBU) and hydrophilic drug bovine serum albumin (BSA) are loaded into the microspheres and scaffold matrix, respectively. In vitro drug release results indicate that BSA has a rapid release while IBU has a sustained release in the dual drug-loaded scaffolds. In vitro cell culture experiments verify that mouse bone mesenchymal stem cells can proliferate on the porous scaffolds well, indicating the good biocompatibility of porous scaffolds. All these results demonstrate that the PLLA microsphere-incorporated ALG-Ca/HAp porous scaffolds have a promising potential for tissue engineering and drug delivery applications.

  18. Sodium Alginate Based Mucoadhesive System for Gatifloxacin and Its In Vitro Antibacterial Activity

    PubMed Central

    Kesavan, Karthikeyan; Nath, Gopal; Pandit, Jayanta K.

    2010-01-01

    The objective of this study was to formulate sodium alginate based ophthalmic mucoadhesive system of gatifloxacin and its in vitro antibacterial potential on pathogenic microorganisms, Staphylococcus aureus and Escherichia coli. Sodium carboxymethylcellulose (NaCMC) was added to the formulations to enhance the gel bioadhesion properties. The prepared formulations were evaluated for their in vitro drug release, gelation behaviour, rheological behavior, and mucoadhesion force. All formulations in non-physiological and physiological condition showed pseudo plastic behavior. Increase in the concentration of sodium alginate and sodium CMC enhanced the mucoadhesive force significantly. In vitro release of gatifloxacin from the system in simulated tear fluid (STF, pH – 7.4), was influenced significantly by the properties and concentration of sodium alginate, NaCMC. Significant reduction in total bacterial count was observed between control and treatment groups with both the test organisms. PMID:21179327

  19. Influence of mechanical properties of alginate-based substrates on the performance of Schwann cells in culture.

    PubMed

    Ning, Liqun; Xu, Yitong; Chen, Xiongbiao; Schreyer, David J

    2016-06-01

    In tissue engineering, artificial tissue scaffolds containing living cells have been studied for tissue repair and regeneration. Notably, the performance of these encapsulated-in-scaffolds cells in terms of cell viability, proliferation, and expression of function during and after the scaffold fabrication process, has not been well documented because of the influence of mechanical, chemical, and physical properties of the scaffold substrate materials. This paper presents our study on the influence of mechanical properties of alginate-based substrates on the performance of Schwann cells, which are the major glial cells of peripheral nervous system. Given the fact that alginate polysaccharide hydrogel has poor cell adhesion properties, in this study, we examined several types of cell-adhesion supplements and found that alginate covalently modified with RGD peptide provided improved cell proliferation and adhesion. We prepared alginate-based substrates for cell culture using varying alginate concentrations for altering their mechanical properties, which were confirmed by compression testing. Then, we examined the viability, proliferation, morphology, and expression of the extracellular matrix protein laminin of Schwann cells that were seeded on the surface of alginate-based substrates (or 2D culture) or encapsulated within alginate-based substrates (3D cultures), and correlated the examined cell performance to the alginate concentration (or mechanical properties) of hydrogel substrates. Our findings suggest that covalent attachment of RGD peptide can improve the success of Schwann cell encapsulation within alginate-based scaffolds, and provide guidance for regulating the mechanical properties of alginate-based scaffolds containing Schwann cells for applications in peripheral nervous system regeneration and repair.

  20. Sequence-specific nucleic acid mobility using a reversible block copolymer gel matrix and DNA amphiphiles (lipid-DNA) in capillary and microfluidic electrophoretic separations.

    PubMed

    Wagler, Patrick; Minero, Gabriel Antonio S; Tangen, Uwe; de Vries, Jan Willem; Prusty, Deepak; Kwak, Minseok; Herrmann, Andreas; McCaskill, John S

    2015-10-01

    Reversible noncovalent but sequence-dependent attachment of DNA to gels is shown to allow programmable mobility processing of DNA populations. The covalent attachment of DNA oligomers to polyacrylamide gels using acrydite-modified oligonucleotides has enabled sequence-specific mobility assays for DNA in gel electrophoresis: sequences binding to the immobilized DNA are delayed in their migration. Such a system has been used for example to construct complex DNA filters facilitating DNA computations. However, these gels are formed irreversibly and the choice of immobilized sequences is made once off during fabrication. In this work, we demonstrate the reversible self-assembly of gels combined with amphiphilic DNA molecules, which exhibit hydrophobic hydrocarbon chains attached to the nucleobase. This amphiphilic DNA, which we term lipid-DNA, is synthesized in advance and is blended into a block copolymer gel to induce sequence-dependent DNA retention during electrophoresis. Furthermore, we demonstrate and characterize the programmable mobility shift of matching DNA in such reversible gels both in thin films and microchannels using microelectrode arrays. Such sequence selective separation may be employed to select nucleic acid sequences of similar length from a mixture via local electronics, a basic functionality that can be employed in novel electronic chemical cell designs and other DNA information-processing systems.

  1. Micropatterning Alginate Substrates for in vitro Cardiovascular Muscle on a Chip**

    PubMed Central

    Agarwal, Ashutosh; Farouz, Yohan; Nesmith, Alexander Peyton; Deravi, Leila F.; McCain, Megan Laura

    2015-01-01

    Soft hydrogels such as alginate are ideal substrates for building muscle in vitro because they have structural and mechanical properties close to the in vivo extracellular matrix (ECM) network. However, hydrogels are generally not amenable to protein adhesion and patterning. Moreover, muscle structures and their underlying ECM are highly anisotropic, and it is imperative that in vitro models recapitulate the structural anisotropy in reconstructed tissues for in vivo relevance due to the tight coupling between sturcture and function in these systems. We present two techniques to create chemical and structural heterogeneities within soft alginate substrates and employ them to engineer anisotropic muscle monolayers: (i) microcontact printing lines of extracellular matrix proteins on flat alginate substrates to guide cellular processes with chemical cues, and (ii) micromolding of alginate surface into grooves and ridges to guide cellular processes with topographical cues. Neonatal rat ventricular myocytes as well as human umbilical artery vascular smooth muscle cells successfully attach to both these micropatterned substrates leading to subsequent formation of anisotropic striated and smooth muscle tissues. Muscular thin film cantilevers cut from these constructs are then employed for functional characterization of engineered muscular tissues. Thus, micropatterned alginate is an ideal substrate for in vitro models of muscle tissue because it facilitates recapitulation of the anisotropic architecture of muscle, mimics the mechanical properties of the ECM microenvironment, and is amenable to evaluation of functional contractile properties. PMID:26213529

  2. Silk sericin loaded alginate nanoparticles: Preparation and anti-inflammatory efficacy.

    PubMed

    Khampieng, Thitikan; Aramwit, Pornanong; Supaphol, Pitt

    2015-09-01

    In this study, silk sericin loaded alginate nanoparticles were prepared by the emulsification method followed by internal crosslinking. The effects of various silk sericin loading concentration on particle size, shape, thermal properties, and release characteristics were investigated. The initial silk sericin loadings of 20, 40, and 80% w/w to polymer were incorporated into these alginate nanoparticles. SEM images showed a spherical shape and small particles of about 71.30-89.50 nm. TGA analysis showed that thermal stability slightly increased with increasing silk sericin loadings. FTIR analysis suggested interactions between alginate and silk sericin in the nanoparticles. The release study was performed in acetate buffer at normal skin conditions (pH 5.5; 32 °C). The release profiles of silk sericin exhibited initial rapid release, consequently with sustained release. These silk sericin loaded alginate nanoparticles were further incorporated into topical hydrogel and their anti-inflammatory properties were studied using carrageenan-induced paw edema assay. The current study confirms the hypothesis that the application of silk sericin loaded alginate nanoparticle gel can inhibit inflammation induced by carrageenan.

  3. Characterization of slow-gelling alginate hydrogels for intervertebral disc tissue-engineering applications.

    PubMed

    Growney Kalaf, Emily A; Flores, Reynaldo; Bledsoe, J Gary; Sell, Scott A

    2016-06-01

    Reversal of intervertebral disc degeneration can have a potentially monumental effect on spinal health. As such, the goal of this research is to create an injectable, cellularized alginate-based nucleus pulposus that will restore disc function; with the primary goal of creating an alginate gel with tailorable rates of gelation to improve functionality over standard CaCl2 crosslinking techniques. Gelation characteristics of 1% sodium alginate were analyzed over various molar concentrations of a 1:2 ratio of CaCO3:glucono-δ-lactone (GDL), with 10% CaCl2 as the control crosslinker. Alginate construct characterization for all concentrations was performed via ultimate and cyclic compressive testing over a 28day degradation period in PBS. Dehydration, swell testing, and albumin release kinetics were determined, and cytotoxicity and cell homogeneity tests showed promise for cellularization strategies. Overall, the 30 and 60mM GDL alginate concentrations presented the most viable option for use in further studies, with a gelation time between 10 and 30min, low hysteresis over control, low percent change in thickness and weight under both PBS degradation and swelling conditions, and stable mechanical properties over 28days in vitro.

  4. Immobilization of Bacillus acidocaldarius whole-cell rhodanese in polysaccharide and insolubilized gelatin gels

    SciTech Connect

    De Riso, L.; Alteriis, E. de; Parascandola, P. |; La Cara, F.; Sada, A.

    1996-04-01

    The presence of rhodanese activity has been investigated in two strains of thermophilic eubacteria and two strains of extremophiles. Bacillus acidocaldarius, a thermoacidophilic eubacterium, showed the highest levels of enzyme activity. Whole cells, previously subjected to one cycle of freeze-thawing, were immobilized by entrapment in the polysaccharide matrices Ca-alginate, {kappa}-carrageenan and chitosan, and in an insolubilized gelatin gel. The results obtained with the different immobilizates in terms of activity yield, possibility of regeneration and operative stability were evaluated with the aim of setting up a continuous system. This was achieved with a system consisting of B. acidocaldarius cells entrapped in an insolubilized gelatin matrix. The latter, in the form of a thin membrane, was employed in a custom-conceived reactor operating as a plug flow reactor. 21 refs., 3 figs., 2 tabs.

  5. Characterization of alginate-like exopolysaccharides isolated from aerobic granular sludge in pilot-plant.

    PubMed

    Lin, Yuemei; de Kreuk, Merle; van Loosdrecht, M C M; Adin, Avner

    2010-06-01

    To understand functional gel-forming exopolysaccharides in aerobic granular sludge, alginate-like exopolysaccharides were specifically extracted from aerobic granular sludge cultivated in a pilot plant treating municipal sewage. The exopolysaccharides were identified by the FAO/WHO alginate identification tests, characterized by biochemical assays, gelation with Ca(2+), blocks fractionation, spectroscopic analysis as UV-visible, FT-IR and MALDI-TOF MS, and electrophoresis. The yield of extractable alginate-like exopolysaccharides was reached 160+/-4mg/g (VSS ratio). They resembled seaweed alginate in UV-visible and MALDI-TOF MS spectra, and distinguished from it in the reactions with acid ferric sulfate, phenol-sulfuric acid and Coomassie brilliant blue G250. Characterized by their high percentage of poly guluronic acid blocks (69.07+/-8.95%), the isolated exopolysaccharides were capable to form rigid, non-deformable gels in CaCl(2). They were one of the dominant exopolysaccharides in aerobic granular sludge. We suggest that polymers play a significant role in providing aerobic granular sludge a highly hydrophobic, compact, strong and elastic structure.

  6. Engineering alginate as bioink for bioprinting

    PubMed Central

    Jia, Jia; Richards, Dylan J.; Pollard, Samuel; Tan, Yu; Rodriguez, Joshua; Visconti, Richard P.; Trusk, Thomas C.; Yost, Michael J.; Yao, Hai; Markwald, Roger R.; Mei, Ying

    2015-01-01

    Recent advances in 3D printing offer an excellent opportunity to address critical challenges faced by current tissue engineering approaches. Alginate hydrogels have been extensively utilized as bioinks for 3D bioprinting. However, most previous research has focused on native alginates with limited degradation. The application of oxidized alginates with controlled degradation in bioprinting has not been explored. Here, we prepared a collection of 30 different alginate hydrogels with varied oxidation percentages and concentrations to develop a bioink platform that can be applied to a multitude of tissue engineering applications. We systematically investigated the effects of two key material properties (i.e. viscosity and density) of alginate solutions on their printabilities to identify a suitable range of material properties of alginates to be applied to bioprinting. Further, four alginate solutions with varied biodegradability were printed with human adipose-derived stem cells (hADSCs) into lattice-structured, cell-laden hydrogels with high accuracy. Notably, these alginate-based bioinks were shown to be capable of modulating proliferation and spreading of hADSCs without affecting structure integrity of the lattice structures (except the highly degradable one) after 8 days in culture. This research lays a foundation for the development of alginate-based bioink for tissue-specific tissue engineering applications. PMID:24998183

  7. Engineering alginate as bioink for bioprinting.

    PubMed

    Jia, Jia; Richards, Dylan J; Pollard, Samuel; Tan, Yu; Rodriguez, Joshua; Visconti, Richard P; Trusk, Thomas C; Yost, Michael J; Yao, Hai; Markwald, Roger R; Mei, Ying

    2014-10-01

    Recent advances in three-dimensional (3-D) printing offer an excellent opportunity to address critical challenges faced by current tissue engineering approaches. Alginate hydrogels have been used extensively as bioinks for 3-D bioprinting. However, most previous research has focused on native alginates with limited degradation. The application of oxidized alginates with controlled degradation in bioprinting has not been explored. Here, a collection of 30 different alginate hydrogels with varied oxidation percentages and concentrations was prepared to develop a bioink platform that can be applied to a multitude of tissue engineering applications. The authors systematically investigated the effects of two key material properties (i.e. viscosity and density) of alginate solutions on their printabilities to identify a suitable range of material properties of alginates to be applied to bioprinting. Further, four alginate solutions with varied biodegradability were printed with human adipose-derived stem cells (hADSCs) into lattice-structured, cell-laden hydrogels with high accuracy. Notably, these alginate-based bioinks were shown to be capable of modulating proliferation and spreading of hADSCs without affecting the structure integrity of the lattice structures (except the highly degradable one) after 8days in culture. This research lays a foundation for the development of alginate-based bioink for tissue-specific tissue engineering applications.

  8. Controlled antiseptic release by alginate polymer films and beads.

    PubMed

    Liakos, Ioannis; Rizzello, Loris; Bayer, Ilker S; Pompa, Pier Paolo; Cingolani, Roberto; Athanassiou, Athanassia

    2013-01-30

    Biodegradable polymeric materials based on blending aqueous dispersions of natural polymer sodium alginate (NaAlg) and povidone iodine (PVPI) complex, which allow controlled antiseptic release, are presented. The developed materials are either free standing NaAlg films or Ca(2+)-cross-linked alginate beads, which properly combined with PVPI demonstrate antibacterial and antifungal activity, suitable for therapeutic applications, such as wound dressing. Glycerol was used as the plasticizing agent. Film morphology was studied by optical and atomic force microscopy. It was found that PVPI complex forms well dispersed circular micro-domains within the NaAlg matrix. The beads were fabricated by drop-wise immersion of NaAlg/PVPI/glycerol solutions into aqueous calcium chloride solutions to form calcium alginate beads encapsulating PVPI solution (CaAlg/PVPI). Controlled release of PVPI was possible when the composite films and beads were brought into direct contact with water or with moist media. Bactericidal and fungicidal properties of the materials were tested against Escherichia coli bacteria and Candida albicans fungi. The results indicated very efficient antibacterial and antifungal activity within 48 h. Controlled release of PVPI into open wounds is highly desired in clinical applications to avoid toxic doses of iodine absorption by the wound. A wide variety of applications are envisioned such as external and internal wound dressings with controlled antiseptic release, hygienic and protective packaging films for medical devices, and polymer beads as water disinfectants.

  9. Nano zinc oxide-sodium alginate antibacterial cellulose fibres.

    PubMed

    Varaprasad, Kokkarachedu; Raghavendra, Gownolla Malegowd; Jayaramudu, Tippabattini; Seo, Jongchul

    2016-01-01

    In the present study, antibacterial cellulose fibres were successfully fabricated by a simple and cost-effective procedure by utilizing nano zinc oxide. The possible nano zinc oxide was successfully synthesized by precipitation technique and then impregnated effectively over cellulose fibres through sodium alginate matrix. XRD analysis revealed the 'rod-like' shape alignment of zinc oxide with an interplanar d-spacing of 0.246nm corresponding to the (101) planes of the hexagonal wurtzite structure. TEM analysis confirmed the nano dimension of the synthesized zinc oxide nanoparticles. The presence of nano zinc oxide over cellulose fibres was evident from the SEM-EDS experiments. FTIR and TGA studies exhibited their effective bonding interaction. The tensile stress-strain curves data indicated the feasibility of the fabricated fibres for longer duration utility without any significant damage or breakage. The antibacterial studies against Escherichia coli revealed the excellent bacterial devastation property. Further, it was observed that when all the parameters remained constant, the variation of sodium alginate concentration showed impact in devastating the E. coli. In overall, the fabricated nano zinc oxide-sodium alginate cellulose fibres can be effectively utilized as antibacterial fibres for biomedical applications.

  10. Chitosan Enriched Three-Dimensional Matrix Reduces Inflammatory and Catabolic Mediators Production by Human Chondrocytes

    PubMed Central

    Oprenyeszk, Frederic; Sanchez, Christelle; Dubuc, Jean-Emile; Maquet, Véronique; Henrist, Catherine; Compère, Philippe; Henrotin, Yves

    2015-01-01

    This in vitro study investigated the metabolism of human osteoarthritic (OA) chondrocytes encapsulated in a spherical matrix enriched of chitosan. Human OA chondrocytes were encapsulated and cultured for 28 days either in chitosan-alginate beads or in alginate beads. The beads were formed by slowly passing dropwise either the chitosan 0.6%–alginate 1.2% or the alginate 1.2% solution through a syringe into a 102 mM CaCl2 solution. Beads were analyzed histologically after 28 days. Interleukin (IL)-6 and -8, prostaglandin (PG) E2, matrix metalloproteinases (MMPs), hyaluronan and aggrecan were quantified directly in the culture supernatant by specific ELISA and nitric oxide (NO) by using a colorimetric method based on the Griess reaction. Hematoxylin and eosin staining showed that chitosan was homogeneously distributed through the matrix and was in direct contact with chondrocytes. The production of IL-6, IL-8 and MMP-3 by chondrocytes significantly decreased in chitosan-alginate beads compared to alginate beads. PGE2 and NO decreased also significantly but only during the first three days of culture. Hyaluronan and aggrecan production tended to increase in chitosan-alginate beads after 28 days of culture. Chitosan-alginate beads reduced the production of inflammatory and catabolic mediators by OA chondrocytes and tended to stimulate the synthesis of cartilage matrix components. These particular effects indicate that chitosan-alginate beads are an interesting scaffold for chondrocytes encapsulation before transplantation to repair cartilage defects. PMID:26020773

  11. Identification of enzymes responsible for extracellular alginate depolymerization and alginate metabolism in Vibrio algivorus.

    PubMed

    Doi, Hidetaka; Tokura, Yuriko; Mori, Yukiko; Mori, Kenichi; Asakura, Yoko; Usuda, Yoshihiro; Fukuda, Hiroo; Chinen, Akito

    2017-02-01

    Alginate is a marine non-food-competing polysaccharide that has potential applications in biorefinery. Owing to its large size (molecular weight >300,000 Da), alginate cannot pass through the bacterial cell membrane. Therefore, bacteria that utilize alginate are presumed to have an enzyme that degrades extracellular alginate. Recently, Vibrio algivorus sp. SA2(T) was identified as a novel alginate-decomposing and alginate-utilizing species. However, little is known about the mechanism of alginate degradation and metabolism in this species. To address this issue, we screened the V. algivorus genomic DNA library for genes encoding polysaccharide-decomposing enzymes using a novel double-layer plate screening method and identified alyB as a candidate. Most identified alginate-decomposing enzymes (i.e., alginate lyases) must be concentrated and purified before extracellular alginate depolymerization. AlyB of V. algivorus heterologously expressed in Escherichia coli depolymerized extracellular alginate without requiring concentration or purification. We found seven homologues in the V. algivorus genome (alyB, alyD, oalA, oalB, oalC, dehR, and toaA) that are thought to encode enzymes responsible for alginate transport and metabolism. Introducing these genes into E. coli enabled the cells to assimilate soluble alginate depolymerized by V. algivorus AlyB as the sole carbon source. The alginate was bioconverted into L-lysine (43.3 mg/l) in E. coli strain AJIK01. These findings demonstrate a simple and novel screening method for identifying polysaccharide-degrading enzymes in bacteria and provide a simple alginate biocatalyst and fermentation system with potential applications in industrial biorefinery.

  12. BANANA GEL.

    PubMed

    McGuire, G; Falk, K G

    1922-03-20

    The conditions for the formation of gels from banana extracts were studied. Gels were obtained with extracts more alkaline than pH 7.0 with very small quantities of calcium, strontium, and barium salts, the gel formation with these salts decreasing in the indicated order. In solutions more acid than pH 6.0, no gels were obtained with these salts. Magnesium, lithium, and sodium salts did not cause gel formation either in acid or alkaline solutions. Pancreatine gave a gel on incubation with banana extract at pH 5.0. The gel-forming property of banana extracts was destroyed on boiling.

  13. Alginate cryogel based glucose biosensor

    NASA Astrophysics Data System (ADS)

    Fatoni, Amin; Windy Dwiasi, Dian; Hermawan, Dadan

    2016-02-01

    Cryogel is macroporous structure provides a large surface area for biomolecule immobilization. In this work, an alginate cryogel based biosensor was developed to detect glucose. The cryogel was prepared using alginate cross-linked by calcium chloride under sub-zero temperature. This porous structure was growth in a 100 μL micropipette tip with a glucose oxidase enzyme entrapped inside the cryogel. The glucose detection was based on the colour change of redox indicator, potassium permanganate, by the hydrogen peroxide resulted from the conversion of glucose. The result showed a porous structure of alginate cryogel with pores diameter of 20-50 μm. The developed glucose biosensor was showed a linear response in the glucose detection from 1.0 to 5.0 mM with a regression of y = 0.01x+0.02 and R2 of 0.994. Furthermore, the glucose biosensor was showed a high operational stability up to 10 times of uninterrupted glucose detections.

  14. Comparison of in-gel and on-membrane digestion methods at low to sub-pmol level for subsequent peptide and fragment-ion mass analysis using matrix-assisted laser-desorption/ionization mass spectrometry.

    PubMed

    Courchesne, P L; Luethy, R; Patterson, S D

    1997-01-01

    The success of the mass spectrometric-based approaches for the identification of gel-separated proteins relies upon recovery of peptides, without high levels of ionization-suppressing contaminants, in solvents compatible with the mass spectrometer being employed. We sought to determine whether in-gel or on-membrane digestion provided a significant advantage when low to sub-pmol quantities of gel-separated proteins were analyzed by matrix-assisted laser-desorption/ionization mass spectrometry (MALDI-MS) with respect to the number and size of released peptides. Serial dilutions of five standard proteins of M(r) 17,000 to 97,000 (from 16 pmol to 125 fmol) were electrophoresed and subjected to in-gel digestion (using a microcolumn clean-up protocol, Courchesne, P.L. and Patterson, S. D., BioTechniques, 1997, in press) or on-membrane digestion following blotting to the PVDF-based membranes, Immobilon-P and Immobilon-CD. Peptide maps were able to be obtained for all proteins at the detection limit of each method (Immobilon-P and Immobilon-CD, 0.5 pmol; and in-gel, 125 fmol), and searches of Swiss-Prot or a non-redundant database (> 193000 entries) successfully identified all of the proteins, except beta-casein. Fragment-ion spectra using a curved-field reflector MALDI-MS were obtained from more than one peptide per protein at loads down to 250 fmol (except beta-casein). Using the uninterpreted data, a search of the nonredundant database and a six-way translation of GenBank dbEST (> 2,208,000 entries total) was able to identify myoglobin, carbonic anhydrase II, and phosphorylase b.

  15. Bacterial community structure and predicted alginate metabolic pathway in an alginate-degrading bacterial consortium.

    PubMed

    Kita, Akihisa; Miura, Toyokazu; Kawata, Satoshi; Yamaguchi, Takeshi; Okamura, Yoshiko; Aki, Tsunehiro; Matsumura, Yukihiko; Tajima, Takahisa; Kato, Junichi; Nishio, Naomichi; Nakashimada, Yutaka

    2016-03-01

    Methane fermentation is one of the effective approaches for utilization of brown algae; however, this process is limited by the microbial capability to degrade alginate, a main polysaccharide found in these algae. Despite its potential, little is known about anaerobic microbial degradation of alginate. Here we constructed a bacterial consortium able to anaerobically degrade alginate. Taxonomic classification of 16S rRNA gene, based on high-throughput sequencing data, revealed that this consortium included two dominant strains, designated HUA-1 and HUA-2; these strains were related to Clostridiaceae bacterium SK082 (99%) and Dysgonomonas capnocytophagoides (95%), respectively. Alginate lyase activity and metagenomic analyses, based on high-throughput sequencing data, revealed that this bacterial consortium possessed putative genes related to a predicted alginate metabolic pathway. However, HUA-1 and 2 did not grow on agar medium with alginate by using roll-tube method, suggesting the existence of bacterial interactions like symbiosis for anaerobic alginate degradation.

  16. Alginate Biosynthesis Factories in Pseudomonas fluorescens: Localization and Correlation with Alginate Production Level.

    PubMed

    Maleki, Susan; Almaas, Eivind; Zotchev, Sergey; Valla, Svein; Ertesvåg, Helga

    2015-12-11

    Pseudomonas fluorescens is able to produce the medically and industrially important exopolysaccharide alginate. The proteins involved in alginate biosynthesis and secretion form a multiprotein complex spanning the inner and outer membranes. In the present study, we developed a method by which the porin AlgE was detected by immunogold labeling and transmission electron microscopy. Localization of the AlgE protein was found to depend on the presence of other proteins in the multiprotein complex. No correlation was found between the number of alginate factories and the alginate production level, nor were the numbers of these factories affected in an algC mutant that is unable to produce the precursor needed for alginate biosynthesis. Precursor availability and growth phase thus seem to be the main determinants for the alginate production rate in our strain. Clustering analysis demonstrated that the alginate multiprotein complexes were not distributed randomly over the entire outer cell membrane surface.

  17. Alginate Biosynthesis Factories in Pseudomonas fluorescens: Localization and Correlation with Alginate Production Level

    PubMed Central

    Maleki, Susan; Almaas, Eivind; Zotchev, Sergey; Valla, Svein

    2015-01-01

    Pseudomonas fluorescens is able to produce the medically and industrially important exopolysaccharide alginate. The proteins involved in alginate biosynthesis and secretion form a multiprotein complex spanning the inner and outer membranes. In the present study, we developed a method by which the porin AlgE was detected by immunogold labeling and transmission electron microscopy. Localization of the AlgE protein was found to depend on the presence of other proteins in the multiprotein complex. No correlation was found between the number of alginate factories and the alginate production level, nor were the numbers of these factories affected in an algC mutant that is unable to produce the precursor needed for alginate biosynthesis. Precursor availability and growth phase thus seem to be the main determinants for the alginate production rate in our strain. Clustering analysis demonstrated that the alginate multiprotein complexes were not distributed randomly over the entire outer cell membrane surface. PMID:26655760

  18. Reduced liver cell death using an alginate scaffold bandage: a novel approach for liver reconstruction after extended partial hepatectomy.

    PubMed

    Shteyer, Eyal; Ben Ya'acov, Ami; Zolotaryova, Lidia; Sinai, Avital; Lichtenstein, Yoav; Pappo, Orit; Kryukov, Olga; Elkayam, Tsiona; Cohen, Smadar; Ilan, Yaron

    2014-07-01

    Extended partial hepatectomy may be needed in cases of large hepatic mass, and can lead to fulminant hepatic failure. Macroporous alginate scaffold is a biocompatible matrix which promotes the growth, differentiation and long-term hepatocellular function of primary hepatocytes in vitro. Our aim was to explore the ability of implanted macroporous alginate scaffolds to protect liver remnants from acute hepatic failure after extended partial hepatectomy. An 87% partial hepatectomy (PH) was performed on C57BL/6 mice to compare non-treated mice to mice in which alginate or collagen scaffolds were implanted after PH. Mice were scarified 3, 6, 24 and 48 h and 6 days following scaffold implantation and the extent of liver injury and repair was examined. Alginate scaffolds significantly increased animal survival to 60% vs. 10% in non-treated and collagen-treated mice (log rank=0.001). Mice with implanted alginate scaffolds manifested normal and prolonged aspartate aminotransferases and alanine aminotransferases serum levels as compared with the 2- to 20-fold increase in control groups (P<0.0001) accompanied with improved liver histology. Sustained normal serum albumin levels were observed in alginate-scaffold-treated mice 48 h after hepatectomy. Incorporation of BrdU-positive cells was 30% higher in the alginate-scaffold-treated group, compared with non-treated mice. Serum IL-6 levels were significantly decreased 3h post PH. Biotin-alginate scaffolds were quickly well integrated within the liver tissue. Collectively, implanted alginate scaffolds support liver remnants after extended partial hepatectomy, thus eliminating liver injury and leading to enhanced animal survival after extended partial hepatectomy.

  19. Synthesis of Thiolated Alginate and Evaluation of Mucoadhesiveness, Cytotoxicity and Release Retardant Properties

    PubMed Central

    Jindal, A. B.; Wasnik, M. N.; Nair, Hema A.

    2010-01-01

    Modification of polymers by covalent attachment of thiol bearing pendant groups is reported to impart many beneficial properties to them. Hence in the present study, sodium alginate–cysteine conjugate was synthesized by carbodiimide mediated coupling under varying reaction conditions and the derivatives characterized for thiol content. The thiolated alginate species synthesized had bound thiol content ranging from 247.8±11.03–324.54±10.107 ΅mol/g of polymer depending on the reaction conditions. Matrix tablets based on sodium alginate-cysteine conjugate and native sodium alginate containing tramadol hydrochloride as a model drug were prepared and mucoadhesive strength and in vitro drug release from the tablets were compared. Tablets containing 75 mg sodium alginate-cysteine conjugate could sustain release of 10 mg of model drug for 3 h, whereas 90% of the drug was released within 1 h from corresponding tablets prepared using native sodium alginate. An approximately 2-fold increase in the minimal detachment force of the tablets from an artificial mucin film was observed for sodium alginate–cysteine conjugate as compared to native sodium alginate. In vitro cytotoxicity studies in L-929 mouse fibroblast cells studied using an MTT assay revealed that at low concentrations of polymer, sodium alginate–cysteine conjugate was less toxic to L-929 mouse fibroblast cell line when compared to native sodium alginate. Hence, thiolation is found to be a simple route to improving polymer performance. The combination of improved controlled drug release and mucoadhesive properties coupled with the low toxicity of these new excipients builds up immense scope for the use of thiolated polymers in mucoadhesive drug delivery systems. PMID:21969750

  20. Magnetite-alginate beads for purification of some starch degrading enzymes.

    PubMed

    Teotia, Sunita; Gupta, M N

    2002-03-01

    Starch degrading enzymes, viz., beta-amylase, glucoamylase, and pullulanase, were purified using magnetite-alginate beads. In each case, the enzyme activity was eluted by using 1.0 M maltose. beta-Amylase (sweet potato), glucoamylase (Aspergillus niger), and pullulanase (Bacillus acidopullulyticus) from their crude preparations were purified 37-, 31-, and 49-fold with 86, 87, and 95% activity recovery, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed single band in each case.

  1. 21 CFR 184.1187 - Calcium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... alginic acid, a natural polyuronide constituent of certain brown algae. Calcium alginate is prepared by... Do. Fats and oils, § 170.3(n)(12) of this chapter 0.5 Do. Gelatins, puddings, § 170.3(n)(22) of...

  2. Nanocrystal/sol-gel nanocomposites

    DOEpatents

    Petruska, Melissa A.; Klimov, Victor L.

    2007-06-05

    The present invention is directed to solid composites including colloidal nanocrystals within a sol-gel host or matrix and to processes of forming such solid composites. The present invention is further directed to alcohol soluble colloidal nanocrystals useful in formation of sol-gel based solid composites.

  3. Nanocrystal/sol-gel nanocomposites

    SciTech Connect

    Petruska, Melissa A; Klimov, Victor L

    2012-06-12

    The present invention is directed to solid composites including colloidal nanocrystals within a sol-gel host or matrix and to processes of forming such solid composites. The present invention is further directed to alcohol soluble colloidal nanocrystals useful in formation of sol-gel based solid composites

  4. Time-Resolved Imaging Study of Jetting Dynamics during Laser Printing of Viscoelastic Alginate Solutions.

    PubMed

    Zhang, Zhengyi; Xiong, Ruitong; Mei, Renwei; Huang, Yong; Chrisey, Douglas B

    2015-06-16

    Matrix-assisted pulsed-laser evaporation direct-write (MAPLE DW) has been successfully implemented as a promising laser printing technology for various fabrication applications, in particular, three-dimensional bioprinting. Since most bioinks used in bioprinting are viscoelastic, it is of importance to understand the jetting dynamics during the laser printing of viscoelastic fluids in order to control and optimize the laser printing performance. In this study, MAPLE DW was implemented to study the jetting dynamics during the laser printing of representative viscoelastic alginate bioinks and evaluate the effects of operating conditions (e.g., laser fluence) and material properties (e.g., alginate concentration) on the jet formation performance. Through a time-resolved imaging approach, it is found that when the laser fluence increases or the alginate concentration decreases, the jetting behavior changes from no material transferring to well-defined jetting to well-defined jetting with an initial bulgy shape to jetting with a bulgy shape to pluming/splashing. For the desirable well-defined jetting regimes, as the laser fluence increases, the jet velocity and breakup length increase while the breakup time and primary droplet size decrease. As the alginate concentration increases, the jet velocity and breakup length decrease while the breakup time and primary droplet size increase. In addition, Ohnesorge, elasto-capillary, and Weber number based phase diagrams are presented to better appreciate the dependence of jetting regimes on the laser fluence and alginate concentration.

  5. Antimicrobial Effects of Silver Nanoparticles Stabilized in Solution by Sodium Alginate

    PubMed Central

    Kubyshkin, Anatoliy; Chegodar, Denis; Katsev, Andrew; Petrosyan, Armen; Krivorutchenko, Yuri; Postnikova, Olga

    2016-01-01

    Background/purpose To investigate the effect of nanosilver particles in solution stabilized in a matrix of sodium alginate on the growth and development of pathogenic bacteria such as Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Proteus vulgaris, Enterobacter cloacae, the antibiotic-resistant strain of Pseudomonas aeruginosa, the yeast-like fungus Candida albicans, and the luminescent bacteria Photobacterium leiognathi Sh1. Methods Isolates of pathogenic bacteria obtained from bronchoalveolar and peritoneal lavage samples from Wistar rats with experimental pneumonia and peritonitis were tested for their susceptibility to silver nanoparticles in solution with an alginate stabilizer. The antifungal activity of silver nanoparticles in sodium alginate was studied for C. albicans (strain CCM885) using the Sabouraud agar method. The biocidal impact of silver nanoparticles in solution with a sodium alginate matrix on the luminescent bacteria P. leiognathi Sh1 was investigated using a BLM 8801 luminometer. Results It was observed that a 0.02-0.05% nanosilver solution with an alginate stabilizer limits the growth and development of pathogenic bacteria within the first 24 hours of exposure. If the concentration of nanosilver solution is 0.0005-0.05%, it inhibits the viability of the fungus C. albicans. A nanosilver solution at a concentration of 0.05-0.2 μg/mL represses bioluminescence in the bacteria P. leiognathi Sh1. From these results, it appears that the biocidal effect of nanosilver is related either to the presence of ions that are formed during dissolution, or to the availability of nanoparticles that interrupt the membrane permeability of bacterial cells. Conclusion Silver nanoparticles stabilized in a solution of sodium alginate possess significant in vitro antimicrobial activity, which is manifested by inhibition of the bioluminescence of P. leiognathi Sh1, and inhibition of the growth and development of the pathogenic bacteria S. aureus, E

  6. Analysis of metal-binding proteins separated by non-denaturating gel electrophoresis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

    PubMed

    Becker, J Susanne; Mounicou, Sandra; Zoriy, Miroslav V; Becker, J Sabine; Lobinski, Ryszard

    2008-09-15

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) have become established as very efficient and sensitive biopolymer and elemental mass spectrometric techniques for studying metal-binding proteins (metalloproteins) in life sciences. Protein complexes present in rat tissues (liver and kidney) were separated in their native state in the first dimension by blue native gel electrophoresis (BN-PAGE). Essential and toxic metals, such as zinc, copper, iron, nickel, chromium, cadmium and lead, were detected by scanning the gel bands using quadrupole LA-ICP-MS with and without collision cell as a microanalytical technique. Several proteins were identified by using MALDI-TOF-MS together with a database search. For example, on one protein band cut from the BN-PAGE gel and digested with the enzyme trypsin, two different proteins - protein FAM44B and cathepsin B precursor - were identified. By combining biomolecular and elemental mass spectrometry, it was possible to characterize and identify selected metal-binding rat liver and kidney tissue proteins.

  7. Identification of differentially expressed proteins of gamma-ray irradiated rat intestinal epithelial IEC-6 cells by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry.

    PubMed

    Bo, Zhang; Yongping, Su; Fengchao, Wang; Guoping, Ai; Yongjiang, Wei

    2005-02-01

    To identify proteins involved in the processes of cellular and molecular response to radiation damage repair in intestinal epithelial IEC-6 cells, we comparatively analyzed the proteome of irradiated IEC-6 cells with that of normal cells. A series of methods were used, including two-dimensional gel electrophoresis (Z-DE), PDQuest software analysis of 2-DE gels, peptide mass fingerprinting based on matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), and Swiss-Prot database searching, to separate and identify differentially expressed proteins. Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) were used to validate the differentially expressed proteins. Image analysis revealed that averages of 608 +/- 39 and 595 +/- 31 protein spots were detected in normal and irradiated IEC-6 cells, respectively. Sixteen differential protein spots were isolated from gels, and measured with MALDI-TOF-MS. A total of 14 spots yielded good spectra, and 11 spots matched with known proteins after database searching. These proteins were mainly involved in anti-oxidation, metabolism, and protein post-translational processes. Western blotting confirmed that stress-70 protein was down-regulated by gamma-irradiation. Up-regulation of ERP29 was confirmed by RT-PCR, indicating that it is involved in ionizing radiation. The clues provided by the comparative proteome strategy utilized here will shed light on molecular mechanisms of radiation damage repair in intestinal epithelial cells.

  8. Biomedical-grade, high mannuronic acid content (BioMVM) alginate enhances the proteoglycan production of primary human meniscal fibrochondrocytes in a 3-D microenvironment

    PubMed Central

    Rey-Rico, Ana; Klich, Angelique; Cucchiarini, Magali; Madry, Henning

    2016-01-01

    Alginates are important hydrogels for meniscus tissue engineering as they support the meniscal fibrochondrocyte phenotype and proteoglycan production, the extracellular matrix (ECM) component chiefly responsible for its viscoelastic properties. Here, we systematically evaluated four biomedical- and two nonbiomedical-grade alginates for their capacity to provide the best three-dimensional (3-D) microenvironment and to support proteoglycan synthesis of encapsulated human meniscal fibrochondrocytes in vitro. Biomedical-grade, high mannuronic acid alginate spheres (BioLVM, BioMVM) were the most uniform in size, indicating an effect of the purity of alginate on the shape of the spheres. Interestingly, the purity of alginates did not affect cell viability. Of note, only fibrochondrocytes encapsulated in BioMVM alginate produced and retained significant amounts of proteoglycans. Following transplantation in an explant culture model, the alginate spheres containing fibrochondrocytes remained in close proximity with the meniscal tissue adjacent to the defect. The results reveal a promising role of BioMVM alginate to enhance the proteoglycan production of primary human meniscal fibrochondrocytes in a 3-D hydrogel microenvironment. These findings have significant implications for cell-based translational studies aiming at restoring lost meniscal tissue in regions containing high amounts of proteoglycans. PMID:27302206

  9. Development of a morphogenetically active scaffold for three-dimensional growth of bone cells: biosilica-alginate hydrogel for SaOS-2 cell cultivation.

    PubMed

    Müller, Werner E G; Schröder, Heinz C; Feng, Qingling; Schlossmacher, Ute; Link, Thorben; Wang, Xiaohong

    2015-11-01

    Polymeric silica is formed from ortho-silicate during a sol-gel formation process, while biosilica is the product of an enzymatically driven bio-polycondensation reaction. Both polymers have recently been described as a template that induces an increased expression of the genes encoding bone morphogenetic protein 2 (BMP-2) and osteoprotegerin in osteoblast-related SaOS-2 cells; simultaneously or subsequently the cells respond with enhanced hydroxyapatite formation. In order to assess whether the biocompatible polymeric silica/biosilica can serve as a morphogenetically active matrix suitable for three-dimensional (3D) cell growth, or even for 3D cell bioprinting, SaOS-2 cells were embedded into a Na-alginate-based hydrogel. Four different gelatinous hydrogel matrices were used for suspending SaOS-2 cells: (a) the hydrogel alone; (b) the hydrogel with 400 μM ortho-silicate; (c) the hydrogel supplemented with 400 μM ortho-silicate and recombinant silicatein to allow biosilica synthesis to occur; and (d) the hydrogel with ortho-silicate and BSA. The SaOS-2 cells showed an increased growth if silica/biosilica components were present in the hydrogel. Likewise intensified was the formation of hydroxyapatite nodules in the silica-containing hydrogels. After an incubation period of 2 weeks, cells present in silica-containing hydrogels showed a significantly higher expression of the genes encoding the cytokine BMP-2, the major fibrillar structural protein collagen 1 and likewise of carbonic anhydrase. It is concluded that silica, and to a larger extent biosilica, retains its morphogenetic/osteogenic potential after addition to Na-alginate-based hydrogels. This property might qualify silica hydrogels to be also used as a matrix for 3D cell printing.

  10. Novel spectrofluorimetric method for measuring the activity of the enzyme alpha-L-fucosidase using the nano composite optical sensor samarium(III)-doxycycline complex doped in sol-gel matrix.

    PubMed

    Attia, M S; Othman, A M; Aboaly, M M; Abdel-Mottaleb, M S A

    2010-07-15

    A novel, simple, sensitive, and precise spectrofluorimetric method was developed for measuring the activity of the enzyme alpha-L-fucosidase (AFU). The method was based upon measuring the quenching of the luminescence intensity of the produced yellow colored complex ion associate of 2-chloro-4-nitrophenol [2-CNP] and a nano composite optical sensor samarium(III)-doxycycline [Sm(3+)-DC](+) complex in a sol-gel matrix at 645 nm. The remarkable quenching of the luminescence intensity of the [Sm(3+)-DC](+) complex doped in a sol-gel matrix by various concentrations of the reagent [2-CNP] was successfully used as an optical sensor for the assessment of AFU activity. The calibration plot was achieved over the concentration range 3.4 x 10(-9)-1.0 x 10(-6) mol L(-1) [2-CNP] with a correlation coefficient of 0.99 and a detection limit of 6.0 x 10(-10) mol L(-1). The method was used satisfactorily for the assessment of the AFU activity in a number of serum samples collected from various patients. A significant correlation between the luminescence activity of the enzyme AFU measured by the proposed procedure and the standard method was applied to patients and controls. The method proceeds without practical artifacts compared to the standard method.

  11. Synergistic effects in semidilute mixed solutions of alginate and lactose-modified chitosan (chitlac).

    PubMed

    Donati, Ivan; Haug, Ingvild J; Scarpa, Tommaso; Borgogna, Massimiliano; Draget, Kurt I; Skjåk-Braek, Gudmund; Paoletti, Sergio

    2007-03-01

    The present study specifically aimed at preparing and characterizing semidilute binary polymer mixtures of alginate and chitlac which might find an application in the field of cell encapsulation. A polyanion, alginate, and a polycation, a lactose-modified chitosan, were mixed under physiological conditions (pH 7.4 and NaCl 0.15) and at a semidilute concentration avoiding associative phase separation. The mutual solubility was found to be dependent on the charge screening effect of the added NaCl salt, being prevented below 0.05 M NaCl. A comparison with the behavior of the polyanion (alginate) under the same experimental conditions revealed that both the viscosity and the relaxation times of the binary polymer solutions are strongly affected by the presence of the polycation. In particular, the occurrence of electrostatic interactions between the two oppositely charged polysaccharides led to a synergistic effect on the zero-shear viscosity of the solution, which showed a 4.2-fold increase with respect to that of the main component of the solution, i.e., alginate. Moreover, the relaxation time, calculated as the reciprocal of the critical share rate, markedly increased upon reducing the alginate fraction in the binary polysaccharide solution. However, the formation of the soluble complexes and the synergistic effect are reduced upon increasing the concentration of the 1:1 electrolyte. By containing a gel-forming polyanion (alginate, e.g., with Ca(2+) ions) and a bioactive polycation (chitlac, bearing a beta-linked D-galactose), the present system can be regarded as a first step toward the development of biologically active scaffold from polysaccharide mixtures.

  12. Agarose gel electrophoresis.

    PubMed

    Smith, D R

    1993-01-01

    After digestion of DNA with a restriction enzyme (Chapter 50), it is usually necessary, for both preparative and analytical purposes, to separate and visualize the products. In most cases, where the products are between 200 and 20,000 bp long, this is achieved by agarose gel electrophoresis. Agarose is a linear polymer that is extracted from seaweed and sold as a white powder. The powder is melted in buffer and allowed to cool, whereby the agarose forms a gel by hydrogen bonding. The hardened matrix contains pores, the size of which depends on the concentration of agarose. The concentration of agarose is referred to as a percentage of agarose to volume of buffer (w/v), and agarose gels are normally in the range of 0.3 to 3%. Many different apparatus arrangements have been devised to run agarose gels; for example, they can be run horizontally or vertically, and the current can be conducted by wicks or the buffer solution. However, today, the "submarine" gel system is almost universally used. In this method, the agarose gel is formed on a supporting plate, and then the plate is submerged into a tank containing a suitable electrophoresis buffer. Wells are preformed in the agarose gel with the aid of a "comb" that is inserted into the cooling agarose before the agarose has gelled. Into these wells are loaded the sample to be analyzed, which has been mixed with a dense solution (a loading buffer) to ensure that the sample sinks into the wells.

  13. Encapsulation of Polymer Colloids in a Sol-Gel Matrix. Direct-Writing of Coassembling Organic-Inorganic Hybrid Photonic Crystals.

    PubMed

    Mikosch, Annabel; Kuehne, Alexander J C

    2016-03-22

    The spontaneous self-assembly of polymer colloids into ordered arrangements provides a facile strategy for the creation of photonic crystals. However, these structures often suffer from defects and insufficient cohesion, which result in flaking and delamination from the substrate. A coassembly process has been developed for convective assembly, resulting in large-area encapsulated colloidal crystals. However, to generate patterns or discrete deposits in designated places, convective assembly is not suitable. Here we experimentally develop conditions for direct-writing of coassembling monodisperse dye-doped polystyrene particles with a sol-gel precursor to form solid encapsulated photonic crystals. In a simple procedure the colloids are formulated in a sol-gel precursor solution, drop-cast on a flat substrate, and dried. We here establish the optimal parameters to form reproducible highly ordered photonic crystals with good optical performance. The obtained photonic crystals interact with light in the visible spectrum with a narrow optical stop-gap.

  14. Coupling sodium dodecyl sulfate-capillary polyacrylamide gel electrophoresis with matrix-assisted laser desorption ionization time-of-flight mass spectrometry via a poly(tetrafluoroethylene) membrane.

    PubMed

    Lu, Joann J; Zhu, Zaifang; Wang, Wei; Liu, Shaorong

    2011-03-01

    Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) is a fundamental analytical technique for proteomic research, and SDS-capillary gel electrophoresis (CGE) is its miniaturized version. Compared to conventional slab-gel electrophoresis, SDS-CGE has many advantages such as increased separation efficiency, reduced separation time, and automated operation. SDS-CGE is not widely accepted in proteomic research primarily due to the difficulties in identifying the well-resolved proteins. MALDI-TOF-MS is an outstanding platform for protein identifications. Coupling the two would solve the problem but is extremely challenging because the MS detector has no access to the SDS-CGE-resolved proteins and the SDS interferes with MS detection. In this work we introduce an approach to address these issues. We discover that poly(tetrafluoroethylene) (PTFE) membranes are excellent materials for collecting SDS-CGE-separated proteins. We demonstrate that we can wash off the SDS bound to the collected proteins and identify these proteins on-membrane with MALDI-TOF-MS. We also show that we can immunoblot and Coomassie-stain the proteins collected on these membranes.

  15. Rheological investigation of specific interactions in Na Alginate and Na MMT suspension.

    PubMed

    Zlopasa, Jure; Norder, Ben; Koenders, Eduard A B; Picken, Stephen J

    2016-10-20

    Here we report on a study of a rheological behavior of sodium alginate and montmorillonite suspension. We find that viscoelastic behavior of this suspension is dramatically affected with increasing volume fraction of montmorillonite platelets. Addition of montmorillonite generally leads to gel formation, which is attributed to interactions of montmorillonite and alginate via H-bonding and attraction between the positive edges of the platelets and the anionic backbone of the biopolymer. A critical concentration for the measured system was observed at 20wt.% montmorillonite, where a crossover to a gel-like structure was detected. The observed gel has a rubber plateau, which develops further with higher montmorillonite concentration. In this physical gel the relaxation maximum was detected, which is associated with the breaking and reformation of the bonds between the platelets and the biopolymer. For this transient behavior, we find that a Maxwell type viscoelasticity quite well describes the relaxation time and the observed G'-G" crossover. We believe that this gel-like behavior plays an important role in formation of highly ordered nanostructures that develop during the drying of these bio-nanocomposite suspensions.

  16. Performance and Biocompatibility of Extremely Tough Alginate/Polyacrylamide Hydrogels

    PubMed Central

    Darnell, Max; Sun, Jeong-Yun; Mehta, Manav; Johnson, Chris; Arany, Praveen; Suo, Zhigang

    2013-01-01

    Although hydrogels now see widespread use in a host of applications, low fracture toughness and brittleness have limited their more broad use. As a recently described interpenetrating network (IPN) of alginate and polyacrylamide demonstrated a fracture toughness of ∼9000 J/m2, we sought to explore the biocompatibility and maintenance of mechanical properties of these hydrogels in cell culture and in vivo conditions. These hydrogels can sustain a compressive strain of over 90% with minimal loss of Young's Modulus as well as minimal swelling for up to 50 days of soaking in culture conditions. Mouse mesenchymal stem cells exposed to the IPN gel-conditioned media maintain high viability, and although cells exposed to conditioned media demonstrate slight reductions in proliferation and metabolic activity (WST assay), these effects are abrogated in a dose-dependent manner. Implantation of these IPN hydrogels into subcutaneous tissue of rats for 8 weeks led to mild fibrotic encapsulation and minimal inflammatory response. These results suggest the further exploration of extremely tough alginate/PAAM IPN hydrogels as biomaterials. PMID:23896005

  17. Gel polymer electrolytes for batteries

    DOEpatents

    Balsara, Nitash Pervez; Eitouni, Hany Basam; Gur, Ilan; Singh, Mohit; Hudson, William

    2014-11-18

    Nanostructured gel polymer electrolytes that have both high ionic conductivity and high mechanical strength are disclosed. The electrolytes have at least two domains--one domain contains an ionically-conductive gel polymer and the other domain contains a rigid polymer that provides structure for the electrolyte. The domains are formed by block copolymers. The first block provides a polymer matrix that may or may not be conductive on by itself, but that can soak up a liquid electrolyte, thereby making a gel. An exemplary nanostructured gel polymer electrolyte has an ionic conductivity of at least 1.times.10.sup.-4 S cm.sup.-1 at 25.degree. C.

  18. Differential effect of the shape of calcium alginate matrices on the physiology of immobilized neuroblastoma N2a and Vero cells: a comparative study.

    PubMed

    Kintzios, S; Yiakoumetis, I; Moschopoulou, G; Mangana, O; Nomikou, K; Simonian, A

    2007-11-30

    In order to investigate the effect of cell immobilization in calcium alginate gels on cell physiology, we immobilized Vero or N2a neuroblastoma cells in gels shaped either as spherical beads or as thin membrane layers. Throughout a culture period of 4 weeks cell viability, RNA and cytoplasmic calcium concentration and glutathione accumulation were assayed by fluorescence microscopy after provision of an appropriate dye. Non-elaborate culture conditions were applied throughout the experimental period in order to evaluate cell viability under less than optimal storage conditions. Vero cell proliferation was observed only in spherical beads, while N2a cell proliferation was observed in both configurations until the third week of culture. Increased [Ca2+]cyt could be associated with cell proliferation only when cells were immobilized in spherical beads, while a considerable decrease in the biosynthesis of reduced glutathione and RNA was observed in cells immobilized in thin membrane layers. The observed effects of the shape of the immobilization matrix may be due to differences in external mass transfer resistance. Therefore, depending on cell type, cell proliferation could have been promoted by either increased (Vero) or decreased (N2a) nutrient and oxygen flow to immobilized cells. The results of the present study could contribute to an improvement of immobilized cell sensor storability.

  19. Adsorption of a cationic surfactant by a magsorbent based on magnetic alginate beads.

    PubMed

    Obeid, Layaly; El Kolli, Nadia; Dali, Noëlle; Talbot, Delphine; Abramson, Sébastien; Welschbillig, Mathias; Cabuil, Valérie; Bée, Agnès

    2014-10-15

    Adsorption of cetylpyridinium chloride (CPC), a cationic surfactant, by magnetic alginate beads (MagAlgbeads) was investigated. The magnetic adsorbent (called magsorbent) was prepared by encapsulation of magnetic functionalized nanoparticles in an alginate gel. The influence on CPC adsorption of several parameters such as contact time, pH and initial surfactant concentration was studied. The equilibrium isotherm shows that adsorption occurs through both electrostatic interactions with charge neutralization of the carboxylate groups of the beads and hydrophobic interactions inducing the formation of surfactant aggregates in the beads. The dosage of calcium ions released in the solution turns out to be a useful tool for understanding the adsorption mechanisms. Adsorption is accompanied by a shrinking of the beads that corresponds to a 45% reduction of the volume. Adsorption kinetic experiments show that equilibrium time is strongly dependent on the surfactant concentration, which monitors the nature of the interactions. On the other hand, since the pH affects the ionization state of adsorption sites, adsorption depends on the pH solution, maximum adsorption being obtained in a large pH range (3.2-12) in agreement with the pKa value of alginate (pKa=3.4-4.2). Finally, due to the formation of micelle-like surfactants aggregates in the magnetic alginate beads, they could be used as a new efficient magsorbent for hydrophobic pollutants.

  20. Fabrication and characterization of macroporous epichlorohydrin cross-linked alginate beads as protein adsorbent.

    PubMed

    Zhang, Weican; Ji, Xiaofei; Sun, Caiyun; Lu, Xuemei

    2013-01-01

    Porous epichlorohydrin cross-linked alginate beads (ECAB) were prepared by the following method. Na-alginate solution containing Na2SO4 was introduced dropwise into CaCl2 solution to simultaneously form CaSO4 precipitate and Ca-alginate gel beads. The resultant beads were cross-linked with epichlorohydrin and then thoroughly washed with ethylenediamine tetraacetic acid (EDTA) solution to remove CaSO4. The structural features of porous ECAB were assessed with scanning electron microscopy (SEM) and experiments on water content and adsorption of bovine serum albumin (BSA). The results showed that macroporous ECAB can be obtained when the mass ratio of sodium sulfate to sodium alginate is 4:1. The adsorption behavior of the macroporous ECAB was well described by the Langmuir isotherm with maximum adsorption capacity equal to 740 mg BSA/g dry weight in 50 mM Na2HPO4-citric acid buffer (pH 4.0). BSA was more effectively adsorbed by macroporous ECAB at around pH 3 and the mechanism of the adsorption of BSA to the ECAB was ion exchange. Finally, experiments of a concentration of 1 mg/mL BSA using macroporous ECAB were performed.

  1. Selective removals of heavy metals (Pb(2+), Cu(2+), and Cd(2+)) from wastewater by gelation with alginate for effective metal recovery.

    PubMed

    Wang, Fei; Lu, Xingwen; Li, Xiao-yan

    2016-05-05

    A novel method that uses the aqueous sodium alginate solution for direct gelation with metal ions is developed for effective removal and recovery of heavy metals from industrial wastewater. The experimental study was conducted on Pb(2+), Cu(2+), and Cd(2+) as the model heavy metals. The results show that gels can be formed rapidly between the metals and alginate in less than 10 min and the gelation rates fit well with the pseudo second-order kinetic model. The optimum dosing ratio of alginate to the metal ions was found to be between 2:1 and 3:1 for removing Pb(2+) and around 4:1 for removing Cu(2+) and Cd(2+) from wastewater, and the metal removal efficiency by gelation increased as the solution pH increased. Alginate exhibited a higher gelation affinity toward Pb(2+) than Cu(2+) and Cd(2+), which allowed a selective removal of Pb(2+) from the wastewater in the presence of Cu(2+) and Cd(2+) ions. Chemical analysis of the gels suggests that the gelation mainly occurred between the metal ions and the -COO(-) and -OH groups on alginate. By simple calcination of the metal-laden gels at 700 °C for 1 h, the heavy metals can be well recovered as valuable resources. The metals obtained after the thermal treatment are in the form of PbO, CuO, and CdO nanopowders with crystal sizes of around 150, 50, and 100 nm, respectively.

  2. Efficient dual-wavelength excitation of Tb3+ emission in rare-earth doped KYF4 cubic nanocrystals dispersed in silica sol-gel matrix

    NASA Astrophysics Data System (ADS)

    del-Castillo, J.; Yanes, A. C.; Santana-Alonso, A.; Méndez-Ramos, J.

    2014-11-01

    Energy transfer from Ce3+ to Tb3+ ions under UV excitation, giving rise to visible emissions, is investigated in sol-gel derived transparent nano-glass-ceramics containing cubic KYF4 nanocrystals, for different doping concentrations of rare-earth ions. Moreover, visible emissions of Tb3+ are also obtained under near-infrared excitation through energy transfer from Yb3+ ions by means of cooperative up-conversion processes. Thus, Ce3+-Tb3+-Yb3+ doped nano-glass-ceramics can be activated in a dual-wavelength mode yielding efficient blue-green emissions of particular interest in photovoltaic silicon solar cells and white-light emitting diodes.

  3. NMR microscopy of heavy metal absorption in calcium alginate beads

    SciTech Connect

    Nestle, N.; Kimmich, R.

    1996-01-01

    In recent years, heavy metal uptake by biopolymer gels, such as Cal-Alginate or chitosan, has been studied by various methods. This is of interest because such materials might be an alternative to synthetical ion-exchange resins in the treatment of industrial waste waters. Most of the work done in this field consisted of studies of equilibrium absorption of different heavy metal ions with dependence on various experimental parameters. In some publications, the kinetics of absorption were studied, too. However, no experiments on the spatial distribution of heavy metals during the absorption process are known to us. Using Cu as an example, it is demonstrated in this article that NMR microscopy is an appropriate tool for such studies. By the method presented here, it is possible to monitor the spatial distribution of heavy metal ions with a time resolution of about 5 min and a spatial resolution of 100 {mu}m or even better. 14 refs., 10 figs.

  4. Bromo-oxidation reaction in enzyme-entrapped alginate hollow microfibers.

    PubMed

    Asthana, Amit; Lee, Kwang Ho; Shin, Su-Jung; Perumal, Jayakumar; Butler, Lauren; Lee, Sang-Hoon; Kim, Dong-Pyo

    2011-06-01

    In this article, the authors present the fabrication of an enzyme-entrapped alginate hollow fiber using a microfluidic device. Further use of enzyme-entrapped alginate hollow fibers as a biocatalytic microchemical reactor for chemical synthesis is also deliberated in this article. To ensure that there is no enzyme leaching from the fiber, fiber surfaces were coated with chitosan. To confine the mobility of reactants and products within the porous hollow fibers the entire fibers were embedded into a transparent polydimethylsiloxane (PDMS) matrix which also works as a support matrix. A vanadium-containing bromoperoxidase enzyme isolated from Corallina confusa was used as a model enzyme to demonstrate the use of these alginate hollow-fiber reactors in bromo-oxidation of phenol red to bromophenol blue at different dye flow rates. Stability of the entrapped enzyme at different temperatures and the effect of the chitosan coating on the reaction conversion were also studied. It was observed that molecules as big as 27 kDa can be retained in the matrix after coating with chitosan while molecules with molecular-weight of around 378 Da can still diffuse in and out of the matrix. The kinetic conversion rate in this microfluidic bioreactor was more than 41-fold faster when compared with the standard test-tube procedure.

  5. Bromo-oxidation reaction in enzyme-entrapped alginate hollow microfibers

    PubMed Central

    Asthana, Amit; Lee, Kwang Ho; Shin, Su-Jung; Perumal, Jayakumar; Butler, Lauren; Lee, Sang-Hoon; Kim, Dong-Pyo

    2011-01-01

    In this article, the authors present the fabrication of an enzyme-entrapped alginate hollow fiber using a microfluidic device. Further use of enzyme-entrapped alginate hollow fibers as a biocatalytic microchemical reactor for chemical synthesis is also deliberated in this article. To ensure that there is no enzyme leaching from the fiber, fiber surfaces were coated with chitosan. To confine the mobility of reactants and products within the porous hollow fibers the entire fibers were embedded into a transparent polydimethylsiloxane (PDMS) matrix which also works as a support matrix. A vanadium-containing bromoperoxidase enzyme isolated from Corallina confusa was used as a model enzyme to demonstrate the use of these alginate hollow-fiber reactors in bromo-oxidation of phenol red to bromophenol blue at different dye flow rates. Stability of the entrapped enzyme at different temperatures and the effect of the chitosan coating on the reaction conversion were also studied. It was observed that molecules as big as 27 kDa can be retained in the matrix after coating with chitosan while molecules with molecular-weight of around 378 Da can still diffuse in and out of the matrix. The kinetic conversion rate in this microfluidic bioreactor was more than 41-fold faster when compared with the standard test-tube procedure. PMID:21799723

  6. Biosynthesis of the Pseudomonas aeruginosa Extracellular Polysaccharides, Alginate, Pel, and Psl

    PubMed Central

    Franklin, Michael J.; Nivens, David E.; Weadge, Joel T.; Howell, P. Lynne

    2011-01-01

    Pseudomonas aeruginosa thrives in many aqueous environments and is an opportunistic pathogen that can cause both acute and chronic infections. Environmental conditions and host defenses cause differing stresses on the bacteria, and to survive in vastly different environments, P. aeruginosa must be able to adapt to its surroundings. One strategy for bacterial adaptation is to self-encapsulate with matrix material, primarily composed of secreted extracellular polysaccharides. P. aeruginosa has the genetic capacity to produce at least three secreted polysaccharides; alginate, Psl, and Pel. These polysaccharides differ in chemical structure and in their biosynthetic mechanisms. Since alginate is often associated with chronic pulmonary infections, its biosynthetic pathway is the best characterized. However, alginate is only produced by a subset of P. aeruginosa strains. Most environmental and other clinical isolates secrete either Pel or Psl. Little information is available on the biosynthesis of these polysaccharides. Here, we review the literature on the alginate biosynthetic pathway, with emphasis on recent findings describing the structure of alginate biosynthetic proteins. This information combined with the characterization of the domain architecture of proteins encoded on the Psl and Pel operons allowed us to make predictive models for the biosynthesis of these two polysaccharides. The results indicate that alginate and Pel share certain features, including some biosynthetic proteins with structurally or functionally similar properties. In contrast, Psl biosynthesis resembles the EPS/CPS capsular biosynthesis pathway of Escherichia coli, where the Psl pentameric subunits are assembled in association with an isoprenoid lipid carrier. These models and the environmental cues that cause the cells to produce predominantly one polysaccharide over the others are subjects of current investigation. PMID:21991261

  7. Immobilization of flax protoplasts in agarose and alginate beads. Correlation between ionically bound cell-wall proteins and morphogenetic response.

    PubMed Central

    Roger, D; David, A; David, H

    1996-01-01

    Linum usitatissimum protoplast-derived colonies that are cultured in auxin-supplemented medium and immobilized in Ca(2+)-alginate matrix form round colonies that develop into polarized, embryo-like structures. On the other hand, protoplast-derived colonies that are immobilized in agarose do not show an organized morphogenetic response, and unique, ionically bound cell-wall protein patterns match this response. Although only slight differences in neosynthesized or total constitutive polypeptides are observed, dramatic changes in ionically bound cell-wall proteins are seen. In protoplasts grown on Ca(2+)-alginate-solidified, auxin-containing medium, several basic polypeptides were strongly induced and were found tightly bound to the cell wall. In contrast, these basic proteins were found only weakly bound to the walls of protoplasts grown on agarose-solidified, auxin-containing medium or on Ca(2+)-alginate-solidified, auxin-free medium, in which they were released into the medium. Our results suggest that plant cells can perceive and respond to the adjacent extracellular matrix, since we show that the growth of flax cells on Ca(2+)-alginate in the presence of auxin-containing medium may promote the binding of specific proteins to the walls. This establishes a direct correlation of an embryo-like morphogenesis with ionically bound cell-wall basic proteins in flax protoplasts grown on Ca(2+)-alginate-solidified, auxin-containing medium. PMID:8938417

  8. Lead removal in rats using calcium alginate.

    PubMed

    Savchenko, Olga V; Sgrebneva, Marina N; Kiselev, Vladimir I; Khotimchenko, Yuri S

    2015-01-01

    Lead (Pb) exposure, even at low levels, causes a variety of health problems. The aims of this study were to investigate the tissue distribution of lead in the bodies of rats, to evaluate lead removal from the internal organs and bones using calcium alginate in doses of 500, 200 and 100 mg/kg per day for 28 days and to assess the impact of calcium alginate on the level of essential elements. Lead (Pb), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu) and zinc (Zn) levels in the blood, hearts, kidneys, livers and femurs of the experimental animals were measured using mass spectrometry with inductively coupled plasma. The results revealed that lead acetate exposure increased the levels of Pb in the blood and organs of the animals and significantly reduced contents of Ca, Mn, Fe, Cu and Zn. Treatment with calcium alginate in dose 500 mg/kg contributed to significant decreases in the amount of lead in the kidney, heart and bones of animals and a slight increase in the content of essential elements in the liver, kidneys and heart, although these changes were not significant. Decreasing of lead was not significant in the internal organs, bones and blood of animals treated with calcium alginate 200 and 100 mg/kg. Consequently, calcium alginate dose of 500 mg/kg more efficiently removes lead accumulated in the body. Calcium alginate does not have negative effect on level of essential elements quite the contrary; reducing the levels of lead, calcium alginate helps normalize imbalances of Ca, Mn, Fe, Cu and Zn. The results of this study suggest that calcium alginate may potentially be useful for the treatment and prevention of heavy metal intoxications.

  9. Mechanical spectroscopy and relaxometry on alginate hydrogels: a comparative analysis for structural characterization and network mesh size determination.

    PubMed

    Turco, Gianluca; Donati, Ivan; Grassi, Mario; Marchioli, Giulia; Lapasin, Romano; Paoletti, Sergio

    2011-04-11

    The structure of calcium-saturated alginate hydrogels has been studied by combining rheological determinations and relaxometry measurements. The mechanical spectroscopy analyses performed on alginate gel cylinders at different polysaccharide concentration allowed estimating their main structural features such as the average mesh size. The calculation was based on the introduction of a front factor in the classical rubber elasticity approach which was correlated to the average length of the Guluronic acid blocks along the polysaccharide chain. Transverse relaxation time (T(2)) determinations performed on the cylinders revealed the presence of two relaxation rates of the water entrapped within the hydrogel network. The cross-correlation of the latter data with the rheological measurements allowed estimating the mesh size distribution of the hydrogel network. The results obtained for the hydrogel cylinders were found to be consistent with the relaxometric analysis performed on the alginate microbeads where, however, only one type of water bound into the network structure was detected. A good correlation was found in the average mesh size determined by means of relaxometric measurements on alginate microbeads and by a statistical analysis performed on TEM micrographs. Finally, the addition of a solution containing calcium ions allowed investigating further the different water relaxation modes within alginate hydrogels.

  10. Annealing behavior of silver, copper, and silver-copper nanoclusters in a silica matrix synthesized by the sol-gel technique

    NASA Astrophysics Data System (ADS)

    De, G.; Gusso, M.; Tapfer, L.; Catalano, M.; Gonella, F.; Mattei, G.; Mazzoldi, P.; Battaglin, G.

    1996-12-01

    Silver, copper, and mixed silver-copper nanocluster-doped silica thin layers were prepared by the sol-gel process. Samples were heat treated in different annealing atmospheres (air, argon, or 5%H2-95%N2) in the temperature range 500-1100 °C. Specimens were characterized by optical absorption spectroscopy, Rutherford backscattering spectrometry, x-ray diffraction, and transmission electron microscopy. Cluster growth and dissolution, as well as migration of metal atoms towards the sample surface, with a subsequent evaporation, were observed to occur at temperatures that depend on the annealing atmosphere. In the mixed silver-copper system, the formation of Ag-Cu phase-separated clusters was observed.

  11. Composite ECM-alginate microfibers produced by microfluidics as scaffolds with biomineralization potential.

    PubMed

    Angelozzi, Marco; Miotto, Martina; Penolazzi, Letizia; Mazzitelli, Stefania; Keane, Timothy; Badylak, Stephen F; Piva, Roberta; Nastruzzi, Claudio

    2015-11-01

    A novel approach to produce artificial bone composites (microfibers) with distinctive features mimicking natural tissue was investigated. Currently proposed inorganic materials (e.g. apatite matrixes) lack self-assembly and thereby limit interactions between cells and the material. The present work investigates the feasibility of creating "bio-inspired materials" specifically designed to overcome certain limitations inherent to current biomaterials. We examined the dimensions, morphology, and constitutive features of a composite hydrogel which combined an alginate based microfiber with a gelatin solution or a particulate form of urinary bladder matrix (UBM). The effectiveness of the composite microfibers to induce and modulate osteoblastic differentiation in three-dimensional (3D) scaffolds without altering the viability and morphological characteristics of the cells was investigated. The present study describes a novel alginate microfiber production method with the use of microfluidics. The microfluidic procedure allowed for precise tuning of microfibers which resulted in enhanced viability and function of embedded cells.

  12. Control of Alginate Core Size in Alginate-Poly (Lactic-Co-Glycolic) Acid Microparticles

    NASA Astrophysics Data System (ADS)

    Lio, Daniel; Yeo, David; Xu, Chenjie

    2016-01-01

    Core-shell alginate-poly (lactic-co-glycolic) acid (PLGA) microparticles are potential candidates to improve hydrophilic drug loading while facilitating controlled release. This report studies the influence of the alginate core size on the drug release profile of alginate-PLGA microparticles and its size. Microparticles are synthesized through double-emulsion fabrication via a concurrent ionotropic gelation and solvent extraction. The size of alginate core ranges from approximately 10, 50, to 100 μm when the emulsification method at the first step is homogenization, vortexing, or magnetic stirring, respectively. The second step emulsification for all three conditions is performed with magnetic stirring. Interestingly, although the alginate core has different sizes, alginate-PLGA microparticle diameter does not change. However, drug release profiles are dramatically different for microparticles comprising different-sized alginate cores. Specifically, taking calcein as a model drug, microparticles containing the smallest alginate core (10 μm) show the slowest release over a period of 26 days with burst release less than 1 %.

  13. Photo-activated ionic gelation of alginate hydrogel: real-time rheological monitoring of the two-step crosslinking mechanism.

    PubMed

    Higham, Alina K; Bonino, Christopher A; Raghavan, Srinivasa R; Khan, Saad A

    2014-07-21

    We examine the gelation of alginate undergoing ionic crosslinking upon ultraviolet (UV) irradiation using in situ dynamic rheology. Hydrogels are formed by combining alginate with calcium carbonate (CaCO3) particles and a photoacid generator (PAG). The PAG is photolyzed upon UV irradiation, resulting in the release of free calcium ions for ionic crosslinking. The viscous and elastic moduli during gelation are monitored as a function of the UV irradiation intensity, exposure time, alginate concentration, and the ratio between alginate and calcium carbonate. Gel time decreases as irradiation intensity increases because a larger concentration of PAG is photolyzed. Interestingly, dark curing, the continuing growth of microstructure in the absence of UV light, is observed. In some instances, the sample transitions from a solution to a gel during the dark curing phase. Additionally, when exposed to constant UV irradiation after the dark curing phase, samples reach the same plateau modulus as samples exposed to constant UV without dark curing, implying that dark curing does not affect the gelation mechanism. We believe the presence of dark curing is the result of the acidic environment persisting within the sample, allowing CaCO3 to dissociate, thereby releasing free Ca(2+) ions capable of binding with the available appropriate ionic blocks of the polymer chains. The growth of microstructure is then detected if the activation barrier has been crossed to release sufficient calcium ions. In this regard, we calculate a value of 30 J that represents the activation energy required to initiate gelation.

  14. Direct deposited porous scaffolds of calcium phosphate cement with alginate for drug delivery and bone tissue engineering.

    PubMed

    Lee, Gil-Su; Park, Jeong-Hui; Shin, Ueon Sang; Kim, Hae-Won

    2011-08-01

    This study reports the preparation of novel porous scaffolds of calcium phosphate cement (CPC) combined with alginate, and their potential usefulness as a three-dimensional (3-D) matrix for drug delivery and tissue engineering of bone. An α-tricalcium phosphate-based powder was mixed with sodium alginate solution and then directly injected into a fibrous structure in a Ca-containing bath. A rapid hardening reaction of the alginate with Ca(2+) helps to shape the composite into a fibrous form with diameters of hundreds of micrometers, and subsequent pressing in a mold allows the formation of 3-D porous scaffolds with different porosity levels. After transformation of the CPC into a calcium-deficient hydroxyapatite phase in simulated biological fluid the scaffold was shown to retain its mechanical stability. During the process biological proteins, such as bovine serum albumin and lysozyme, used as model proteins, were observed to be effectively loaded onto and released from the scaffolds for up to more than a month, proving the efficacy of the scaffolds as a drug delivering matrix. Mesenchymal stem cells (MSCs) were isolated from rat bone marrow and then cultured on the CPC-alginate porous scaffolds to investigate the ability to support proliferation of cells and their subsequent differentiation along the osteogenic lineage. It was shown that MSCs increasingly actively populated and also permeated into the porous network with time of culture. In particular, cells cultured within a scaffold with a relatively high porosity level showed favorable proliferation and osteogenic differentiation. An in vivo pilot study of the CPC-alginate porous scaffolds after implantation into the rat calvarium for 6 weeks revealed the formation of new bone tissue within the scaffold, closing the defect almost completely. Based on these results, the newly developed CPC-alginate porous scaffolds could be potentially useful as a 3-D matrix for drug delivery and tissue engineering of bone.

  15. Alginate/polyoxyethylene and alginate/gelatin hydrogels: preparation, characterization, and application in tissue engineering.

    PubMed

    Aroguz, Ayse Z; Baysal, Kemal; Adiguzel, Zelal; Baysal, Bahattin M

    2014-05-01

    Hydrogels are attractive biomaterials for three-dimensional cell culture and tissue engineering applications. The preparation of hydrogels using alginate and gelatin provides cross-linked hydrophilic polymers that can swell but do not dissolve in water. In this work, we first reinforced pure alginate by using polyoxyethylene as a supporting material. In an alginate/PEO sample that contains 20 % polyoxyethylene, we obtained a stable hydrogel for cell culture experiments. We also prepared a stable alginate/gelatin hydrogel by cross-linking a periodate-oxidized alginate with another functional component such as gelatin. The hydrogels were found to have a high fluid uptake. In this work, preparation, characterization, swelling, and surface properties of these scaffold materials were described. Lyophilized scaffolds obtained from hydrogels were used for cell viability experiments, and the results were presented in detail.

  16. Controlled microfluidic production of alginate beads for in situ encapsulation of microbes

    SciTech Connect

    Kalyanaraman, Meenaa; Retterer, Scott T; McKnight, Timothy E; Ericson, Milton Nance; Allman, Steve L; Elkins, James G; Palumbo, Anthony Vito; Keller, Martin; Doktycz, Mitchel John

    2009-01-01

    The development and refinement of a microfluidic-based alginate bead generator system for bacterial encapsulation is presented. The resulting microgels have application for the encapsulation of single cells, and can allow for small scale, clonal expansion of thousands of isolated cells in parallel. PDMS based microfluidic chips were fabricated using conventional lithography techniques to produce both externally gelled and directly gelled alginate microspheres using a controlled, water-in-oil emulsion system. The production of directly gelled beads, formed by the in-chip mixing of aqueous alginate and calcium chloride solutions dispersed within an organic carrier flowstream is qualitatively compared to a system, which produces beads and relies on diffusion of a crosslinking agent from the carrier fluid to cause gelation (external gelation). While the direct gelation scheme allows the use of biocompatible oils as the organic carrier, it also has a detrimental effect on device stability often resulting in clogging and gel-streaming at the microfluidic interface of these solutions. A design for the continuous production of directly gelled beads was evaluated in terms of the threshold flow conditions and reagent concentrations that did not result in clogging or streaming. Monodisperse alginate microgels of 30 mum diameter were produced at frequencies of over 500 beads per second. The beads could be completely dispersed into aqueous media using an off-chip washing protocol to remove the organic phase. The microgels effectively encapsulated individual or small numbers of GFP-expressing Escherichia. coli, which could be subsequently clonally expanded. The described microfluidic platform is a robust front-end sample preparation technology that shows strong potential for use in drug delivery systems, biosensors, and other cell-based microcompartmentalization applications. The co-culturing of microbial colonies in a large population of alginate beads will allow for functional

  17. Cytoprotective alginate/polydopamine core/shell microcapsules in microbial encapsulation.

    PubMed

    Kim, Beom Jin; Park, Taegyun; Moon, Hee Chul; Park, So-Young; Hong, Daewha; Ko, Eun Hyea; Kim, Ji Yup; Hong, Jong Wook; Han, Sang Woo; Kim, Yang-Gyun; Choi, Insung S

    2014-12-22

    Chemical encapsulation of microbes in threedimensional polymeric microcapsules promises various applications, such as cell therapy and biosensors, and provides a basic platform for studying microbial communications. However, the cytoprotection of microbes in the microcapsules against external aggressors has been a major challenge in the field of microbial microencapsulation, because ionotropic hydrogels widely used for microencapsulation swell uncontrollably, and are physicochemically labile. Herein, we developed a simple polydopamine coating for obtaining cytoprotective capability of the alginate capsule that encapsulated Saccharomyces cerevisiae. The resulting alginate/ polydopamine core/shell capsule was mechanically tough, prevented gel swelling and cell leakage, and increased resistance against enzymatic attack and UV-C irradiation. We believe that this multifunctional core/shell structure will provide a practical tool for manipulating microorganisms inside the microcapsules.

  18. Development of novel alginate based hydrogel films for wound healing applications.

    PubMed

    Pereira, Rúben; Carvalho, Anabela; Vaz, Daniela C; Gil, M H; Mendes, Ausenda; Bártolo, Paulo

    2013-01-01

    Alginate and Aloe vera are natural materials widely investigated and used in the biomedical field. In this research work, thin hydrogel films composed by alginate and Aloe vera gel in different proportions (95:5, 85:15 and 75:25, v/v) were prepared and characterized. The films were evaluated regarding the light transmission behavior, contact angle measurements, and chemical, thermal and mechanical properties. These thin hydrogel films, prepared by crosslinking reaction using 5% calcium chloride solution, were also investigated relatively to their water solubility and swelling behavior. Results showed that Aloe vera improved the transparency of the films, as well their thermal stability. The developed films present adequate mechanical properties for skin applications, while the solubility studies demonstrated the insolubility of the films after 24h of immersion in distilled water. The water absorption and swelling behavior of these films were greatly improved by the increase in Aloe vera proportion.

  19. Hybrid Gel Composed of Native Heart Matrix and Collagen Induces Cardiac Differentiation of Human Embryonic Stem Cells without Supplemental Growth Factors

    PubMed Central

    Duan, Yi; Liu, Zen; O'Neill, John; Wan, Leo Q.; Freytes, Donald O.; Vunjak-Novakovic, Gordana

    2011-01-01

    Our goal was to assess the ability of native heart extracellular matrix (ECM) to direct cardiac differentiation of human embryonic stem cells (hESCs) in vitro. In order to probe the effects of cardiac matrix on hESC differentiation, a series of hydrogels was prepared from decellularized ECM from porcine hearts by mixing ECM and collagen type I at varying ratios. Maturation of cardiac function in embryoid bodies formed from hESCs was documented in terms of spontaneous contractile behavior and the mRNA and protein expression of cardiac markers. Hydrogel with high ECM content (75% ECM, 25% collagen, no supplemental soluble factors) increased the fraction of cells expressing cardiac marker troponin T, when compared with either hydrogel with low ECM content (25% ECM, 75% collagen, no supplemental soluble factors) or collagen hydrogel (100% collagen, with supplemental soluble factors). Furthermore, cardiac maturation was promoted in high-ECM content hydrogels, as evidenced by the striation patterns of cardiac troponin I and by upregulation of Cx43 gene. Consistently, high-ECM content hydrogels improved the contractile function of cardiac cells, as evidenced by increased numbers of contracting cells and increased contraction amplitudes. The ability of native ECM hydrogel to induce cardiac differentiation of hESCs without the addition of soluble factors makes it an attractive biomaterial system for basic studies of cardiac development and potentially for the delivery of therapeutic cells into the heart. PMID:21744185

  20. Effects of Chitin Whiskers on Physical Properties and Osteoblast Culture of Alginate Based Nanocomposite Hydrogels.

    PubMed

    Huang, Yao; Yao, Mengyu; Zheng, Xing; Liang, Xichao; Su, Xiaojuan; Zhang, Yu; Lu, Ang; Zhang, Lina

    2015-11-09

    Novel nanocomposite hydrogels composed of polyelectrolytes alginate and chitin whiskers with biocompatibility were successfully fabricated based on the pH-induced charge shifting behavior of chitin whiskers. The chitin whiskers with mean length and width of 300 and 20 nm were uniformly dispersed in negatively charged sodium alginate aqueous solution, leading to the formation of the homogeneous nanocomposite hydrogels. The experimental results indicated that their mechanical properties were significantly improved compared to alginate hydrogel and the swelling trends were inhibited as a result of the strong electrostatic interactions between the chitin whiskers and alginate. The nanocomposite hydrogels exhibited certain crystallinity and hierarchical structure with nanoscale chitin whiskers, similar to the structure of the native extracellular matrix. Moreover, the nanocomposite hydrogels were successfully applied as bone scaffolds for MC3T3-E1 osteoblast cells, showing their excellent biocompatibility and low cytotoxicity. The results of fluorescent micrographs and scanning electronic microscope (SEM) images revealed that the addition of chitin whiskers into the nanocomposite hydrogels markedly promoted the cell adhesion and proliferation of the osteoblast cells. The biocompatible nanocomposite hydrogels have potential application in bone tissue engineering.

  1. Carboxymethyl starch/alginate microspheres containing diamine oxidase for intestinal targeting

    PubMed Central

    Blemur, Lindsay; Le, Tien Canh; Marcocci, Lucia; Pietrangeli, Paola

    2015-01-01

    Abstract The association of carboxymethyl starch (CMS) and alginate is proposed as a novel matrix for the entrapment of bioactive agents in microspheres affording their protection against gastrointestinal degradation. In this study, the enzyme diamine oxidase (DAO) from white pea (Lathyrus sativus) was immobilized by inclusion in microspheres formed by ionotropic gelation of CMS/alginate by complexation with Ca2+. The association of CMS to alginate generated a more compact structure presenting a lesser porosity, thus decreasing the access of gastric fluid inside the microspheres and preventing the loss of entrapped enzyme. Moreover, the immobilized enzyme remained active and was able to oxidize the polyamine substrates even in the presence of degrading proteases of pancreatin. The inclusion yield in terms of entrapped protein was of about 82%–95%. The DAO entrapped in calcium CMS/alginate beads retained up to 70% of its initial activity in simulated gastric fluid (pH 2.0). In simulated intestinal fluid (pH 7.2) with pancreatin, an overall retention of 65% of activity for the immobilized DAO was observed over 24 H, whereas in similar conditions the free enzyme was totally inactivated. Our project proposes the vegetal DAO as an antihistaminic agent orally administered to treat food histaminosis and colon inflammation. PMID:25779356

  2. Dental mesenchymal stem cells encapsulated in alginate hydrogel co-delivery microencapsulation system for cartilage regeneration

    PubMed Central

    Moshaverinia, Alireza; Xu, Xingtian; Chen, Chider; Akiyama, Kentaro; Snead, Malcolm L; Shi, Songtao

    2013-01-01

    Dental-derived MSCs are promising candidates for cartilage regeneration, with high chondrogenic differentiation capacity. This property contributes to making dental MSCs an advantageous therapeutic option compared to current treatment modalities. The MSC delivery vehicle is the principal determinant for the success of MSC-mediated cartilage regeneration therapies. The objectives of this study were to: (1) develop a novel co-delivery system based on TGF-β1 loaded RGD-coupled alginate microspheres encapsulating Periodontal Ligament Stem Cells (PDLSCs) or Gingival Mesenchymal Stem Cells (GMSCs); and (2) investigate dental MSC viability and chondrogenic differentiation in alginate microspheres. The results revealed the sustained release of TGF-β1 from the alginate microspheres. After 4 weeks of chondrogenic differentiation in vitro, PDLSCs, GMSCs as well as human bone marrow mesenchymal stem cells (hBMMSC) (as positive control) revealed chondrogenic gene expression markers (Col II and Sox-9) via qPCR, as well as matrix positively stained by toluidine blue and safranin-O. In animal studies, ectopic cartilage tissue regeneration was observed inside and around the transplanted microspheres, confirmed by histochemical and immunofluorescent staining. Interestingly, PDLSCs showed more chondrogenesis than GMSCs and hBMMSCs (P<0.05). Taken together, these results suggest that RGD-modified alginate microencapsulating dental MSCs make a promising candidate for cartilage regeneration. Our results highlight the vital role played by the microenvironment, as well as value of presenting inductive signals for viability and differentiation of MSCs. PMID:23891740

  3. Electrophoretic deposition of antibiotic loaded PHBV microsphere-alginate composite coating with controlled delivery potential.

    PubMed

    Chen, Qiang; Li, Wei; Goudouri, Ourania-Menti; Ding, Yaping; Cabanas-Polo, Sandra; Boccaccini, Aldo R

    2015-06-01

    Electrophoretic deposition (EPD) technique has been developed for the fabrication of antibiotic-loaded PHBV microsphere (MS)-alginate antibacterial coatings. The composite coatings deposited from suspensions with different MS concentrations were produced in order to demonstrate the versatility of the proposed method for achieving functional coatings with tailored drug loading and release profiles. Linearly increased deposit mass with increasing MS concentrations was obtained, and MS were found to be homogeneously stabilized in the alginate matrix. Chemical composition, surface roughness and wettability of the deposited coatings were measured by Fourier transform infrared (FTIR) spectroscopy, laser profilometer and water contact angle instruments, respectively. The co-deposition mechanism was described by two separate processes according to the results of relevant measurements: (i) the deposition of alginate-adsorbed MS and (ii) the non-adsorbed alginate. Qualitative antibacterial tests indicated that MS containing coatings exhibit excellent inhibition effects against E. coli (gram-negative bacteria) after 1h of incubation. The proposed coating system combined with the simplicity of the EPD technique can be considered a promising surface modification approach for the controlled in situ delivery of drug or other biomolecules.

  4. Carboxymethyl starch/alginate microspheres containing diamine oxidase for intestinal targeting.

    PubMed

    Blemur, Lindsay; Le, Tien Canh; Marcocci, Lucia; Pietrangeli, Paola; Mateescu, Mircea Alexandru

    2016-05-01

    The association of carboxymethyl starch (CMS) and alginate is proposed as a novel matrix for the entrapment of bioactive agents in microspheres affording their protection against gastrointestinal degradation. In this study, the enzyme diamine oxidase (DAO) from white pea (Lathyrus sativus) was immobilized by inclusion in microspheres formed by ionotropic gelation of CMS/alginate by complexation with Ca(2+) . The association of CMS to alginate generated a more compact structure presenting a lesser porosity, thus decreasing the access of gastric fluid inside the microspheres and preventing the loss of entrapped enzyme. Moreover, the immobilized enzyme remained active and was able to oxidize the polyamine substrates even in the presence of degrading proteases of pancreatin. The inclusion yield in terms of entrapped protein was of about 82%-95%. The DAO entrapped in calcium CMS/alginate beads retained up to 70% of its initial activity in simulated gastric fluid (pH 2.0). In simulated intestinal fluid (pH 7.2) with pancreatin, an overall retention of 65% of activity for the immobilized DAO was observed over 24 H, whereas in similar conditions the free enzyme was totally inactivated. Our project proposes the vegetal DAO as an antihistaminic agent orally administered to treat food histaminosis and colon inflammation.

  5. Ni{sub 0.5}Zn{sub 0.5}Fe{sub 2}O{sub 4} nanoparticles dispersed in a SiO{sub 2} matrix synthesized by sol-gel processing

    SciTech Connect

    Pozo Lopez, G.; Condo, A.M.; Urreta, S.E.; Silvetti, S.P.; Aguirre, M. del C.

    2012-12-15

    (Ni{sub 0.5}Zn{sub 0.5}Fe{sub 2}O{sub 4})x/(SiO{sub 2})(100 - x) (x = 5, 20 and 50 wt.%) nanocomposites are synthesized by a sol-gel method using tetraethylorthosilicate (TEOS) and metallic nitrates as precursors, and by further annealing the powders for 1 h at 1273 K. X-ray diffraction (XRD), transmission electron microscopy (TEM), room temperature vibrating sample magnetometry (VSM) and SQUID measurements are employed for structural, morphological and magnetic sample characterization. For all the concentrations analyzed, the powder nanocomposites actually consist of spinel NiZn ferrite nanoparticles, dispersed in an amorphous silica matrix. TEM studies reveal different particle size distributions and particle morphologies for the three ferrite contents. The 20 wt.%-NiZn ferrite samples consist of nearly spherical nanoparticles, of about 8 nm, mainly superparamagnetic, well-dispersed in the amorphous silica matrix, while the 5 wt.%-NiZn ferrite samples exhibit a bimodal particle size distribution (5 and 30 nm) of single-domain nanoparticles embedded in the silica. In the 50 wt.%-NiZn ferrite samples, two particle families are observed: small round superparamagnetic nanoparticles of about 8 nm embedded in the amorphous silica matrix and large, non-spherical, ferrimagnetic ones, forming agglomerates outside the matrix. In all the synthesized samples, thickness fringes are observed inside some of the ferrite nanoparticles in dark field images. This contrast is explained using the theory of electron diffraction in a weak beam dark field (WBDF) condition and considering spherical ferrite nanoparticles. A large range of tailored magnetic properties varying the fraction, dispersion and mean size of the ferrimagnetic NiZn ferrite particles is obtained. Room temperature saturation magnetization values are found in the range 3.0-30.4 Am{sup 2}/kg for the different concentration samples. Coercivity values, between 1.9 and 7.6 mT, are more than 50% higher than those measured

  6. Alginate increases water stability whilst maintaining diet digestibility in farmed saltwater crocodiles ().

    PubMed

    Francis, Magdalene; Morel, Patrick C H; Wilkinson, Brian H P; Wester, Timothy J

    2017-02-01

    Saltwater crocodile () farming in Papua New Guinea is an emerging industry that supplies high-quality skins to the fashion industry. Crocodiles are semiaquatic and fed high-quality feed made from extrudated animal byproducts (i.e., forced through a die at low pressure but not heat treated); however, it disintegrates on contact with water, and this leads to low utilization. Alginate is used extensively in food and pharmaceutical processes because it quickly forms a gel at room temperature; however, its effects on nutrient availability are equivocal, and its utility in crocodile diets is unknown. Extrudated chicken byproduct-based crocodile diets were formulated (as-fed) with and without 1.7 and 3.3% Na alginate with either CaCl or CaCO to cross-link. After immersion in water at 30°C for 24 h, feed retained on a 0.5-mm screen was measured to determine DM retention (DMR). Regardless of inclusion level, alginate addition resulted in a 13-fold increase in DMR ( < 0.05) when CaCO was used as a Ca source; however, CaCl use resulted in a much lower DMR. In a digestibility trial, 10 juvenile crocodiles (2.2 to 2.4 yr of age; 1.2 to 1.9 kg BW) were chosen from farm-raised stocks and fed extrudated chicken byproduct-based diets with and without 1.5% Na alginate and 1.9% CaCO. Animals fed 2% BW for 12 d and with excreta collected the last 5 d were slaughtered and had digesta sampled from the ileum. There were no differences in apparent ileal digestibilities of any AA, N (65.0 vs. 55.8%, SE = 12.2%), and OM (46.8 vs. 39.6%, SE = 12.8%) between diets with and without alginate, respectively. Total-tract digestibilities of OM (69.8 vs. 39.2%, SE = 9.1%) and energy (72.2 vs. 44.4%, SE = 8.3%), however, were greater in alginate-containing diets ( < 0.05). Our study showed that alginate addition to crocodile feed improved its stability in water and did not impair nutrient digestion. Application of these findings should greatly decrease feed wastage, which ultimately will benefit

  7. Electrophoretic deposition of ZnO/alginate and ZnO-bioactive glass/alginate composite coatings for antimicrobial applications.

    PubMed

    Cordero-Arias, L; Cabanas-Polo, S; Goudouri, O M; Misra, S K; Gilabert, J; Valsami-Jones, E; Sanchez, E; Virtanen, S; Boccaccini, A R

    2015-10-01

    Two organic/inorganic composite coatings based on alginate, as organic matrix, and zinc oxide nanoparticles (n-ZnO) with and without bioactive glass (BG), as inorganic components, intended for biomedical applications, were developed by electrophoretic deposition (EPD). Different n-ZnO (1-10 g/L) and BG (1-1.5 g/L) contents were studied for a fixed alginate concentration (2 g/L). The presence of n-ZnO was confirmed to impart antibacterial properties to the coatings against gram-negative bacteria Escherichia coli, while the BG induced the formation of hydroxyapatite on coating surfaces thereby imparting bioactivity, making the coating suitable for bone replacement applications. Coating composition was analyzed by thermogravimetric analysis (TG), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and energy dispersive X-ray spectroscopy (EDS) analyses. Scanning electron microscopy (SEM) was employed to study both the surface and the cross section morphology of the coatings. Polarization curves of the coated substrates made in cell culture media at 37 °C confirmed the corrosion protection function of the novel organic/inorganic composite coatings.

  8. Sargassum filipendula alginate from Brazil: seasonal influence and characteristics.

    PubMed

    Bertagnolli, Caroline; Espindola, Ana Paula D M; Kleinübing, Sirlei Jaiana; Tasic, Ljubica; da Silva, Meuris Gurgel Carlos

    2014-10-13

    The aim of this work is focused on the extraction and characterization of the Brazilian seaweed Sargassum filipendula alginate. Alginates obtained at different seasons were characterized by liquid state nuclear magnetic resonance spectroscopy and scanning electron microscopy. The alginate extraction efficiency was about 20%. Different seasons of the year and different stages in the life cycle of Sargassum sp. in southeastern Brazil influenced the M/G and, consequently, the technological properties of extracted alginates.

  9. Adsorption and enzyme (beta-galactosidase and alpha-chymotrypsin): immobilization properties of gel fiber prepared by the gel formation of cellulose acetate and titanium iso-propoxide

    PubMed

    Kurokawa; Suzuki; Tamai

    1998-09-05

    We prepared a new composite gel fiber by the gel formation of cellulose acetate and titanium iso-propoxide. The fiber is harder than alginate gel; it is also stable in common solvents, phosphate solution, and electrolyte solutions over a wide range of pH from 3 to 10. The fiber shows amphoretic adsorption properties depending on pH, namely, it acts anionic with decreasing pH and cationic with increasing pH. However, the fiber had no adsorption property for a pyrogen endotoxin. The beta-galactosidase and alpha-chymotrypsin not retained in alginate gel were immobilized on the fibers by this method. The pH, temperature, and repeated run stabilities of the immobilized enzyme were compared to those of the native one. Copyright 1998 John Wiley & Sons, Inc.

  10. Copolymers For Capillary Gel Electrophoresis

    SciTech Connect

    Liu, Changsheng; Li, Qingbo

    2005-08-09

    This invention relates to an electrophoresis separation medium having a gel matrix of at least one random, linear copolymer comprising a primary comonomer and at least one secondary comonomer, wherein the comonomers are randomly distributed along the copolymer chain. The primary comonomer is an acrylamide or an acrylamide derivative that provides the primary physical, chemical, and sieving properties of the gel matrix. The at least one secondary comonomer imparts an inherent physical, chemical, or sieving property to the copolymer chain. The primary and secondary comonomers are present in a ratio sufficient to induce desired properties that optimize electrophoresis performance. The invention also relates to a method of separating a mixture of biological molecules using this gel matrix, a method of preparing the novel electrophoresis separation medium, and a capillary tube filled with the electrophoresis separation medium.

  11. Enhancing cell migration in shape-memory alginate-collagen composite scaffolds: In vitro and ex vivo assessment for intervertebral disc repair.

    PubMed

    Guillaume, Olivier; Naqvi, Syeda Masooma; Lennon, Kerri; Buckley, Conor Timothy

    2015-04-01

    Lower lumbar disc disorders pose a significant problem in an aging society with substantial socioeconomic consequences. Both inner tissue (nucleus pulposus) and outer tissue (annulus fibrosus) of the intervertebral disc are affected by such debilitating disorders and can lead to disc herniation and lower back pain. In this study, we developed an alginate-collagen composite porous scaffold with shape-memory properties to fill defects occurring in annulus fibrosus tissue of degenerated intervertebral discs, which has the potential to be administered using minimal invasive surgery. In the first part of this work, we assessed how collagen incorporation on preformed alginate scaffolds influences the physical properties of the final composite scaffold. We also evaluated the ability of annulus fibrosus cells to attach, migrate, and proliferate on the composite alginate-collagen scaffolds compared to control scaffolds (alginate only). In vitro experiments, performed in intervertebral disc-like microenvironmental conditions (low glucose and low oxygen concentrations), revealed that for alginate only scaffolds, annulus fibrosus cells agglomerated in clusters with limited infiltration and migration capacity. In comparison, for alginate-collagen scaffolds, annulus fibrosus cells readily attached and colonized constructs, while preserving their typical fibroblastic-like cell morphology with spreading behavior and intense cytoskeleton expression. In a second part of this study, we investigated the effects of alginate-collagen scaffold when seeded with bone marrow derived mesenchymal stem cells. In vitro, we observed that alginate-collagen porous scaffolds supported cell proliferation and extracellular matrix deposition (collagen type I), with secretion amplified by the local release of transforming growth factor-β3. In addition, when cultured in ex vivo organ defect model, alginate-collagen scaffolds maintained viability of transplanted mesenchymal stem cells for up to 5

  12. 21 CFR 172.858 - Propylene glycol alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Propylene glycol alginate. 172.858 Section 172.858... Propylene glycol alginate. The food additive propylene glycol alginate (CAS Reg. No. 9005-37-2) may be used... the act: (1) The name of the additive, “propylene glycol alginate” or “propylene glycol ester...

  13. Oral sustained delivery of paracetamol from in situ-gelling gellan and sodium alginate formulations.

    PubMed

    Kubo, Wataru; Miyazaki, Shozo; Attwood, David

    2003-06-04

    The purpose of this study was to evaluate the potential for the oral sustained delivery of paracetamol of two formulations with in situ gelling properties. Oral administration of aqueous solutions of either gellan gum (1.0%, w/v) or sodium alginate (1.5%, w/v) containing calcium ions in complexed form resulted in the formation of gel depots in rabbit and rat stomachs as a consequence of the release of the calcium ions in the acidic environment. In vitro studies demonstrated diffusion-controlled release of paracetamol from the gels over a period of 6h. The bioavailability of paracetamol from the gels formed in situ in the stomachs of rabbits following oral administration of the liquid formulations was similar to that of a commercially available suspension containing an identical dose of paracetamol.

  14. Living bacteria in silica gels

    NASA Astrophysics Data System (ADS)

    Nassif, Nadine; Bouvet, Odile; Noelle Rager, Marie; Roux, Cécile; Coradin, Thibaud; Livage, Jacques

    2002-09-01

    The encapsulation of enzymes within silica gels has been extensively studied during the past decade for the design of biosensors and bioreactors. Yeast spores and bacteria have also been recently immobilized within silica gels where they retain their enzymatic activity, but the problem of the long-term viability of whole cells in an inorganic matrix has never been fully addressed. It is a real challenge for the development of sol-gel processes. Generic tests have been performed to check the viability of Escherichia coli bacteria in silica gels. Surprisingly, more bacteria remain culturable in the gel than in an aqueous suspension. The metabolic activity of the bacteria towards glycolysis decreases slowly, but half of the bacteria are still viable after one month. When confined within a mineral environment, bacteria do not form colonies. The exchange of chemical signals between isolated bacteria rather than aggregates can then be studied, a point that could be very important for 'quorum sensing'.

  15. Biodegradable alginate microspheres as a delivery system for naked DNA.

    PubMed Central

    Aggarwal, N; HogenEsch, H; Guo, P; North, A; Suckow, M; Mittal, S K

    1999-01-01

    Sodium alginate is a naturally occurring polysaccharide that can easily be polymerized into a solid matrix to form microspheres. These biodegradable microspheres were used to encapsulate plasmid DNA containing the bacterial beta-galactosidase (LacZ) gene under the control of either the cytomegalovirus (CMV) immediate-early promoter or the Rous sarcoma virus (RSV) early promoter. Mice inoculated orally with microspheres containing plasmid DNA expressed LacZ in the intestine, spleen and liver. Inoculation of mice with microspheres containing both the plasmid DNA and bovine adenovirus type 3 (BAd3) resulted in a significant increase in LacZ expression compared to those inoculated with microspheres containing only the plasmid DNA. Our results suggest that adenoviruses are capable of augumenting transgene expression by plasmid DNA both in vitro and in vivo. Images Figure 3. PMID:10369574

  16. Alginate lyases from alginate-degrading Vibrio splendidus 12B01 are endolytic.

    PubMed

    Badur, Ahmet H; Jagtap, Sujit Sadashiv; Yalamanchili, Geethika; Lee, Jung-Kul; Zhao, Huimin; Rao, Christopher V

    2015-03-01

    Alginate lyases are enzymes that degrade alginate through β-elimination of the glycosidic bond into smaller oligomers. We investigated the alginate lyases from Vibrio splendidus 12B01, a marine bacterioplankton species that can grow on alginate as its sole carbon source. We identified, purified, and characterized four polysaccharide lyase family 7 alginates lyases, AlyA, AlyB, AlyD, and AlyE, from V. splendidus 12B01. The four lyases were found to have optimal activity between pH 7.5 and 8.5 and at 20 to 25°C, consistent with their use in a marine environment. AlyA, AlyB, AlyD, and AlyE were found to exhibit a turnover number (kcat) for alginate of 0.60 ± 0.02 s(-1), 3.7 ± 0.3 s(-1), 4.5 ± 0.5 s(-1), and 7.1 ± 0.2 s(-1), respectively. The Km values of AlyA, AlyB, AlyD, and AlyE toward alginate were 36 ± 7 μM, 22 ± 5 μM, 60 ± 2 μM, and 123 ± 6 μM, respectively. AlyA and AlyB were found principally to cleave the β-1,4 bonds between β-d-mannuronate and α-l-guluronate and subunits; AlyD and AlyE were found to principally cleave the α-1,4 bonds involving α-l-guluronate subunits. The four alginate lyases degrade alginate into longer chains of oligomers.

  17. Alginate Lyases from Alginate-Degrading Vibrio splendidus 12B01 Are Endolytic

    PubMed Central

    Badur, Ahmet H.; Jagtap, Sujit Sadashiv; Yalamanchili, Geethika; Lee, Jung-Kul; Zhao, Huimin

    2015-01-01

    Alginate lyases are enzymes that degrade alginate through β-elimination of the glycosidic bond into smaller oligomers. We investigated the alginate lyases from Vibrio splendidus 12B01, a marine bacterioplankton species that can grow on alginate as its sole carbon source. We identified, purified, and characterized four polysaccharide lyase family 7 alginates lyases, AlyA, AlyB, AlyD, and AlyE, from V. splendidus 12B01. The four lyases were found to have optimal activity between pH 7.5 and 8.5 and at 20 to 25°C, consistent with their use in a marine environment. AlyA, AlyB, AlyD, and AlyE were found to exhibit a turnover number (kcat) for alginate of 0.60 ± 0.02 s−1, 3.7 ± 0.3 s−1, 4.5 ± 0.5 s−1, and 7.1 ± 0.2 s−1, respectively. The Km values of AlyA, AlyB, AlyD, and AlyE toward alginate were 36 ± 7 μM, 22 ± 5 μM, 60 ± 2 μM, and 123 ± 6 μM, respectively. AlyA and AlyB were found principally to cleave the β-1,4 bonds between β-d-mannuronate and α-l-guluronate and subunits; AlyD and AlyE were found to principally cleave the α-1,4 bonds involving α-l-guluronate subunits. The four alginate lyases degrade alginate into longer chains of oligomers. PMID:25556193

  18. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry analysis.

    PubMed

    Hu, S; Qiu, N; Liu, Y; Zhao, H; Gao, D; Song, R; Ma, M

    2016-05-01

    A proteomic study of egg white proteins from 2 major poultry species, namely quail (Coturnix coturnix) and duck (Anas platyrhynchos), was performed with comparison to those of chicken (Gallus gallus) through 2-dimensional polyacrylamide gel electrophoresis (2-DE) analysis. By using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS), 29 protein spots representing 10 different kinds of proteins as well as 17 protein spots designating 9 proteins were successfully identified in quail and duck egg white, respectively. This report suggested a closer relationship between quail and chicken egg white proteome patterns, whereas the duck egg white protein distribution on the 2-DE map was more distinct. In duck egg white, some well-known major proteins, such as ovomucoid, clusterin, extracellular fatty acid-binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), were not detected, while two major protein spots identified as "deleted in malignant brain tumors 1" protein (DMBT1) and vitellogenin-2 were found specific to duck in the corresponding range on the 2-DE gel map. These interspecies diversities may be associated with the egg white protein functions in cell defense or regulating/supporting the embryonic development to adapt to the inhabiting environment or reproduction demand during long-term evolution. The findings of this work will give insight into the advantages involved in the application on egg white proteins from various egg sources, which may present novel beneficial properties in the food industry or related to human health.

  19. Role of Calcium Alginate and Mannitol in Protecting Bifidobacterium

    PubMed Central

    Dianawati, Dianawati; Mishra, Vijay

    2012-01-01

    Fourier transform infrared (FTIR) spectroscopy was carried out to ascertain the mechanism of Ca-alginate and mannitol protection of cell envelope components and secondary proteins of Bifidobacterium animalis subsp. lactis Bb12 after freeze-drying and after 10 weeks of storage at room temperature (25°C) at low water activities (aw) of 0.07, 0.1, and 0.2. Preparation of Ca-alginate and Ca-alginate-mannitol as microencapsulants was carried out by dropping an alginate or alginate-mannitol emulsion containing bacteria using a burette into CaCl2 solution to obtain Ca-alginate beads and Ca-alginate-mannitol beads, respectively. The wet beads were then freeze-dried. The aw of freeze-dried beads was then adjusted to 0.07, 0.1, and 0.2 using saturated salt solutions; controls were prepared by keeping Ca-alginate and Ca-alginate-mannitol in aluminum foil without aw adjustment. Mannitol in the Ca-alginate system interacted with cell envelopes during freeze-drying and during storage at low aws. In contrast, Ca-alginate protected cell envelopes after freeze-drying but not during 10-week storage. Unlike Ca-alginate, Ca-alginate-mannitol was effective in retarding the changes in secondary proteins during freeze-drying and during 10 weeks of storage at low aws. It appears that Ca-alginate-mannitol is more effective than Ca-alginate in preserving cell envelopes and proteins after freeze-drying and after 10 weeks of storage at room temperature (25°C). PMID:22843535

  20. A three-dimensional culture system using alginate hydrogel prolongs hatched cattle embryo development in vitro.

    PubMed

    Zhao, Shuan; Liu, Zhen-Xing; Gao, Hui; Wu, Yi; Fang, Yuan; Wu, Shuai-Shuai; Li, Ming-Jie; Bai, Jia-Hua; Liu, Yan; Evans, Alexander; Zeng, Shen-Ming

    2015-07-15

    No successful method exists to maintain the three-dimensional architecture of hatched embryos in vitro. Alginate, a linear polysaccharide derived from brown algae, has characteristics that make it an ideal material as a three-dimensional (3D) extracellular matrix for in vitro cell, tissue, or embryo culture. In this study, alginate hydrogel was used for IVC of posthatched bovine embryos to observe their development under the 3D system. In vitro-fertilized and parthenogenetically activated posthatched bovine blastocysts were cultured in an alginate encapsulation culture system (AECS), an alginate overlay culture system (AOCS), or control culture system. After 18 days of culture, the survival rate of embryos cultured in AECS was higher than that in the control group (P < 0.05), and the embryos were expanded and elongated in AECS with the maximal length of 1.125 mm. When the AECS shrinking embryos were taken out of the alginate beads on Day 18 and cultured in the normal culture system, 9.09% of them attached to the bottoms of the plastic wells and grew rapidly, with the largest area of an attached embryo being 66.00 mm(2) on Day 32. The embryos cultured in AOCS developed monovesicular or multivesicular morphologies. Total cell number of the embryos cultured in AECS on Day 19 was significantly higher than that of embryos on Day 8. Additionally, AECS and AOCS supported differentiation of the embryonic cells. Binuclear cells were visible in Day-26 adherent embryos, and the messenger RNA expression patterns of Cdx2 and Oct4 in AOCS-cultured embryos were similar to those in vivo embryos, whereas IFNT and ISG15 messenger RNA were still expressed in Day-26 and Day-32 prolong-cultured embryos. In conclusion, AECS and AOCS did support cell proliferation, elongation, and differentiation of hatched bovine embryos during prolonged IVC. The culture system will be useful to further investigate the molecular mechanisms controlling ruminant embryo elongation and implantation.

  1. New insights into Pseudomonas fluorescens alginate biosynthesis relevant for the establishment of an efficient production process for microbial alginates.

    PubMed

    Maleki, Susan; Mærk, Mali; Hrudikova, Radka; Valla, Svein; Ertesvåg, Helga

    2017-07-25

    Alginate denotes a family of linear polysaccharides with a wide range of industrial and pharmaceutical applications. Presently, all commercially available alginates are manufactured from brown algae. However, bacterial alginates have advantages with regard to compositional homogeneity and reproducibility. In order to be able to design bacterial strains that are better suited for industrial alginate production, defining limiting factors for alginate biosynthesis is of vital importance. Our group has been studying alginate biosynthesis in Pseudomonas fluorescens using several complementary approaches. Alginate is synthesised and transported out of the cell by a multiprotein complex spanning from the inner to the outer membrane. We have developed an immunogold labelling procedure in which the porin AlgE, as a part of this alginate factory, could be detected by transmission electron microscopy. No time-dependent correlation between the number of such factories on the cell surface and alginate production level was found in alginate-producing strains. Alginate biosynthesis competes with the central carbon metabolism for the key metabolite fructose 6-phosphate. In P. fluorescens, glucose, fructose and glycerol, are metabolised via the Entner-Doudoroff and pentose phosphate pathways. Mutational analysis revealed that disruption of the glucose 6-phosphate dehydrogenase gene zwf-1 resulted in increased alginate production when glycerol was used as carbon source. Furthermore, alginate-producing P. fluorescens strains cultivated on glucose experience acid stress due to the simultaneous production of alginate and gluconate. The combined results from our studies strongly indicate that the availability of fructose 6-phosphate and energy requires more attention in further research aimed at the development of an optimised alginate production process.

  2. Production and characterization of engineered alginate-based microparticles containing ECM powder for cell/tissue engineering applications.

    PubMed

    Mazzitelli, Stefania; Luca, Giovanni; Mancuso, Francesca; Calvitti, Mario; Calafiore, Riccardo; Nastruzzi, Claudio; Johnson, Scott; Badylak, Stephen F

    2011-03-01

    A method for the production of engineered alginate-based microparticles, containing extracellular matrix and neonatal porcine Sertoli cells (SCs), is described. As a source for extracellular matrix, a powder form of isolated and purified urinary bladder matrix (UBM) was employed. We demonstrated that the incorporation of UBM does not significantly alter the morphological and dimensional characteristics of the microparticles. The alginate microparticles were used for SC encapsulation as an immunoprotective barrier for transplant purposes, while the co-entrapped UBM promoted retention of cell viability and function. These engineered microparticles could represent a novel approach to enhancing immunological acceptance and increasing the functional life-span of the entrapped cells for cell/tissue engineering applications. In this respect, it is noteworthy that isolated neonatal porcine SCs, administered alone in highly biocompatible microparticles, led to diabetes prevention and reversion in nonobese diabetic (NOD) mice.

  3. Influence of calcination temperature on structural and magnetic properties of nanocomposites formed by Co-ferrite dispersed in sol-gel silica matrix using tetrakis(2-hydroxyethyl) orthosilicate as precursor

    PubMed Central

    2011-01-01

    Effects of calcination temperatures varying from 400 to 1000°C on structural and magnetic properties of nanocomposites formed by Co-ferrite dispersed in the sol-gel silica matrix using tetrakis(2-hydroxyethyl) orthosilicate (THEOS) as water-soluble silica precursor have been investigated. Studies carried out using XRD, FT-IR, TEM, STA (TG-DTG-DTA) and VSM techniques. Results indicated that magnetic properties of samples such as superparamagnetism and ferromagnetism showed great dependence on the variation of the crystallinity and particle size caused by the calcination temperature. The crystallization, saturation magnetization Ms and remenant magnetization Mr increased as the calcination temperature increased. But the variation of coercivity Hc was not in accordance with that of Ms and Mr, indicating that Hc is not determined only by the crystallinity and size of CoFe2O4 nanoparticles. TEM images showed spherical nanoparticles dispersed in the silica network with sizes of 10-30 nm. Results showed that the well-established silica network provided nucleation locations for CoFe2O4 nanoparticles to confinement the coarsening and aggregation of nanoparticles. THEOS as silica matrix network provides an ideal nucleation environment to disperse CoFe2O4 nanoparticles and thus to confine them to aggregate and coarsen. By using THEOS as water-soluble silica precursor over the currently used TEOS and TMOS, the organic solvents are not needed owing to the complete solubility of THEOS in water. Synthesized nanocomposites with adjustable particle sizes and controllable magnetic properties make the applicability of Co-ferrite even more versatile. PMID:21486494

  4. Highly sensitive analysis of nucleic acids using capillary gel electrophoresis with ultraviolet detection based on the combination of matrix field-amplified and head-column field-amplified stacking injection.

    PubMed

    Lian, Dong-Sheng; Zhao, Shu-Jin

    2015-01-26

    To develop a highly sensitive method for analyzing nucleic acids using capillary gel electrophoresis with ultraviolet detection (CGE-UV), we combined matrix field-amplified with head-column field-amplified stacking injection (C-FASI) to employ the advantages of two methods. Without diminishing the resolution, a limit of detection of 0.13 ng/ml (signal/noise=3) in a 300,000-fold diluted sample was obtained, the sensitivity is 102,308 times higher than that achieved with normal pressure injection, 3077 times that with normal electrokinetic injection, 154 times that with pressure field-amplified sample stacking injection, and 31 times that with matrix field-amplified stacking injection. After establishing the method, we tested the detection of a φX174-Hae III digest DNA product without purification and with a high ionic strength. At the lowest dilution of 5000-fold, sample at a concentration of 10 ng/ml was enriched and detected. The relative standard deviations for migration time and peak area (n=3) were 0.03-1.15 and 0.72-6.42, respectively. To further validate C-FASI was applicable for real sample, a 400 bp PCR product without purification was directly detected with a limit of detection at the concentration of 6000-fold dilution (signal/noise=3), The relative standard deviations for migration time and peak area (n=6) were 0.44 and 4.8, respectively. These results indicated that C-FASI had good qualitative and quantitative detection abilities and CGE-UV based on C-FASI is easy to perform, practical, highly-sensitive and robust for nucleic acid detection, which makes it a highly valuable tool for genetic diagnostics based on nucleic acid analysis.

  5. Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers

    NASA Astrophysics Data System (ADS)

    Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

    2016-10-01

    Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5–100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed.

  6. Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers.

    PubMed

    Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

    2016-10-03

    Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5-100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed.

  7. Diffusion Retardation by Binding of Tobramycin in an Alginate Biofilm Model

    PubMed Central

    Cao, Bao; Christophersen, Lars; Jensen, Peter Østrup; Sneppen, Kim; Høiby, Niels; Moser, Claus

    2016-01-01

    Microbial cells embedded in a self-produced extracellular biofilm matrix cause chronic infections, e. g. by Pseudomonas aeruginosa in the lungs of cystic fibrosis patients. The antibiotic killing of bacteria in biofilms is generally known to be reduced by 100–1000 times relative to planktonic bacteria. This makes such infections difficult to treat. We have therefore proposed that biofilms can be regarded as an independent compartment with distinct pharmacokinetics. To elucidate this pharmacokinetics we have measured the penetration of the tobramycin into seaweed alginate beads which serve as a model of the extracellular polysaccharide matrix in P. aeruginosa biofilm. We find that, rather than a normal first order saturation curve, the concentration of tobramycin in the alginate beads follows a power-law as a function of the external concentration. Further, the tobramycin is observed to be uniformly distributed throughout the volume of the alginate bead. The power-law appears to be a consequence of binding to a multitude of different binding sites. In a diffusion model these results are shown to produce pronounced retardation of the penetration of tobramycin into the biofilm. This filtering of the free tobramycin concentration inside biofilm beads is expected to aid in augmenting the survival probability of bacteria residing in the biofilm. PMID:27100887

  8. Electrosprayed Multi-Core Alginate Microcapsules as Novel Self-Healing Containers

    PubMed Central

    Hia, Iee Lee; Pasbakhsh, Pooria; Chan, Eng-Seng; Chai, Siang-Piao

    2016-01-01

    Alginate microcapsules containing epoxy resin were developed through electrospraying method and embedded into epoxy matrix to produce a capsule-based self-healing composite system. These formaldehyde free alginate/epoxy microcapsules were characterized via light microscope, field emission scanning electron microscope, fourier transform infrared spectroscopy and thermogravimetric analysis. Results showed that epoxy resin was successfully encapsulated within alginate matrix to form porous (multi-core) microcapsules with pore size ranged from 5–100 μm. The microcapsules had an average size of 320 ± 20 μm with decomposition temperature at 220 °C. The loading capacity of these capsules was estimated to be 79%. Under in situ healing test, impact specimens showed healing efficiency as high as 86% and the ability to heal up to 3 times due to the multi-core capsule structure and the high impact energy test that triggered the released of epoxy especially in the second and third healings. TDCB specimens showed one-time healing only with the highest healing efficiency of 76%. The single healing event was attributed by the constant crack propagation rate of TDCB fracture test. For the first time, a cost effective, environmentally benign and sustainable capsule-based self-healing system with multiple healing capabilities and high healing performance was developed. PMID:27694922

  9. Development and evaluation of in situ gel of pregabalin

    PubMed Central

    Madan, Jyotsana R; Adokar, Bhushan R; Dua, Kamal

    2015-01-01

    Aim and Background: Pregabalin (PRG), an analog of gamma-aminobutyric acid, reduces the release of many neurotransmitters, including glutamate, and noradrenaline. It is used for the treatment of epilepsy; simple and complex partial convulsion. The present research work aims to ensure a high drug absorption by retarding the advancement of PRG formulation through the gastrointestinal tract. The work aims to design a controlled release PRG formulation which is administered as liquid and further gels in the stomach and floats in gastric juice. Materials and Methods: In situ gelling formulations were prepared using sodium alginate, calcium chloride, sodium citrate, hydroxypropyl methylcellulose (HPMC) K100M, and sodium bicarbonate. The prepared formulations were evaluated for solution viscosity, drug content, in vitro gelling studies, gel strength, and in vitro drug release. The final formulation was optimized using a 32 full factorial design. Results: The formulation containing 2.5% w/v sodium alginate and 0.2% w/v calcium chloride were considered optimum since it showed minimum floating lag time (18 s), optimum viscosity (287.3 cps), and gel strength (4087.17 dyne/cm2). The optimized formulation follows Korsmeyer-Peppas kinetic model with n value 0.3767 representing Fickian diffusion mechanism of drug release. Conclusion: Floating in situ gelling system of PRG can be formulated using sodium alginate as a gelling polymer and calcium chloride as a complexing agent to control the drug release for about 12 h for the treatment of epilepsy. PMID:26682193

  10. Alginate Particles as Platform for Drug Delivery by the Oral Route: State-of-the-Art

    PubMed Central

    2014-01-01

    Pharmaceutical research and development aims to design products with ensured safety, quality, and efficacy to treat disease. To make the process more rational, coherent, efficient, and cost-effective, the field of Pharmaceutical Materials Science has emerged as the systematic study of the physicochemical properties and behavior of materials of pharmaceutical interest in relation to product performance. The oral route is the most patient preferred for drug administration. The presence of a mucus layer that covers the entire gastrointestinal tract has been exploited to expand the use of the oral route by developing a mucoadhesive drug delivery system that showed a prolonged residence time. Alginic acid and sodium and potassium alginates have emerged as one of the most extensively explored mucoadhesive biomaterials owing to very good cytocompatibility and biocompatibility, biodegradation, sol-gel transition properties, and chemical versatility that make possible further modifications to tailor their properties. The present review overviews the most relevant applications of alginate microparticles and nanoparticles for drug administration by the oral route and discusses the perspectives of this biomaterial in the future. PMID:25101184

  11. Acid-labile formylation of amino terminal proline of human immunodeficiency virus type 1 p24(gag) was found by proteomics using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

    PubMed

    Fuchigami, Takashi; Misumi, Shogo; Takamune, Nobutoki; Takahashi, Ichiro; Takama, Michiho; Shoji, Shozo

    2002-05-10

    HIV-1(LAV-1) particles were collected by ultracentrifugation, treated with subtilisin, and then purified by Sepharose CL-4B column chromatography to remove microvesicles. The lysate of the purified human immunodeficiency virus type 1 (HIV-1) particles was subjected to two-dimensional (2D) gel electrophoresis and stained, and the stained spots were excised and digested with trypsin. The resulting peptide fragments were characterized by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Twenty-five proteins were identified as the proteins inside the virion and the acid-labile formyl group of an amino terminal proline residue of HIV-1(LAV-1) p24(gag) was determined by MALDI-TOF MS before and after weak-acid treatments (0.6 N hydrochloric acid) and confirmed by post-source decay (PSD) of the N-formylated N-terminal tryptic peptide (N-formylated Pro(1)-Arg(18)). The role of formylation has been unclear so far, but it is surmised that the acid-labile formylation of HIV-1(LAV-1) p24(gag) may play a critical role in the formation of the HIV-1 core for conferring HIV-1 infectivity.

  12. The Effect of Autologous Platelet-Rich Gel on the Dynamic Changes of the Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 Expression in the Diabetic Chronic Refractory Cutaneous Ulcers.

    PubMed

    Li, Lan; Chen, Dawei; Wang, Chun; Liu, Guanjian; Ran, Xingwu

    2015-01-01

    Aim. To investigate the dynamic changes on the expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) in the diabetic chronic refractory cutaneous ulcers after the autologous platelet-rich gel (APG) treatment. Methods. The study was developed at the Diabetic Foot Care Centre, West China Hospital. The granulation tissues from the target wounds were taken before and within 15 days after APG application. The expression of MMP-2 and TIMP-2 as well as transforming growth factor-β1 (TGF-β1) in the granulation tissue was detected by q TR-PCR and IHC. The relationship between the expression level of MMP-2 and TIMP-2 and their ratio and that of TGF-β1 was analyzed. Results. The expression of MMP-2 (P < 0.05) was suppressed, and the expression of TIMP-2 (P < 0.05) was promoted, while the ratio of MMP-2/TIMP-2 (P < 0.05) was decreased after APG treatments. The expression of TGF-β1 had negative correlation with the ratio of MMP-2/TIMP-2 (P < 0.05) and positive correlation with the expression of TIMP-2 (P < 0.05). Conclusions. APG treatment may suppress the expression of MMP-2, promoting that of the TIMP-2 in the diabetic chronic refractory cutaneous wounds. TGF-β1 may be related to these effects.

  13. Effect of nutrients on alginate synthesis in Azotobacter vinelandii and characterization of the produced alginate.

    PubMed

    Sabry, S A; Ghanem, K M; Sabra, W A

    1996-12-01

    The role of nutrients on alginate production by Azotobacter vinelandii was studied in batch cultures. The largest amount of bacterial alginate was obtained in presence of: 0.3 g/l MgSO4.7H2O. 0.4 g/l NaCl, 42 mg/l CaCl2.2H2O,.4 mg/l KH2PO4, 16 mg/l K2HPO4, 2.5 mg/l FeSO4.7H2O, 2.9 mg/l H3BO3, 2 mg/l ZnSO4.7H2O, 2 mg/l Na2MoO4.2H2O, 0.3 mg/l CuSO4.5H2O, 0.2 mg/l MnCl2.4H2O. Alginate production was not enhanced by natural additives or inducing agents, except for acetate, which increased alginate yield. The pure alginate contained 0.36% ash and 0.4% protein. It is similar to algal alginate, but it has an extra acetyl group. It contains 69.5% M-M block, 27.5% M-G block and 3% G-G block.

  14. 21 CFR 184.1187 - Calcium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... use in food (as served) (percent) Functional use Baked goods, § 170.3(n)(1) of this chapter 0.002... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Calcium alginate. 184.1187 Section 184.1187 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD...

  15. 21 CFR 184.1133 - Ammonium alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections, frostings... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Ammonium alginate. 184.1133 Section 184.1133 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD...

  16. 21 CFR 184.1133 - Ammonium alginate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... prepared by the neutralization of purified alginic acid with appropriate pH control agents. (b) The ingredient meets the specifications of the Food Chemicals Codex, 3d Ed. (1981), p. 18, which is incorporated... food Maximum level of use in food (as served) (percent) Functional use Confections, frostings, §...

  17. 21 CFR 184.1187 - Calcium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...-35-0) is the calcium salt of alginic acid, a natural polyuronide constituent of certain brown algae... this chapter 0.6 Do. Fats and oils, § 170.3(n)(12) of this chapter 0.5 Do. Gelatins, puddings, §...

  18. 21 CFR 184.1133 - Ammonium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...-34-9) is the ammonium salt of alginic acid, a natural polyuronide constituent of certain brown algae..., § 170.3(n)(9) of this chapter 0.4 Stabilizer, thickener, § 170.3(o)(28) of this chapter. Fats and...

  19. 21 CFR 184.1187 - Calcium alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...-35-0) is the calcium salt of alginic acid, a natural polyuronide constituent of certain brown algae... this chapter 0.6 Do. Fats and oils, § 170.3(n)(12) of this chapter 0.5 Do. Gelatins, puddings, §...

  20. 21 CFR 184.1133 - Ammonium alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...-34-9) is the ammonium salt of alginic acid, a natural polyuronide constituent of certain brown algae..., § 170.3(n)(9) of this chapter 0.4 Stabilizer, thickener, § 170.3(o)(28) of this chapter. Fats and...

  1. 21 CFR 184.1133 - Ammonium alginate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections, frostings... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium alginate. 184.1133 Section 184.1133 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD...

  2. Ultra-structural changes and expression of chondrogenic and hypertrophic genes during chondrogenic differentiation of mesenchymal stromal cells in alginate beads

    PubMed Central

    Dashtdar, Havva; Selvaratnam, Lakshmi; Balaji Raghavendran, Hanumantharao; Suhaeb, Abdulrazzaq Mahmod; Ahmad, Tunku Sara

    2016-01-01

    Chondrogenic differentiation of mesenchymal stromal cells (MSCs) in the form of pellet culture and encapsulation in alginate beads has been widely used as conventional model for in vitro chondrogenesis. However, comparative characterization between differentiation, hypertrophic markers, cell adhesion molecule and ultrastructural changes during alginate and pellet culture has not been described. Hence, the present study was conducted comparing MSCs cultured in pellet and alginate beads with monolayer culture. qPCR was performed to assess the expression of chondrogenic, hypertrophic, and cell adhesion molecule genes, whereas transmission electron microscopy (TEM) was used to assess the ultrastructural changes. In addition, immunocytochemistry for Collagen type II and aggrecan and glycosaminoglycan (GAG) analysis were performed. Our results indicate that pellet and alginate bead cultures were necessary for chondrogenic differentiation of MSC. It also indicates that cultures using alginate bead demonstrated significantly higher (p < 0.05) chondrogenic but lower hypertrophic (p < 0.05) gene expressions as compared with pellet cultures. N-cadherin and N-CAM1 expression were up-regulated in second and third weeks of culture and were comparable between the alginate bead and pellet culture groups, respectively. TEM images demonstrated ultrastructural changes resembling cell death in pellet cultures. Our results indicate that using alginate beads, MSCs express higher chondrogenic but lower hypertrophic gene expression. Enhanced production of extracellular matrix and cell adhesion molecules was also observed in this group. These findings suggest that alginate bead culture may serve as a superior chondrogenic model, whereas pellet culture is more appropriate as a hypertrophic model of chondrogenesis. PMID:26966647

  3. Injectable hydrogels embedded with alginate microspheres for controlled delivery of bone morphogenetic protein-2.

    PubMed

    Zhu, Youjia; Wang, Jiulong; Wu, Jingjing; Zhang, Jun; Wan, Ying; Wu, Hua

    2016-03-23

    Some delivery carriers with injectable characteristics were built using the thermosensitive chitosan/dextran-polylactide/glycerophosphate hydrogel and selected alginate microspheres for the controllable release of bone morphogenetic protein-2 (BMP-2). BMP-2 was first loaded into the microspheres with an average size of around 20 μm and the resulting microspheres were then embedded into the gel in order to achieve well-controlled BMP-2 release. The microsphere-embedded gels show their incipient gelation temperature at around 32 °C and pH at about 7.1. Some gels had their elastic modulus close to 1400 Pa and the ratio of elastic modulus to viscous modulus at around 34, revealing that they behaved like mechanically strong gels. Optimized microsphere-embedded gels were found to be able to administer the BMP-2 release without significant initial burst release in an approximately linear manner over a period of time longer than four weeks. The release rate and the released amount of BMP-2 from these gels could be regulated individually or cooperatively by the initial BMP-2 load and the dextran-polylactide content in the gels. Measurements of the BMP-2 induced alkaline phosphatase activity in C2C12 cells confirmed that C2C12 cells responded to BMP-2 in a dose-dependent way and the released BMP-2 from the microsphere-embedded gels well retained their bioactivity. In vivo assessment of some gels revealed that the released BMP-2 maintained its osteogenesis functions.

  4. Inhibitory effects of laminaran and alginate on production of putrefactive compounds from soy protein by intestinal microbiota in vitro and in rats.

    PubMed

    Nakata, Toru; Kyoui, Daisuke; Takahashi, Hajime; Kimura, Bon; Kuda, Takashi

    2016-06-05

    Soybean is one of the major components of the Japanese diet. In traditional Japanese cuisine, soybean-based food items are often consumed with brown algae. In this study, we examined the effect of water-soluble and fermentable polysaccharides, laminaran and sodium alginate, from brown algae, on putrefactive compound production, by human faecal microbiota in broth containing 3% (w/v) soy protein. We also investigated the effect of 2% laminaran or alginate diet on caecal putrefactive compounds in rats maintained on diets containing 20% (w/w) soy protein. The caecal microbiota was also analysed using denaturing gradient gel electrophoresis and pyrosequencing with primers targeting the bacterial 16S rRNA gene. The polysaccharides, particularly laminaran, inhibited ammonia, phenol, and indole production by human faecal microbiota. Both the algal polysaccharides lowered the caecal indole content. Laminaran was found to increase the number of Coprobacter, whereas Helicobacter was found to decrease in the presence of both laminaran and sodium alginate.

  5. Adsorption of CO2 by alginate immobilized zeolite beads

    NASA Astrophysics Data System (ADS)

    Suratman, A.; Kunarti, E. S.; Aprilita, N. H.; Pamurtya, I. C.

    2017-03-01

    Immobilized zeolit in alginate beads for adsorption of CO2 was developed. Alginate immobilized zeolit beads was generated by dropping the mixture of Na-alginate and zeolite solution into Ca2+ solution. The adsorption efficacy such as the influence of contact time, mass of zeolite, flowrate of CO2, and mass of adsorbent was evaluated. The adsorption of CO2 onto alginate immobilized zeolit beads was investigated by performing both equilibrium and kinetic batch test. Bead was characterized by FTIR and SEM. Alginate immobilized zeolit beads demonstrated significantly higher sorption efficacy compared to plain alginate beads and zeolite with 0.25 mmol CO2 adsorbed /g adsorbent. Optimum condition was achieved with mass composition of alginate:zeolite (3:1), flowrate 50 mL/min for 20 minutes. The alginate immobilized zeolit beads showed that adsorption of CO2 followed Freundlich isotherm and pseudo second order kinetic model. Adsorption of CO2 onto alginate immobilized zeolite beads is a physisorption with adsorption energy of 6.37 kJ/mol. This results indicates that the alginate immobilized zeolit beads can be used as promising adsorbents for CO2.

  6. The Effect of Chondroitin Sulphate and Hyaluronic Acid on Chondrocytes Cultured within a Fibrin-Alginate Hydrogel.

    PubMed

    Little, Christopher J; Kulyk, William M; Chen, Xiongbiao

    2014-09-18

    Osteoarthritis is a painful degenerative joint disease that could be better managed if tissue engineers can develop methods to create long-term engineered articular cartilage tissue substitutes. Many of the tissue engineered cartilage constructs currently available lack the chemical stimuli and cell-friendly environment that promote the matrix accumulation and cell proliferation needed for use in joint cartilage repair. The goal of this research was to test the efficacy of using a fibrin-alginate hydrogel containing hyaluronic acid (HA) and/or chondroitin sulphate (CS) supplements for chondrocyte culture. Neonatal porcine chondrocytes cultured in fibrin-alginate hydrogels retained their phenotype better than chondrocytes cultured in monolayer, as evidenced by analysis of their relative expression of type II versus type I collagen mRNA transcripts. HA or CS supplementation of the hydrogels increased matrix glycosaminoglycan (GAG) production during the first week of culture. However, the effects of these supplements on matrix accumulation were not additive and were no longer observed after two weeks of culture. Supplementation of the hydrogels with CS or a combination of both CS and HA increased the chondrocyte cell population after two weeks of culture. Statistical analysis indicated that the HA and CS treatment effects on chondrocyte numbers may be additive. This research suggests that supplementation with CS and/or HA has positive effects on cartilage matrix production and chondrocyte proliferation in three-dimensional (3D) fibrin-alginate hydrogels.

  7. Falsirhodobacter sp. alg1 Harbors Single Homologs of Endo and Exo-Type Alginate Lyases Efficient for Alginate Depolymerization

    PubMed Central

    Takahashi, Mami; Tanaka, Reiji; Miyake, Hideo; Shibata, Toshiyuki; Chow, Seinen; Kuroda, Kouichi; Ueda, Mitsuyoshi; Takeyama, Haruko

    2016-01-01

    Alginate-degrading bacteria play an important role in alginate degradation by harboring highly efficient and unique alginolytic genes. Although the general mechanism for alginate degradation by these bacteria is fairly understood, much is still required to fully exploit them. Here, we report the isolation of a novel strain, Falsirhodobacter sp. alg1, the first report for an alginate-degrading bacterium from the family Rhodobacteraceae. Genome sequencing reveals that strain alg1 harbors a primary alginate degradation pathway with only single homologs of an endo- and exo-type alginate lyase, AlyFRA and AlyFRB, which is uncommon among such bacteria. Subsequent functional analysis showed that both enzymes were extremely efficient to depolymerize alginate suggesting evolutionary interests in the acquirement of these enzymes. The exo-type alginate lyase, AlyFRB in particular could depolymerize alginate without producing intermediate products making it a highly efficient enzyme for the production of 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH). Based on our findings, we believe that the discovery of Falsirhodobacter sp. alg1 and its alginolytic genes hints at the potentiality of a more diverse and unique population of alginate-degrading bacteria. PMID:27176711

  8. Aerosol gels

    NASA Technical Reports Server (NTRS)

    Sorensen, Christopher M. (Inventor); Chakrabarti, Amitabha (Inventor); Dhaubhadel, Rajan (Inventor); Gerving, Corey (Inventor)

    2010-01-01

    An improved process for the production of ultralow density, high specific surface area gel products is provided which comprises providing, in an enclosed chamber, a mixture made up of small particles of material suspended in gas; the particles are then caused to aggregate in the chamber to form ramified fractal aggregate gels. The particles should have a radius (a) of up to about 50 nm and the aerosol should have a volume fraction (f.sub.v) of at least 10.sup.-4. In preferred practice, the mixture is created by a spark-induced explosion of a precursor material (e.g., a hydrocarbon) and oxygen within the chamber. New compositions of matter are disclosed having densities below 3.0 mg/cc.

  9. Antibacterial performance of alginic acid coating on polyethylene film.

    PubMed

    Karbassi, Elika; Asadinezhad, Ahmad; Lehocký, Marian; Humpolíček, Petr; Vesel, Alenka; Novák, Igor; Sáha, Petr

    2014-08-21

    Alginic acid coated polyethylene films were examined in terms of surface properties and bacteriostatic performance against two most representative bacterial strains, that is, Escherichia coli and Staphylococcus aureus. Microwave plasma treatment followed by brush formation in vapor state from three distinguished precursors (allylalcohol, allylamine, hydroxyethyl methacrylate) was carried out to deposit alginic acid on the substrate. Surface analyses via various techniques established that alginic acid was immobilized onto the surface where grafting (brush) chemistry influenced the amount of alginic acid coated. Moreover, alginic acid was found to be capable of bacterial growth inhibition which itself was significantly affected by the brush type. The polyanionic character of alginic acid as a carbohydrate polymer was assumed to play the pivotal role in antibacterial activity. The cell wall composition of two bacterial strains along with the substrates physicochemical properties accounted for different levels of bacteriostatic performance.

  10. A novel alternative to cryopreservation for the short-term storage of stem cells for use in cell therapy using alginate encapsulation.

    PubMed

    Chen, Bo; Wright, Bernice; Sahoo, Rashmita; Connon, Che J

    2013-07-01

    Efficient transport of stem/progenitor cells without affecting their survival and function is a key factor in any practical cell-based therapy. However, the current approach using liquid nitrogen for the transfer of stem cells requires a short delivery time window is technically challenging and financially expensive. The present study aims to use semipermeable alginate hydrogels (crosslinked by strontium) to encapsulate, store, and release stem cells, to replace the conventional cryopreservation method for the transport of therapeutic cells within world-wide distribution time frame. Human mesenchymal stem cell (hMSC) and mouse embryonic stem cells (mESCs) were successfully stored inside alginate hydrogels for 5 days under ambient conditions in an air-tight environment (sealed cryovial). Cell viability, of the cells extracted from alginate gel, gave 74% (mESC) and 80% (hMSC) survival rates, which compared favorably to cryopreservation. More importantly, the subsequent proliferation rate and detection of common stem cell markers (both in mRNA and protein level) from hMSCs and mESCs retrieved from alginate hydrogels were also comparable to (if not better than) results gained following cryopreservation. In conclusion, this new and simple application of alginate hydrogel encapsulation may offer a cheap and robust alternative to cryopreservation for the transport and storage of stem cells for both clinical and research purposes.

  11. Synthesis of photoresponsive hybrid alginate hydrogel with photo-controlled release behavior.

    PubMed

    Chiang, Chien-Ying; Chu, Chih-Chien

    2015-03-30

    A photoresponsive hybrid alginate hydrogel was successfully prepared by Ca(2+)-mediated crosslinking reaction with a mixture of β-cyclodextrin-grafted alginate (β-CD-Alg) and diazobenzene-modified poly(ethylene glycol) (Az2-PEG). The water-soluble Az2-PEG exhibits efficient trans-to-cis isomerization of the terminal azobenzene moieties under UV-light irradiation and readily switched back to the initial trans state under visible light. Because of low affinity between β-CD and cis-Az, the host-guest inclusion complex formed by β-CD and trans-Az gradually dissociates under UV-light exposure. Accordingly, the bulk gel exhibits substantial photo-induced transformation in gel morphology by the appearance of significant comb-like cavities. This photosensitive behavior accompanied by the structural degradation enables the rapid release of entrapped dye molecules under UV light stimulus. Moreover, an incident light with higher power and mild acidic environment are capable of accelerating the photo-triggered release, thus allowing the potential applications toward acute wound healing.

  12. Network generation enhances interpretation of proteomics data sets by a combination of two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    PubMed

    Wang, Xijun; Zhang, Aihua; Sun, Hui; Wu, Gelin; Sun, Wenjun; Yan, Guangli

    2012-10-21

    Recent advances in proteomic technologies have enabled us to create detailed protein-protein interaction maps in diseases. As the size of the interaction dataset increases, powerful computational methods are required in order to effectively interpret network models from large scale interactome data. In this study, we carried out comparative proteomics to construct and identify the proteins networks associated with hepatic injury (HI) which are largely unknown, as a case study. All proteins expressed were separated and identified by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight-time-of-flight mass spectrometry (MALDI-TOF/TOF MS). Protein-interacting networks and pathways were mapped using STRING analysis program. We have performed for the first time a comprehensive profiling of changes in protein expression of HI rats, to uncover the networks altered by treated with CCl(4). Identification of fifteen spots (seven over-expressed and eight under-expressed) were established by MALDI-TOF/TOF MS. These proteins were subjected to functional pathway analysis using STRING software for better understanding of the biological context of the identified proteins. It suggested that modulation of multiple vital physiological pathways including DNA repair process, cell apoptosis, oxidation reduction, signal transduction, metabolic process, intracellular signaling cascade, regulation of biological processes, cell communication, regulation of cellular process, and molecular transport. In summary, the present study provides the first protein-interacting network maps and novel insights into the biological responses and potential pathways of HI. The generation of protein interaction networks clearly enhances the interpretation of proteomic data, particularly in respect of understanding molecular mechanisms of panel protein biomarkers.

  13. Covalently antibacterial alginate-chitosan hydrogel dressing integrated gelatin microspheres containing tetracycline hydrochloride for wound healing.

    PubMed

    Chen, Huinan; Xing, Xiaodong; Tan, Huaping; Jia, Yang; Zhou, Tianle; Chen, Yong; Ling, Zhonghua; Hu, Xiaohong

    2017-01-01

    An antibacterial and biodegradable composite hydrogel dressing integrated with microspheres is developed for drug delivery and wound healing. The mechanism of gelation is attributed to the Schiff-base reaction between aldehyde and amino groups of oxidized alginate (OAlg) and carboxymethyl chitosan (CMCS). To enhance antibacterial and mechanical properties, tetracycline hydrochloride (TH) loaded gelatin microspheres (GMs) were fabricated by an emulsion cross-linking method, followed by integrating into the OAlg-CMCS hydrogel to produce a composite gel dressing. In vitro gelation time, swelling, degradation, compressive modulus and rheological properties of the gel dressing were investigated as the function of microsphere ratios. With increasing ratios of microspheres from 10 to 40mg/mL, the composite dressing manifested shorter gelation time and lower swelling ratios, as well as higher mechanical strength. Comparing to other formulations, the gel dressing with 30mg/mL microspheres showed more suitable stabilities and mechanical properties for wound healing. Also, in vitro drug release results showed that the loaded TH could be sustained release from the composite gel dressing by contrast with pure hydrogels and microspheres. Furthermore, powerful bacteria growth inhibition effects against Escherichia coli and Staphylococcus aureus suggested that the composite gel dressing, especially the one with 30mg/mL GMs containing TH, has a promising future in treatment of bacterial infection.

  14. All-natural composite wound dressing films of essential oils encapsulated in sodium alginate with antimicrobial properties.

    PubMed

    Liakos, Ioannis; Rizzello, Loris; Scurr, David J; Pompa, Pier Paolo; Bayer, Ilker S; Athanassiou, Athanassia

    2014-03-25

    We present natural polymeric composite films made of essential oils (EOs) dispersed in sodium alginate (NaAlg) matrix, with remarkable anti-microbial and anti-fungal properties. Namely, elicriso italic, chamomile blue, cinnamon, lavender, tea tree, peppermint, eucalyptus, lemongrass and lemon oils were encapsulated in the films as potential active substances. Glycerol was used to induce plasticity and surfactants were added to improve the dispersion of EOs in the NaAlg matrix. The topography, chemical composition, mechanical properties, and humidity resistance of the films are presented analytically. Antimicrobial tests were conducted on films containing different percentages of EOs against Escherichia coli bacteria and Candida albicans fungi, and the films were characterized as effective or not. Such diverse types of essential oil-fortified alginate films can find many applications mainly as disposable wound dressings but also in food packaging, medical device protection and disinfection, and indoor air quality improvement materials, to name a few.

  15. Detoxification of Hg(II) from aqueous and enzyme media: Pristine vs. tailored calcium alginate hydrogels.

    PubMed

    Sarkar, Kangkana; Ansari, Zarina; Sen, Kamalika

    2016-10-01

    Calcium alginate (CA) hydrogels were tailored using phenolic compounds (PC) like, thymol, morin, catechin, hesperidin, during their preparation. The PC incorporated gels show modified surface features as indicated by scanning electron microscopic images (SEM). The rheological studies show that excepting the hesperidin incorporated gels all the other kinds including calcium alginate pristine have similar mechanical strength. The hesperidine incorporated CA gels had the maximum capacity to adsorb Hg. The Freundlich adsorption isotherms show higher values of adsorption capacity for all PC incorporated CA beads than the pristine CA (PCA). The hesperidin incorporated CA gels were found to show the best adsorption condition at neutral pH and an optimum contact time of 2.5h at 25°C. Considering the possibility of ingested Hg detoxification from human alimentary tract, the hesperidin and morin incorporated CA beads were further modified through incorporation of cod liver oil as the digestion time of fat in stomach is higher. In vitro uptake capacities of Hg in pepsin and pancreatin containing enzyme media were studied with hesperidin and morin incorporated beads and their corresponding fat incorporated beads also. In the pepsin medium, there was no uptake by hesperidin and fat-hesperidin incorporated beads, which is possibly due to the higher acidity of the medium. But in pancreatin medium Hg was taken up by both kinds of beads. Morin and morin-fat incorporated beads were efficient to uptake Hg from both the pepsin and pancreatin medium. The tailored CA beads may therefore serve as efficient scaffolds to rescue Hg ingested individuals.

  16. Versatile click alginate hydrogels crosslinked via tetrazine-norbornene chemistry.

    PubMed

    Desai, Rajiv M; Koshy, Sandeep T; Hilderbrand, Scott A; Mooney, David J; Joshi, Neel S

    2015-05-01

    Alginate hydrogels are well-characterized, biologically inert materials that are used in many biomedical applications for the delivery of drugs, proteins, and cells. Unfortunately, canonical covalently crosslinked alginate hydrogels are formed using chemical strategies that can be biologically harmful due to their lack of chemoselectivity. In this work we introduce tetrazine and norbornene groups to alginate polymer chains and subsequently form covalently crosslinked click alginate hydrogels capable of encapsulating cells without damaging them. The rapid, bioorthogonal, and specific click reaction is irreversible and allows for easy incorporation of cells with high post-encapsulation viability. The swelling and mechanical properties of the click alginate hydrogel can be tuned via the total polymer concentration and the stoichiometric ratio of the complementary click functional groups. The click alginate hydrogel can be modified after gelation to display cell adhesion peptides for 2D cell culture using thiol-ene chemistry. Furthermore, click alginate hydrogels are minimally inflammatory, maintain structural integrity over several months, and reject cell infiltration when injected subcutaneously in mice. Click alginate hydrogels combine the numerous benefits of alginate hydrogels with powerful bioorthogonal click chemistry for use in tissue engineering applications involving the stable encapsulation or delivery of cells or bioactive molecules.

  17. PLGA/alginate composite microspheres for hydrophilic protein delivery.

    PubMed

    Zhai, Peng; Chen, X B; Schreyer, David J

    2015-11-01

    Poly(lactic-co-glycolic acid) (PLGA) microspheres and PLGA/alginate composite microspheres were prepared by a novel double emulsion and solvent evaporation technique and loaded with bovine serum albumin (BSA) or rabbit anti-laminin antibody protein. The addition of alginate and the use of a surfactant during microsphere preparation increased the encapsulation efficiency and reduced the initial burst release of hydrophilic BSA. Confocal laser scanning microcopy (CLSM) of BSA-loaded PLGA/alginate composite microspheres showed that PLGA, alginate, and BSA were distributed throughout the depths of microspheres; no core/shell structure was observed. Scanning electron microscopy revealed that PLGA microspheres erode and degrade more quickly than PLGA/alginate composite microspheres. When loaded with anti-laminin antibody, the function of released antibody was well preserved in both PLGA and PLGA/alginate composite microspheres. The biocompatibility of PLGA and PLGA/alginate microspheres were examined using four types of cultured cell lines, representing different tissue types. Cell survival was variably affected by the inclusion of alginate in composite microspheres, possibly due to the sensitivity of different cell types to excess calcium that may be released from the calcium cross-linked alginate.

  18. Graphene oxide increases the viability of C2C12 myoblasts microencapsulated in alginate.

    PubMed

    Ciriza, J; Saenz del Burgo, L; Virumbrales-Muñoz, M; Ochoa, I; Fernandez, L J; Orive, G; Hernandez, R M; Pedraz, J L

    2015-09-30

    Cell microencapsulation represents a great promise for long-term drug delivery, but still several challenges need to be overcome before its translation into the clinic, such as the long term cell survival inside the capsules. On this regard, graphene oxide has shown to promote proliferation of different cell types either in two or three dimensions. Therefore, we planned to combine graphene oxide with the cell microencapsulation technology. We first studied the effect of this material on the stability of the capsules and next we analyzed the biocompatibility of this chemical compound with erythropoietin secreting C2C12 myoblasts within the microcapsule matrix. We produced 160 μm-diameter alginate microcapsules with increasing concentrations of graphene oxide and did not find modifications on the physicochemical parameters of traditional alginate microcapsules. Moreover, we observed that the viability of encapsulated cells within alginate microcapsules containing specific graphene oxide concentrations was enhanced. These results provide a relevant step for the future clinical application of graphene oxide on cell microencapsulation.

  19. Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation.

    PubMed

    Hoesli, Corinne A; Raghuram, Kamini; Kiang, Roger L J; Mocinecová, Dušana; Hu, Xiaoke; Johnson, James D; Lacík, Igor; Kieffer, Timothy J; Piret, James M

    2011-02-01

    Alginate has been used to protect transplanted pancreatic islets from immune rejection and as a matrix to increase the insulin content of islet progenitor cells. The throughput of alginate bead generation by the standard extrusion and external gelation method is limited by the rate of droplet formation from nozzles. Alginate bead generation by emulsion and internal gelation is a scaleable alternative that has been used with biological molecules and microbial cells, but not mammalian cells. We describe the novel adaptation of this process to mammalian cell immobilization. After optimization, the emulsion process yielded 90 ± 2% mouse insulinoma 6 (MIN6) cell survival, similar to the extrusion process. The MIN6 cells expanded at the same rate in both bead types to form pseudo-islets with increased glucose stimulation index compared to cells in suspension. The emulsion process was suitable for primary pancreatic exocrine cell immobilization, leading to 67 ± 32 fold increased insulin expression after 10 days of immobilized culture. Due to the scaleability and broad availability of stirred mixers, the emulsion process represents an attractive option for laboratories that are not equipped with extrusion-based cell encapsulators, as well as for the production of immobilized or encapsulated cellular therapeutics on a clinical scale.

  20. Development and characterization of novel alginate-based hydrogels as vehicles for bone substitutes.

    PubMed

    Morais, D S; Rodrigues, M A; Silva, T I; Lopes, M A; Santos, M; Santos, J D; Botelho, C M

    2013-06-05

    In this work three different hydrogels were developed to associate, as vehicles, with the synthetic bone substitute GR-HAP. One based on an alginate matrix (Alg); a second on a mixture of alginate and chitosan (Alg/Ch); and a third on alginate and hyaluronate (Alg/HA), using Ca(2+) ions as cross-linking agents. The hydrogels, as well as the respective injectable bone substitutes (IBSs), were fully characterized from the physical-chemical point of view. Weight change studies proved that all hydrogels were able to swell and degrade within 72 h at pH 7.4 and 4.0, being Alg/HA the hydrogel with the highest degradation rate (80%). Rheology studies demonstrated that all hydrogels are non-Newtonian viscoelastic fluids, and injectability tests showed that IBSs presented low maximum extrusion forces, as well as quite stable average forces. In conclusion, the studied hydrogels present the necessary features to be successfully used as vehicles of GR-HAP, particularly the hydrogel Alg/HA.

  1. Ultrasoft Alginate Hydrogels Support Long-Term Three-Dimensional Functional Neuronal Networks.

    PubMed

    Palazzolo, Gemma; Broguiere, Nicolas; Cenciarelli, Orlando; Dermutz, Harald; Zenobi-Wong, Marcy

    2015-08-01

    Neuron development and function are exquisitely sensitive to the mechanical properties of their surroundings. Three-dimensional (3D) cultures are therefore being explored as they better mimic the features of the native extracellular matrix. Limitations of existing 3D culture models include poorly defined composition, rapid degradation, and suboptimal biocompatibility. Here we show that ionically cross-linked ultrasoft hydrogels made from unmodified alginate can potently promote neuritogenesis. Alginate hydrogels were characterized mechanically and a remarkable range of stiffness (10-4000 Pa) could be produced by varying the macromer content (0.1-0.4% w/v) and CaCl2 concentration. Dissociated rat embryonic cortical neurons encapsulated within the softest of the hydrogels (0.1% w/v, 10 mM CaCl2) showed excellent viability, extensive formation of axons and dendrites, and long-term activity as determined by calcium imaging. In conclusion, alginate is an off-the-shelf, easy to handle, and inexpensive material, which can be used to make ultrasoft hydrogels for the formation of stable and functional 3D neuronal networks. This 3D culture system could have important applications in neuropharmacology, toxicology, and regenerative medicine.

  2. Effects of surface functionalized graphene oxide on the behavior of sodium alginate.

    PubMed

    Nie, Ling; Liu, Changhua; Wang, Jia; Shuai, Ying; Cui, Xiaoyan; Liu, Li

    2015-03-06

    The aim of this study was to improve the miscibility between fillers and polymer through modifying the face of graphene oxide (GO). In order to compare the effects of GO and modified graphene oxide (MGO) to sodium alginate (SA), sodium alginate/graphene oxide (SA/GO-n) and sodium alginate/modified graphene oxide (SA/MGO-n) biocomposite films were prepared, then the interaction between nanofillers and matrix was evaluated. The structure, morphologies and properties of biocomposites were characterized by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), thermal gravimetric analysis (TGA), Ultraviolet-visible (UV-vis), scanning electron microscopy (SEM) and mechanical tests. The results revealed that strong interactions existed between GO (MGO) and SA. Compared with neat SA film, the maximum level of Young's moduli (E), tensile strength (σ(b)) and elongation at break (ɛ(b)) of the SA/MGO biocomposites improved by 37.8%, 68.4% and 44.9%, while that of the SA/GO biocomposites improved by 19.6%, 44% and 36.5%.

  3. Electrophoresis of DNA in agarose gels, polyacrylamide gels and in free solution.

    PubMed

    Stellwagen, Nancy C

    2009-06-01

    This review describes the electrophoresis of curved and normal DNA molecules in agarose gels, polyacrylamide gels and in free solution. These studies were undertaken to clarify why curved DNA molecules migrate anomalously slowly in polyacrylamide gels but not in agarose gels. Two milestone papers are cited, in which Ferguson plots were used to estimate the effective pore size of agarose and polyacrylamide gels. Subsequent studies on the effect of the electric field on agarose and polyacrylamide gel matrices, DNA interactions with the two gel matrices, and the effect of curvature on the free solution mobility of DNA are also described. The combined results suggest that the anomalously slow mobilities observed for curved DNA molecules in polyacrylamide gels are primarily due to preferential interactions of curved DNAs with the polyacrylamide gel matrix; the restrictive pore size of the matrix is of lesser importance. In free solution, DNA mobilities increase with increasing molecular mass until leveling off at a plateau value of (3.17 +/- 0.01) x 10(-4) cm2/V s in 40 mM Tris-acetate-EDTA buffer at 20 degrees C. Curved DNA molecules migrate anomalously slowly in free solution as well as in polyacrylamide gels, explaining why the Ferguson plots of curved and normal DNAs containing the same number of base pairs extrapolate to different mobilities at zero gel concentration.

  4. Stability testing of alginate-chitosan films.

    PubMed

    Rabisková, Miloslava; Dvorácková, Katerina; Kofronvá, Lenka

    2012-02-01

    Pellets containing rutin prepared by the extrusion/spheronization method were coated with sodium alginate-chitosan film. Important quality parameters in the pellets before coating were determined, and after coating the dissolution profiles of the drug were evaluated in dissolution media of the pH corresponding to the conditions in the gastrointestinal tract. Samples of coated pellets were located in the boxes for stability testing under different conditions, i.e. 25 degrees C and 60% of relative humidity (RH); 30 degrees C and 65% RH and 40 degrees C and 75% RH. After 1, 3, 6, 9 and 12 months (or 1, 3 and 6 months), the dissolution test was repeated and compared with the original profiles using similarity factors. All similarity factor values above 50 indicate excellent stability of alginate-chitosan films.

  5. [Determination of proximal chemical composition of squid (dosidicus gigas) and development of gel products].

    PubMed

    Abugoch, L; Guarda, A; Pérez, L M; Paredes, M P

    1999-06-01

    The good nutritional properties of meat from big squid (Dosidicus gigas) living on the Chilean coast, was determined through its proximal composition 70 cal/100 g fresh meat; 82.23 +/- 0.98% moisture; 15.32 +/- 0.93% protein; 1.31 +/- 0.12% ashes; 0.87 +/- 0.18% fat and 0.27% NNE (non-nitrogen extract). The big squid meat was used to develop a gel product which contained NaCl and TPP. It was necessary to use additives for gel preparation, such as carragenin or alginate or egg albumin, due to the lack of gelation properties of squid meat. Formulations containing egg albumin showed the highest gel force measured by penetration as compared to those that contained carragenin or alginate.

  6. Biocompatibility of microcapsules for cell immobilization elaborated with different type of alginates.

    PubMed

    Orive, G; Ponce, S; Hernández, R M; Gascón, A R; Igartua, M; Pedraz, J L

    2002-09-01

    The biocompatibility of alginate-PLL-alginate (APA) microcapsules has been evaluated with respect to impurity levels. The impurity content of three different alginates (a raw high M-alginate, a raw high G-alginate and a purified high G-alginate) has been determined and the in vivo antigenic response of APA beads made with each alginate assessed. Results show that purification of the alginate not only reduces the total amount of impurities (63% less in polyphenols, 91.45% less in endotoxins and 68.5% less in protein in relation to raw high M-alginate), but also avoids an antibody response when microcapsules of this material are implanted in mice. In contrast, raw alginates produced a detectable antibody response though the differences in their impurity content. Consequently, this work revealed that purity of the alginate rather than their chemical composition, is probably of greater importance in determining microcapsule biocompatibility.

  7. Three-dimensional electrospun polycaprolactone (PCL)/alginate hybrid composite scaffolds.

    PubMed

    Kim, Min Seong; Kim, GeunHyung

    2014-12-19

    Micro/nanofibrous scaffolds have been used widely in biomedical applications because the micro/nano-scale fibres resemble natural extracellular matrix and the high surface-to-volume ratio encourages cellular activities (attachment and proliferation). However, poor mechanical properties, low controllability of various shapes and difficulties in obtaining controllable pore structure have been obstacles to their use in hard-tissue regeneration. To overcome these shortcomings, we suggest a new composite system, which uses a combination method of wet electrospinning, rapid prototyping and a physical punching process. Using the process, we obtained polycaprolactone (PCL)/alginate composite scaffolds, consisting of electrospun PCL/alginate fibres and micro-sized PCL struts, with mean pore sizes of 821 ± 55 μm. To show the feasibility of the scaffolds for hard-tissue regeneration, the scaffolds were assessed not only for physical properties, including hydrophilicity, water absorption, and tensile and compressive strength, but also in vitro cellular responses (cell viability and proliferation) and osteogenic differentiation (alkaline phosphatase (ALP) activity, and mineralisation) by culturing with pre-osteoblasts (MC3T3-E1 cells). With the reinforcing micro-sized PCL struts, the elastic modulus of the PCL/alginate scaffold was significantly improved versus a pure PCL scaffold. Additionally, due to the alginate component in the fibrous scaffold, they showed significantly enhanced hydrophilic behaviour, water absorption (∼8-fold) and significant biological activities (∼1.6-fold for cell viability at 7 days, ∼2.3-fold for ALP activity at 14 days and ∼6.4-fold for calcium mineralisation at 14 days) compared with those of a pure PCL fibrous scaffold.

  8. Multipotent stromal cells derived from common marmoset Callithrix jacchus within alginate 3D environment: Effect of cryopreservation procedures.

    PubMed

    Gryshkov, Oleksandr; Hofmann, Nicola; Lauterboeck, Lothar; Pogozhykh, Denys; Mueller, Thomas; Glasmacher, Birgit

    2015-08-01

    Multipotent stromal cells derived from the common marmoset monkey Callithrix jacchus (cjMSCs) possess high phylogenetic similarity to humans, with a great potential for preclinical studies in the field of regenerative medicine. Safe and effective long-term storage of cells is of great significance to clinical and research applications. Encapsulation of such cell types within alginate beads that can mimic an extra-cellular matrix and provide a supportive environment for cells during cryopreservation, has several advantages over freezing of cells in suspension. In this study we have analysed the effect of dimethyl sulfoxide (Me2SO, 2.5-10%, v/v) and pre-freeze loading time of alginate encapsulated cjMSCs in Me2SO (0-45 min) on the viability and metabolic activity of the cells after freezing using a slow cooling rate (-1°C/min). It was found that these parameters affect the stability and homogeneity of alginate beads after thawing. Moreover, the cjMSCs can be frozen in alginate beads with lower Me2SO concentration of 7.5% after 30 min of loading, while retaining high cryopreservation outcome. We demonstrated the maximum viability, membrane integrity and metabolic activity of the cells under optimized, less cytotoxic conditions. The results of this study are another step forward towards the application of cryopreservation for the long-term storage and subsequent applications of transplants in cell-based therapies.

  9. Surface detail reproduction and dimensional accuracy of stone models: influence of disinfectant solutions and alginate impression materials.

    PubMed

    Guiraldo, Ricardo Danil; Borsato, Thaís Teixeira; Berger, Sandrine Bittencourt; Lopes, Murilo Baena; Gonini-Jr, Alcides; Sinhoreti, Mário Alexandre Coelho

    2012-01-01

    This study compared the surface detail reproduction and dimensional accuracy of stone models obtained from molds disinfected with 2% sodium hypochlorite, 2% chlorhexidine digluconate or 0.2% peracetic acid to models produced using molds which were not disinfected, with 3 alginate materials (Cavex ColorChange, Hydrogum 5 and Jeltrate Plus). The molds were prepared over matrix containing 20-, 50-, and 75-µm lines, performed under pressure with perforated metal tray. The molds were removed following gelation and either disinfected (using one of the solutions by spraying followed by storage in closed jars for 15 min) or not disinfected. The samples were divided into 12 groups (n=5). Molds were filled with dental gypsum Durone IV and 1 h after the start of the stone mixing the models were separated from the tray. Surface detail reproduction and dimensional accuracy were evaluated using optical microscopy on the 50-µm line with 25 mm in length, in accordance with the ISO 1563 standard. The dimensional accuracy results (%) were subjected to ANOVA. The 50 µm-line was completely reproduced by all alginate impression materials regardless of the disinfection procedure. There was no statistically significant difference in the mean values of dimensional accuracy in combinations between disinfectant procedure and alginate impression material (p=0.2130) or for independent factors. The disinfectant solutions and alginate materials used in this study are no factors of choice regarding the surface detail reproduction and dimensional accuracy of stone models.

  10. Dental mesenchymal stem cells encapsulated in an alginate hydrogel co-delivery microencapsulation system for cartilage regeneration.

    PubMed

    Moshaverinia, Alireza; Xu, Xingtian; Chen, Chider; Akiyama, Kentaro; Snead, Malcolm L; Shi, Songtao

    2013-12-01

    Dental-derived mesenchymal stem cells (MSCs) are promising candidates for cartilage regeneration, with a high capacity for chondrogenic differentiation. This property helps make dental MSCs an advantageous therapeutic option compared to current treatment modalities. The MSC delivery vehicle is the principal determinant for the success of MSC-mediated cartilage regeneration therapies. The objectives of this study were to: (1) develop a novel co-delivery system based on TGF-β1 loaded RGD-coupled alginate microspheres encapsulating periodontal ligament stem cells (PDLSCs) or gingival mesenchymal stem cells (GMSCs); and (2) investigate dental MSC viability and chondrogenic differentiation in alginate microspheres. The results revealed the sustained release of TGF-β1 from the alginate microspheres. After 4 weeks of chondrogenic differentiation in vitro, PDLSCs and GMSCs as well as human bone marrow mesenchymal stem cells (hBMMSCs) (as positive control) revealed chondrogenic gene expression markers (Col II and Sox-9) via qPCR, as well as matrix positively stained by Toluidine Blue and Safranin-O. In animal studies, ectopic cartilage tissue regeneration was observed inside and around the transplanted microspheres, confirmed by histochemical and immunofluorescent staining. Interestingly, PDLSCs showed more chondrogenesis than GMSCs and hBMMSCs (p<0.05). Taken together, these results suggest that RGD-modified alginate microencapsulating dental MSCs make a promising candidate for cartilage regeneration. Our results highlight the vital role played by the microenvironment, as well as value of presenting inductive signals for viability and differentiation of MSCs.

  11. Effect of nutrients on the biodegradation of tributyltin (TBT) by alginate immobilized microalga, Chlorella vulgaris, in natural river water.

    PubMed

    Jin, Jing; Yang, Lihua; Chan, Sidney M N; Luan, Tiangang; Li, Yan; Tam, Nora F Y

    2011-01-30

    The removal and degradation of tributyltin (TBT) by alginate immobilized Chlorella vulgaris has been evidenced in our previously published work. The present study was further to investigate the effect of spiked nutrient concentrations on the TBT removal capacity and degradation in the same alginate immobilized C. vulgaris. During the 14-d experiment, compared to the control (natural river water), the spiked nutrient groups (50% or 100% nutrients of the commercial Bristol medium as the reference, marked as 1/2N or 1N) showed more rapid cell proliferation of microalgae and higher TBT removal rate. Moreover, significantly more TBT was adsorbed onto the alginate matrix, but less TBT was taken up by the algal cells of the nutrient groups than that of the control. Mass balance data showed that TBT was lost as inorganic tin in the highest degree in 1N group, followed by 1/2N group and the least was in the control, but the relative abundance of the intermediate products of debutylation (dibutyltin and monobutyltin) were comparable among three groups. In conclusion, the addition of nutrients in contaminated water stimulated the growth and physiological activity of C. vulgaris immobilized in alginate beads and improved its TBT degradation efficiency.

  12. Production and evaluation of dry alginate-chitosan microcapsules as an enteric delivery vehicle for probiotic bacteria.

    PubMed

    Cook, Michael T; Tzortzis, George; Charalampopoulos, Dimitris; Khutoryanskiy, Vitaliy V

    2011-07-11

    This study investigates the production of alginate microcapsules, which have been coated with the polysaccharide chitosan, and evaluates some of their properties with the intention of improving the gastrointestinal viability of a probiotic ( Bifidobacterium breve ) by encapsulation in this system. The microcapsules were dried by a variety of methods, and the most suitable was chosen. The work described in this Article is the first report detailing the effects of drying on the properties of these microcapsules and the viability of the bacteria within relative to wet microcapsules. The pH range over which chitosan and alginate form polyelectrolyte complexes was explored by spectrophotometry, and this extended into swelling studies on the microcapsules over a range of pHs associated with the gastrointestinal tract. It was shown that chitosan stabilizes the alginate microcapsules at pHs above 3, extending the stability of the capsules under these conditions. The effect of chitosan exposure time on the coating thickness was investigated for the first time by confocal laser scanning microscopy, and its penetration into the alginate matrix was shown to be particularly slow. Coating with chitosan was found to increase the survival of B. breve in simulated gastric fluid as well as prolong its release upon exposure to intestinal pH.

  13. Biopolymer matrix for nano-encapsulation of urease - A model protein and its application in urea detection.

    PubMed

    Saxena, Abhishek; Bhattacharya, Arpita; Kumar, Satish; Epstein, Irving R; Sahney, Rachana

    2017-03-15

    Alginate microparticles and nanoparticles crosslinked with Ca(+2) ions are frequently employed in biomedical applications. Here we use microemulsion polymerization to prepare alginate nanoparticles (nanogels) using different crosslinking ions (Ca(+2), Sr(+2), Ba(+2)) to encapsulate a model protein, urease enzyme (jackbeans). With alginate concentrations of 0.2wt% in the aqueous phase, emulsion droplets showed good stability and narrow, monomodal distributions with radii ∼65±10nm. The size of the nanogel varies with the crosslinking cation and its affinity for the mannuronate and guluronate units in the linear alginate chain. The nanogels were further characterized using dynamic light scattering, scanning electron microscopy, energy dispersive X-ray spectrometry and zeta potential. This work demonstrates the potential application of Ba-alginate as an alternative matrix for nano-encapsulation of proteins and its use for biomedical applications.

  14. Stress-relaxation behavior in gels with ionic and covalent crosslinks.

    PubMed

    Zhao, Xuanhe; Huebsch, Nathaniel; Mooney, David J; Suo, Zhigang

    2010-03-15

    Long-chained polymers in alginate hydrogels can form networks by either ionic or covalent crosslinks. This paper shows that the type of crosslinks can markedly affect the stress-relaxation behavior of the gels. In gels with only ionic crosslinks, stress relaxes mainly through breaking and subsequent reforming of the ionic crosslinks, and the time scale of the relaxation is independent of the size of the sample. By contrast, in gels with only covalent crosslinks, stress relaxes mainly through migration of water, and the relaxation slows down as the size of the sample increases. Implications of these observations are discussed.

  15. Screening of Alginate Lyase-Producing Bacteria and Optimization of Media Compositions for Extracellular Alginate Lyase Production

    PubMed Central

    Tavafi, Hadis; Abdi- Ali, Ahya A; Ghadam, Parinaz; Gharavi, Sara

    2017-01-01

    Background: Alginate is a linear polysaccharide consisting of guluronate (polyG) and mannuronate (polyM) subunits. Methods: In the initial screening of alginate-degrading bacteria from soil, 10 isolates were able to grow on minimal medium containing alginate. The optimization of cell growth and alginate lyase (algL) production was carried out by the addition of 0.8% alginate and 0.2-0.3 M NaCl to the culture medium. Of 10 isolates, one was selected based on its fast growth rate on minimal 9 medium containing 0.4% sodium alginate. The selected bacterium, identified based on morphological and biochemical characteristics, as well as 16S rDNA sequence data, was confirmed to be an isolate belonging to the genus Bacillus and designated as Bacillus sp. TAG8. Results: The results showed the ability of Bacillus sp. TAG8 in utilizing alginate as a sole carbon source. Bacillus sp. TAG8 growth and algL production were augmented with an increase in sodium alginate concentration and also by the addition of 0.2-0.3 M NaCl. Molecular analysis of TAG8 algL gene showed 99% sequence identity with algL of Pseudomonas aeruginosa PAO1. The algL produced by Bacillus sp. TAG8 cleaved both polyM and polyG blocks in alginate molecule, as well as acetylated alginate residues, confirming the bifunctionality of the isolated lyase. Conclusion: The identification of novel algL genes from microbial communities constitutes a new approach for exploring lyases with specific activity against bacterial alginates and may thus contribute to the eradication of persistent biofilms from clinical samples. PMID:27432784

  16. Use of laboratory-grown bacterial alginate in copper removal.

    PubMed

    Kivilcimdan Moral, Ç; Doğan, Ö; Sanin, F D

    2012-01-01

    Industrial production leads to toxic heavy metal pollution in water bodies. Copper is one of the examples that requires removal from effluents before being discharged. It is difficult and sometimes very expensive to remove toxic heavy metals by conventional treatment techniques. This study aims to remove copper by the use of bacterial alginate as a non-conventional technique. Bacterial alginates (natural polymers composed of mannuronic and guluronic acid monomers) were synthesized by Azotobacter vinelandii ATCC(®) 9046 in a laboratory fermentor under controlled environmental conditions. The alginates produced, with a range of different characteristics in terms of monomer distribution and viscosity, were investigated for maximum copper uptake capacities. The average copper uptake capacities of alginates produced were found to be about 1.90 mmol/L Cu(2+)/g alginate. Although the GG-block amount of alginates was varied from 12 to 87% and culture broth viscosities were changed within the range of 1.47 and 14 cP, neither the block distribution nor viscosities of alginate samples considerably affected the copper uptake of alginates.

  17. A Controlled Drug-Delivery Experiment Using Alginate Beads

    ERIC Educational Resources Information Center

    Farrell, Stephanie; Vernengo, Jennifer

    2012-01-01

    This paper describes a simple, cost-effective experiment which introduces students to drug delivery and modeling using alginate beads. Students produce calcium alginate beads loaded with drug and measure the rate of release from the beads for systems having different stir rates, geometries, extents of cross-linking, and drug molecular weight.…

  18. 21 CFR 172.858 - Propylene glycol alginate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Propylene glycol alginate. 172.858 Section 172.858... CONSUMPTION Multipurpose Additives § 172.858 Propylene glycol alginate. The food additive propylene glycol... information required by the act: (1) The name of the additive, “propylene glycol alginate” or...

  19. 21 CFR 172.858 - Propylene glycol alginate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Propylene glycol alginate. 172.858 Section 172.858... CONSUMPTION Multipurpose Additives § 172.858 Propylene glycol alginate. The food additive propylene glycol... information required by the act: (1) The name of the additive, “propylene glycol alginate” or...

  20. 21 CFR 172.858 - Propylene glycol alginate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Propylene glycol alginate. 172.858 Section 172.858... CONSUMPTION Multipurpose Additives § 172.858 Propylene glycol alginate. The food additive propylene glycol... information required by the act: (1) The name of the additive, “propylene glycol alginate” or...

  1. Structurally stable gel bead containing entrapped enzyme and method for manufacture thereof

    DOEpatents

    Woodward, J.

    1998-12-08

    This research provides a structurally stable gel bead containing an entrapped enzyme and a method for its manufacture. The enzyme is covalently cross-linked to gelatin in the presence of glutaraldehyde prior to the formation of the gel bead, to prevent leakage of the enzyme. Propylene glycol alginate is then added to the mixture. Once the gel beads are formed, they are then soaked in glutaraldehyde, which imparts structural stability to the gel beads. This method can be used with many types of enzymes, such as proteases, carbohydrases, proteases, ligases, isomerases, oxidoreductases, and specialty enzymes. These and other enzymes can be immobilized in the gel beads and utilized in a number of enzymatic processes. Exogenously added ions are not required to maintain the structural stability of these gel beads. 7 figs.

  2. Structurally stable gel bead containing entrapped enzyme and method for manufacture thereof

    DOEpatents

    Woodward, Jonathan

    1998-01-01

    A structurally stable gel bead containing an entrapped enzyme and a method for its manufacture. The enzyme is covalently cross-linked to gelatin in the presence of glutaraldehyde prior to the formation of the gel bead, to prevent leakage of the enzyme. Propylene glycol alginate is then added to the mixture. Once the gel beads are formed, they are then soaked in glutaraldehyde, which imparts structural stability to the gel beads. This method can be used with many types of enzymes, such as proteases, carbohydrases, proteases, ligases, isomerases, oxidoreductases, and specialty enzymes. These and other enzymes can be immobilized in the gel beads and utilized in a number of enzymatic processes. Exogenously added ions are not required to maintain the structural stability of these gel beads.

  3. Applications of Alginate-Based Bioinks in 3D Bioprinting

    PubMed Central

    Axpe, Eneko; Oyen, Michelle L.

    2016-01-01

    Three-dimensional (3D) bioprinting is on the cusp of permitting the direct fabrication of artificial living tissue. Multicellular building blocks (bioinks) are dispensed layer by layer and scaled for the target construct. However, only a few materials are able to fulfill the considerable requirements for suitable bioink formulation, a critical component of efficient 3D bioprinting. Alginate, a naturally occurring polysaccharide, is clearly the most commonly employed material in current bioinks. Here, we discuss the benefits and disadvantages of the use of alginate in 3D bioprinting by summarizing the most recent studies that used alginate for printing vascular tissue, bone and cartilage. In addition, other breakthroughs in the use of alginate in bioprinting are discussed, including strategies to improve its structural and degradation characteristics. In this review, we organize the available literature in order to inspire and accelerate novel alginate-based bioink formulations with enhanced properties for future applications in basic research, drug screening and regenerative medicine. PMID:27898010

  4. Maximization of volatile fatty acids production from alginate in acidogenesis.

    PubMed

    Pham, Hong Duc; Seon, Jiyun; Lee, Seong Chan; Song, Minkyung; Woo, Hee-Chul

    2013-11-01

    In this study, the response surface methodology (RSM) was applied to determine the optimum fermentative condition of alginate with the respect to the simultaneous effects of alginate concentration and initial pH to maximize the production of total volatile fatty acids (TVFAs) and alcohols. The results showed that the alginate fermentation was significantly affected by initial pH than by alginate concentration and there was no interaction between the two variables. The optimum condition was 6.2g alginate/L and initial pH 7.6 with a maximum TVFAs yield of 37.1%. Acetic acids were the main constituents of the TVFAs mixtures (i.e., 71.9-95.5%), while alcohols (i.e., ethanol, butanol, and propanol) were not detected.

  5. The influence of polymeric component of bioactive glass-based nanocomposite paste on its rheological behaviors and in vitro responses: hyaluronic acid versus sodium alginate.

    PubMed

    Sohrabi, Mehri; Hesaraki, Saeed; Kazemzadeh, Asghar

    2014-04-01

    Different biocomposite pastes were prepared from a solid phase that was nanoparticles of sol-gel-derived bioactive glass and different liquid phases including 3% hyaluronic acid solution, sodium alginate solutions (3% and 10 %) or mixtures of hyaluronic acid and sodium alginate (3% or 10 %) solutions in 50:50 volume ratio. Rheological properties of the pastes were measured in both rotatory and oscillatory modes. The washout behavior and in vitro apatite formation of the pastes were determined by soaking them in simulated body fluid under dynamic situation for 14 days. The proliferation and alkaline phosphatase activity of MG-63 osteoblastic cells were also determined using extracts of the pastes. All pastes could be easily injected from the standard syringes with different tip diameters. All pastes exhibited visco-elastic character, but a nonthixotropic paste was obtained using hyaluronic acid in which the loss modulus was higher than the storage modulus. The thixotropy and storage modulus were increasingly improved by adding/using sodium alginate as mixing liquid. Moreover, the pastes in which the liquid phase was sodium alginate or mixture of hyaluronic acid and 10% sodium alginate solution revealed better apatite formation ability and washout resistance than that made of hyaluronic acid alone. No cytotoxicity effects were observed by extracts of the pastes on osteoblasts but better alkaline phosphatase activity was found for the pastes containing hyaluronic acid. Overall, injectable biocomposites can be produced by mixing bioactive glass nanoparticles and sodium alginate/hyaluronic acid polymers. They are potentially useful for hard and even soft tissues treatments.

  6. Molecular and biopharmaceutical investigation of alginate-inulin synbiotic co-encapsulation of probiotic to target the colon.

    PubMed

    Atia, Abdelbasset; Gomma, Ahmed I; Fliss, Ismail; Beyssac, Eric; Garrait, Ghiselain; Subirade, Muriel

    2017-03-28

    Colon targeting, as a site-specific delivery for oral formulation, remains a major challenge, especially for sensitive bioactive components such as therapeutic forms of phages, live attenuated virus, and prebiotics-probiotics association. Synbiotics could be used to protect encapsulated probiotics during the gastrointestinal tract and control their release in the colon. To achieve these goals, effective prebiotics such as inulin could be combined with alginate-the most exploited polymer used for probiotic encapsulation-in the form of beads. This work aimed to study the biopharmaceutical behavior of alginate beads (A) and inulin-Alginate beads of different inulin concentrations (5 or 20%) in 2% alginate (AI5, AI20). Beads were loaded with three probiotic strains (Pediococcus acidilactici Ul5, Lactobacillus reuteri and Lactobacillus salivarius). Dissolution of beads was studied by USP4 under conditions simulating the gastrointestinal condition. The survival rates of the bacterial strains were measured by a specific qPCR bacterial count. Mucoadhesiveness of beads was studied by an ex-vivo method using intestinal mucosa. To understand the behavior of each formulation, the ultrastructure of the polymeric network was studied using scanning electron microscopy (SEM). Molecular interactions between alginate and inulin were studied by Fourier transform infrared spectroscopy (FTIR). Dissolution results suggested that the presence of inulin in beads provided more protection for the tested bacterial strains against the acidic pH. AI5 was the most effective formulation to deliver probiotics to the colon simulation conditions. FTIR and SEM investigations explained the differences in behavior of each formula. The developed symbiotic form provided a promising matrix for the development of colonic controlled release systems.

  7. The Influence of Arginine on the Response of Enamel Matrix Derivative (EMD) Proteins to Thermal Stress: Towards Improving the Stability of EMD-Based Products

    PubMed Central

    Bolisetty, Sreenath; Marascio, Matteo; Gemperli Graf, Anja; Garamszegi, Laszlo; Mezzenga, Raffaele; Fischer, Peter; Månson, Jan-Anders

    2015-01-01

    In a current procedure for periodontal tissue regeneration, enamel matrix derivative (EMD), which is the active component, is mixed with a propylene glycol alginate (PGA) gel carrier and applied directly to the periodontal defect. Exposure of EMD to physiological conditions then causes it to precipitate. However, environmental changes during manufacture and storage may result in modifications to the conformation of the EMD proteins, and eventually premature phase separation of the gel and a loss in therapeutic effectiveness. The present work relates to efforts to improve the stability of EMD-based formulations such as Emdogain™ through the incorporation of arginine, a well-known protein stabilizer, but one that to our knowledge has not so far been considered for this purpose. Representative EMD-buffer solutions with and without arginine were analyzed by 3D-dynamic light scattering, UV-Vis spectroscopy, transmission electron microscopy and Fourier transform infrared spectroscopy at different acidic pH and temperatures, T, in order to simulate the effect of pH variations and thermal stress during manufacture and storage. The results provided evidence that arginine may indeed stabilize EMD against irreversible aggregation with respect to variations in pH and T under these conditions. Moreover, stopped-flow transmittance measurements indicated arginine addition not to suppress precipitation of EMD from either the buffers or the PGA gel carrier when the pH was raised to 7, a fundamental requirement for dental applications. PMID:26670810

  8. Dimensional changes of alginate dental impression materials.

    PubMed

    Nallamuthu, N; Braden, M; Patel, M P

    2006-12-01

    The weight loss and corresponding dimensional changes of two dental alginate impression materials have been studied. The weight loss kinetics indicate this to be a diffusion controlled process, but with a boundary condition at the surface of the concentration decreasing exponentially with time. This is in marked contrast to most desorption processes, where the surface concentration becomes instantaneously zero. The appropriate theory has been developed for an exponential boundary condition, and its predictions compared with experimental data; the agreement was satisfactory. The diffusion coefficients for two thicknesses of the same material were not identical as predicted by theory; the possible reasons for this are discussed.

  9. In vivo bioengineered ovarian tumors based on collagen, matrigel, alginate and agarose hydrogels: a comparative study.

    PubMed

    Zheng, Li; Hu, Xuefeng; Huang, Yuanjie; Xu, Guojie; Yang, Jinsong; Li, Li

    2015-01-29

    Scaffold-based tumor engineering is rapidly evolving the study of cancer progression. However, the effects of scaffolds and environment on tumor formation have seldom been investigated. In this study, four types of injectable hydrogels, namely, collagen type I, Matrigel, alginate and agarose gels, were loaded with human ovarian cancer SKOV3 cells and then injected into nude mice subcutaneously. The growth of the tumors in vitro was also investigated. After four weeks, the specimens were harvested and analyzed. We found that tumor formation by SKOV3 cells was best supported by collagen, followed by Matrigel, alginate, control (without scaffold) and agarose in vivo. The collagen I group exhibited a larger tumor volume with increased neovascularization and increased necrosis compared with the other materials. Further, increased MMP activity, upregulated expression of laminin and fibronectin and higher levels of HIF-1α and VEGF-A in the collagen group revealed that the engineered tumor is closer to human ovarian carcinoma. In order, collagen, Matrigel, alginate, control (without scaffold) and agarose exhibited decreases in tumor formation. All evidence indicated that the in vivo engineered tumor is scaffold-dependent. Bioactive hydrogels are superior to inert hydrogels at promoting tumor regeneration. In particular, biomimetic hydrogels are advantageous because they provide a microenvironment that mimics the ECM of natural tumors. On the other hand, typical features of cancer cells and the expression of genes related to cancer malignancy were far less similar to the natural tumor in vitro, which indicated the importance of culture environment in vivo. Superior to the in vitro culture, nude mice can be considered satisfactory in vivo 'bioreactors' for the screening of favorable cell vehicles for tumor engineering in vitro.

  10. Supported Molecular Matrix Electrophoresis.

    PubMed

    Matsuno, Yu-Ki; Kameyama, Akihiko

    2015-01-01

    Mucins are difficult to separate using conventional gel electrophoresis methods such as SDS-PAGE and agarose gel electrophoresis, owing to their large size and heterogeneity. On the other hand, cellulose acetate membrane electrophoresis can separate these molecules, but is not compatible with glycan analysis. Here, we describe a novel membrane electrophoresis technique, termed "supported molecular matrix electrophoresis" (SMME), in which a porous polyvinylidene difluoride (PVDF) membrane filter is used to achieve separation. This description includes the separation, visualization, and glycan analysis of mucins with the SMME technique.

  11. Molecular interaction in alginate beads reinforced with sodium starch glycolate or magnesium aluminum silicate, and their physical characteristics.

    PubMed

    Puttipipatkhachorn, Satit; Pongjanyakul, Thaned; Priprem, Aroonsri

    2005-04-11

    Diclofenac calcium-alginate (DCA) beads were reinforced with different amounts of sodium starch glycolate (SSG) or magnesium aluminum silicate (MAS) and were prepared using ionotropic gelation method. Complex formation of sodium alginate (SA) and SSG or MAS in calcium-alginate beads was revealed using FTIR spectroscopy. Differential scanning calorimetric study indicated that diclofenac sodium (DS) in amorphous form was dispersed in the matrix of DCA beads. The thermal behavior of SSG-DCA and MAS-DCA beads was similar to the control bead. Both additives can improve the entrapment efficiency of DCA beads. The swelling and water uptake of the beads depended on the properties of incorporated additives. The SSG-DCA beads showed a higher water uptake and swelling than MAS-DCA beads. Moreover, the swelling of the beads showed a good correlation with the square root of time. The release kinetic of the beads in pH 6.8 phosphate buffer was swelling controlled mechanism, while that in distilled water followed Higuchi's model. The slower release rate and the longer lag time in pH 6.8 phosphate buffer was obtained from the SSG-DCA and MAS-DCA beads because of complex formation between SA and SSG or MAS. However, SSG in the beads could increase the release of DS from the beads in distilled water because it acted as a channeling agent. In contrast, MAS retarded the release of DS from the beads in distilled water due to the stronger matrix formation.

  12. HAp granules encapsulated oxidized alginate-gelatin-biphasic calcium phosphate hydrogel for bone regeneration.

    PubMed

    Sarker, Avik; Amirian, Jhaleh; Min, Young Ki; Lee, Byong Taek

    2015-11-01

    Bone repair in the critical size defect zone using 3D hydrogel scaffold is still a challenge in tissue engineering field. A novel type of hydrogel scaffold combining ceramic and polymer materials, therefore, was fabricated to meet this challenge. In this study, oxidized alginate-gelatin-biphasic calcium phosphate (OxAlg-Gel-BCP) and spherical hydroxyapatite (HAp) granules encapsulated OxAlg-Gel-BCP hydrogel complex were fabricated using freeze-drying method. Detailed morphological and material characterizations of OxAlg-Gel-BCP hydrogel (OGB00), 25wt% and 35wt% granules encapsulated hydrogel (OGB25 and OGB35) were carried out for micro-structure, porosity, chemical constituents, and compressive stress analysis. Cell viability, cell attachment, proliferation and differentiation behavior of rat bone marrow-derived stem cell (BMSC) on OGB00, OGB25 and OGB35 scaffolds were confirmed by MTT assay, Live-Dead assay, and confocal imaging in vitro experiments. Finally, OGB00 and OGB25 hydrogel scaffolds were implanted in the critical size defect of rabbit femoral chondyle for 4 and 8 weeks. The micro-CT analysis and histological studies conducted by H&E and Masson's trichrome demonstrated that a significantly higher (***p<0.001) and earlier bone formation happened in case of 25% HAp granules encapsulated OxAlg-Gel-BCP hydrogel than in OxAlg-Gel-BCP complex alone. All results taken together, HAp granules encapsulated OxAlg-Gel-BCP system can be a promising 3D hydrogel scaffold for the healing of a critical bone defect.

  13. The effects of bulking, viscous and gel-forming dietary fibres on satiation.

    PubMed

    Wanders, Anne J; Jonathan, Melliana C; van den Borne, Joost J G C; Mars, Monica; Schols, Henk A; Feskens, Edith J M; de Graaf, Cees

    2013-04-14

    The objective was to determine the effects of dietary fibre with bulking, viscous and gel-forming properties on satiation, and to identify the underlying mechanisms. We conducted a randomised crossover study with 121 men and women. Subjects were healthy, non-restrained eaters, aged 18-50 years and with normal BMI (18.5-25 kg/m²). Test products were cookies containing either: no added fibre (control), cellulose (bulking, 5 g/100 g), guar gum (viscous, 1.25 g/100 g and 2.5 g/100 g) or alginate (gel forming, 2.5 g/100 g and 5 g/100 g). Physico-chemical properties of the test products were confirmed in simulated upper gastrointestinal conditions. In a cinema setting, ad libitum intake of the test products was measured concurrently with oral exposure time per cookie by video recording. In a separate study with ten subjects, 4 h gastric emptying rate of a fixed amount of test products was assessed by ¹³C breath tests. Ad libitum energy intake was 22 % lower for the product with 5 g/100 g alginate (3.1 (sd 1.6) MJ) compared to control (4.0 (sd 2.2) MJ, P< 0.001). Intake of the other four products did not differ from control. Oral exposure time for the product with 5 g/100 g alginate (2.3 (sd 1.9) min) was 48 % longer than for control (1.6 (sd 0.9) min, P= 0.01). Gastric emptying of the 5 g/100 g alginate product was faster compared to control (P< 0.05). We concluded that the addition of 5 g/100 g alginate (i.e. gel-forming fibre) to a low-fibre cookie results in earlier satiation. This effect might be due to an increased oral exposure time.

  14. 3D Bioprinting of Heterogeneous Aortic Valve Conduits with Alginate/Gelatin Hydrogels

    PubMed Central

    Duan, Bin; Hockaday, Laura A.; Kang, Kevin H.; Butcher, Jonathan T.

    2013-01-01

    Heart valve disease is a serious and growing public health problem for which prosthetic replacement is most commonly indicated. Current prosthetic devices are inadequate for younger adults and growing children. Tissue engineered living aortic valve conduits have potential for remodeling, regeneration, and growth, but fabricating natural anatomical complexity with cellular heterogeneity remain challenging. In the current study, we implement 3D bioprinting to fabricate living alginate/gelatin hydrogel valve conduits with anatomical architecture and direct incorporation of dual cell types in a regionally constrained manner. Encapsulated aortic root sinus smooth muscle cells (SMC) and aortic valve leaflet interstitial cells (VIC) were viable within alginate/gelatin hydrogel discs over 7 days in culture. Acellular 3D printed hydrogels exhibited reduced modulus, ultimate strength, and peak strain reducing slightly over 7-day culture, while the tensile biomechanics of cell-laden hydrogels were maintained. Aortic valve conduits were successfully bioprinted with direct encapsulation of SMC in the valve root and VIC in the leaflets. Both cell types were viable (81.4±3.4% for SMC and 83.2±4.0% for VIC) within 3D printed tissues. Encapsulated SMC expressed elevated alpha-smooth muscle actin when printed in stiff matrix, while VIC expressed elevated vimentin in soft matrix. These results demonstrate that anatomically complex, heterogeneously encapsulated aortic valve hydrogel conduits can be fabricated with 3D bioprinting. PMID:23015540

  15. Calcium alginate nanoparticles synthesized through a novel interfacial cross-linking method as a potential protein drug delivery system.

    PubMed

    Nesamony, Jerry; Singh, Priti R; Nada, Shadia E; Shah, Zahoor A; Kolling, William M

    2012-06-01

    The goal of this research work was to develop a novel technique to synthesize calcium alginate nanoparticles using pharmaceutically relevant microemulsions. Stable microemulsion-based reactors were prepared using aqueous sodium alginate, aqueous calcium chloride, dioctyl sodium sulfosuccinate (DOSS), and isopropyl myristate. The reactor microemulsions were characterized via conductivity and dynamic light scattering (DLS) experiments. The conductivity data indicated composition- and reagent-dependent variations in electrical conductivity when the aqueous phase containing reagents were present at or above a Wo (Wo = [DOSS]/[water]) value of 14. The reactor microemulsions were of approximately 6 nm sized droplets. When the reactor microemulsions were mixed and sonicated for 1 h approximately, 350-nm-sized calcium alginate nanoparticles were produced, as indicated by DLS measurements. The particles were isolated and characterized via low-vacuum scanning electron microscopy. The electron micrographs corroborate the DLS results. The nanoparticles were evaluated as a drug delivery system by incorporating bovine serum albumin (BSA) and performing in vitro release and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) studies. The BSA release profile was characterized by an initial burst release followed by a sustained-release phase. SDS-PAGE studies indicated that the incorporated protein did not suffer covalent aggregation or degradation via fragmentation.

  16. Calcium alginate particles for the combined delivery of platelet lysate and vancomycin hydrochloride in chronic skin ulcers.

    PubMed

    Mori, Michela; Rossi, Silvia; Bonferoni, Maria Cristina; Ferrari, Franca; Sandri, Giuseppina; Riva, Federica; Del Fante, Claudia; Perotti, Cesare; Caramella, Carla

    2014-01-30

    The aim of the present work was the development of a powder formulation for the combined delivery of platelet lysate and of a model antibiotic drug, vancomycin hydrochloride (VCM), in chronic skin ulcers. Calcium alginate particles were prepared by freeze-drying beads obtained by ionic gelation method. The experimental conditions adopted permitted the complete loading of VCM and of PDGF AB, the growth factor chosen as representative of those contained in PL. Such particles where able to absorb PBS (mimicking wound exudate), to form a gel and to modulate the release of VCM and of PDGF AB. They are characterized by enhancement properties of human fibroblast proliferation due to PL presence. In particular, PL, when loaded in alginate particles, was able not only to increase the number of viable cells, but also the number of cells in proliferative phase. Such properties were comparable to those of fresh PL indicating the capability of calcium alginate particles to load PL bioactive substances without altering their activity. The formulation developed is characterized by an easier and a less painful administration with respect to traditional gauzes and semisolid preparations and permits the loading in the same dosage form of active substances of different nature avoiding eventual incompatibility problems.

  17. Molecular engineering of manipulated alginate-based polyurethanes.

    PubMed

    Daemi, Hamed; Barikani, Mehdi

    2014-11-04

    The novel soluble alginate-based polyurethanes in organic solvents were synthesized by the reaction of NCO-terminated prepolymers and tributylammonium alginate (TBA-Alg) for the first time. The chemical structures of synthesized polyurethanes were characterized using FTIR, (1)H NMR and TGA. The reaction completion was confirmed by disappearing of NCO band in FTIR spectra. Furthermore, a peak at 4.71 ppm and some small peaks at a range of 4.12-4.37 ppm in the (1)H NMR of alginate-based polyurethanes were assigned to the backbone of alginate. The results of both FTIR and (1)H NMR were remarkably confirmed by TGA data. The ionic nature of polyurethane backbone not only affects on thermal properties of samples, but it also changes the chemically-bonded alginate morphology. Both polyether and polyester based non-ionic polyurethanes extended by TBA-Alg illustrated the distinct alginate, whereas those ionomers extended by alginate were appeared as the continuous systems at nanoscale.

  18. Oyster mushroom’s lipase enzyme entrapment on calcium alginate as biocatalyst in the synthesis of lauryl diethanolamide

    NASA Astrophysics Data System (ADS)

    Wijayati, N.; Masubah, K.; Supartono

    2017-02-01

    Lipase is an enzyme with large biotechnology applications, such as hydrolysis in the food industry, applications in chemical industry, synthesis of polymers and surfactants. Lipase was isolated from oyster mushroom with activity 0,93 U/mg and protein content 1,1234 mg/mL. Lipase was immobilized by entrapment method in a matrix of Ca-alginate. This report describes that we have developed for the synthesis of lauryl diethanolamide The result showed that the optimum condition of lipase immobilization was achieved on 3% Na-alginate solution with protein content 0,84 mg/mL and the activity 3,33 U/mg. An amide (22.911%) formed from the amidation of lauric acid and diethanolamine.

  19. One pot synthesis of metal ion anchored alginate-gelatin binary biocomposite for efficient Cr(VI) removal.

    PubMed

    Gopalakannan, Venkatrajan; Viswanathan, Natrayasamy

    2016-02-01

    Biopolymers are widely used for the removal of chromium from aqueous medium but it possesses limitations like poor sorption capacity and low stability. To overcome the limitations of biopolymers and to improve their properties, the present study was designed in such a way to develop a novel sorbent with enhanced chromium sorption capacity and better stability by synthesizing metal ion cross-linked binary biocomposites using biopolymers like alginate and gelatin cross-linked with Ca2+, Ce3+ and Zr4+ ions namely Ca@AlgGel, Ce@AlgGel and Zr@AlgGel composites. The functional groups, agglomeration, surface area, surface morphology, elemental analysis and thermal stability of the composites were investigated by FTIR, TEM, BET, SEM with EDAX and TGA analysis. The chromium removal studies of the biocomposites were carried out in batch mode. The sorption process was optimized by varying the influencing aspects like contact time, dosage, presence of common ions, pH, initial chromium concentration and temperature. The maximum sorption capacity of Ca@AlgGel, Ce@AlgGel and Zr@AlgGel composites were found to be 19.40, 24.50 and 25.40 mg/g, respectively. The sorption data was fitted by using Freundlich, Langmuir and Dubinin-Radushkevich (D-R) isotherms. Thermodynamic parameters indicate the nature of chromium sorption. The suitability of the composite materials was also tested under the field conditions.

  20. Alginate-based hybrid aerogel microparticles for mucosal drug delivery.

    PubMed

    Gonçalves, V S S; Gurikov, P; Poejo, J; Matias, A A; Heinrich, S; Duarte, C M M; Smirnova, I

    2016-10-01

    The application of biopolymer aerogels as drug delivery systems (DDS) has gained increased interest during the last decade since these structures have large surface area and accessible pores allowing for high drug loadings. Being biocompatible, biodegradable and presenting low toxicity, polysaccharide-based aerogels are an attractive carrier to be applied in pharmaceutical industry. Moreover, some polysaccharides (e.g. alginate and chitosan) present mucoadhesive properties, an important feature for mucosal drug delivery. This feature allows to extend the contact of DDS with biological membranes, thereby increasing the absorption of drugs through the mucosa. Alginate-based hybrid aerogels in the form of microparticles (<50μm) were investigated in this work as carriers for mucosal administration of drugs. Low methoxyl pectin and κ-carrageenan were co-gelled with alginate and further dried with supercritical CO2 (sc-CO2). Spherical mesoporous aerogel microparticles were obtained for alginate, hybrid alginate/pectin and alginate/κ-carrageenan aerogels, presenting high specific surface area (370-548m(2)g(-1)) and mucoadhesive properties. The microparticles were loaded with ketoprofen via adsorption from its solution in sc-CO2, and with quercetin via supercritical anti-solvent precipitation. Loading of ketoprofen was in the range between 17 and 22wt% whereas quercetin demonstrated loadings of 3.1-5.4wt%. Both the drugs were present in amorphous state. Loading procedure allowed the preservation of antioxidant activity of quercetin. Release of both drugs from alginate/κ-carrageenan aerogel was slightly faster compared to alginate/pectin. The results indicate that alginate-based aerogel microparticles can be viewed as promising matrices for mucosal drug delivery applications.

  1. Quantitative Assessment of Islets of Langerhans Encapsulated in Alginate

    PubMed Central

    Johnson, Amy S.; O'Sullivan, Esther; D'Aoust, Laura N.; Omer, Abdulkadir; Bonner-Weir, Susan; Fisher, Robert J.; Weir, Gordon C.

    2011-01-01

    Improved methods have recently been developed for assessing islet viability and quantity in human islet preparations for transplantation, and these measurements have proven useful for predicting transplantation outcome. The objectives of this study were to adapt these methods for use with microencapsulated islets, to verify that they provide meaningful quantitative measurements, and to test them with two model systems: (1) barium alginate and (2) barium alginate containing a 70% (w/v) perfluorocarbon (PFC) emulsion, which presents challenges to use of these assays and is of interest in its own right as a means for reducing oxygen supply limitations to encapsulated tissue. Mitochondrial function was assessed by oxygen consumption rate measurements, and the analysis of data was modified to account for the increased solubility of oxygen in the PFC-alginate capsules. Capsules were dissolved and tissue recovered for nuclei counting to measure the number of cells. Capsule volume was determined from alginate or PFC content and used to normalize measurements. After low oxygen culture for 2 days, islets in normal alginate lost substantial viable tissue and displayed necrotic cores, whereas most of the original oxygen consumption rate was recovered with PFC alginate, and little necrosis was observed. All nuclei were recovered with normal alginate, but some nuclei from nonrespiring cells were lost with PFC alginate. Biocompatibility tests revealed toxicity at the islet periphery associated with the lipid emulsion used to provide surfactants during the emulsification process. We conclude that these new assay methods can be applied to islets encapsulated in materials as complex as PFC-alginate. Measurements made with these materials revealed that enhancement of oxygen permeability of the encapsulating material with a concentrated PFC emulsion improves survival of encapsulated islets under hypoxic conditions, but reformulation of the PFC emulsion is needed to reduce toxicity

  2. A simple strategy to fabricate poly (acrylamide-co-alginate)/gold nanocomposites for inactivation of bacteria

    NASA Astrophysics Data System (ADS)

    Zhang, Yanan; Lou, Zhichao; Zhang, Xiaohong; Hu, Xiaodan; Zhang, Haiqian

    2014-12-01

    A facile and efficient approach to prepare uniform gold nanoparticles (Au NPs) in hybrid hydrogel consisting of acrylamide (AM) and alginate (SA) for antibacterial applications is reported. In this study, reduction of gold ions by acrylamide and alginate (AM-SA) occurred before the polymerization and as-obtained gold colloids are stabilized by AM-SA immediately in the absence of commonly used reducing agents and protective reagents. Via transmittance electron microscopy results, we can conclude that the obtained gold nanoparticles in hydrogel are well dispersed. Furthermore, ultraviolet-visible absorption spectroscopy, Fourier transform infrared and thermogravimetric analysis were used to characterize the structure and composition of the synthetic nanocomposites. Our approach provides well-dispersed nanoparticles around 8 mm in size. It is important to underline that nanoparticle aggregation was not observed during and after gel formation. The prepared Au NPs exhibited remarkable stability in the presence of high pH s, and a range of salt concentrations. Importantly, the hydrogel/gold nanocomposites showed a non-compromised activity to inhibit the growth of a model bacterium, Escherichia coli. With their excellent mechanical behavior, as well as the remained antibacterial activity, the nanocomposites should get various potential applications in the fields of pharmaceutical science and tissue engineering.

  3. Effects of chain conformation and entanglement on the electrospinning of pure alginate.

    PubMed

    Nie, Huarong; He, Aihua; Zheng, Jianfen; Xu, Shanshan; Li, Junxing; Han, Charles C

    2008-05-01

    As a natural biopolymer, sodium alginate (SA) has been widely used in the biomedical field in the form of powder, liquid, gel, and compact solid, but not in the form of nanofiber. Electrospinning is an effective method to fabricate nanofibers. However, electrospinning of SA from its aqueous solution is still a challenge. In this study, an effort has been made to solve this problem and find the key reasons that hinder the electrospinning of alginate aqueous solution. Through this research, it was found that pure SA nanofibers could be fabricated successfully by introducing a strong polar cosolvent, glycerol, into the SA aqueous solutions. The study on the properties of the modified SA solution showed that increasing glycerol content increased the viscosity of the SA solution greatly and, meanwhile, decreased the surface tension and the conductivity of the SA solution. The rheological results indicated that the increase in glycerol content could result in the enhanced entanglements of SA chains. Two schematic molecular models were proposed to depict the change of SA chain conformation in aqueous solution with and without glycerol. The main contribution of glycerol to the electrospinning process is to improve the flexibility and entanglement of SA chains by disrupting the strong inter- and intramolecular hydrogen bondings among SA chains, then forming new hydrogen bondings with SA chains.

  4. Equilibrium, kinetic and thermodynamic studies of uranium biosorption by calcium alginate beads.

    PubMed

    Bai, Jing; Fan, Fangli; Wu, Xiaolei; Tian, Wei; Zhao, Liang; Yin, Xiaojie; Fan, Fuyou; Li, Zhan; Tian, Longlong; Wang, Yang; Qin, Zhi; Guo, Junsheng

    2013-12-01

    Calcium alginate beads are potential biosorbent for radionuclides removal as they contain carboxyl groups. However, until now limited information is available concerning the uptake behavior of uranium by this polymer gel, especially when sorption equilibrium, kinetics and thermodynamics are concerned. In present work, batch experiments were carried out to study the equilibrium, kinetics and thermodynamics of uranium sorption by calcium alginate beads. The effects of initial solution pH, sorbent amount, initial uranium concentration and temperature on uranium sorption were also investigated. The determined optimal conditions were: initial solution pH of 3.0, added sorbent amount of 40 mg, and uranium sorption capacity increased with increasing initial uranium concentration and temperature. Equilibrium data obtained under different temperatures were fitted better with Langmuir model than Freundlich model, uranium sorption was dominated by a monolayer way. The kinetic data can be well depicted by the pseudo-second-order kinetic model. The activation energy derived from Arrhenius equation was 30.0 kJ/mol and the sorption process had a chemical nature. Thermodynamic constants such as ΔH(0), ΔS(0) and ΔG(0) were also evaluated, results of thermodynamic study showed that the sorption process was endothermic and spontaneous.

  5. Hydrogels of sodium alginate in cationic surfactants: Surfactant dependent modulation of encapsulation/release toward Ibuprofen.

    PubMed

    Jabeen, Suraya; Chat, Oyais Ahmad; Maswal, Masrat; Ashraf, Uzma; Rather, Ghulam Mohammad; Dar, Aijaz Ahmad

    2015-11-20

    The interaction of cetyltrimethylammoium bromide (CTAB) and its gemini homologue (butanediyl-1,4-bis (dimethylcetylammonium bromide), 16-4-16 with biocompatible polymer sodium alginate (SA) has been investigated in aqueous medium. Addition of K2CO3 influences viscoelastic properties of surfactant impregnated SA via competition between electrostatic and hydrophobic interactions. Viscosity of these polymer-surfactant systems increases with increase in concentration of K2CO3, and a cryogel is formed at about 0.5M K2CO3 concentration. The thermal stability of gel (5% SA+0.5M K2CO3) decreases with increase in surfactant concentration, a minimum is observed with increase in 16-4-16 concentration. The impact of surfactant addition on the alginate structure vis-à-vis its drug loading capability and release thereof was studied using Ibuprofen (IBU) as the model drug. The hydrogel with 16-4-16 exhibits higher IBU encapsulation and faster release in comparison to the one containing CTAB. This higher encapsulation-cum-faster release capability has been related to micelle mediated solubilization and greater porosity of the hydrogel with gemini surfactant.

  6. Tapioca starch blended alginate mucoadhesive-floating beads for intragastric delivery of Metoprolol Tartrate.

    PubMed

    Biswas, Nikhil; Sahoo, Ranjan Kumar

    2016-02-01

    The objective of the study was to develop tapioca starch blended alginate mucoadhesive-floating beads for the intragastric delivery of Metoprolol Tartrate (MT). The beads were prepared by ionotropic gelation method using calcium chloride as crosslinker and gas forming calcium carbonate (CaCO3) as floating inducer. The alginate gel beads having 51-58% entrapped MT showed 90% release within 45 min in gastric medium (pH 1.2). Tapioca starch blending markedly improved the entrapment efficiency (88%) and sustained the release for 3-4 h. A 12% w/w HPMC coating on these beads extended the release upto 9-11 h. In vitro wash off and buoyancy test in gastric media revealed that the beads containing CaCO3 has gastric residence of more than 12 h. In vitro optimized multi-unit formulation consisting of immediate and sustained release mucoadhesive-floating beads (40:60) showed good initial release of 42% MT within 1h followed by a sustained release of over 90% for 11 h. Pharmacokinetic study performed in rabbit model showed that the relative oral bioavailability of MT after administration of oral solution, sustain release and optimized formulation was 51%, 67% and 87%, respectively. Optimized formulation showed a higher percent inhibition of isoprenaline induced heart rate in rabbits for almost 12 h.

  7. Rheological behavior and Ibuprofen delivery applications of pH responsive composite alginate hydrogels.

    PubMed

    Jabeen, Suraya; Maswal, Masrat; Chat, Oyais Ahmad; Rather, Ghulam Mohammad; Dar, Aijaz Ahmad

    2016-03-01

    Synthesis and structural characterization of hydrogels composed of sodium alginate, polyethylene oxide and acrylic acid with cyclodextrin as the hydrocolloid prepared at different pH values is presented. The hydrogels synthesized show significant variations in rheological properties, drug encapsulation capability and release kinetics. The hydrogels prepared at lower pH (pH 1) are more elastic, have high tensile strength and remain almost unaffected by varying temperature or frequency. Further, their Ibuprofen encapsulation capacity is low and releases it slowly. The hydrogel prepared at neutral pH (pH 7) is viscoelastic, thermo-reversible and also exhibits sol-gel transition on applying frequency and changing temperature. It shows highest Ibuprofen encapsulation capacity and also optimum drug release kinetics. The hydrogel prepared at higher pH (pH 12) is more viscous, has low tensile strength, is unstable to change in temperature and has fast drug release rate. The study highlights the pH responsiveness of three composite alginate hydrogels prepared under different conditions to be employed in drug delivery applications.

  8. Chitosan-alginate membranes accelerate wound healing.

    PubMed

    Caetano, Guilherme Ferreira; Frade, Marco Andrey Cipriani; Andrade, Thiago Antônio Moretti; Leite, Marcel Nani; Bueno, Cecilia Zorzi; Moraes, Ângela Maria; Ribeiro-Paes, João Tadeu

    2015-07-01

    The purpose of this study was to evaluate the efficacy of chitosan-alginate membrane to accelerate wound healing in experimental cutaneous wounds. Two wounds were performed in Wistar rats by punching (1.5 cm diameter), treated with membranes moistened with saline solution (CAM group) or with saline only (SL group). After 2, 7, 14, and 21 days of surgery, five rats of each group were euthanized and reepithelialization was evaluated. The wounds/scars were harvested for histological, flow cytometry, neutrophil infiltrate, and hydroxyproline analysis. CAM group presented higher inflammatory cells recruitment as compared to SL group on 2(nd) day. On the 7(th) day, CAM group showed higher CD11b(+) level and lower of neutrophils than SL group. The CAM group presented higher CD4(+) cells influx than SL group on 2(nd) day, but it decreased during the follow up and became lower on 14(th) and 21(st) days. Higher fibroplasia was noticed on days 7 and 14 as well as higher collagenesis on 21(st) in the CAM group in comparison to SL group. CAM group showed faster reepithelialization on 7(th) day than SL group, although similar in other days. In conclusion, chitosan-alginate membrane modulated the inflammatory phase, stimulated fibroplasia and collagenesis, accelerating wound healing process in rats.

  9. Enantioselective Recognition by Chiral Supramolecular Gels.

    PubMed

    Zhang, Li; Jin, Qingxian; Liu, Minghua

    2016-10-06

    Chiral supramolecular gels, in which small organic molecules self-assemble into chiral nanostructures and entangle each other to immobilize solvents through various noncovalent interactions, can work as a matrix for enantioselective recognition on chiral analytes. Through gelation and the formation of well-defined nanostructures, the chiral sense of the component molecules can be accumulated or amplified, and thus, the enantioselective recognition ability can be enhanced. Furthermore, a chiral microenvironment formed in the gel networks could provide additional stereochemical recognition geometry and attribute to efficient recognition. In this focus review, enantioselective recognition on chiral analytes through chiral supramolecular gels, with either amplified signals or the gel-sol phase transition, is discussed. This review is expected to provide useful insights into the design and fabrication of supramolecular gel systems with chiral features and high enantioselectivity.

  10. Thermogelling Bioadhesive Scaffolds for Intervertebral Disc Tissue Engineering: Preliminary In Vitro Comparison of Aldehyde-Based Versus Alginate Microparticle-Mediated Adhesion

    PubMed Central

    Wiltsey, C.; Christiani, T.; Williams, J.; Scaramazza, J.; Van Sciver, C.; Toomer, K.; Sheehan, J.; Branda, A.; Nitzl, A.; England, E.; Kadlowec, J.; Iftode, C.; Vernengo, J.

    2015-01-01

    Tissue engineering of certain load-bearing parts of the body can be dependent on scaffold adhesion or integration with the surrounding tissue to prevent dislocation. One such area is the regeneration of the intervertebral disc (IVD). In this work, poly(N-isopropylacrylamide) (PNIPAAm) was grafted with chondroitin sulfate (CS) (PNIPAAm-g-CS) and blended with aldehyde-modified CS to generate an injectable polymer that can form covalent bonds with tissue upon contact. However, the presence of the reactive aldehyde groups can compromise the viability of encapsulated cells. Thus, liposomes were encapsulated in the blend, designed to deliver the ECM derivative, gelatin, after the polymer has adhered to tissue and reached physiological temperature. This work is based on the hypothesis that the discharge of gelatin will enhance the biocompatibility of the material by covalently reacting with, or “end-capping”, the aldehyde functionalities within the gel that did not participate in bonding with tissue upon contact. As a comparison, formulations were also created without CS aldehyde and with an alternative adhesion mediator, mucoadhesive calcium alginate particles. Gels formed from blends of PNIPAAm-g-CS and CS aldehyde exhibited increased adhesive strength compared to PNIPAAm-g-CS alone (p<0.05). However, the addition of gelatin-loaded liposomes to the blend significantly decreased the adhesive strength (p<0.05). The encapsulation of alginate microparticles within PNIPAAm-g-CS gels caused the tensile strength to increase two-fold over that of PNIPAAm-g-CS blends with CS aldehyde (p<0.05). Cytocompatibility studies indicate that formulations containing alginate particles exhibit reduced cytotoxicity over those containing CS aldehyde. Overall, the results indicated that the adhesives composed of alginate microparticles encapsulated in PNIPAAm-g-CS have the potential to serve as a scaffold for IVD regeneration. PMID:25641647

  11. Thermogelling bioadhesive scaffolds for intervertebral disk tissue engineering: preliminary in vitro comparison of aldehyde-based versus alginate microparticle-mediated adhesion.

    PubMed

    Wiltsey, C; Christiani, T; Williams, J; Scaramazza, J; Van Sciver, C; Toomer, K; Sheehan, J; Branda, A; Nitzl, A; England, E; Kadlowec, J; Iftode, C; Vernengo, J

    2015-04-01

    Tissue engineering of certain load-bearing parts of the body can be dependent on scaffold adhesion or integration with the surrounding tissue to prevent dislocation. One such area is the regeneration of the intervertebral disc (IVD). In this work, poly(N-isopropylacrylamide) (PNIPAAm) was grafted with chondroitin sulfate (CS) (PNIPAAm-g-CS) and blended with aldehyde-modified CS to generate an injectable polymer that can form covalent bonds with tissue upon contact. However, the presence of the reactive aldehyde groups can compromise the viability of encapsulated cells. Thus, liposomes were encapsulated in the blend, designed to deliver the ECM derivative, gelatin, after the polymer has adhered to tissue and reached physiological temperature. This work is based on the hypothesis that the discharge of gelatin will enhance the biocompatibility of the material by covalently reacting with, or "end-capping", the aldehyde functionalities within the gel that did not participate in bonding with tissue upon contact. As a comparison, formulations were also created without CS aldehyde and with an alternative adhesion mediator, mucoadhesive calcium alginate particles. Gels formed from blends of PNIPAAm-g-CS and CS aldehyde exhibited increased adhesive strength compared to PNIPAAm-g-CS alone (p<0.05). However, the addition of gelatin-loaded liposomes to the blend significantly decreased the adhesive strength (p<0.05). The encapsulation of alginate microparticles within PNIPAAm-g-CS gels caused the tensile strength to increase twofold over that of PNIPAAm-g-CS blends with CS aldehyde (p<0.05). Cytocompatibility studies indicate that formulations containing alginate particles exhibit reduced cytotoxicity over those containing CS aldehyde. Overall, the results indicated that the adhesives composed of alginate microparticles encapsulated in PNIPAAm-g-CS have the potential to serve as a scaffold for IVD regeneration.

  12. Hydrogen Photoproduction by Nutrient-Deprived Chalamydomonas reinhardtii Cells Immobilized Within Thin Alginate Films Under Aerobic and Anaerobic Conditions

    SciTech Connect

    Kosourov, S. N.; Seibert, M.

    2009-01-01

    A new technique for immobilizing H{sub 2}-photoproducing green algae within a thin (<400 {micro}m) alginate film has been developed. Alginate films with entrapped sulfur/phosphorus-deprived Chlamydomonas reinhardtii, strain cc124, cells demonstrate (a) higher cell density (up to 2,000 {micro}g Chl mL{sup -1} of matrix), (b) kinetics of H{sub 2} photoproduction similar to sulfur-deprived suspension cultures, (c) higher specific rates (up to 12.5 {micro}mol mg{sup -1} Chl h{sup -1}) of H{sub 2} evolution, (d) light conversion efficiencies to H{sub 2} of over 1% and (e) unexpectedly high resistance of the H{sub 2}-photoproducing system to inactivation by atmospheric O{sub 2}. The algal cells, entrapped in alginate and then placed in vials containing 21% O{sub 2} in the headspace, evolved up to 67% of the H{sub 2} gas produced under anaerobic conditions. The results indicate that the lower susceptibility of the immobilized algal H{sub 2}-producing system to inactivation by O{sub 2} depends on two factors: (a) the presence of acetate in the medium, which supports higher rates of respiration and (b) the capability of the alginate polymer itself to effectively separate the entrapped cells from O{sub 2} in the liquid and headspace and restrict O{sub 2} diffusion into the matrix. The strategy presented for immobilizing algal cells within thin polymeric matrices shows the potential for scale-up and possible future applications.

  13. Chemical modification of alginic acid by ultrasonic irradiation

    NASA Astrophysics Data System (ADS)

    Murdzheva, Dilyana; Denev, Panteley

    2016-03-01

    Abstract: Chemical modification of alginic acid has been done by ultrasonic irradiation to obtain its methylated, ethylated and isopropylated derivatives. The influence of ultrasonic frequency and power on esterification process of alginic acid has been investigated. Alginate derivatives have been characterized by degree of esterification (DE) and IR-FT spectroscopy. It has been found that 45 kHz ultrasonic frequency accelerated modification process as reduced the reaction time from 16 hours to 2 hours. The obtained results showed that ultrasound irradiation increased the reaction efficiency in methanol and depended on the ratio of the M/G.

  14. Highly Conductive and Uniform Alginate/Silver Nanowire Composite Transparent Electrode by Room Temperature Solution Processing for Organic Light Emitting Diode.

    PubMed

    Lian, Lu; Dong, Dan; Yang, Shuai; Wei, Bingwu; He, Gufeng

    2017-04-05

    A novel transparent electrode composed of alginate/silver nanowire (AgNW) with high conductivity and low roughness is fabricated via a solution process at room temperature. The sol-gel transition of the alginate triggered by CaCl2 solution bonds the AgNWs to the substrate tightly. Meanwhile, Cl(-) in the solution can renovate the cracks on the AgNW surfaces created during the mechanical pressing, resulting in a great increase of the electrical conductivity. The alginate/AgNW composite film can reach a sheet resistance of 2.3 Ω/sq with a transmittance of 83% at 550 nm. The conductivity of the composite film remains stable after bending and tape tests, demonstrating excellent flexibility and great adhesion of AgNWs to the substrate. Moreover, the composite film shows better stability to resist longtime storage than conventional annealed-AgNW film. The organic light emitting diode using such alginate/AgNW composite film as anode presents current densities and luminances comparable to those of indium tin oxide (ITO) anode, and higher efficiencies are obtained due to the better charge balance.

  15. Microfluidic vascular channels in gels using commercial 3D printers

    NASA Astrophysics Data System (ADS)

    Selvaganapathy, P. Ravi; Attalla, Rana

    2016-03-01

    This paper details the development of a three dimensional (3D) printing system with a modified microfluidic printhead used for the generation of complex vascular tissue scaffolds. The print-head features an integrated coaxial nozzle that allows the fabrication of hollow, calcium-polymerized alginate tubes that can easily be patterned using 3Dbioprinting techniques. This microfluidic design allows the incorporation of a wide range of scaffold materials as well as biological constituents such as cells, growth factors, and ECM material. With this setup, gel constructs with embedded arrays of hollow channels can be created and used as a potential substitute for blood vessel networks.

  16. Alginate synthesis in Pseudomonas aeruginosa: the role of AlgL (alginate lyase) and AlgX.

    PubMed Central

    Monday, S R; Schiller, N L

    1996-01-01

    Previous studies localized an alginate lyase gene (algL) within the alginate biosynthetic gene cluster at 34 min on the Pseudomonas aeruginosa chromosome. Insertion of a Tn501 polar transposon in a gene (algX) directly upstream of algL in mucoid P. aeruginosa FRD1 inactivated expression of algX, algL, and other downstream genes, including algA. This strain is phenotypically nonmucoid; however, alginate production could be restored by complementation in trans with a plasmid carrying all of the genes inactivated by the insertion, including algL and algX. Alginate production was also recovered when a merodiploid that generated a complete alginate gene cluster on the chromosome was constructed. However, alginate production by merodiploids formed in the algX::Tn501 mutant using an alginate cluster with an algL deletion was not restored to wild-type levels unless algL was provided on a plasmid in trans. In addition, complementation studies of Tn501 mutants using plasmids containing specific deletions in either algL or algX revealed that both genes were required to restore the mucoid phenotype. Escherichia coli strains which expressed algX produced a unique protein of approximately 53 kDa, consistent with the gene product predicted from the DNA sequencing data. These studies demonstrate that AlgX, whose biochemical function remains to be defined, and AlgL, which has alginate lyase activity, are both involved in alginate production by P. aeruginosa. PMID:8550492

  17. Engineered yeast whole-cell biocatalyst for direct degradation of alginate from macroalgae and production of non-commercialized useful monosaccharide from alginate.

    PubMed

    Takagi, Toshiyuki; Yokoi, Takahiro; Shibata, Toshiyuki; Morisaka, Hironobu; Kuroda, Kouichi; Ueda, Mitsuyoshi

    2016-02-01

    Alginate is a major component of brown macroalgae. In macroalgae, an endolytic alginate lyase first degrades alginate into oligosaccharides. These oligosaccharides are further broken down into monosaccharides by an exolytic alginate lyase. In this study, genes encoding various alginate lyases derived from alginate-assimilating marine bacterium Saccharophagus degradans were isolated, and their enzymes were displayed using the yeast cell surface display system. Alg7A-, Alg7D-, and Alg18J-displaying yeasts showed endolytic alginate lyase activity. On the other hand, Alg7K-displaying yeast showed exolytic alginate lyase activity. Alg7A, Alg7D, Alg7K, and Alg18J, when displayed on yeast cell surface, demonstrated both polyguluronate lyase and polymannuronate lyase activities. Additionally, polyguluronic acid could be much easily degraded by Alg7A, Alg7K, and Alg7D than polymannuronic acid. In contrast, polymannuronic acid could be much easily degraded by Alg18J than polyguluronic acid. We further constructed yeasts co-displaying endolytic and exolytic alginate lyases. Degradation efficiency by the co-displaying yeasts were significantly higher than single alginate lyase-displaying yeasts. Alg7A/Alg7K co-displaying yeast had maximum alginate degrading activity, with production of 1.98 g/L of reducing sugars in a 60-min reaction. This system developed, along with our findings, will contribute to the efficient utilization and production of useful and non-commercialized monosaccharides from alginate by Saccharomyces cerevisiae.

  18. Novel zinc alginate hydrogels prepared by internal setting method with intrinsic antibacterial activity.

    PubMed

    Straccia, Maria Cristina; d'Ayala, Giovanna Gomez; Romano, Ida; Laurienzo, Paola

    2015-07-10

    In this paper, a controlled gelation of alginate was performed for the first time using ZnCO3 and GDL. Uniform and transparent gels were obtained and investigated as potential wound dressings. Homogeneity, water content, swelling capability, water evaporation rate, stability in normal saline solution, mechanical properties and antibacterial activity were assessed as a function of zinc concentration. Gelation rate increased at increasing zinc content, while a decrease in water uptake and an improvement of stability were found. Release of zinc in physiological environments showed that concentration of zinc released in solution lies below the cytotoxicity level. Hydrogels showed antimicrobial activity against Escherichia coli. The hydrogel with highest zinc content was stabilized with calcium by immersion in a calcium chloride solution. The resulting hydrogel preserved homogeneity and antibacterial activity. Furthermore, it showed even an improvement of stability and mechanical properties, which makes it suitable as long-lasting wound dressing.

  19. Removal of copper ions from aqueous solution by calcium alginate immobilized kaolin.

    PubMed

    Li, Yanhui; Xia, Bing; Zhao, Quansheng; Liu, Fuqiang; Zhang, Pan; Du, Qiuju; Wang, Dechang; Li, Da; Wang, Zonghua; Xia, Yanzhi

    2011-01-01

    Kaolin has been widely used as an adsorbent to remove heavy metal ions from aqueous solutions. However, the lower heavy metal adsorption capacity of kaolin limits its practical application. A novel environmental friendly material, calcium alginate immobilized kaolin (kaolin/CA), was prepared using a sol-gel method. The effects of contact time, pH, adsorbent dose, and temperature on Cu2+ adsorption by kaolin/CA were investigated. The Langmuir isotherm was used to describe the experimental adsorption, the maximum Cu2+ adsorption capacity of the kaolin/CA reached up to 53.63 mg/g. The thermodynamic studies showed that the adsorption reaction was a spontaneous and endothermic process.

  20. Polyurethane and alginate immobilized algal biomass for the removal of aqueous toxic metals

    SciTech Connect

    Fry, I.V.; Mehlhorn, R.J.

    1992-12-01

    We describe the development of immobilized, processed algal biomass for use as an adsorptive filter in the removal of toxic metals from waste water. To fabricate an adsorptive filter from precessed biomass several crucial criteria must be met, including: (1) high metal binding capacity, (2) long term stability (both mechanical and chemical), (3) selectivity for metals of concern (with regard to ionic competition), (4) acceptable flow capacity (to handle large volumes in short time frames), (5) stripping/regeneration (to recycle the adsorptive filter and concentrate the toxic metals to manageable volumes). This report documents experiments with processed algal biomass (Spirulina platensis and Spirulina maxima) immobilized in either alginate gel or preformed polyurethane foam. The adsorptive characteristics of these filters were assessed with regard to the criteria listed above.

  1. Functionalized magnetic iron oxide/alginate core-shell nanoparticles for targeting hyperthermia

    PubMed Central

    Liao, Shih-Hsiang; Liu, Chia-Hung; Bastakoti, Bishnu Prasad; Suzuki, Norihiro; Chang, Yung; Yamauchi, Yusuke; Lin, Feng-Huei; Wu, Kevin C-W

    2015-01-01

    Hyperthermia is one of the promising treatments for cancer therapy. However, the development of a magnetic fluid agent that can selectively target a tumor and efficiently elevate temperature while exhibiting excellent biocompatibility still remains challenging. Here a new core-shell nanostructure consisting of inorganic iron oxide (Fe3O4) nanoparticles as the core, organic alginate as the shell, and cell-targeting ligands (ie, D-galactosamine) decorated on the outer surface (denoted as Fe3O4@Alg-GA nanoparticles) was prepared using a combination of a pre-gel method and coprecipitation in aqueous solution. After treatment with an AC magnetic field, the results indicate that Fe3O4@Alg-GA nanoparticles had excellent hyperthermic efficacy in a human hepatocellular carcinoma cell line (HepG2) owing to enhanced cellular uptake, and show great potential as therapeutic agents for future in vivo drug delivery systems. PMID:26005343

  2. Formulation and Coating of Alginate and Alginate-Hydroxypropylcellulose Pellets Containing Ranolazine.

    PubMed

    Segale, Lorena; Mannina, Paolo; Giovannelli, Lorella; Muschert, Susanne; Pattarino, Franco

    2016-11-01

    The formulation and the coating composition of biopolymeric pellets containing ranolazine were studied to improve their technological and biopharmaceutical properties. Eudragit L100 (EU L100) and Eudragit L30 D-55-coated alginate and alginate-hydroxypropylcellulose (HPC) pellets were prepared by ionotropic gelation using 3 concentrations of HPC (0.50%, 0.65%, and 1.00% wt/wt) and applying different percentages (5%, 10%, 20%, and 30% wt/wt) of coating material. The uncoated pellets were regular in shape and had mean diameter between 1490 and 1570 μm. The rate and the entity of the swelling process were affected by the polymeric composition: increasing the HPC concentration, the structure of the pellets became more compact and slowed down the penetration of fluids. Coated alginate-HPC formulations were able to control the drug release at neutral pH: a higher quantity of HPC in the system determined a slower release of the drug. The nature of the coating polymer and the coating level applied affected the drug release in acidic environment: EU L100 gave better performance than Eudragit L30 D-55 and the best coating level was 20%. The pellets containing 0.65% of HPC and coated with 20% EU L100 represented the best formulation, able to limit the drug release in acidic environment and to control it at pH 6.8.