Kurt I. Draget

Alginate based new materials

Present and future applications of alginates are mainly linked to the most striking feature of the alginate molecule; i.e. a sol/gel transition in the presence of multivalent cations, e.g. Ca2+, almost independent on temperature. These very mild conditions, combined with the fact that alginates are highly characterised and understood both in the liquid and in the gel phase, makes this biopolymer unique compared to other gelling polysaccharides. Only pectins resemble alginate in the sol/gel transition behaviour, but this system can hardly be said to be as well characterised and understood as the alginates. The properties of alginate solutions and gels suggest biomedical and pharmaceutical uses. In this paper, the question of the specifications required by a polymer for applications in some biomedical areas will be discussed.

Homogeneous alginate gels: A technical approach

Abstract Homogeneous alginate gels can be made by internal liberation of calcium ions. Non-toxic gels at neutral pH were achieved by mixing particles of CaCO 3 with the slowly hydrolysing proton donor d -glucono-δ-lactone into the alginate solutions. This paper summarizes the essential factors controlling the properties of the resulting gel, such as pH, optical clarity, gel homogeneity, gel strength and syneresis.

Ionic and acid gel formation of epimerised alginates; the effect of AlgE4.

AlgE4 is a mannuronan C5 epimerase converting homopolymeric sequences of mannuronate residues in alginates into mannuronate/guluronate alternating sequences. Treating alginates of different biological origin with AlgE4 resulted in different amounts of alternating sequences. Both ionically cross-linked alginate gels as well as alginic acid gels were prepared from the epimerised alginates. Gelling kinetics and gel equilibrium properties were recorded and compared to results obtained with the original non-epimerised alginates. An observed reduced elasticity of the alginic acid gels following epimerisation by AlgE4 seems to be explained by the generally increased acid solubility of the alternating sequences. Ionically (Ca(2+)) cross-linked gels made from epimerised alginates expressed a higher degree of syneresis compared to the native samples. An increase in the modulus of elasticity was observed in calcium saturated (diffusion set) gels whereas calcium limited, internally set alginate gels showed no change in elasticity. An increase in the sol-gel transitional rate of gels made from epimerised alginates was also observed. These results suggest an increased possibility of creating new junction zones in the epimerised alginate gel due to the increased mobility in the alginate chain segments caused by the less extended alternating sequences.

Alginate-based solid media for plant tissue culture

SummaryA new method for solid medium plant tissue culture based on in situ gelation of alginate is proposed as an alternative to agar-based media. In situ gelation by the use of dispersed CaCO3 and the slowly hydrolysing acid glucono-δ-lactone (GDL) was the basis for the use of alginate as a gelling agent. Inexpensive alginate-based media can be made in a wide range of pH values. Biological tests of these gels, concerning sterile seed growth and microcalli plating of Brassica napus (cv. Topas) and biomass production of Panax ginseng callus, showed results equal to those achieved with agar-based gels.

Swelling and partial solubilization of alginic acid gel beads in acidic buffer

Abstract Swelling behaviour of alginic acid gel beads with different chemical composition, molecular weight and size was studied in acetate buffer at pH4. A correlation was observed between the swelling behaviour in this buffer and the equilibrium properties of alginic acid gels. High contents of long l -guluronic acid blocks (G-blocks), known to give a high acid gel strength, reduced the rate of swelling and also the amount of solubilized alginate molecules leaching out of the gel beads. Compared to the original alginate, the leaching molecules had a lower average molecular weight, higher content of mannuronic acid residues and a reduced average length of G-blocks. Swelling capacity, rate of swelling and solubility of alginic acid seemed to depend on a balance between the tendency of homopolymeric blocks to form intermolecular junction zones, and the tendency of alginate to reduce the chemical potential of water. As expected, swelling rate increased with increasing temperature and decreasing bead size.

Regeneration, Cultivation and Differentiation of Plant Protoplasts Immobilized in Ca-alginate Beads

Summary A method for growing plant protoplasts immobilized in Ca-alginate is presented in detail. Two types of protoplasts, isolated from hypocotyls of Brassica napus cv. Niklas and leaf mesophyll tissue of Nicotiana sanderae cv. Crimson Bedder, have been used as examples to illustrate the general applicability of the method. The major advantage of the method is improved cellular protection during the first critical weeks after isolation. This is a protection both against mechanical strain and strong gradients in environmental conditions. Higher plating efficiency may thus be achieved and handling simplified, particularly when changing nutrient medium. Both regeneration and differentiation have repeatedly been achieved.

Oligosaccharides As Modulators of Rheology in Complex Mucous Systems

Mucus rheology is integral to physiological function with the exact secretion rheology resulting from the macromolecular components, both mucin and nonmucin, and the interactions between these macromolecules. Here we present data demonstrating that low-molecular-weight guluronic acid oligomers extracted from alginate are able to disrupt intermolecular interactions in both purified and native mucous systems, resulting in rheological changes that are compatible with a lower cross-link density and thus reduced resistance to deformation. Additionally, these changes are associated with altered physiological function, raising the possibility of the use of such oligomers in biomedical applications.

Controlling the elastic modulus of cellulose nanofibril hydrogels—scaffolds with potential in tissue engineering

AbstractCellulose nanofibrils (CNF) form hydrogels at low concentrations. These hydrogels are held together by transient interactions such as entanglement of fibrils, non-specific ionic interactions and hydrogen bonds; and are thus vulnerable for changes in the chemical environment or the influence of mechanical forces. By a covalent crosslinking of the fibrils, stable permanent gels can be formed. In this study we have produced CNF by using TEMPO mediated oxidation followed by fibrillation. During this procedure, carboxyl and aldehyde groups are introduced on the CNF surfaces. The aldehyde groups are suitable sites for crosslinking, as aldehydes readily form covalent bonds to primary amines through formation of Schiff bases. For this purpose the diamines ethylenediamine and hexamethylenediamine, differing with four carbon atoms in the chain, were used as crosslinker molecules. The results show that by varying the concentration and length of the crosslinker molecules, the elastic modulus of the gels could be controlled. The reversible gels were in this way transformed to irreversible gels by a simple water based reaction. Controlling gel strength is one important premise for the use of CNF in applications such as tissue engineering.

Gelling concept combining chitosan and alginate-proof of principle.

Biocompatible hydrogels are very interesting for applications in, e.g., tissue engineering and for immobilization of cells, such as calcium-alginate gels where the calcium ions form specific interactions with the guluronic acid units. We here report on a new gelling system of chitosan and alginate containing only mannuronic acid (poly-M), which are prepared using the following steps: (i) mixing at a pH well above 7 where the chitosan is mainly uncharged; (ii) controlled lowering of the pH by adding the slowly hydrolyzing d-glucono-δ-lactone (GDL); (iii) formation of a homogeneous chitosan-alginate gel upon leaving the mixture at room temperature. Some properties of the new gelling system are demonstrated herein by adding controlled amounts of GDL to (i) a mixture of a polymeric and neutral-soluble chitosan with poly-M oligomers (MO) and (ii) a mixture of poly-M and neutral-soluble chitosan oligomers. The neutral-solubility of the polymeric chitosan is achieved by selecting a polymeric chitosan with an intermediate degree of acetylation of 40%, while the neutral-solubility of the fully de-N-acetylated chitosan oligomers (CO) is obtained by selecting oligomers with a chain length below 10. A proof of concept of the new gelling system is demonstrated by measuring the gel strengths of the polymeric chitosan-MO, and a poly-M-CO. The results show that the gel strength increases with decreasing the pH from neutral to 5, and that the gel strength decreases with increasing ionic strength, indicative of an ionic gel formation. Poly-M formed relatively strong gels with CO while an alginate highly enriched in Guluronic acid formed gels of very limited mechanical strength, suggesting the importance of the match in charge distances in the poly-M and chitosan, both with diequatorially linked sugar units in the (4)C1 conformation.

Alterations in Mucus Barrier Function and Matrix Structure Induced by Guluronate Oligomers

The effect of guluronate oligomers on the barrier properties of mucous matrices was investigated in terms of the mobility of nanoparticles in mucous matrices by fluorescence recovery after photobleaching (FRAP), cellular uptake of nanoparticles in mucus secreting cells (HT29-MTX), and mucin matrix architecture by scanning electron microscopy (SEM). Guluronate oligomers improved nanoparticle mobility in both native and highly purified mucus matrices and improved cellular uptake of nanoparticles through a mucus layer. Addition of guluronate oligomers to mucin matrices resulted in a decrease in the density of network cross-links and an increase in matrix pore size. Based on these data, we conclude that guluronate oligomers are able to improve nanoparticle mobility in several mucus matrices and alter network architecture in mucin matrices in a manner that suggests a reduction in barrier function. As such, there may be a potential application for guluronate oligomers in mucosal delivery of nanomedicines.