Everything is from the AN INTRODUCTION TO BIOMATERIALS unless otherwise cited Glass Transition Temperature The glass transition temperature

is defined as the approximate midpoint of the temperature range at which a non-crystalline (amorphous) polymer changes from brittle, glass state to a rubbery, soft state. In Roger et al., powder samples of alginate exhibit Tg ranging from 95C to 136C.AZ6 In the same experiment, no significant effect on Tg was observed for different molecular weight samples. AZ6 There was, however, an increase in Tg with increase G content. AZ6 This effect can be explained by the presence of residual Ca 2+ ions in the alginate powder, crosslinking oligomeric G-rich chains. AZ6 Roger et al. also determined the Tg of sodium alginate films to be 158C. They mentioned that it was higher than glass transition temperatures mentioned previously in literature for sodium alginate films ranging from 95C to 110C. AZ6 The source of this discrepancy is hypothesized to be residual Ca2+ content in the alginate sources used experimentally, highlighting the importance of source variability in determining the mechanical and thermal properties of alginates. Experimentally, substituting sodium with calcium in alginate films increases Tg concurrent with the theory that divalent cations have strong binding effects at junction points between chains. AZ6 Tg of alginate films also varies according to the crosslinking cation. Experimentally, the Tg of alginate films having various cations decreased in the order A13+ > Ca2+ > Fe2+ > Cu2+ > Na+.AZ7

Figure The relationship between Tg, ionic radius of crosslinking cation, and heat capacity in alginate films (Cp)(Nakumura et al., 1995). Figure demonstrates the relationship between Tg, ionic radius of crosslinking cation, and heat capacity in alginate films. AZ7 Molecular Weight

Commercial alginates are polydisperse with respect to molecular weight. The most common method to calculate molecular weight is based upon intrinsic viscosity which can only give an estimate of the molecular weight. Most producers blend alginates to target a given viscosity specification product with the same intrinsic viscosity which may have different molecular weight populations. Therefore, in biomedical applications, equivalent products may have different molecular weight populations and therefore have differing biomaterial interactions. Solutions The rheological properties of a pure alginate solution are strongly dependent on the molecular weight of the alginate, while the structural composition may play only a minor role, such as with viscosity. Solubility The solubility of alginate is related to the rate of dissociation of the alginate molecule. At low pH (pH<3), M-structures and G-structures will precipitate as alginic acid, while alternating M,G structures will remain in solution despite being fully protonated. This pH dependent physical property of alginates is used in pharmaceutical applications as an antireflux remedy which, when swallowed, forms an alginic acid raft on top of stomach contents (Mandel et al., 2000). Thickening The thickening properties of alginate are related to the molecular weight and conformation of the alginate molecule in solution. Alginates flow properties will be affected by interactions with other molecules as well as competition for water at high concentrations. Therefore, increasing alginate concentration corresponds to increased viscosity (Kong and Mooney, 2002). Cross-linking materials, such as calcium, present in even small concentrations will induce artificial viscosities higher than real ones. Solutions like these have thixothropic flow properties. Swellability Swellability is associated with the rate of hydration, and will strongly depend on the form of alginate. Cross-linked alginates will swell slower than pure sodium alginate. Swelling properties of alginates have been used in dietary products and sustained release tablets (Miyazaki et al., 200). Furthermore, swelling properties of partially neutralized alginic acid has a long tradition as a tablet disintegrant. Film-forming Properties

The film forming properties of alginate are a result of entanglements of alginate molecules when water is removed. When cross-linked gels are dried, they exhibit this they exhibit this type of behavior. Therefore, the molecular weight of alginate needs to be above a lower limit to achieve film formation and avoid brittleness. In situ, alginate films can be formed by spraying alginate solution to a binding surface.

Cross-linked Gels In general, the stiffness of alginate hydrogels can be enhanced by increasing alginate concentration (Kong and Mooney, 2002). However, this will also increase viscosity which may be unfavorable in particular processing applications. Therefore, to increase post-gel stiffness with minimal pre-gel viscosity increase, increasing quantities of low molecular weight (MW) alginate (M n ~ 3.3x104 g/mol) can be incorporated into 1% (w/w) alginate solutions composed of high MW alginate (M n ~ 2.2x105 g/mol) (Kong and Mooney, 2002). The experimental results seen below are evidence of this method:

Fig - (Left)Effects of molecular weight distribution on stiffness of cross-linked hydrogels. (Right) The effects of molecular weight distribution on the viscosity of pre-gelled solution. (-- unary system, -o- binary system).

Gelling properties of alginate are related to both M/G composition and the sequential organization of M and G along the chain. Alginate forms gels with many di and multivalent cations including, . G-block structures, with consecutive gluronic residues, can cross-link with multivalent cations, with the exception of magnesium. Therefore, increases in G-content and molecular weight generally produce a stronger gel (Smidsrod and Skjak-Braek, 1990).

(INSERT IMAGE from book)(M-blocks and G-Blocks) The gelling process is of particular importance to the gelling properties. Gelling can occur externally, through the diffusion of cross-linking agent from the outside, or internally, by homogeneously releasing a cross-linking agent from the inside. In internal gelling, ratio between the cross-linking agents and alginate is important in determination of gel strength. Furthermore, strength properties in high-G alginate is much different than in high-M alginate at a particular cross-linking agent/alginate ratio. Evidence of this is seen in the experimental graph below: (INSERT PICTURE pg. 269 (15.6)) High-G alginates gels (G>60%) cross-linked with Ca2+, exhibit maximum gel strength, measured by the force needed (in grams) to rotate a metal plate immersed into the gel by 30, when the amount of calcium stoichiometrically matches the amount of calcium-binding guluronic acid blocks. As seen in the figure, when the alginate concentration in the gelling solution is higher than the optimal ratio, calcium will not bind available G-sites optimally, and will have lower gel strength. Contrarily, high-M alginate have shorter and fewer G-blocks and therefore show a continuous increase in gel strength for higher alginate concentrations. This is because the addition of high-M alginate does not present a surplus of G-sites, but a relative increase in calcium ion binding G-sites below threshold that facilitate increased gel strength.

An important consideration when measuring gel strength is to differentiate between elastic and viscous moduli. In particular, the viscous modulus is often misinterpreted as gel strength in high viscosity solutions.

Applications Sodium alginate is the main form of alginate used (McHugh, 2003). Other types of alginate include alginic acid, calcium, ammonium and potassium salts, and propylene glycol alginate, an ester of alginic acid. Alginic acid and calcium alginate can be removed during the calcium alginate process for making sodium alginate, and dried and milled to appropriate particle size. Ammonium and potassium salts are made by neutralization of moist alginic acid with ammonium hydroxide and

potassium carbonate (McHugh, 2003). Propylene glycol alginate can be produced in a pressure vessel by treating moist alginic acid, partially reacted with sodium carbonate, with liquid propylene oxide (McHugh, 2003). Majority of the applications of alginates are based on three intrinsic properties. The first property is an ability to increase the viscosity of an aqueous solution when dissolved. The second property is alginates ability to form gels. This gelation process occurs when calcium salt is added to a solution of sodium alginate in water. Chemically, calcium displaces the sodium from the alginate, and holds the long alginate molecules together. This gelling process is independent of a heat requirement which contrasts traditional agar gels which require heating water to approximately 80C before the gel forms when cooled below 40C (McHugh, 2003). The third important property is the ability to form films of sodium/calcium alginate and calcium alginates fibres (McHugh, 2003). Due to the large variability of alginates, sellers offer a range of alginates with different viscosities and mechanical properties. The arrangement and overall M/G ratio of alginate chains varies from one species to another. Therefore, different alginates produce a range of viscosity values when dissolved in water. Biomedical Applications Good quality stable fibers made from mixed salts of sodium and calcium alginate and processed into non-woven fabric are used in wound dressings. AZ1 These dressing have good wound healing and haemostatic properties. AZ1 Furthermore, they can be absorbed by body fluids when the calcium in the fibers is exchanged by sodium from body fluids converting it into a soluble sodium alginate. This property makes it easy to remove from wounds as well.AZ1 Alergic and inflammatory reactions to alginate dressings are rare. AZ12 Pharmaceutically, the swelling properties of alginic acid powder in water has led to its use as tablet disintegrant. AZ1 Alginic acid has also been used in dietary products and foods to give an artificial full feeling after swelling. To reduce irritating reflux, alginic acid has also been used to keep gastric contents in place. AZ1 Alginate is also widely used for the controlled release of medicinal drugs and chemicals. In certain applications, the chemicals of interest are placed in calcium alginate beads, with tight control of size distributions, and are slowly released when exposed to the appropriate environment. Alginate microspheres, coated with chitosan to improve mechanical strength, have been used in oral controlled drug release systems as well. A successful example of this application is the use of alginate microspheres, prepared by an emulsion based process, for oral insulin delivery in rats.AZ11 In tissue engineering, alginate is a biomaterial that can be used in scaffolds. Alginate has been used to encapsulate cells in the solid freeform fabrication of

tissue constructs.AZ7 Successful examples of this application include the ability of hepatocytes to grow, proliferate, and maintain hepatocyte specific function in porous alginate scaffolds and tissue constructs. AZ7,AZ10 Furthermore, the time dependent degradation of alginate microspheres has been used to better tissue engineering processes. Calcium alginate microcapsules are used to encapsulate signaling molecules, nucleotides, and biofluids in specific scaffolds.AZ8 Here, they offer the advantage of controlled release kinetics. AZ9 Industry In the food industry, the thickening properties of alginate are used in syrups, sauces, and ice cream.AZ1 However, sodium alginate is not useful when the emulsion is acidic due to the formation of insoluble alginic acid forms. Propylene glycol alginate is used instead because of its stability in mild acidic conditions. Sodium alginates improve the texture and body of yogurt, but propylene glycol alginate is used in the stabilization of milk proteins under acidic conditions. In ice cream, alginates are used as stabilizers, reducing the formation of ice crystals during freezing and producing a smoother product. Edible instant dessert jellies can be made from alginate-calcium mixtures because they are formed by simply mixing powders with water or milk without the addition of heat. Alginate gels, formed from powders composed of sodium alginate, calcium carbonate, lactic acid and calcium lactate are used to in re-formed/re-structured food products including meats. Calcium alginate coatings are used to preserve meats including fish to prevent oxidation and bacterial contamination. AZ1 Industrially, beads of calcium alginate are used to immobilize biocatalysts and active whole cells that are responsible industrially for the conversion of glucose to fructose, starch to ethanol, and the continuous production of yogurt. The material entraps the enzyme and is still penetrable enough to allow for the diffusion of the substance that needs to be converted. In textiles, alginates are used to thicken for pastes containing dyes. Advantages of using alginates of traditional starch include lower reactivity with the dyes and easier removal out of finished textiles. AZ1 Textiles account for a significant portion of the global alginate market (approx: 50%). AZ1 The paper industry uses the excellent film forming properties of alginate for surface sizing. The oil resistance of alginate improves the oil resistance and greaseproof properties of paper surfaces. In the welding industry, alginate coatings are applied in the immediate vicinity of the weld to control temperature, oxygen, and hydrogen availability. Alginates intrinsic properties also make them very useful in the molding industry. The poor adhesion of alginate films to many surfaces in combination with insolubility in non-aqueous environments have made them ideal candidates for mold-release agents.AZ1 The extremely accurate, flexible, and fast setting medium properties of alginate molding compounds have led to their common use in dentistry, prosthetics, and life casting.

(AZ1) McHugh, Dennis J. A Guide to the Seaweed Industry. New York: Food and Agriculture Organization of the United Nations, 2003. (AZ2) Mandel, K.G., et al. Review article: alginate-raft formulations in the treatment of heartburn and acid reflux. Ailment Pharm Therap, 14, 2000, 669. (AZ3) Miyazaki, S., Kubo, W., Attwood, D. Oral sustained delivery of theophylline using in-situ gelation of sodium alginate. J Controlled Release, 67, 2000, 275. (AZ4) Smidsrod, O., Skjak-Braek, G. Alginate as immobilization matrix for cells. TIBTECH 8, 1990, 71. (AZ5) Kong, H., Mooney, D.J. Controlling material properties of ionically cross-linked alginate hydrogels by varying molecular weight distribution. Mat Res Soc Symp Proc 711, 2002, 227. (AZ6) Roger, S., Bee, A., Balnois, E., Bourmaud, A., Le Deit, H., Grohens, Y., Cabuil, V. Physical and mechanical properties of alginate films containing calcium cations and ferrofluids. Fifth International Conference Polymer-Solvent Complexes & Intercalates. July 11-14, 2004. Lorient, France. (AZ7) Chang R, Nam J, and Sun W. Direct cell writing of 3D microorgan for in vitro pharmacokinetic model. Tissue Engineering 14, 157, 2008. (AZ8) Bidanda, Bopaya, and Paulo Bartolo, eds. Bio-Materials and Prototyping Applications in Medicine. New York: Springer, 2007. 17. (AZ9) Smith, M.K., Riddle, K.W., and Mooney, D.J. Delivery of Hepatotrophic Factors Fails to Enhance Longer-Term Survival of Subcutaneously Transplanted Hepatocytes. Tissue Engineering 12, 235, 2006 (AZ10) Fisher, John P., Antonios G. Mikos, and Joseph D. Bronzino. Tissue Engineering. New York: C R C P LLC, 2007. (AZ11) Jerry, N., Anitha, Y., Sharma, C.P., Sony, P. In vivo absorption studies from an oral delivery system. Drug Del 8, 2001, 19. (AZ12) Batchelor, Andrew W., and Margam Chandrasekaran. Service Characteristics of Biomedical Materials and Implants. New York: Imperial College P, 2004. 178.