Ionically cross-linked alginate hydrogels as drug delivery systems for analgesics in broiler chickens : thesis presented in partial fulfilment of the requirement for the degree of Masters of Science in Chemistry at Massey University, Palmerston North, Manawatu, New Zealand

Treating birds with analgesic drugs requires continuous injections of near lethal concentrations to maintain the therapeutic dose in the blood plasma. This is due to birds having higher metabolic rates than mammals. Therefore, there is a need to develop drug delivery systems that can control and slow down the release of analgesics in birds. This study was designed to analyse the sustained release of the model analgesics, sodium salicylate and sodium aspirin, from ionically cross-linked alginate hydrogels, in in vitro and in vivo experiments using broiler chickens as the model bird. Analgesic loaded hydrogels separated into two layers, unlike the homogeneous blank hydrogels. This was labelled as the separation effect. Swelling studies indicated the absence of the insoluble cross-linked alginate material in the hydrogels where the separation effect occurred, with most of the hydrogels dissolving back into the medium. The highest equilibrium swelling percentage achieved in the loaded hydrogels was 68 %. In comparison, the highest equilibrium swelling percentage in the blank hydrogels was 622 %. In vitro drug release profiles showed that the hydrogels released up to 100% of the sodium salicylate within 3.33 hours. In contrast, the hydrogels containing sodium aspirin released only 35 % of the encapsulated drug. Hydrogels containing a drug concentration of 150 mg/mL were injected into the model birds at a dose rate of 150 mg/Kg. No chicken reacted negatively to the hydrogel injection. In vivo results indicate sustained release of the model analgesic from the hydrogels compared to the release from the aqueous solutions of the drug. The effective concentration for an analgesic effect of sodium salicylate was maintained by the group injected with an aqueous solution of sodium salicylate 18 hours after the injection. The groups injected with the hydrogel with the maximum calcium chloride content saw the largest sustained release, with the plasma concentration of sodium salicylate remaining over the effective concentration for up to 36 hours after the injection. Keywords: Sodium salicylate, sodium aspirin, hydrogel, analgesia, sustained release, broiler chicken

Mechanical behaviors of hydrogel-impregnated sand

Hydrogel has been widely used in medical studies due to their unique integration of solid and liquid properties. There is limited studies of using hydrogel in construction materials. The goal of this study was to investigate the effect of hydrogel on mechanical behaviors of sandy materials. The effects of reaction time, sodium alginate content, and curing temperature on mechanical behaviors of hydrogel-impregnated sand were studied through unconfined compression tests, falling head permeability tests, consolidated and undrained triaxial tests, scanning electron microscopy, and durability tests. The unconfined compression strength (UCS) increased with sodium alginate content, but the hydraulic conductivity of hydrogel-impregnated sand decreased with sodium alginate content. The optimum reaction time and curing temperature were found to be 3 days and 50 °C, respectively, for the hydrogel-impregnated sand. The stress-strain curves of hydrogel-impregnated sand indicated that the ductility of hydrogel-impregnated sand was significantly improved compared with the traditional cementitious method. Moreover, the results of durability tests indicated that approximately 60% of the original UCS of hydrogel-impregnated sand still remained after 12 wet-dry and freeze-thaw cycles

Stimulus-responsive Injectable Polysaccharide Scaffolds for Soft Tissue Engineering Prepared by O/W High Internal Phase Emulsion

This thesis describes work on the development of several novel stimuli-responsive porous hydrogels prepared from oil-in-water (o/w) high internal phase emulsion (HIPE) as injectable scaffolds for soft tissue engineering. Firstly, by copolymerising glycidyl methacrylate (GMA) derivatised dextran and N-isopropylacrylamide (NIPAAm) in the aqueous phase of a toluene-in-water HIPE, thermo-responsive polyHIPE hydrogels were obtained. The temperature depended modulus of these porous hydrogels, as revealed by oscillatory mechanical measurements, indicated improvements of the mechanical properties of these hydrogels when heated from room temperature to human body temperature, as the polyNIPAAm copolymer segments starts to phase separate from the aqueous phase and causes the hydrogel to form a more compact structure within the aqueous phase of the polyHIPE. Secondly ion responsive methacrylate modified alginate polyHIPE hydrogels were prepared. The physical dimensions, pore and pore throat sizes as well as water uptakes of these ion responsive hydrogels can be controllably decreased in the presence of Ca2+ ions and are fully recovered after disruption of the ionic crosslinking using a chelating agent (sodium citrate). These ion-responsive polyHIPE hydrogels also possess good mechanical properties (modulus up to 20 kPa). Both of these polyHIPE hydrogels could be easily extruded through a hypodermic needle while breaking into small fragments (about 0.5 to 3.0 mm in diameter), but the interconnected porous morphology was maintained after injection as revealed by SEM characterisation. Furthermore, the hydrogel fragments produced during injection can be crosslinked into a coherent scaffold under very mild condition using Ca2+ salts and alginate aqueous solution as the ionically crosslinkable adhesive. In order to increase the pore size of these covalently crosslinked polyHIPE hydrogels and also find a biocompatible nontoxic emulsifier as substitution to traditional surfactants, methyl myristate-in-water and soybean oil-in-water HIPEs solely stabilised by hydroxyapatite (HAp) nanoparticle were prepared. These Pickering- HIPEs were used as template to prepare polyHIPE hydrogels. Dextran-GMA, a water soluble monomer, was polymerised in the continuous phase of the HAp Pickering HIPEs leading to porous hydrogels with a tunable pore size varying from 1.5 μm to 41.0 μm. HAp is a nontoxic biocompatible emulsifier, which potentially provides extra functions, such as promoting hard tissue cell proliferation. HIPE-templated materials whose porous structure is maintained solely by the reversible physical aggregation between thermo-responsive dextran-b-polyNIPAAm block polymer chains in an aqueous environment (for this type of HIPE templated material we coined the name thermo-HIPEs) were prepared. No chemical reaction is required for the solidification of this porous material. This particular feature should provide a safer route to injectable scaffolds as issues of polymerisation/crosslinking chemistry or residual initiator fragments or monomers potentially being cytotoxic do not arise in our case, as all components are purified polymers prior to HIPE formation. Thermo-HIPEs with soybean oil or squalene as dispersed oil phase were prepared. Also in this HIPE system it was possible to replace the original surfactant Triton X405 with colloidal HAp nanoparticles or pH/thermo-responsive polyNIPAAm-co- AA microgel particles. The pore sizes and the mechanical properties of colloidal particles stabilised thermo-HIPEs showed improvement compared with thermo-HIPEs stabilised by Triton X405. In summary new injectable polyHIPEs have been prepared which retain their pore morphology during injection and can be solidified by either a thermal or ion (Ca2+) or chelating ion (Ca2+) stimulus. The materials used are intrinsically biocompatible and thus makes these porous injectable scaffolds excellent candidates for soft tissue engineering

Design and in Vitro Evaluation of a New Nano-Microparticulate System for Enhanced Aqueous-Phase Solubility of Curcumin

Curcumin, a yellow polyphenol derived from the turmeric Curcuma longa, has been associated with a diverse therapeutic potential including anti-inflammatory, antioxidant, antiviral, and anticancer properties. However, the poor aqueous solubility and low bioavailability of curcumin have limited its potential when administrated orally. In this study, curcumin was encapsulated in a series of novel nano-microparticulate systems developed to improve its aqueous solubility and stability. The nano-microparticulate systems are based entirely on biocompatible, biodegradable, and edible polymers including chitosan, alginate, and carrageenan. The particles were synthesized via ionotropic gelation. Encapsulating the curcumin into the hydrogel nanoparticles yielded a homogenous curcumin dispersion in aqueous solution compared to the free form of curcumin. Also, the in vitro release profile showed up to 95% release of curcumin from the developed nano-microparticulate systems after 9 hours in PBS at pH 7.4 when freeze-dried particles were used.CONACYTCUPIAPharmac

Multi-Nozzle Biopolymer Deposition for Freeform Fabrication of Tissue Constructs

Advanced freeform fabrication techniques have been recently used for the construction of tissue scaffolds because of the process repeatability and capability of high accuracy in fabrication resolution at the macro and micro scales. Among many applicable tissue scaffolding materials, polymeric materials have unique properties in terms of the biocompatibility and degradation, and have thus been widely utilized in tissue engineering applications. Hydrogels, such as alginate, has been one of the most important polymer scaffolding materials because of its biocompatibility and internal structure similarity to that of the extracellular matrix of many tissues, and its relatively moderate processing. Three-dimensional deposition has been an entreating freeform fabrication method of biopolymer and particularly hydrogel scaffolds because of its readiness to deposit fluids at ambient temperatures. This paper presents a recent development of biopolymer deposition based freeform fabrication for 3-diemnsinal tissue scaffolds. The system configuration of multi-nozzles used in the deposition of sodium alginate solutions and Poly-?- Caprolactone (PCL) are described. Studies on polymer deposition feasibility and structural formability are conducted, and the preliminary results are presented.Mechanical Engineerin

Artificial leaf device for hydrogen generation from immobilised C. reinhardtii microalgae

We developed a fully biomimetic leaf-like device for hydrogen production which allows incorporated fabric-immobilised microalgae culture to be simultaneously hydrated with media and harvested from the produced hydrogen in a continuous flow regime without the need to replace the algal culture. Our leaf device produces hydrogen by direct photolysis of water resulting from redirecting the photosynthetic pathways in immobilised microalgae due to the lack of oxygen. In contrast to the many other reports in the literature on batch photobioreactors producing hydrogen from suspension culture of microalgae, we present the first report where this is done in a continuous manner from a fabric-immobilised microalgae culture. The reported artificial leaf device maximises the sunlight energy utilisation per gram of algae and can be upscaled cheaply and easily to cover large areas. We compared the production of hydrogen from both immobilised and suspended cultures of C. reinhardtii microalgae under sulphur, phosphorus and oxygen deprived conditions. The viability and potential of this approach is clearly demonstrated. Even though this is a first prototype, the hydrogen yield of our artificial leaf device is twenty times higher per gram of algae than in previously the reported batch reactors. Such leaf-like devices could potentially be made from flexible plastic sheets and installed on roofs and other sun-exposed surfaces that are inaccessible by photovoltaic cells. The ability to continuously produce inexpensive hydrogen by positioning inexpensive sheets onto any surface could have an enormous importance in the field of biofuels. The proposed new concept can provide a cleaner and very inexpensive way of bio-hydrogen generation by flexible sheet-like devices

Alginate hydrogel has a negative impact on in vitro collagen 1 deposition by fibroblasts

Hydrogels have been widely investigated as 3D culture substrates because of their reported structural similarity to the extracellular matrix (ECM). Limited ECM deposition, however, occurs within these materials, so the resulting “tissues” bear little resemblance to those found in the body. Here matrix deposition by fibroblasts encapsulated within a calcium alginate (Ca-alg) hydrogel was investigated. Although the cells transcribed mRNA for coll Iα over a period of 3 weeks, very little collagen protein deposition was observed within the gel by histology or immunohistochemistry (IHC). Although molecular diffusion demonstrated charge dependency, this did not prevent the flux of both positively and negative charged amino acids through the gel, suggesting that the absence of ECM could not be attributed to substrate limitation. The flux of protein, however, was charge-dependent as proteins with a net negative charge passed quickly through the Ca-alg into the medium. The minimal collagen deposition within the Ca-alg was attributed to a combination of rapid movement of negatively charged procollagen through the gel and steric hindrance of fibril formation

Utilizing osteocyte derived factors to enhance cell viability and osteogenic matrix deposition within IPN hydrogels

Many bone defects arising due to traumatic injury, disease, or surgery are unable to regenerate, requiring intervention. More than four million graft procedures are performed each year to treat these defects making bone the second most commonly transplanted tissue worldwide. However, these types of graft suffer from a limited supply, a second surgical site, donor site morbidity, and pain. Due to the unmet clinical need for new materials to promote skeletal repair, this study aimed to produce novel biomimetic materials to enhance stem/stromal cell osteogenesis and bone repair by recapitulating aspects of the biophysical and biochemical cues found within the bone microenvironment. Utilizing a collagen type I-alginate interpenetrating polymer network we fabricated a material which mirrors the mechanical and structural properties of unmineralized bone, consisting of a porous fibrous matrix with a young's modulus of 64 kPa, both of which have been shown to enhance mesenchymal stromal/stem cell (MSC) osteogenesis. Moreover, by combining this material with biochemical paracrine factors released by statically cultured and mechanically stimulated osteocytes, we further mirrored the biochemical environment of the bone niche, enhancing stromal/stem cell viability, differentiation, and matrix deposition. Therefore, this biomimetic material represents a novel approach to promote skeletal repair

The Use of Poly(vinyl alcohol)-based Hydrogels in Biomedical Applications

Polymers have found increasing favor in biomedical applications due to the greater control that researchers can exert over their properties. Researchers have focused on the development of therapies using biologically compatible polymers due to their ability to limit potentially harmful interactions with the body. This research has led to advances in tissue engineering, controlled and targeted drug delivery, and other biomedical fields, with the goal of improving both the effectiveness and accessibility of health care. Poly(vinyl alcohol) (PVA) hydrogels possess several chemical properties that make them well suited for biomedical applications. These include inertness and stability, biocompatibility, and pH-responsiveness. As a result, PVA based materials have been studied for potential applications in areas of biomedicine such as targeted drug delivery, tissue engineering, and wound healing. This thesis examines the properties of PVA and seeks to understand how the chemical and physical structure affects their properties. It then examines how these properties enhance their utility in potential biomedical applications. Finally, it reviews the research into development of PVA based materials for three different biomedical applications.Chemical Engineerin