Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

The association between tridimensional scaffolds to cells of interest has provided excellent perspectives for obtaining viable complex tissues in vitro, such as skin, resulting in impressive advances in the field of tissue engineering applied to regenerative therapies. The use of multipotent mesenchymal stromal cells in the treatment of dermo-epidermal wounds is particularly promising due to several relevant properties of these cells, such as high capacity of proliferation in culture, potential of differentiation in multiple skin cell types, important paracrine and immunomodulatory effects, among others. Membranes of chitosan complexed with xanthan may be potentially useful as scaffolds for multipotent mesenchymal stromal cells, given that they present suitable physico-chemical characteristics and have adequate tridimensional structure for the adhesion, growth, and maintenance of cell function. Therefore, the purpose of this work was to assess the applicability of bioactive dressings associating dense and porous chitosan-xanthan membranes to multipotent mesenchymal stromal cells for the treatment of skin wounds. The membranes showed to be non-mutagenic and allowed efficient adhesion and proliferation of the mesenchymal stromal cells in vitro. In vivo assays performed with mesenchymal stromal cells grown on the surface of the dense membranes showed acceleration of wound healing in Wistar rats, thus indicating that the use of this cell-scaffold association for tissue engineering purposes is feasible and attractive.

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Obtaining of xanthan gum impregnated of cellulose microfibrils derived from sugarcane bagasse

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Drag coefficient in a crystal suspension

A single air bubble rising in xanthan gum crystal suspension has been studied experimentally. The suspension was made by different concentrations of xanthan gum solutions with 0.23 mm polystyrene crystal particles. Drag co-efficient data and a new correlation of drag coefficient is presented for spherical and nonspherical bubbles in non-Newtonian crystal suspension. The correlation is developed in terms of the Reynolds number, Re and the bubble shape factor, � (the ratio between the surface equivalent sphere diameter to the volume equivalent sphere diameter). The experimental drag coefficient was found to be consistent with this new predicted correlation and published data over the ranges, 0.1

Crushed tablets : does the administration of food vehicles and thickened fluids to aid medication swallowing alter drug release?

Purpose. To evaluate the influence of co-administered vehicles on in vitro dissolution in simulated gastric fluid of crushed immediate release tablets as an indicator for potential drug bioavailability compromise. Methods. Release and dissolution of crushed amlodipine, atenolol, carbamazepine and warfarin tablets were tested with six foods and drinks that are frequently used in the clinical setting as mixers for crushed medications (water, orange juice, honey, yoghurt, strawberry jam and water thickened with Easythick powder) in comparison to whole tablets. Five commercial thickening agents (Easythick Advanced, Janbak F, Karicare, Nutilis, Viscaid) at three thickness levels were tested for their effect on the dissolution of crushed atenolol tablets. Results. Atenolol dissolution was unaffected by mixing crushed tablets with thin fluids or food mixers in comparison to whole tablets or crushed tablets in water, but amlodipine was delayed by mixing with jam. Mixing crushed warfarin and carbamazepine tablets with honey, jam or yoghurt caused them to resemble the slow dissolution of whole tablets rather than the faster dissolution of crushed tablets in water or orange juice. Crushing and mixing any of the four medications with thickened water caused a significant delay in dissolution. When tested with atenolol, all types of thickening agents at the greatest thickness significantly restricted dissolution, and products that are primarily based on xanthan gum also delayed dissolution at the intermediate thickness level. Conclusions. Dissolution testing, while simplistic, is a widely used and accepted method for comparing drug release from different formulations as an indicator for in vivo bioavailability. Thickened fluids have the potential to retard drug dissolution when used at the thickest levels. These findings highlight potential clinical implications of the addition of these agents to medications for the purpose of dose delivery and indicate that further investigation of thickened fluids and their potential to influence therapeutic outcomes is warranted.

Delivering crushed tablets using thickened fluids : is drug diffusion into gastric fluids restricted?

Solid medications are often crushed and mixed with food or thickened water to aid drug delivery for those who cannot or prefer not to swallow whole tablets or capsules. Dysphagic patients have the added problem of being unable to safely swallow thin fluids so water thickened with polysaccharides is used to deliver crushed medications and ensure safe swallowing. It is postulated that these polysaccharide systems may restrict drug release by reducing the diffusion of the drug into gastric fluids. METHODS By using a vertical diffusion cell separated with a synthetic membrane, the diffusion of a model drug (atenolol) was studied from a donor system containing the drug dispersed into thickened water with xanthan gum (concentration range from 0.005%-2.2%) into a receptor system containing simulated gastric fluid (SGF) at 37°C. The amount of drug transferred was measured over 8 hours and diffusion coefficients estimated using the Higuchi model approach. RESULTS Atenolol diffusion decreased with increasing xanthan gum concentration up to 1.0%, above which diffusion remained around 300 μ2s-1. The rheological measurements captured the influence of the structure and conformation of the polysaccharide in water on the movement and availability of the drug in SGF. DISCUSSION Dose form administration for dysphagic patients’ needs special attention from general practitioners, pharmacist and patients. Improving drug release of crushed tablets from thickening agents requires a reduction in the diffusion pathway (e.g. by decreasing drop size radius). This approach could make the drug available in SGF in a short time without compromising the mechanical aspects of thickening agents that guarantee safe swallowing.

Oral medication delivery in impaired swallowing: Thickening liquid medications for safe swallowing alters dissolution characteristics

Acetaminophen (paracetamol) is available in a wide range of oral formulations designed to meet the needs of the population across the age-spectrum, but for people with impaired swallowing, i.e. dysphagia, both solid and liquid medications can be difficult to swallow without modification. The effect of a commercial polysaccharide thickener, designed to be added to fluids to promote safe swallowing by dysphagic patients, on rheology and acetaminophen dissolution was tested using crushed immediate-release tablets in water, effervescent tablets in water, elixir and suspension. The inclusion of the thickener, comprised of xanthan gum and maltodextrin, had a considerable impact on dissolution; acetaminophen release from modified medications reached 12-50% in 30 minutes, which did not reflect the pharmacopeia specification for immediate release preparations. Flow curves reflect the high zero-shear viscosity and the apparent yield stress of the thickened products. The weak gel nature, in combination with high G’ values compared to G” (viscoelasticity) and high apparent yield stress, impact drug release. The restriction on drug release from these formulations is not influenced by the theoretical state of the drug (dissolved or dispersed), and the approach typically used in clinical practice (mixing crushed tablets into pre-prepared thickened fluid) cannot be improved by altering the order of incorporation or mixing method.

Scalable weak aligning medium for enantiodiscrimination of water soluble chiral molecules

The study reports the first indication of a lyotropic liquid crystalline phase of an aqueous solution of polysaccharide xanthan gum, as a physical parameter dependent scalable and reversible weak alignment medium, for enantiodiscrimination of water soluble chiral molecules.

Study of physicochemical, microbial and sensory properties of surimi produced from black mouth croaker (Atrobucca nibe) at freezing condition (-18ºC)

Black mouth croaker (Atrobucca nibe) is considered as a new valuable fish stock in the Oman Sea. In this study, surimi was manufactured from nonmarket size of the fish, manually and different cryoprotectant agents were added to the surimi. Finally changes in physiochemical, microbiological and sensory quality, characteristics of the surimi and kamaboko gel samples were assessed during 6 months at freezing storage (-18ºC). Surimi samples with the addition of Iranian tragacanth gum (TG), xanthan gum (XG), chitosan (CS) and whey protein concentrate (WPC) at 1% (w/w) were prepared to evaluate their impacts as a cryoprotectant on the surimi, individually. The results showed that the whiteness and lightness indexes in all surimi samples were gradually decreased during frozen storage. This trend of decreasing was more intensity in the control sample from 61.08±0.131 to 54.21±0.067 was recorded (p<0.05). Water holding capacity (WHC) in all treatments was decreased during 6 months. The lowest WHC (g/g) was obtained in the surimi without cryoprotectants and maximum WHC was measured in Tcs and Twpc samples, respectively (p<0.05). The lowest breaking force was calculated in Txg (166.00±22.627 g) and Tc (271.50±263.16 g) during 6 months at frozen storage, respectively (p<0.05), while Twpc treatment with slight variations showed the highest breaking force (p<0.05). Also, the lowest gel strength was obtained in Txg (68.22±6.740 g.cm) after 6 month of frozen storage (p<0.05). All Kamaboko surimi gels texture profile analysis parameters decreaced with increasing shelf life. This decreasing trend in the control sample was more severe. Floding results were reduced in all samples during storage (p<0.05). The best protective results probably were obtained in WPC, chitosan and commercial cryoprotectant agents, respectively due to protein stabilization of myofibrillar proteins and the protein-protein network structure, leading to the formation of surimi gel with strong textural properties during frozen conditions. The average number of surimi polygonal structures were significantly decreased (number per mm2) and their area were significantly increased (μm2) in all treatments (p<0.05). With increasing storage time, moisture, protein contents and pH were decreaced. Maximun TVB-N index was calculated in Tc (7.93±0.400 mg/100g) and Txg (7.88±0.477), respectively (p<0.05). TBRAs index was increased in all treatments during frozen storage, while this trend was reached in maximum value in Tc (p<0.05). Sensory evaluation of the fish finger quality characteristics (color, odor, texture and overall acceptability) preapare from frozen black mouth croaker surimi was decreaced during 6 month frozen storage. After the period of frozen storage the highest quality scores were measured in Twpc, Tcs and Tcc samples, respectively (p<0.05). In this study, coliform bacteria were not found in all treatments during frozen storage. The surimi sample containing chitosan showed lower mesophilic and psychrotropic bacteria (log cfu/g) than other treatments during frozen storage (p<0.05). Salt-soluble proteins extractions of all treatments were decreased during frozen storage. This decreacing trend was highest in Tcs (45.74±0.176%) and lowest in Tc treatments after 6 month of frozen storage (29.92±0.224%) (p<0.05). Although commercial cryoprotectant agents were successful in limiting the denaturation of proteins but sugar contents were not accepted for diabetics or those who disagree with the sweet taste and high calorie food. Hence, commercial cryoprotectant agents can be replaced with whey protein concentrate and chitosan at 1% level (w/w) consider that they were showed proper protection of the surimi myofibrillar proteins during storage.

Cereal products for specific dietary requirements. Evaluation and improvement of technological and nutritional properties of gluten free raw materials and end products

Coeliac disease is one of the most common food intolerances worldwide and at present the gluten free diet remains the only suitable treatment. A market overview conducted as part of this thesis on nutritional and sensory quality of commercially available gluten free breads and pasta showed that improvements are necessary. Many products show strong off-flavors, poor mouthfeel and reduced shelf-life. Since the life-long avoidance of the cereal protein gluten means a major change to the diet, it is important to also consider the nutritional value of products intending to replace staple foods such as bread or pasta. This thesis addresses this issue by characterising available gluten free cereal and pseudocereal flours to facilitate a better raw material choice. It was observed that especially quinoa, buckwheat and teff are high in essential nutrients, such as protein, minerals and folate. In addition the potential of functional ingredients such as inulin, β-glucan, HPMC and xanthan to improve loaf quality were evaluated. Results show that these ingredients can increase loaf volume and reduce crumb hardness as well as rate of staling but that the effect diverges strongly depending on the bread formulation used. Furthermore, fresh egg pasta formulations based on teff and oat flour were developed. The resulting products were characterised regarding sensory and textural properties as well as in vitro digestibility. Scanning electron and confocal laser scanning microscopy was used throughout the thesis to visualise structural changes occurring during baking and pasta making

Development and mechanical characterization of solvent-cast polymeric films as potential drug delivery systems to mucosal surfaces

Solvent-cast films from three polymers, carboxymethylcellulose (CMC), sodium alginate (SA), and xanthan gum, were prepared by drying the polymeric gels in air. Three methods, (a) passive hydration, (b) vortex hydration with heating, and (c) cold hydration, were investigated to determine the most effective means of preparing gels for each of the three polymers. Different drying conditions [relative humidity - RH (6-52%) and temperature (3-45 degrees C)] were investigated to determine the effect of drying rate on the films prepared by drying the polymeric gels. The tensile properties of the CMC films were determined by stretching dumbbell-shaped films to breaking point, using a Texture Analyser. Glycerol was used as a plasticizer, and its effects on the drying rate, physical appearance, and tensile properties of the resulting films were investigated. Vortex hydration with heating was the method of choice for preparing gels of SA and CMC, and cold hydration for xanthan gels. Drying rates increased with low glycerol content, high temperature, and low relative humidity. The residual water content of the films increased with increasing glycerol content and high relative humidity and decreased at higher temperatures. Generally, temperature affected the drying rate to a greater extent than relative humidity. Glycerol significantly affected the toughness (increased) and rigidity (decreased) of CMC films. CMC films prepared at 45 degrees C and 6% RH produced suitable films at the fastest rate while films containing equal quantities of glycerol and CMC possessed an ideal balance between flexibility and rigidity.

Protection of cells from salinity stress by extracellular polymeric substances in diatom biofilms

Diatom biofilms are abundant in the marine environment. It is assumed (but untested) that extracellular polymeric substances(EPS), produced by diatoms, enable cells to cope with fluctuating salinity. To determine the protective role of EPS, Cylindrotheca closterium was grown in xanthan gum at salinities of 35, 50, 70 and 90 ppt. A xanthan matrix significantly increased cell viability (determined by SYTOX-Green), growth rate and population density by up to 300, 2, 300 and 200%, respectively. Diatoms grown in 0.75% w/v xanthan, subjected to acute salinity shock treatments (at salinities 17.5, 50, 70 and 90 ppt) maintained photosynthetic capacity, F_q′/F_m′, within 4% of pre-shock values, whereas F_q′/F_m′ in cells grown without xanthan declined by up to 64% with hypersaline shock. Biofilms that developed in xanthan at standard salinity helped cells to maintain function during salinity shock. These results provide evidence of the benefits of living in an EPS matrix for biofilm diatoms.

A comparative study of biopolymers and alum in the separation and recovery of pulp fibres from paper mill effluent by flocculation

Recovery of cellulose fibres from paper mill effluent has been studied using common polysaccharides or biopolymers such as Guar gum, Xanthan gum and Locust bean gum as flocculent. Guar gum is commonly used in sizing paper and routinely used in paper making. The results have been compared with the performance of alum, which is a common coagulant and a key ingredient of the paper industry. Guar gum recovered about 3.86 mg/L of fibre and was most effective among the biopolymers. Settling velocity distribution curves demonstrated that Guar gum was able to settle the fibres faster than the other biopolymers; however, alum displayed the highest particle removal rate than all the biopolymers at any of the settling velocities. Alum, Guar gum, Xanthan gum and Locust bean gum removed 97.46%, 94.68%, 92.39% and 92.46% turbidity of raw effluent at a settling velocity of 0.5 cm/min, respectively. The conditions for obtaining the lowest sludge volume index such as pH, dose and mixing speed were optimised for guar gum which was the most effective among the biopolymers. Response surface methodology was used to design all experiments, and an optimum operational setting was proposed. The test results indicate similar performance of alum and Guar gum in terms of floc settling velocities and sludge volume index. Since Guar gum is a plant derived natural substance, it is environmentally benign and offers a green treatment option to the paper mills for pulp recycling.

Preparation and characterization of novel nanocomposites of inorganic/polysaccharide type

O uso de polímeros naturais no âmbito da preparação de nanocompósitos não tem sido tão amplamente estudado quando comparado com os polímeros sintéticos. Assim, esta tese tem como objectivo estudar metodologias para a preparação de novos materiais nanocompósitos sob a forma de dispersões e filmes utilizando polissacarídeos como matriz. A tese está dividida em cinco capítulos sendo o último capítulo dedicado às conclusões gerais e a sugestões para trabalhos futuros. Inicialmente é apresentada uma breve revisão bibliográfica sobre os principais temas colocando esta tese em contexto. Considerações sobre o uso de polímeros naturais e a sua combinação com a utilização de nanopartículas inorgânicas para a fabricação de novos bionanocomposites são descritas e os objectivos e outline da tese são também apresentados. No segundo capítulo, a preparação de partículas de sílica puras ou modificadas bem como a sua caracterização por FTIR, SEM, TEM, TGA, DLS (tamanho e potencial zeta) e medições de ângulo de contacto são discutidas. De modo a melhorar a compatibilidade da sílica com os polissacarídeos, as partículas SiO2 foram modificados com dois compostos do tipo organosilano: 3- metacril-oxipropil-trimetoxissilano (MPS) e 3-aminopropil-trimetoxissilano (APS). As partículas SiO2@MPS foram posteriormente encapsuladas com de poli(metacrilato de glicidilo) utilizando a técnica de polimerização em emulsão. A utilização dos nanocompósitos resultantes na preparação de dispersões de bionanocompósitos não foi bem sucedida e por esse motivo não os estudos não foram prosseguidos. O uso de SiO2@APS na preparação de dispersões bionanocomposite foi eficiente. No terceiro capítulo é apresentada uma revisão sobre dispersões bionanocompósitas e respectiva caracterização destacando aspectos fundamentais sobre reologia e microestrutura. Em seguida, é discutido o estudo sistemático realizado sobre o comportamento reológico de dispersões de SiO2 utilizando três polissacarídeos distintos no que concerne a carga e as características gelificantes: a goma de alfarroba (não iónica), o quitosano (catiónico) e a goma xantana (aniónica) cujas propriedades reológicas são amplamente conhecidas. Os estudos reológicos realizados sob diferentes condições demonstraram que a formação de géis frágeis e/ou bem estruturados depende do tamanho SiO2, da concentração, do pH e da força iónica. Estes estudos foram confirmados por análises microestruturais usando a microscopia electrónica a baixas temperaturas (Cryo-SEM). No quarto capítulo, são apresentados os estudos relativos à preparação e caracterização de filmes bionanocompósitos utilizando quitosano como matriz. Primeiramente é apresentada uma revisão sobre filmes de bionanocompósitos e os aspectos fundamentais das técnicas de caracterização utilizadas. A escolha do plasticizante e da sua concentração são discutidas com base nas propriedades de filmes de quitosano preparados. Em seguida, o efeito da concentração de sílica e dos métodos utilizados para a dispersar na matriz de polissacarídeo, bem como o efeito da modificação da superfície da sílica é avaliado. As características da superfície e as propriedades de barreira, mecânicas e térmicas são discutidas para cada conjunto de filmes preparados antes e após a sua neutralização. Os resultados obtidos mostraram que a dispersão das cargas no plasticizante e posterior adição à matriz polissacarídica resultaram apenas em pequenas melhorias já que o problema da agregação de sílica não foi ultrapassado. Por esse motivo foram preparados filmes com SiO2@APS os quais apresentaram propriedades melhores apesar da agregação das partículas não ter sido completamente impedida. Tal pode estar relacionado com o processo de secagem dos filmes. Finalmente, no capítulo 5, são apresentadas as principais conclusões obtidas e algumas sugestões para trabalho futuro.