Microencapsulação de oleoresina de cúrcuma (curcuma longa l.) em misturas de goma arábica, maltodextrina e amido modificado

Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

Nanocompósitos com atividade antioxidante de amido de milho/isolado proteico de soro de leite/ óleo essencial de alecrim obtidos por extrusão termoplástica

In order to improve the quality and safety of food, the active packaging emerges as a new technology based on the release of composites beneficial to food products. Thus, biodegradable films incorporated with active substances have the function of acting as a barrier to external elements, protecting the product and increasing its shelf life. They are formulated from proteins, polysaccharides, lipids or from the combination of these compounds. However, there is a need to improve the performance properties of these packages. Nanotechnologies, then, emerges with the study of many nanoparticles as additives to modify the performance of biodegradable polymers. With this, we aimed at developing and active antioxidant film of corn starch blenders and whey protein isolate with rosemary essential oil or microcapsules of rosemary essential oil reinforced with sodium montmorillonite (MMTNa + ) nanoparticles by extrusion. The films were developed and characterized in a first stage for the selection of the best polymeric blender using the following analyses: water vapor permeability (WVP), machanical properties; optical, thermogravimetry (TG), differential scanning calorimetry (DSC), x-ray diffraction (XRD) and scanning electron microscopy (SEM). In the second stage, montmorillonite clay nanoparticles and rosemary essential oil were added as reinforcement to evaluate its antioxidant effect. In a third stage, we studied the addition of microcapsules of rosemary essential oil (MR) as a form of protecting the active agent and its antioxidant potential in the films. The results indicate that the development of p olymeric blender with 30% of corn starch substitution is the most indicated for future work. The addition of rosemary essential oil or microcapsule of rosemary essential oil allowed for the obtaining of nanocomposites with antioxidant potential for application in food packages.

Uranium-lead dating method at the Pará-Iso isotope geology laboratory, UFPA, Belém – Brazil

ABSTRACT: Analytical procedures for U-Pb isotope dilution analyses at the Pará-Iso isotope geology laboratory of the Federal University of Pará (UFPA) are described in detail. The procedures are applied to zircon, titanite, rutile, apatite, columbite-tantalite and whole rock. Reagent preparation and chemical processing are done in clean-room conditions. Samples are dissolved using Teflon™ microcapsules in steel jacket Teflon™ Parr Instrument™ bomb or Teflon™ screw cap containers. U and Pb are separated using anion exchange AG 1x8 resin columns. Typical blanks for mineral sample amounts of 0.01 to 1.0 mg are less than 1 pg U and 20-30 pg Pb. Isotope analysis of the U and Pb from the same filament are carried out using a Finnigan MAT 262 mass-spectrometer in static and dynamic modes. The current analytical level is demonstrated on analyses of international standard zircon 91500 with three different ²³⁵U-²⁰⁵Pb and ²³⁵U-²⁰⁸Pb isotope tracers and whole rock standards. Results of analyses of two zircon samples are also presented.

Microencapsulação de óleo de palma por coacervação complexa em matrizes de gelatina/goma arábica e gelatina/alginato

Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

Stability of curcumin microencapsulated by spray and freeze drying in binary and ternary matrices of maltodextrin, gum arabic and modified starch

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

Microencapsulação por gelificação iônica e interação eletrostática do corante de buriti (Mauritia flexuosa L. f.)

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

Microencapsulation of lycopene by gelatin-pectin complex coacervation

The aim of this work was to produce and characterize microcapsules of lycopene and to evaluate their stability in comparison with free lycopene. An oily dispersion of lycopene was encapsulated by complex coacervation using gelatin and pectin. Samples were analyzed at four different pH values (3, 3.5, 4 and 4.5) and three proportions of core (25, 50 and 100%). The moisture, water activity, solubility, hygroscopicity, encapsulation efficiency and stability of lycopene microcapsules kept at 10 and 25C were determined. The amount of lycopene in the microcapsule did not have a significant (P < 0.05) effect on water activity, hygroscopic characteristics or the efficiency of microencapsulation. The degradation of lycopene was linear, with an average loss of 14% per week. Therefore, despite the formation of microcapsules and the high values of encapsulation efficiency, the encapsulation method and the wall materials used in this work did not provide effective protection of the lycopene from degradation during storage.

Morteros acumuladores con parafinas microencapsuladas para el aprovechamiento de la energía solar en suelos radiantes

Esta Tesis plantea la pregunta de si el uso de morteros con parafinas microencapsuladas combinado con colectores solares térmicos puede reducir el consumo de energías convencionales, en un sistema tradicional de suelo radiante. Se pretende contribuir al conocimiento acerca del efecto que produce en el edificio, el calor latente acumulado en suelos radiantes, utilizando morteros de cemento Portland con material de cambio de fase (PCM), en conjunto con la energía solar. Para cumplir con este propósito, la investigación se desarrolla considerando diversos aspectos. En primer lugar, se revisa y analiza la documentación disponible en la actualidad, de almacenamiento de energía mediante calor latente en la construcción, y en particular la aplicación de microcápsulas de PCM en morteros y suelos radiantes. También se revisa la documentación relacionada con la aplicación de la energía solar térmica y en suelo radiante. Se analiza la normativa vigente respecto al material, a los colectores solares y al suelo radiante. Se verifica que no hay normativa relacionada con mortero-PCM, debido a esto se aplica en la investigación una adaptación de la existente. La fase experimental desarrollada esta principalmente dirigida a la cuantificación, caracterización y evaluación de las propiedades físicas, mecánicas y térmicas del mortero de cemento Portland con parafinas microencapsuladas. Los resultados obtenidos y su análisis, permiten conocer el comportamiento de este tipo de morteros, con las diferentes variables aplicadas en la investigación. Además, permite disponer de la información necesaria, para crear una metodología para el diseño de morteros con parafina microencapsulada, tanto del punto de vista de su resistencia a la compresión y contenido de PCM, como de su comportamiento térmico como acumulador de calor. Esto se logra procesando la información obtenida y generando modelos matemáticos, para dosificar mezclas, y predecir la acumulación de calor en función de su composición. Se determinan los tipos y cantidades de PCM, y el cemento más adecuado. Se obtienen importantes conclusiones respecto a los aspectos constructivos a considerar en la aplicación de morteros con PCM, en suelo radiante. Se analiza y evalúa la demanda térmica que se puede cubrir con el suelo radiante, utilizando morteros con parafina microencapsulada, a través de la acumulación de energía solar producida por colectores solares, para condiciones climáticas, técnicas y tipologías constructivas específicas. Se determina que cuando los paneles cubren más de 60 % de la demanda por calefacción, se puede almacenar en los morteros con PCM, el excedente generado durante el día. Se puede cubrir la demanda de acumulación de energía con los morteros con PCM, en la mayoría de los casos analizados. Con esto, se determina que el uso de morteros con PCM, aporta a la eficiencia energética de los edificios, disminuyendo el consumo de energías convencionales, reemplazándola por energía solar térmica. En esta investigación, el énfasis está en las propiedades del material mortero de cemento-PCM y en poder generar metodologías que faciliten su uso. Se aborda el uso de la energía solar, para verificar que es posible su acumulación en morteros con PCM aplicados en suelo radiante, posibilitando el reemplazo de energías convencionales. Quedan algunos aspectos de la aplicación de energía solar a suelo radiante con morteros con PCM, que no han sido tratados con la profundidad que requieren, y que resultan interesantes de evaluar en este tipo de aplicaciones constructivas, como entre otros, los relacionados con la cuantificación de los ahorros de energía en las diferentes estaciones del año, de la estabilización de temperaturas internas, su análisis de costo y la optimización de este tipo de sistemas para utilización en verano, los que dan pie para otras Tesis o proyectos de investigación. ABSTRACT This Thesis proposes the question of whether the use of mortars with microencapsulated paraffin combined with solar thermal collectors can reduce conventional energy consumption in a traditional heating floor system. It aims to contribute to knowledge about the effect that it has on the building, the latent heat accumulated in heating floor, using Portland cement mortars with phase change material (PCM), in conjunction with solar energy. To fulfill this purpose, the research develops it considering various aspects. First, it reviews and analyzes the documentation available today, about energy storage by latent heat in the building, and in particular the application of PCM microcapsules in mortars and heating floors. It also reviews the documentation related to the application of solar thermal energy and heating floor. Additionally, it analyzes the current regulations regarding to material, solar collectors and heating floors. It verifies that there aren’t regulations related to PCM mortar, due to this, it applies an adaptation in the investigation. The experimental phase is aimed to the quantification, mainly, characterization and evaluation of physical, mechanical and thermal properties of Portland cement mortar with microencapsulated paraffin. The results and analysis, which allow us to know the behavior of this type of mortars with different variables applied in research. It also allows having the information necessary to create a methodology for designing mortars with microencapsulated paraffin, both from the standpoint of its resistance to compression and PCM content, and its thermal performance as a heat accumulator. This accomplishes by processing the information obtained, and generating mathematical models for dosing mixtures, and predicting heat accumulation depending on their composition. The research determines the kinds and amounts of PCM, and the most suitable cement. Relevant conclusions obtain it regarding constructive aspects to consider in the implementation of PCM mortars in heating floor. Also, it analyzes and evaluates the thermal demand that it can be covered in heating floor using microencapsulated paraffin mortars, through the accumulation of solar energy produced by solar collectors to weather conditions, technical and specific building typologies. It determines that if the panels cover more than 60% of the demand for heating, the surplus generated during the day can be stored in PCM mortars. It meets the demand of energy storage with PCM mortars, in most of the cases analyzed. With this, it determines that the use of PCM mortars contributes to building energy efficiency, reducing consumption of conventional energy, replacing it with solar thermal energy. In this research approaches the use of solar energy to determine that it’s possible to verify its accumulation in PCM mortars applied in heating floor, enabling the replacement of conventional energy. The emphasis is on material properties of PCM mortar and, in order to generate methodologies to facilitate their use. There are some aspects of solar energy application in PCM mortars in heating floor, which have not been discussed with the depth required, and that they are relevant to evaluate in this kind of construction applications, including among others: the applications related to the energy savings quantification in different seasons of the year, the stabilizing internal temperatures, its cost analysis and optimization of these systems for use in summer, which can give ideas for other thesis or research projects.

Co-encapsulation of enzymes and antibodies for chemical deactivation of pathogens on paper

Le papier bioactif est obtenu par la modification de substrat du papier avec des biomolécules et des réactifs. Ce type de papier est utilisé dans le développement de nouveaux biocapteurs qui sont portables, jetables et économiques visant à capturer, détecter et dans certains cas, désactiver les agents pathogènes. Généralement les papiers bioactifs sont fabriqués par l’incorporation de biomolécules telles que les enzymes et les anticorps sur la surface du papier. L’immobilisation de ces biomolécules sur les surfaces solides est largement utilisée pour différentes applications de diagnostic comme dans immunocapteurs et immunoessais mais en raison de la nature sensible des enzymes, leur intégration au papier à grande échelle a rencontré plusieurs difficultés surtout dans les conditions industrielles. Pendant ce temps, les microcapsules sont une plate-forme intéressante pour l’immobilisation des enzymes et aussi assez efficace pour permettre à la fonctionnalisation du papier à grande échelle car le papier peut être facilement recouvert avec une couche de telles microcapsules. Dans cette étude, nous avons développé une plate-forme générique utilisant des microcapsules à base d’alginate qui peuvent être appliquées aux procédés usuels de production de papier bioactif et antibactérien avec la capacité de capturer des pathogènes à sa surface et de les désactiver grâce à la production d’un réactif anti-pathogène. La conception de cette plate-forme antibactérienne est basée sur la production constante de peroxyde d’hydrogène en tant qu’agent antibactérien à l’intérieur des microcapsules d’alginate. Cette production de peroxyde d’hydrogène est obtenue par oxydation du glucose catalysée par la glucose oxydase encapsulée à l’intérieur des billes d’alginate. Les différentes étapes de cette étude comprennent le piégeage de la glucose oxydase à l’intérieur des microcapsules d’alginate, l’activation et le renforcement de la surface des microcapsules par ajout d’une couche supplémentaire de chitosan, la vérification de la possibilité d’immobilisation des anticorps (immunoglobulines G humaine comme une modèle d’anticorps) sur la surface des microcapsules et enfin, l’évaluation des propriétés antibactériennes de cette plate-forme vis-à-vis l’Escherichia coli K-12 (E. coli K-12) en tant qu’un représentant des agents pathogènes. Après avoir effectué chaque étape, certaines mesures et observations ont été faites en utilisant diverses méthodes et techniques analytiques telles que la méthode de Bradford pour dosage des protéines, l’électroanalyse d’oxygène, la microscopie optique et confocale à balayage laser (CLSM), la spectrométrie de masse avec désorption laser assistée par matrice- temps de vol (MALDI-TOF-MS), etc. Les essais appropriés ont été effectués pour valider la réussite de modification des microcapsules et pour confirmer à ce fait que la glucose oxydase est toujours active après chaque étape de modification. L’activité enzymatique spécifique de la glucose oxydase après l’encapsulation a été évaluée à 120±30 U/g. Aussi, des efforts ont été faits pour immobiliser la glucose oxydase sur des nanoparticules d’or avec deux tailles différentes de diamètre (10,9 nm et 50 nm) afin d’améliorer l’activité enzymatique et augmenter l’efficacité d’encapsulation. Les résultats obtenus lors de cette étude démontrent les modifications réussies sur les microcapsules d’alginate et aussi une réponse favorable de cette plate-forme antibactérienne concernant la désactivation de E. coli K-12. La concentration efficace de l’activité enzymatique afin de désactivation de cet agent pathogénique modèle a été déterminée à 1.3×10-2 U/ml pour une concentration de 6.7×108 cellules/ml de bactéries. D’autres études sont nécessaires pour évaluer l’efficacité de l’anticorps immobilisé dans la désactivation des agents pathogènes et également intégrer la plate-forme sur le papier et valider l’efficacité du système une fois qu’il est déposé sur papier.

Co-encapsulation of enzymes and antibodies for chemical deactivation of pathogens on paper

Le papier bioactif est obtenu par la modification de substrat du papier avec des biomolécules et des réactifs. Ce type de papier est utilisé dans le développement de nouveaux biocapteurs qui sont portables, jetables et économiques visant à capturer, détecter et dans certains cas, désactiver les agents pathogènes. Généralement les papiers bioactifs sont fabriqués par l’incorporation de biomolécules telles que les enzymes et les anticorps sur la surface du papier. L’immobilisation de ces biomolécules sur les surfaces solides est largement utilisée pour différentes applications de diagnostic comme dans immunocapteurs et immunoessais mais en raison de la nature sensible des enzymes, leur intégration au papier à grande échelle a rencontré plusieurs difficultés surtout dans les conditions industrielles. Pendant ce temps, les microcapsules sont une plate-forme intéressante pour l’immobilisation des enzymes et aussi assez efficace pour permettre à la fonctionnalisation du papier à grande échelle car le papier peut être facilement recouvert avec une couche de telles microcapsules. Dans cette étude, nous avons développé une plate-forme générique utilisant des microcapsules à base d’alginate qui peuvent être appliquées aux procédés usuels de production de papier bioactif et antibactérien avec la capacité de capturer des pathogènes à sa surface et de les désactiver grâce à la production d’un réactif anti-pathogène. La conception de cette plate-forme antibactérienne est basée sur la production constante de peroxyde d’hydrogène en tant qu’agent antibactérien à l’intérieur des microcapsules d’alginate. Cette production de peroxyde d’hydrogène est obtenue par oxydation du glucose catalysée par la glucose oxydase encapsulée à l’intérieur des billes d’alginate. Les différentes étapes de cette étude comprennent le piégeage de la glucose oxydase à l’intérieur des microcapsules d’alginate, l’activation et le renforcement de la surface des microcapsules par ajout d’une couche supplémentaire de chitosan, la vérification de la possibilité d’immobilisation des anticorps (immunoglobulines G humaine comme une modèle d’anticorps) sur la surface des microcapsules et enfin, l’évaluation des propriétés antibactériennes de cette plate-forme vis-à-vis l’Escherichia coli K-12 (E. coli K-12) en tant qu’un représentant des agents pathogènes. Après avoir effectué chaque étape, certaines mesures et observations ont été faites en utilisant diverses méthodes et techniques analytiques telles que la méthode de Bradford pour dosage des protéines, l’électroanalyse d’oxygène, la microscopie optique et confocale à balayage laser (CLSM), la spectrométrie de masse avec désorption laser assistée par matrice- temps de vol (MALDI-TOF-MS), etc. Les essais appropriés ont été effectués pour valider la réussite de modification des microcapsules et pour confirmer à ce fait que la glucose oxydase est toujours active après chaque étape de modification. L’activité enzymatique spécifique de la glucose oxydase après l’encapsulation a été évaluée à 120±30 U/g. Aussi, des efforts ont été faits pour immobiliser la glucose oxydase sur des nanoparticules d’or avec deux tailles différentes de diamètre (10,9 nm et 50 nm) afin d’améliorer l’activité enzymatique et augmenter l’efficacité d’encapsulation. Les résultats obtenus lors de cette étude démontrent les modifications réussies sur les microcapsules d’alginate et aussi une réponse favorable de cette plate-forme antibactérienne concernant la désactivation de E. coli K-12. La concentration efficace de l’activité enzymatique afin de désactivation de cet agent pathogénique modèle a été déterminée à 1.3×10-2 U/ml pour une concentration de 6.7×108 cellules/ml de bactéries. D’autres études sont nécessaires pour évaluer l’efficacité de l’anticorps immobilisé dans la désactivation des agents pathogènes et également intégrer la plate-forme sur le papier et valider l’efficacité du système une fois qu’il est déposé sur papier.

Electrodeposition of composite copper/liquid-containing microcapsule coatings

This paper presents results on the preparation of microcapsules containing liquid organosilica, and their co-deposition with copper in an acidic copper electrolyte onto a carbon steel cathode to form a copper/microcapsule composite coating. Microscopic analyses of the surface and the cross-section of the coating confirm the incorporation of the liquid-containing microcapsules in the coating layer. The influence of microcapsules in the electrolyte on the cathode polarization, as well as that of process conditions on the microcapsule inclusion, is also discussed. (C) 2004 Kluwer Academic Publishers.

Emulsion strategies in the microencapsulation of cells: Pathways to thin coherent membranes

Microencapsulation of cell spheroids in an immunoselective, highly biocompatible, biomembrane offers a way to create viable implantation options in the treatment of insulin-dependent diabetes mellitus (IDDM). Traditionally the encapsulation process has been achieved through the injection/extrusion of alginate/cell mixtures into a calcium chloride solution to produce calcium alginate capsules around the cells. A novel alternative is explored here through a procedure using an emulsion process to produce thin adherent calcium alginate membranes around cell spheroids. In this study, a thorough investigation has been used to establish the emulsion process parameters that are critical to the formation of a coherent alginate coat both on a model spheroid system and subsequently on cell spheroids. Optical and fluorescence microscopy are used to assess the morphology and coherence of the calcium alginate/ poly-L-ornithine/alginate (APA) capsules produced. (c) 2005 Wiley Periodicals, Inc.

Microencapsulation studies with P(HB-HV) polymers

Microencapsulation processes, based upon the concept of solvent evaporation, have been employed within these studies to prepare microparticles from poly--hydroxybutyrate homopolymers and copolymers thereof with 3-hydroxyvalerate [P(HB-HV) polymers]. Variations in the preparative technique have facilitated the manufacture of two structurally distinct forms of microparticle. Thus, monolithic microspheres and reservoir-type microcapsules have been respectively fabricated by single and double emulsion-solvent evaporation processes. The objective of the studies reported in chapter three is to asses how a range of preparative variables affect the yield, shape and surface morphology of P(HB-HV) microcapsules. The following chapter then describes how microcapsule morphology in general, and microcapsule porosity in particular, can be regulated by blending the fabricating P(HB-HV) polymer with poly--caprolactone [PCL]. One revelation of these studies is the ability to generate uniformly microporous microcapsules from blends of various high molecular weight P(HB-HV) polymers with a low molecular weight form of PCL. These microcapsules are of particular interest because they may have the potential to facilitate the release of an encapsulated macromolecule via an aqueous diffusion mechanism which is not reliant on polymer degradation. In order to investigate this possibility, one such formulation is used in chapter five to encapsulate a wide range of different macromolecules, whose in vitro release behaviour is subsequently evaluated. The studies reported in chapter six centre on the preparation and characterization of hydrocortisone-loaded microspheres, prepared from a range of P(HB-HV) polymers, using a single emulsion-solvent evaporation process. In this chapter, the influence of the organic phase viscosity on the efficiency of drug encapsulation is the focus of initial investigations. Thereafter, it is shown how the strategies previously adopted for the regulation of microcapsule morphology can also be applied to single emulsion systems, with profound implications for the rate of drug release.

Incorporation and release of macromolecules from biodegradable polymer vehicles

The initial objective of this work was to evaluate and introduce fabrication techniques based on W/0/W double emulsion and 0/W single emulsion systems with solvent evaporation for the incorporation of a surrogate macromolecule (BSA) into microspheres and microcapsules fabricated using P(HB-HV}, PEA and their blends. Biodegradation, expressed as changes in the gross and ultrastructural morphology of BSA loaded microparticulates with time was monitored using SEM concomitant with BSA release. Spherical microparticulates were successfully fabricated using both the W/0/W and 0/W emulsion systems. Both microspheres and microcapsules released BSA over a period of 24 to 26 days. BSA release from P(HB-HV)20% PCL 11 microcapsules increased steadily with time, while BSA release from all other microparticulates was characterised by an initial lag phase followed by exponential release lasting 6-11 days. Microcapsules were found to biodegrade more rapidly than microspheres fabricated from the same polymer. The incubation of microparticulates in newborn calf serum; synthetic gastric juice and pancreatin solution showed that microspheres and microcapsules were susceptible to enzymatic biodegradation. The in vitro incubation of microparticulates in Hank's buffer demonstrated limited biodegradation of microspheres and microcapsules by simple chemical hydrolysis. BSA release was thought to ocurr as a result of the macromolecule diffusing through either inherent micropores or via pores and channels generated in situ by previously dissolved BSA. However, in all cases, irrespective of percentage loading or fabrication polymer, low encapsulation efficiencies were obtained with W/0/W and 0/W techniques (4.2±0.9%- 15.5±0.5%,n=3), thus restricting the use of these techniques for the generation of microparticulate sustained drug delivery devices. In order to overcome this low encapsulation efficiency, a W/0 single emulsion technique was developed and evaluated in an attempt to minimise the loss of the macromolecule into the continuous aqueous phase and increase encapsulation efficiency. Poly(lactide-co-glycolide) [PLCG] 75:25 and 50:50, PEA alone and PEA blended with PLCG 50:50 to accelerate biodegradation, were used to microencapsulate the water soluble antibiotic vancomycin, a putative replacement for gentamicin in the control of bacterial infection in orthopaedic surgery especially during total hip replacement. Spherical microspheres (17.39±6.89~m,n=74-56.5±13.8~m,n=70) were successfully fabricated with vancomycin loadings of 10, 25 and 50%, regardless of the polymer blend used. All microspheres remained structurally intact over the period of vancomycin release and exhibited high percentage yields( 40. 75±2 .86%- 97.16±4.3%,n=3)and encapsulation efficiencies (47.75±9.0%- 96.74±13.2%,n=12). PLCG 75:25 microspheres with a vancomycin loading of 50% were judged to be the most useful since they had an encapsulation efficiency of 96.74+13.2%, n=12 and sustained therapeutically significant vancomycin release (15-25μg/ml) for up to 26 days. This work has provided the means for the fabrication of a spectrum of prototype biodegradable microparticulates, whose biodegradation has been characterised in physiological media and which have the potential for the sustained delivery of therapeutically useful macromolecules including water soluble antibiotics for orthopaedic applications.

Microencapsulation strategies for islet transplantation

A variety of islet microencapsulation techniques have been investigated to establish which method provides the least occlusive barrier to net insulin release in vitro, and optimum biocompatibility for islet implantation in vivo. NMRI mouse islets have been microencapsulated with Na+ -alginate-poly-L-lysine (PLL)/poly-L-ornithine (PLO)-alginate, Ba2+ -alginate and agarose gels. Both free and microencapsulated islets responded to glucose challenge in static incubation and perifusion by significantly increasing their rate of insulin release and theophylline significantly potentiated the insulin response to glucose. While little insulin was released from microencapsulated islets after short term (2 hours) static incubation, significantly greater amounts were released in response to glucose challenge after extended (8-24 hours) incubation. However, insulin release from all types of microencapsulated islets was significantly reduced compared with free islets. Na+ -alginate-PLO-alginate microencapsulated islets were significantly more responsive to elevated glucose than Na+ -alginate-PLL-alginate microencapsulated islets, due to the enhanced porosity of PLO membranes. The outer alginate layer created a significant barrier to glucose/insulin exchange and reduced the insulin responsiveness of microencapsulated islets to glucose. Ba2+ -alginate membrane coated islets, generated by the density gradient method, were the most responsive to glucose challenge. Low concentrations of NG-monomethyl L-arginine (L-NMMA) had no significant effect on glucose stimulated insulin release from either free or microencapsulated islets. However, 1.0 mmol/1 L-NMMA significantly inhibited the insulin response of both free and microencapsulated islets to glucose challenge. In vivo work designed to evaluate the extent of pericapsular fibrosis after 28 days ip. and sc. implantation of microencapsulated islets into STZ-diabetic recipients, revealed that the inclusion of islets within microcapsules increased their immunogenicity and markedly increased the extent of pericapsular fibrosis. When the outer alginate layer was omitted from microcapsules, little or no pericapsular mononuclear cell deposition was observed. The subcutaneous site was not suitable for microencapsulated islet transplantation in NMRI recipient mice. Systemic immunosuppression using cyclosporin A was effective in preventing pericapsular mononuclear cell deposition, while L-NMMA loading into microcapsules had no significant effect on pericapsular fibrosis, although it did maintain the integrity of microencapsulated islets.