Additive manufacturing of non plastic porcelain material by direct writing and freeze casting.

Two direct consolidation methods usually used for advanced ceramics have been combined in this project in order to develop a novel fabrication route for traditional ceramics. Specifically the method used is based on the Additive Manufacturing extrusion process using direct writing of high solid loading ceramic pastes and then freeze-casting to solidify the deposited material. This novel fabrication method, for which a patent has been granted, has been christened “Direct Writing Freeze-Casting” (DWFC). Although the DWFC process is the subject of investigation by other researchers for a range of different applications, including the production of medical implants with alumina, the research presented in this thesis focuses on its use in the manufacture of white wares, giftware, and applied arts and crafts in general. This new system will provide designers, potters, artists, craft makers and manufacturers with a flexible and automated way of manufacturing porcelain objects. One of the major challenges to be overcome to exploit the DWFC process is the development of suitable slurry material formulations. Initial trials demonstrated that it is not possible to use conventional clay based porcelain materials with a platelet shaped microstructure which inhibits freeze casting. In this thesis the development and characterisation of non plastic porcelain slurry, based on substitution of kaolin (clay) with a calcined clay material (molochite), which can be processed using this new method is presented. The new non plastic porcelain formulation, which has a high solid load of 75.47% wt., has been subjected to detailed analysis to assess its suitability at each stage of the process; extrusion, freeze-casting (solidification) and firing.National Consul of Sciences and Technology Mexico, CONACY

Development of novel functional thickened drinks enriched with vitamin D3 for the older adult population behaviour under dynamic in vitro digestion

The older adult population is greatly affected by malnutrition. This condition can arise from inappropriate micro and micronutrient intake, as well as due to experienced age-related changes. For instance, a significant number of older adults suffer from dysphagia. Thus, the development of foods with adequate consistency/viscosity are vital for overcome this condition. Within this framework, novel functional thickened drinks were developed and tailored for the older adult population, by using chia seed gum as the thickening agent enriched with vitamin D3- loaded solid lipid nanoparticles (SLN). Rheological properties of different thickened drinks formulated with water, semi-skimmed milk or orange juice were optimized using a central composite rotational design. Results showed that SLN delivered high vitamin D3 encapsulation efficiency with high concentration (i.e., 32 IU/mg). The different drink matrices attained distinct rheological properties, being observed that the viscosity and consistency attained with the same thickening agent concentration was higher in more complex matrices. An optimized formulation containing the tolerable upper intake level (ca. 4000 IU) was selected to study the vitamin D3 release and bioaccessibility on older adults population using a dynamic in vitro gastrointestinal digestion system. Once more, strong matrix effects were observed and significantly affected vitamin D3 release profile, bioaccessibility and recovery during the in vitro digestion. Additionally, the drinks enrichment with SLN-VitD3 did not display a negative effect on cells viability. The development of novel functional thickened drinks tailored for older adults could provide a safer swallowing and improved vitamin D3 intake.Jean-Michel Fernandes and Joana T. Martins acknowledge the Portuguese Foundation for Science and Technology (FCT) for his fellowship with reference SFRH/BD/147286/2019 (https://doi.org/10.54499/SFRH/BD/147286/2019 and for her Assistant Research contract obtained under the scope of Scientific Stimulus Employment with reference 2022.00788.CEECIND/CP1718/CT0024 (https://doi.org/10.54499/2022.00788.CEECIND/CP1718/CT0024), respectively. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, with DOI 10.54499/UIDB/04469/2020, and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020. The authors are thankful to Lipoid for kindly donating LIPOID S 75 samples.info:eu-repo/semantics/publishedVersio

Tissue Bioprinting:A granular approach

The field of bioprinting aims to fabricate tissues and organs that can be used to model drug efficacy and disease mechanisms, repair and ultimate replace dysfunctional parts in living beings. It utilizes computer aided design to create a construct that mimics native tissue, by adding step-by-step every detail of the complex multiscale hierarchical structure. This approach, demands the ability to be able to a) design a-priori the features of the tissue, and b) have the ability to actually create the features of that resolution. In extrusion bioprinting, hydrogels containing cells are used as the bioink, and then the desired structure is extruded through a nozzle forming the desired pattern. The hydrogel matrix attempts to mimic the extracellular matrix (ECM), and the cells attempt to grow within that matrix. However, creating a hydrogel that can mimic the dynamic nature of ECM, which changes in response to cellular behavior is non-trivial. Moreover, the resolution necessary is in the order of 10-100μm which leads to the challenge of fabricating very small nozzles, and ensuring that the shear stress during extrusion remains minimal.This thesis describes i) an approach where particle suspensions are 3D printed in anembedding bath and result in self assembled complex tissue fibers that are a fraction of a nozzle, utilizing a first time measured phenomenon of diffusive packing, ii) the invention of a platform where controlling the volume fraction of the particle suspensions allows them to transition reversibly between granular bioinks and embedding baths, as well as a newly observed intermediate state of freestanding embedding baths, iii) the 3D printing of bioactive microgel inks into bioactive embedding baths that cells can populate the granular interstitial space iv) theinvention of a freeze-dying method that allows those granular architectures to formmodular blocks that can be used as tissue blueprints, and have an extended shelf life, a necessary component for clinical translation. Together, this thesis presents how to 3D print cells and particles that can self-assemble into tissues, how to make blueprints for tissues, surpassing current customization and resolution limits and how to preserve them into a shelf steady state so that one day they can be translated into a clinical application

Exopolysaccharide production by Enterobacter A47: optimization of cultivation conditions and study of polymer functional properties

Dissertação para obtenção do Grau de Mestre em BiotecnologiaFucoPol is a fucose-containing biopolymer produced by the bacterium Gram-negative, Enterobacter A47 DSM 23139. It is an exopolysaccharide (EPS) composed of neutral sugars (fucose, galactose, glucose), an acidic sugar (glucuronic acid), and also non-saccharide substituents (acetate, pyruvate and succinate). In this work, two primary objectives were considered: the optimization of EPS production by varying two different parameters, and the functional properties’ assessment of different EPS produced by the bacterium. In the optimization study, the influence of different dissolved oxygen concentrations (controlled at 10, 30 and 60% of air saturation), and the impact of phosphate concentration reduction in the culture medium (5.25 and 3.81 g L-1) were assessed. The objective was to evaluate not only the effect of both parameters on cellular growth and exopolysaccharide-synthesis, but also to determine their influence in EPS chemical composition. The best results were obtained with DO at 10%, with the highest EPS production (6.11 g L-1), and although a reduction of phosphate concentration didn’t affect the EPS production, it reduced the exopolysaccharide’s fucose content to 24%mol. Secondly, the functional properties of nine distinct exopolysaccharides synthesized by Enterobacter A47 under different cultivation conditions were evaluated. Rheologically, most of the EPS polymer solutions showed shear-thinning behavior, wherein EPS-s, EPS-g and GNEX achieved the highest apparent viscosity (0.2 - 0.3 Pa.s) at lower shear rates (0.3 to 1 s-1). Also, most of the polymers presented emulsifying capacity at different temperatures. Concerning the film-forming capacity, EPS-s, EPS-g and EPS-x films demonstrated to have good mechanical properties and high water vapour permeability. And, all polymers described flocculating activity at 0.01% of EPS, which was significantly diminished at lower concentration, 0.001%

Development of non-ionic surfactant vesicles for inhaled drug delivery

Previously held under moratorium from 1 December 2016 until 6 December 2021Non-ionic surfactant vesicles (NIVs) provide similar bioavailability as liposomes but they are associated with higher stability profiles and reduced production cost. However, NIVs formulations are limited by the stability of the encapsulated drug and little is know about their use for inhaled delivery. Therefore, in this study, NIVs for pulmonary delivery were developed and factors, which affected their efficacy and stability, were determined. The addition of the cryoprotectant sucrose to NIVs containing amphotericin B (AmB-NIVs) allowed the physicochemical characteristics of the vesicles to be maintained upon freeze-drying and rehydration. Those were proven stable up to 4 months and up to 21 days when reconstituted and stored at 4°C. Pulmonary administration of AmB-NIVs was able to supress the progress of visceral leishmaniasis in vivo with larger particles being superior to smaller sizes (p ≤ 0.05 in the liver and spleen). Incursion of hydroxypropyl-γ-cyclodextrin (HPγCD), rather than sucrose (p ≤ 0.01), protected empty-NIVs when freeze-dried and rehydrated with different drugs. Therefore, empty-NIVs rehydrated with drugs such as cisplatin or gemcitabine, could be formulated in situ evading undesirable drug instability issues. The effect of using different types of vibrating-mesh nebulisers (active and passive) on the stability of AmB-NIVs was determined. AmB-NIVs properties, like drug entrapment efficiency, were not critically impaired upon nebulisation. The NIVs viscosity, the type of drug nebulised and the inhaler used affected the final aerosolised outcome i.e. aerosol output and aerosol droplet size. In vitro screening of novel minor binders (MGBs) against B16 F10 cells and L. donovani identified potential lead compounds. Studies in a murine model of V visceral leishmaniasis showed that intravenous administration of MGB58 encapsulated within NIVs reduced the parasite burdens in the liver (p ≤ 0.05). In conclusion, NIVs have been optimised to provide a stable and successful inhaled drug delivery platform with enhanced bioavailability, especially to target intracellular diseases such as leishmaniasis.Non-ionic surfactant vesicles (NIVs) provide similar bioavailability as liposomes but they are associated with higher stability profiles and reduced production cost. However, NIVs formulations are limited by the stability of the encapsulated drug and little is know about their use for inhaled delivery. Therefore, in this study, NIVs for pulmonary delivery were developed and factors, which affected their efficacy and stability, were determined. The addition of the cryoprotectant sucrose to NIVs containing amphotericin B (AmB-NIVs) allowed the physicochemical characteristics of the vesicles to be maintained upon freeze-drying and rehydration. Those were proven stable up to 4 months and up to 21 days when reconstituted and stored at 4°C. Pulmonary administration of AmB-NIVs was able to supress the progress of visceral leishmaniasis in vivo with larger particles being superior to smaller sizes (p ≤ 0.05 in the liver and spleen). Incursion of hydroxypropyl-γ-cyclodextrin (HPγCD), rather than sucrose (p ≤ 0.01), protected empty-NIVs when freeze-dried and rehydrated with different drugs. Therefore, empty-NIVs rehydrated with drugs such as cisplatin or gemcitabine, could be formulated in situ evading undesirable drug instability issues. The effect of using different types of vibrating-mesh nebulisers (active and passive) on the stability of AmB-NIVs was determined. AmB-NIVs properties, like drug entrapment efficiency, were not critically impaired upon nebulisation. The NIVs viscosity, the type of drug nebulised and the inhaler used affected the final aerosolised outcome i.e. aerosol output and aerosol droplet size. In vitro screening of novel minor binders (MGBs) against B16 F10 cells and L. donovani identified potential lead compounds. Studies in a murine model of V visceral leishmaniasis showed that intravenous administration of MGB58 encapsulated within NIVs reduced the parasite burdens in the liver (p ≤ 0.05). In conclusion, NIVs have been optimised to provide a stable and successful inhaled drug delivery platform with enhanced bioavailability, especially to target intracellular diseases such as leishmaniasis

Sunnah foods for the treatment of infertility in women with PCOS: a literature review

Infertility is a global health issue that is on the rise. The leading cause of female infertility, PCOS, is considered incurable and lifelong in conventional medicine. Challenging and delayed diagnosis, ineffective treatment modalities and long-term treatment has left both physicians and patients disgruntled. Islamically however, it is believed that every illness has a cure. This paper is therefore aimed at reviewing the effect of Sunnah foods in treating infertility among patients with PCOS. The study will also provide future recommendation for researchers and practitioners

La diffusion sélective lors de la déshydratation osmotique de la mangue : impact de la solution hypertonique et des prétraitements

La déshydratation osmotique est une technique de séchage partiel ayant lieu à de basses températures (moins de 50°C), permettant de préserver la qualité des produits comparativement au séchage conventionnel. De plus, une formulation du produit peut être réalisée en utilisant des solutions riches en composés bénéfiques, comme le sirop d'agave qui contient des prébiotiques (inuline). Elle est pratique pour conserver les fruits saisonniers comme la mangue dont les caractéristiques organoleptiques et nutritionnelles la placent parmi les fruits les plus consommés au monde. Cependant, l'imprégnation de solutés (sucres ou sels) augmente la teneur calorique des produits après la déshydratation osmotique. De plus le sucrose, le soluté le plus utilisé, n'est pas adapté à certains consommateurs qui y sont intolérants. Enfin, la teneur élevée en sucres dans les aliments est incriminée dans les maladies cardiovasculaires et l'obésité. C'est pourquoi, ce projet avait pour but d'optimiser le procédé de déshydratation osmotique de la mangue afin de produire des mangues déshydratées osmotiquement avec du sirop d'agave et ayant une teneur ajoutée en sucres réduite. Dans un premier temps, la viscosité et la rhéologie des solutions osmotiques de composition différentes ont été caractérisées, suivi de la déshydratation osmotique des morceaux de mangues Tommy Atkins d'épaisseurs 0.4 cm et 1.5 cm. Les résultats ont montré que l'augmentation de la viscosité, de la taille des molécules de solutés ainsi que de l'épaisseur de la mangue peuvent permettre de réduire le gain en sucres ajoutés tout en maintenant une quantité suffisante de perte en eau. En second lieu, des analyses par chromatographie liquide haute performance de la quantité et du profil en sucres individuels des mangues déshydratées dans les différentes solutions osmotiques, ont montré que la composition initiale de la mangue en différents sucres ainsi que la composition de la solution osmotique influencent le profil final en sucres. Une perte en sucrose et un gain en fructose et glucose ont été observés dans la mangue lorsque des solutions pauvres en sucrose ont été utilisées, permettant ainsi de moduler le profil final de sucres du produit. La présence d'inuline a été détectée dans la mangue après la déshydratation osmotique, ce composé prébiotique est bénéfique pour la flore intestinale et est une valeur ajoutée dans le produit final. Une analyse par microscopie électronique à balayage a permis d'observer le mode de dépôt des différents solutés sur la mangue au cours de la déshydratation osmotique, et ainsi déterminer les mécanismes par lesquels une réduction d'entrée de solides est possible. Finalement, des prétraitements de congélation/décongélation et de champ électrique pulsé ont permis de modifier la structure microscopique de la mangue avant de la soumettre à la déshydratation osmotique. Cette étape a montré que le type de prétraitement impacte l'effet sur le transfert de matières. La congélation/décongélation a augmenté le gain en sucres au détriment de la perte en eau, et l'effet du champ électrique pulsé (dans les écarts des variables utilisées dans cette étude) était négligeable sur le transfert de matières en général. Cependant, l'utilisation de solutions osmotiques à viscosité élevée a permis de réduire le gain en sucres dans le cas des mangues dont la structure cellulaire a été sévèrement endommagée par la congélation/décongélation. Cette thèse constitue une contribution dans la production de mangues déshydratées (et de fruits en général) ayant une teneur en sucres ajoutés réduite et des ingrédients fonctionnels tels que l'inuline qui est bénéfique pour l'organisme.Osmotic dehydration is a partial drying technique which necessitates low temperatures (less than 50°C), allowing product quality to be preserved compared to conventional drying. It allows product formulation throughout solutions rich in beneficial compounds, such as agave syrup which contains prebiotics (inulin). It is practical for preserving seasonal fruits such as mango which organoleptic and nutritional characteristics rank it among the most consumed fruits in the world. However, the impregnation of solutes (sugars or salts) increases the caloric content of products after osmotic dehydration. In addition, sucrose, the most used solute, is not suitable for certain consumers who are sucrose intolerant. Finally, the high content of sugars in food is incriminated in cardiovascular diseases and obesity. Therefore, this project aimed at optimizing mango osmotic dehydration process to produce osmotically dehydrated mangoes in agave syrup and with low sugar content. Firstly, viscosity and rheology of osmotic solutions of different compositions were characterized, followed by the osmotic dehydration of Tommy Atkins mangoes with thicknesses of 0.4 cm and 1.5 cm. The results showed that increasing solution viscosity, solutes molecules size, as well as mango thickness can reduce sugar gain while maintaining enough water loss. Secondly, high performance liquid chromatography results showed that initial composition of mango sugars as well as composition of osmotic solution influence the final sugar profiles of dehydrated mango. A loss in sucrose together with a gain in fructose and glucose have been reported in mango when osmotic solutions with low concentration of sucrose were used. Inulin was found in mango after osmotic dehydration in solutions containing inulin, this prebiotic compound is beneficial for gut microbiota and is therefore an added value in the final product. An analysis by scanning electron microscopy demonstrated the behavior of different solutes on the mango surface during osmotic dehydration allowing the understanding of the mechanisms by which solids gain could be reduced. Finally, freeze-thawing, and pulsed electric field pretreatments were applied to mango to modify its tissue structure before osmotic dehydration. Results indicated that the type of pretreatment impacts the mass transfer differently. Freeze-thawing increased sugar gain and negatively affects water loss, where as pulsed electric field effect was negligible on mass transfer in general. However, high viscosity osmotic solutions reduced sugar gain for frozen-thawed mango. This thesis contributes to the research field of processed mangoes and in general, processed fruits, with low sugar content together with added functional ingredients such as inulin which is beneficial for the gut microbiota

Effect of high pressure homogenization (HPH) on the rheological properties of tomato juice

Orientadores: Marcelo Cristianini, Albert IbarzTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: A homogeneização a alta pressão (HAP) tem sido estudada por diversos autores como metodologia não térmica para a conservação de alimentos, especialmente produtos de frutas. Trabalhos realizados com esses produtos e testes industriais indicam aumento de consistência devido a HAP. No entanto, poucos trabalhos da literatura estudam alterações físico-químicas em produtos de frutas devido à HAP, especialmente em relação às características reológicas. A avaliação de tais alterações é essencial não só para o entendimento e dimensionamento do processo, mas também permite a utilização do método para provocar alterações intencionais e desejáveis em alimentos, como aumento de consistência. O presente trabalho estudou o efeito da homogeneização a alta pressão (HAP) nas características reológicas de suco de tomate. Na primeira parte do trabalho, avaliaram-se as propriedades reológicas dependentes do tempo, em estado estacionário e de viscoelasticidade linear de sucos não processados, determinando assim as técnicas e os modelos matemáticos mais adequados para utilização nas demais etapas do projeto. Em seguida avaliou-se o efeito da HAP nas características reológicas de um modelo de soro de suco de tomate. A HAP reduziu a viscosidade do modelo, sendo tal redução modelada como função da pressão de homogeneização (PH). Avaliou-se então o efeito da HAP no suco. A HAP reduziu o tamanho das partículas em suspensão, assim como sua distribuição, promovendo maior interação entre partículas e entre partículas e o soro. Como consequência, resultou em aumento da consistência, tixotropía, comportamento viscoso e elástico dos produtos. O efeito da PH se mostrou assintótico, sendo discutidas as razões para tal, modelando-se os parâmetros obtidos como função da PH. Por fim, avaliou-se o efeito da HAP nas propriedades de deformação e relaxação do suco de tomate. Utilizando o modelo de Burger, identificaram-se os componentes estruturais do suco de tomate associados aos comportamentos reológicos. Dessa forma, o presente trabalho descreveu as mudanças estruturais associadas ao processo que afetam a reologia do produto. Conclui-se que a HAP é uma poderosa ferramenta para promoção de alterações físicas em produtos de frutasAbstract: High pressure homogenization (HPH) has been widely studied as a non-thermal method for food preservation. Industrial assessments and scientific works have demonstrated the consistency increasing in those products due to HPH. However, just a few works in literature have studied the physico-chemical changes in fruit products due HPH processes. The evaluation of rheological changes due to HPH is important not only for process design, but also for using this technology in order to promote desirable changes in food products, such as improvement of its consistency. The present work studied the effect of high pressure homogenization (HPH) on the rheological properties of tomato juice. Firstly, it was evaluated the tomato juice time-dependent, steady-state shear and viscoelastic properties. This first part was carried out in order to evaluate the rheological methods and mathematical models to be used in the work. Then, it was evaluated the effect of HPH on the rheological properties of a tomato juice serum model. The HPH decreased the serum model viscosity, which was modelled as function of the homogenization pressure (PH). The HPH effect on tomato juice was then evaluated. The HPH decreased the suspended particle dimensions and distribution, resulting in higher particle-particle and serum-particle interaction. As a result, it was observed an increasing in product consistency, thixotropy, viscous and elastic behaviour. The effect of PH showed an asymptotic behaviour, which was described and modelled. Finally, it was evaluated the effect of HPH on tomato juice creep and recovery properties. The mechanical Burger model well explained the juice creep compliance, and its parameters were associated to the juice structural components. Therefore, the present work described the tomato juice structural and rheological changes due to HPH processing. It was concluded that the HPH process is a valuable tool to promote physical properties changes in food productsDoutoradoTecnologia de AlimentosDoutor em Tecnologia de Alimento

Biomimetic synthetic lubricants (Biosurfactants)

Poly(styrene-co-maleic anhydride) (PSMA) based copolymers are known to undergo conformational transition in response to environmental stimuli. This smart behaviour makes it possible to mimic the behaviour of native apoproteins. The primary aim of this study was to develop a better understanding of the structure-property relationships of various PSMA-based copolymers sought. The work undertaken in this thesis has revealed that the responsive behaviour of PSMA-based copolymers can be tailored by varying the molecular weight, hydrophobic (styrene) and hydrophilic (maleic acid) balance, and more so in the presence of additional hydrophobic, mono-partial ester moieties. Novel hydrophilic and hydrophobic synthetic surfactant protein analogues have successfully been prepared. These novel lipid solubilising agents possess a broad range of HLB (hydrophilic-lipophilic balance) values that have been estimated. NMR spectroscopy was utilised to confirm the structures for PSMA-based copolymers sought and proved useful in furthering understanding of the structure-property relationships of PSMA-based copolymers. The association of PSMA with the polar phospholipid, 2-dilauryl-sn-glycero-3- phosphocholine (DLPC) produces polymer-lipid complexes analogous to lipoprotein assemblies present in the blood plasma. NMR analysis reveals that the PSMA-based copolymers are not perfectly alternating. Regio-irregular structures, atactic and random monomer sequence distribution have been identified for all materials studied. Novel lipid solubilising agents (polyanionic surfactants) have successfully been synthesised from a broad range of PSMA-based copolymers with desired estimated HLB values that interact with polar phospholipids (DLPC/DPPC) uniquely. Very low static and dynamic surface tensions have been observed via the du Noϋy ring method and Langmuir techniques and correlate well with the estimated HLB values. Synthetic protein-lipid analogues have been successfully synthesised, that mimic the unique surface properties of native biological lubricants without the use of solvents. The novel PSMA-DLPC complexes have successfully been combined with hyaluronan (hyaluronic acid, HA). Today, the employment of HA is economically feasible, because it is readily available from bacterial fermentation processes in a thermally stable form - HyaCare®. The work undertaken in this thesis highlights the usage of HA in biolubrication applications and how this can be optimised and thus justified by carefully selecting the biological source, concentration, molecular weight, purity and most importantly by combining it with compatible boundary lubricating agents (polar phospholipids). Experimental evidence supports the belief that the combined HA and PSMA-DLPC complexes provide a balance of rheological, biotribological and surface properties that are composition dependent, and show competitive advantage as novel synthetic biological lubricants (biosurfactants)

Proliposome formulations for delivery via medical nebulisers

This study aims to investigate the ability of proliposomes to generate liposomes for delivery using air-jet, ultrasonic and vibrating-mesh nebulisers. Particulate-based proliposomes successfully generated liposomes under static conditions. Manually dispersed proliposomes generated multilamellar vesicles, with formulation having a small effect on the liposome size. Using sucrose as a carrier, liposomes were generated or dispersed in situ from proliposomes within the medical nebulisers investigated. The Pari (air-jet) and the Omron (vibrating-mesh) nebulisers produced large mass and lipid outputs with a large lipid fraction deposited in the lower stage of a two stage impinger. The Liberty (Ultrasonic) nebuliser failed to deliver more than 6% of the lipid employed. Multilamellar liposomes were generated from ethanol-based proliposomes. The resultant vesicles entrapped 62% of the available salbutamol sulphate compared to only 1.23% entrapped by liposomes made by the thin film method. Aeroneb Pro or Aeroneb Go vibrating-mesh nebulisers generated aerosol droplets of larger volume median diameter and narrower size distribution than the Pari (air-jet) nebuliser. Unlike the vibrating-mesh nebulisers, the performance of the jet nebuliser was largely independent of formulation. A nebuliser-dependent significant loss of the originally entrapped drug was demonstrated. A customised large mesh Aeroneb Pro reduced the drug losses during nebulisation. High sensitivity differential scanning calorimetry showed that the phospholipid phase transitions and liposomal bilayer interaction with beclometasone dipropionate were dependent on the method of liposome manufacture. Ethanol-based proliposomes produced liposomes having no pretransition, with a very low incorporation of the steroid (max. 1 mole%). This was attributed to an alcohol-induced interdigitation of the bilayers. 1 to 2.5 mole% steroid seemed to be optimal for incorporation in liposomes manufactured by the thin film or particulate-based proliposome method. Jet-nebulisation of particulate-based proliposomes delivered vesicles with enhanced steroid incorporation compared to liposomes generated by manual dispersion of these proliposomes