Gotowa bibliografia na temat „Bioactive Thin Film”

Porous bioactive thin film on commercially pure titanium substrate was prepared by micro-arc oxidation (MAO) in electrolytic solution, which contained calcium acetate, β-glycerol phosphate disodium salt pentahydrate (β-GP) and lanthanum nitrate. The phases and microstructure of the bioactive films were examined by X-ray diffraction, scanning electron microscopy with energy dispersive X-ray spectrometer and electron probe microanalysis. The results showed that: (1) porous bioactive films with about 10μm were formed on titanium substrate by MAO; (2) phases of the thin films were hydroxyapatite, anatase and rutile; (3) elements of Ca, P, and Ti of films were identified by EDS.

Poly methyl methacrylate(PMMA) with mar resistant or poly ethtylene(PE) substrates were surface-treated by applying plasma discharge or alkaline solution. Ceramic thin film comprised of silicon oxide, titanium oxide and zirconium oxide was formed on these surface treated substrates respectively from an aqueous solution, and after the formation of ceramic thin film, titanium oxide thin film was also coated on them from aqueous solution at ordinary temperature and pressure. The thin film coated polymer material was tested in mechanical property. The thin film was hard and the adhesion strength to the organic polymer substrate was very high. The substrate was soaked in SBF and apatite was formed on the substrate. This method is promising for developing hard and soft tissue implants with various mechanical properties as well as high bioactivity.

Pulsed laser deposition (PLD) is a relatively new technique for producing thin films. It presents unique advantages for the deposition of bioactive ceramics. The mechanism and characteristics of the technique PLD are introduced. Its applications and current research status in hydroxyapatite and bioglass thin films are reviewed. The effect of processing parameters of PLD, including atmosphere, substrate temperature, laser wavelength and target properties, on the structures and the properties of the hydroxyapatite film, is analyzed in detail. Future application trends are also analyzed.

Effect of the energy density on the composition, morphology and deposition rate of the bioglass thin films deposited by pulsed laser was studied by energy dispersive spectroscopy (EDS), scanning electron microscopy (SEM), X-ray diffraction (XRD) and crystal lattice monitor. There is slight compositional difference between the film and the target at lower energy density. Morphology of the films is compact with the particles on the surface of them. Structure of the films is amorphous glass and the size of the particles increases with the energy density. Deposition rate increases with the energy density and the energy density threshold of the film growth is about 2.5J/cm2.

We explored the potential of biomimetic thin films fabricated by means of matrix-assisted pulsed laser evaporation (MAPLE) for releasing combinations of active substances represented by flavonoids (quercetin dihydrate and resveratrol) and antifungal compounds (amphotericin B and voriconazole) embedded in a polyvinylpyrrolidone biopolymer; the antifungal activity of the film components was evaluated using in vitro microbiological assays. Thin films were deposited using a pulsed KrF* excimer laser source which were structurally characterized using atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR). High-quality thin films with chemical structures similar to dropcast ones were created using an optimum laser fluence of ~80 mJ/cm2. Bioactive substances were included within the polymer thin films using the MAPLE technique. The results of the in vitro microbiology assay, which utilized a modified disk diffusion approach and were performed using two fungal strains (Candida albicans American Type Culture Collection (ATCC) 90028 and Candida parapsilosis American Type Culture Collection (ATCC) 22019), revealed that voriconazole was released in an active form from the polyvinylpyrrolidone matrix. The results of this study show that the MAPLE-deposited bioactive thin films have a promising potential for use in designing combination devices, such as drug delivery devices, and medical device surfaces with antifungal activity.

Antibacterial efficacy of silver nanoparticles (Ag NPs) deposited alternatively layer by layer (LBL) on chitosan polymer in the form of a thin film over a quartz plate and stainless steel strip has been studied. An eight-bilayer chitosan/silver (Cs/Ag)8 hybrid was prepared having a known concentration of silver. Techniques such as UV-visible spectroscopy, inductively coupled plasma optical emission spectroscopy (ICP-OES), and atomic force microscopy (AFM) were carried out to understand and elucidate the physical nature of the film. Gram-negative bacteria, Escherichia coli (E. coli), were used as a test sample in saline solution for antibacterial studies. The growth inhibition at different intervals of contact time and, more importantly, the antibacterial properties of the hybrid film on repeated cycling in saline solution have been demonstrated. AFM studies are carried out for the first time on the microbe to know the morphological changes affected by the hybrid film. The hybrid films on aging (3 months) are found to be as bioactive as before. Cytotoxicity experiments indicated good biocompatibility. The hybrid can be a promising bioactive material for the prevention of biofilms specific to E. coli and in purification of water for safe drinking.

Mesoporous materials bear great potential for biotechnological applications due to their biocompatibility and versatility. Their high surface area and pore interconnection allow the immobilization of molecules and their subsequent controlled delivery. Modifications of the mesoporous material with the addition of different chemical species, make them particularly suitable for the production of bioactive coatings. Functionalized thin films of mesoporous silica and titania can be used as scaffolds with properties as diverse as promotion of cell growth, inhibition of biofilms formation, or development of sensors based on immobilized enzymes. The possibility to pattern them increase their appeal as they can be incorporated into devices and can be tailored both with respect to architecture and functionalization. In fact, selective surface manipulation is the ground for the fabrication of advanced micro devices that combine standard micro/nanofluids with functional materials. In this review, we will present the advantages of the functionalization of silica and titania mesoporous materials deposited in thin film. Different functional groups used to modify their properties will be summarized, as well as functionalization methods and some examples of applications of modified materials, thus giving an overview of the essential role of functionalization to improve the performance of such innovative materials.

Pulsed laser deposition (PLD) was used to deposit hydroxyapatite (HA) thin films on various substrates, including silicon (100) and titanium (Ti-6Al-4V) alloy. Thin films of amorphous HA were deposited at room temperature and then annealed over a range of temperatures. The microstructure and composition of the films were determined using scanning electron microscopy (SEM), energy dispersive x-ray spectroscopy (EDS), and X-ray diffraction (XRD). The HA films were found to achieve total crystallinity at 350㮠The mechanical properties of the films were studied by means of nanoindentation and scratch adhesion testing. Crystalline and adherent HA thin films prepared using PLD and post deposition annealing have many potential medical and dental applications.

Les applications présentées dans cette thèse valorisent de diverses manières le principe d'ablation laser, c'est à dire l'arrachement de la matière d'une surface solide suite à l'irradiation avec un faisceau laser. Le plasma généré par irradiation laser impulsionnel a été utilisé pour le dépôt de couches minces ou de nanoparticules et pour l'analyse compositionnelle des couches d'épaisseur nanométrique. Nous avons synthétisé par dépôt laser combinatoire des librairies compositionnelles d'un oxyde mixte transparent d'In et Zn. En utilisant le plasma d'ablation pour le diagnostic compositionnel, nous avons déterminé les concentrations d'indium et de zinc dans les couches minces par spectroscopie laser. Des couches minces de bioverre ont été synthétisées par dépôt laser impulsionnel sur des substrats de titane. En contact avec des cellules ostéoblastes, les bioverres ont stimulé la prolifération et ont augmenté la viabilité. La prolifération des ostéoblastes cultivés sur les couches de bioverre a été 30% supérieure a l'échantillon de contrôle. On a déposé par PLD des couches minces ou nanoparticules adhérentes de ZnO sur des substrats textiles hydrophiles dans un flux d'oxygène ou sous vide pour obtenir des structures avec différentes mouillabilités. En augmentant le nombre d'impulsions laser de 10 à 100 nous avons observé la transition du recouvrement par des nanoparticules isolées vers des couches minces. En fonction de l'atmosphère environnant lors du dépôt, les couches minces et les nanoparticules ont changé leur mouillabilité, passant d'hydrophile en flux d'oxygène à un comportement superhydrophobe (angle de contact de 157°) en cas de dépôt sous vide
The applications presented in this thesis exploit in different modes the principle of laser ablation, i.e. the material removal from a solid surface following irradiation with a pulsed laser beam. The plasma generated by laser ablation was used for thin films or nanoparticles deposition and for the compositional analysis of nanometric thin films. We synthesized by combinatorial pulsed laser deposition, thin film libraries of a complex oxide of In and Zn. Using the ablation plasma for compositional diagnostic, we determined the In and Zn concentrations in films by Laser Induced Breakdown Spectroscopy using a procedure based on the spectral luminance calculation of a plasma in local thermodynamic equilibrium. Thin films of bioactive glass were synthesized by pulsed laser deposition, magnetron sputtering and MAPLE on Ti substrates and tested the transfer accuracy by physico-chemical tests and their functionality in vitro. In contact with human osteoblast cells, the bioactive glasses stimulated their proliferation and enhanced their viability. The proliferation of osteoblasts cultivated on bioactive films was 30% superior to the control sample. ZnO thin films or nanoparticles were deposited on hydrophilic textile substrates in oxygen flux or in vacuum in order to obtain structures with different wetting behavior. Increasing the number of laser pulses from 10 to 100, we observed a coating transition from isolated nanoparticles to thin films fully coating the textile fibers. Function of the ambient atmosphere during experiments, the structures changed their wetting behavior, passing from hydrophilic in oxygen flux to superhydrophobic (157°) in case of deposition in vacuum

Capítulo 1. A investigação teve como objetivo desenvolver filmes finos de sílica micropadronizada (FFSM) contendo micro-agregados de nanohidroxiapatita (nanoHA) que não ficassem totalmente cobertos pela sílica e assim pudessem interagir diretamente com as células vizinhas e o objetivo específico foi avaliar o efeito da presença de dois filmes (FFSM com ou sem a adição de nanohidroxiapatita) na resistência característica (?0) e no módulo de Weibull (m) de uma Y-TZP. Processamento sol-gel e litografia foram usados para aplicar o FFSM sobre os espécimes de Y-TZP. Três grupos experimentais foram produzidos: Y-TZP, Y-TZP+FFSM e Y-TZP+FFSM+nanoHA borrifada. Todas as superfícies foram caracterizadas por MEV/EDS e testadas em resistência à flexão em quatro pontos (n=30) em água a 37°C. A análise de Weibull foi usada para determinar ?0 e m (método de probabilidade máxima). A Y-TZP foi recoberta com êxito com o FFSM e FFSM+nanoHA. Micrografias indicaram que os micro-agregados de nanoHA não foram totalmente cobertos pela sílica. Não houve diferença estatisticamente significativa entre os grupos experimentais para ?0 e m. Essa investigação obteve sucesso em produzir filmes finos de sílica micropadronizada contendo micro-agregados de nanoHA que permaneceram expostos ao meio ambiente. Os filmes desenvolvidos não prejudicaram a confiabilidade estrutural da Y-TZP comercial, como confirmado pela estatística de Weibull. Capítulo 2. Objetivos: avaliar o efeito da concentração de vidro bioativo (VB), de zero e 10% em massa e da temperatura de sinterização (1.200°C e 1.300°C) na microestrutura, densidade relativa e resistência à flexão do compósito Y-TZP/VB. Material e métodos: os pós de Y-TZP e Y-TZP/VB foram prensados uniaxialmente e sinterizados a 1.200°C e 1.300°C por 1 hora. A microestrutura foi caracterizada pela análise de difração de raios X, microscopia eletrônica de varredura e espectroscopia de energia dispersiva (EDS). A densidade relativa foi calculada por meio dos valores de densidade obtidos pelo princípio de Archimedes. Para a resistência à flexão, espécimes (n=6) foram fraturados no teste de resistência à flexão biaxial usando um dispositivo de pistão sobre 3 esferas em uma máquina universal de ensaios. Resultados: a adição de VB diminuiu o tamanho de grão do compósito, aumentou as porosidades e causou uma diminuição significativa na densidade relativa (Y-TZP/1.300°C=97,7%a; Y-TZP/1.200°C=91,1%b; Y-TZP/VB/1.300°C 79,7%c e Y-TZP/VB/1.200°C 77,4%d) e diminuiu também significativamente a resistência à flexão (em MPa, Y-TZP/1.300°C=628,3a; Y-TZP/1.200°C=560,8b; Y-TZP/VB=1.300°C=189,1c e Y-TZP/VB/1.200°C=153,0c). As fases cristalinas de zircônia cúbica estabilizada por cálcio e silicato de sódio zircônio foram formadas após a adição de VB. Conclusão: a adição de vidro bioativo na Y-TZP aumentou a porosidade e resultou na formação de zircônia cúbica estabilizada com cálcio e silicato de sódio zircônio. A adição de vidro também resultou na diminuição do tamanho de grão, densidade e resistência à flexão. Os espécimes sinterizados a 1.300°C mostraram valores de densidade superior e grãos maiores quando comparados ao grupo sinterizado a 1.200°C.
Chapter 1. This investigation aimed at developing micropatterned silica thin films (MSTF) containing nanoHA micro-aggregates that were not completely covered by silica so that they could directly interact with the surrounding cells and the specific objectives was to evaluate the effect of the presence of two films (MSTF with or without nanoHA addition) on the characteristic strength (?0) and Weibull modulus (m) of a Y-TZP. Sol-gel process and soft-lithography were used to apply the MSTF onto the Y-TZP specimens. Three experimental groups were produced: Y-TZP, Y-TZP+MSTF and Y-TZP+MSTF+sprayed nanoHA. All surfaces were characterized by SEM/EDS and tested for four-point flexural strength (n=30) in water at 37°C. Weibull analysis was used to determine m and ?0 (maximum likelihood method). Y-TZP was successfully coated with MSFT and MSFT+nanoHA. SEM micrographs indicated that the micro-aggregates of nanoHA were not entirely covered by the silica. There was no statistically significant difference among the experimental groups for ?0 and m. This investigation was successful in producing a micropatterned silica thin film containing nanoHA micro-aggregates that remained exposed to the environment. The developed films did not jeopardize the structural reliability of a commercial Y-TZP, as confirmed by the Weibull statistics. Chapter 2. Objectives: to evaluate the effect the bioactive glass (BG) concentration (0 and 10wt%) and the sintering temperature (1.200°C and 1.300°C) on the microstructure, relative density and flexural strength of the composite Y-TZP/BG. Methods: The Y-TZP and Y-TZP/BG powders were uniaxially pressed and sintered at 1.200°C or 1.300°C for 1 h. The microstructure was characterized by X-ray diffraction analysis, scanning electron microscopy and Energy Dispersive X-ray Spectroscopy. Relative density was calculated from density values obtained using the Archimedes\' principle. For the flexural strength, specimens (n=6) were fractured in a biaxial flexural setup using a piston-on-3-balls fixture in a universal testing machine. Results: BG addition decreased the grain size of the composite, increased porosity and caused a significant decrease in the relative density (Y-TZP/1.300°C=97.7%a; Y-TZP/1.200°C=91.1%b; Y-TZP/BG/1.300°C 79.7%c and Y-TZP/BG/1.200°C 77.4%d) and flexural strength (in MPa, Y-TZP/1.300°C=628.3a; Y-TZP/1.200°C=560.8b; Y-TZP/BG=1.300°C=189.1c and Y-TZP/BG/1.200°C=153.0c). The crystalline phases of calcium stabilized cubic zirconia and sodium zirconium silicate were formed after the addition of BG. Conclusion: Addition of bioactive glass to Y-TZP increased porosity and resulted in the formation of calcium stabilized cubic zirconia and sodium zirconium silicate. Also, glass addition resulted in decrease in grain size, density and flexural strength. Composite specimens sintered at 1.300°C showed the highest density values and larger grains compared to those sintered at 1.200°C

博士
國立成功大學
材料科學(工程)學系
86
First part of writing, the purer, crystalline hydroxyapatite (HA) films with thickness of roughly 10 um and 1 um have been deposited on titanium substrate using pulsed-laser deposition (PLD) technique. Experimental results indicate that structure and properties of the PLD-HA films varied with deposition parameters. The PLD process used in the present study did not induce significant amounts of other calcium phosphate phases than apatite, or significant changes in the behavior of hydroxyl or phosphate functional group. The EDS-determined Ca/P ratios of as-calcined HA powder (1.78) and sintered HA target for PLD (1.79) were very close. After PLD process, the Ca/P ratio of HA films increased to 1.99 and 2.71 for ~10 um and ~1um HA film. Cross-section SEM-EDS point analysis indicated ~10um HA film that the value of Ca/P ratio was significantly higher in the region near surface, particularly near coating-substrate interface, than in the coating interior. Broad face SEM showed that HA coating was comprised of numerous essentially spheroidal-shaped particles of different sizes, while the lateral morphology indicated that columnar and dome-shaped structures both existed in the film. The morphology significant change and recrystalline appeared at above 600℃ heat treatment. Adhesion strength of the coating, mostly in the range of 30-40 MPa for ~10 um thickness which was found closely related to the fractography of the tested specimen. However, the higher adhesion strength (70 MPa) have occurred at as-coated ~1 um thickness PLD-HA on lower roughness Ti substrate. The HA crystals may precipitate on the surface of HA coatings and the HA film no significant dissolve in same Hank''s solution tank. The second part, used two kind commercial HA, one was Merck''s HA (MHA) and the other was Ferak''s HA (FHA), the former was more thermal stable than the later. The effects of doped bioactive glass (BG) on the structure and properties of sintered hydroxyapatite (HA) have been studied. The result showed that the apatite structure of MHA will keep stable to 1350℃, and the FHA have occurred Beta-TCP at above 700℃. A 900℃ calcination treatment would increase the degree of crystallinity and amount of hydroxyl group in HA, but resulted in loss of a small amount of phosphorus. Anyway, the 900℃ calcination did not significant affect the densification of sintered body, for example, the final body density of MHA can reach to 98.5% of theoretically. Addition of BG in HA phase made decomposition and hydroxyl group breakdown processes decreased HA density and promoted microcracking when sintered at high temperatures. The optimal sintered properties were obtained at 1200-1300℃ for 100-1200 min. Practically, MHA, 2.5wt%BG-MHA, FHA, 2.5wt%BG-FHA sintered at 1250℃for 100 min, the FHA (120 MPa) bonding strength appeared more than MHA (93.5 MPa), it were higher about 30%. Have decomposed of sintered bodies (2.5wt%BG-MHA/ FHA/ 2.5wt%BG- FHA composition) may regain apatite structure and OH- group by post-heat treatment at 900℃ in air. Better mechanical properties performance of 900℃ post-heat treatment often appeared at 12-18 h holding time, as it is the bending strength of MHA and FHA were 123 MPa and 124 MPa. Although the better bending strength of sintered bodies for 2.5wt%BG-MHA (100 MPa) and 2.5wt%BG-FHA (119 MPa) have increased gradually by 900℃ heat treatment time, but still lower than MHA and FHA . The un- heat treatment group of MHA/ 2.5wt%BG-MHA/ FHA/ 2.5wt%BG-FHA sintered bodies may precipitate the HA crystals on the surface of all samples in Hank''s solution. The 900℃-18h heat treatment group of MHA/ 2.5wt%BG-MHA/ FHA/ 2.5wt%BG-FHA sintered bodies just odds and ends particles on the surface of all samples in Hank''s solution. Among of them, the addition of BG could dissolve more in same Hank''s solution tank.

Visando o aumento da biodisponibilidade dos compostos bioactivos administrados por via oral (com especial ênfase para a absorção bucal), o plano de trabalhos deste programa doutoral visou a optimização de formulações de filmes orais e de micro- e nanopartículas que, conjugados constituam sistemas inovadores com atividade sinérgica. As moléculas bioactivas seleccionadas para estudar o comportamento e eficácia das formulações optimizadas foram a cafeína e dois péptidos presentes na proteína do soro do leite com actividades antihipertensora (sequência: KGYGGVSLPEW) e relaxante (sequência: YLGYLEQLLR). O processo de optimização das formulações foi iniciado pela seriação e comparação preliminar dos excipientes para a produção de filmes e micro/nanopartículas. No primeiro estudo realizado, foram optimizadas e comparadas duas formulações de filmes (usando-se os polímeros carboximetilcelulose sódica e gelatina tipo A) como veículos para libertação oral de cafeína. Verificou-se, através da análise por espectroscopia no infravermelho por transformada de Fourier com reflectância total atenuada (FTIR-ATR), que a estrutura química da cafeína não fora alterada durante o processo de produção dos filmes. Concluiu-se também, através do ensaio de dissolução estabelecido pela Farmacopeia Americana (USP), que os filmes produzidos com gelatina tipo A permitiram uma libertação mais lenta da cafeína ao passo que os filmes de carboximetilcelulose apresentaram um perfil de libertação imediata. Em concordância, o valor de permeabilidade aparente da cafeína, determinada através do ensaio de permeabilidade ex vivo, através de excisões de intestino delgado de origem porcina, verificou-se superior quando esta foi veiculada pelos filmes de carboximetilcelulose, comparativamente com os filmes de gelatina tipo A. O tempo de desintegração de ambas as formulações mostrou-se, contudo, demasiado alto para formulações orodispersíveis, não ocorrendo desintegração completa após 30 segundos. Ainda na sequência da escolha do polímero com melhores características para integrar a composição de filmes orais, uma nova formulação contendo goma-guar foi optimizada por desenho factorial. Os filmes de goma-guar apresentaram características mecânicas e físico-químicas superiores às verificadas para os filmes de carboximetilcelulose e gelatina, tomando-se como factores de decisão a capacidade de absorção de água, a erosão em saliva artificial e o tempo de desintegração apresentados pelos filmes de goma-guar. Procedeu-se também à optimização (por desenho fatorial) de uma formulação de micropartículas de alginato que garantissem, em conjunto com os filmes de goma-guar, uma libertação controlada de cafeína, assim como uma maior biodisponibilidade da mesma. A associação de micropartículas de alginato aos filmes de gomaguar – GfB - não induziu alterações das características químicas da cafeína, de acordo com o verificado por FTIR-ATR, nem toxicidade para as linhas celulares usadas para mimetizar as mucosas bucal (TR146) e intestinal (Caco-2/HT29-MTX), de acordo com os resultados obtidos pelo ensaio de viabilidade celular MTT (Brometo de 3-(4,5-Dimethylthiazol-2-yl)-2,5- Diphenyltetrazólio). Adicionalmente, os perfis de libertação e permeabilidade in vitro (através das linhas celulares TR146 e Caco-2/HT29-MTX cultivadas em camada) e ex vivo (através de epitélio intestinal de origem porcina) mostraram-se mais lentos que os observados para as micropartículas de alginato, filmes de goma-guar ou com a solução controlo de cafeína. A formulação GfB promoveu o aumento do contacto efectivo entre a cafeína e o epitélio bucal, oferecendo uma permeação mais completa ao longo do tempo. De forma a incrementar também a biodisponibilidade do péptido KGYGGVSLPEW com actividade anti-hipertensora, as micropartículas de alginato foram substituídas por nanopartículas de ácido poli(láctico-co-glicólico) – PLGA - por estas oferecerem uma eficácia de associação superior, assim como um maior potencial de permeação das membranas biológicas, dado o tamanho de partícula ser significativamente inferior. A formulação de nanopartículas de PLGA foi optimizada por desenho factorial. O sistema compreendido pelas nanopartículas de PLGA associadas aos filmes de goma-guar (GfNp) não comprometeu a viabilidade das linhas celulares TR146 e Caco-2/HT29-MTX às concentrações testadas. O sistema desenvolvido promoveu a libertação e permeabilidade controladas do péptido, através das células TR146 e Caco-2/HT29- MTX cultivadas em camada, comparativamente com os filmes e nanopartículas isoladamente, assim como com a solução de péptido livre (controlo). Contudo, a permeabilidade aparente verificou-se superior para a formulação GfNp, comparativamente com as restantes formulações. Estes resultados deveram-se ao contacto íntimo entre o péptido e o epitélio absorptivo, promovido pela formulação GfNp. Verificou-se ainda, através da realização do ensaio in vitro da capacidade de inibição da enzima conversora da angiotensina I, que o péptido transportado por GfNp apresentava a maior actividade anti-hipertensora após ser sujeito à simulação do tracto gastrointestinal, comparativamente com o péptido transportado pelas nanopartículas ou filme, isoladamente, ou com a solução de péptido livre. O sistema previamente optimizado para a libertação do péptido antihipertensor foi também usado de forma a incrementar a biodisponibilidade do péptido relaxante alfa-casozepina (sequência: YLGYLEQLLR). Através do ensaio MTT, foi possível concluir que nenhuma das formulações comprometeu a viabilidade da linha celular TR146 e da co-cultura Caco-2 /HT29- MTX. Por isso, a permeabilidade do péptido, sujeito às condições do tracto gastrointestinal simulado, através dos modelos in vitro bucal e intestinal foi estudada. Verificou-se que a associação de nanopartículas de PLGA com filmes de goma-guar promoveu um aumento da permeabilidade face às nanopartículas e filmes não conjugados, assim como com o péptido em solução (controlo). Estes resultados estão correlacionados com o incremento da mucoadesão conferida pela associação das nanopartículas de PLGA com os filmes de goma-guar, verificada através da análise da adesividade e trabalho de adesão à língua de vaca. Validada a efectividade das formulações para a libertação e permeabilidade de cafeína e péptidos bioactivos, foram realizados estudos preliminares de modo a verificar a estabilidade da formulação GfB e a compreender a opinião de potenciais futuros consumidores face aos produtos desenvolvidos. As formulações foram sujeitas a condições de degradação acelerada (i.e. 40 ºC e 75% de humidade relativa) de acordo com a International Conference of Harmonization (ICH), não se verificando alterações químicas da cafeína em nenhum dos tempos de amostragem (imediatamente após a preparação da formulação e após 3, 6 e 9 meses) através da análise do espectro obtido por ATR-FTIR, assim como dos tempos de retenção em HPLC-UV. Verificou-se ainda um aumento significativo do conteúdo em água de GfB ao longo dos tempos de amostragem. Por fim, um estudo por focus group e um estudo de análise sensorial com um painel naive permitiram compreender a adequabilidade dos sabores propostos, assim como a tolerância à acidez e amargor por parte do consumidor. Observou-se uma ligeira tendência para a aceitação do sabor a menta e alguma tolerância ao amargor e acidez quando a menta foi usada na formulação. Os sistemas para libertação oral de compostos bioactivos, desenvolvidos e optimizados no âmbito desta tese, induziram melhorias significativas no comportamento farmacocinético in vitro dos compostos veiculados. De facto, a associação de filmes orais com micro- ou nanopartículas pode representar um novo sistema de libertação que ofereça maior efectividade e adesão por parte do consumidor/utente.
Aiming for the protection and absorption enhancement of bioactive compounds administered by oral route (with special focus on buccal absorption), this thesis had as goals, the optimization of oral films and micro/nanoparticles which, through conjugation of both, worked as innovative oral delivery systems with synergic activity. The bioactive molecules selected to study the behaviour and efficacy of the optimized formulations were caffeine and two whey-derived peptides with antihypertensive and relaxing activities (sequences: KGYGGVSLPEW and YLGYLEQLLR, respectively). The optimization process began with the selection and preliminary comparison of the characteristics of the excipients to be used to prepare films. Indeed, the first study included the optimization and comparison of two film formulations as carriers for the oral release of caffeine, prepared using sodium carobymethylcellullose and type A gelatine as polymers. It was observed by the analysis of the spectra obtained by Fourier-transformed infrared spectroscopy with attenuated total reflectance (FTIR-ATR) that caffeine chemical structure was not altered during the film production process. It was also observed, through the dissolution assay established by the United States Pharmacopoeia (USP), that type A gelatine films offered a slow caffeine release, whereas caroboxymethylcellulose films offered a burst release profile. Accordingly, the apparent permeability of caffeine observed from the ex vivo permeability assay, across small intestine tissues from porcine origin, was higher for carboxymethylcellullose films than for type A gelatine films. Nonetheless, disintegration time of both formulations was too high to meet the criteria of orodispersible formulations, taking longer than 30 s to achieve total disintegration. Still in the process of choosing the best polymer to integrate the composition of oral films, a new formulation containing guar gum as polymer was optimized by factorial design. Guar gum films presented superior mechanical and physico-chemical characteristics than carobxymethylcellullose or type A gelatine films, mainly regarding water-uptake capacity, erosion in artificial saliva and disintegration time. Furthermore, a formulation of alginate microparticles was also optimized by factorial design to associate with guar gum films and guarantee the controlled release of caffeine, as well as an increased bioavailability. The association of alginate beads to guar gum films – GfB – did not induce alterations in the chemical characteristics of caffeine, as outlined in the data obtained by FTIR-ATR, nor cytotoxicity to the cell lines used to mimic the buccal (TR146) or intestinal (Caco-2/HT29-MTX) mucosa, as determined by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide). Additionally, the release and in vitro (through TR146 and Caco-2/HT29-MTX cell lines) and ex vivo (through porcine intestinal mucosa) permeability profiles of caffeine from GfB was slower when compared with alginate microparticles, guar gum films or caffeine control solution. GfB also increased the effective contact between caffeine and buccal epithelia, offering a more complete permeation along time. Further, aiming to increment the bioavailability of the peptide KGYGGVSLPEW with antihypertensive activity, alginate beads were replaced with poly(lactic-co-glycolic) acid (PLGA) nanoparticles, since the last offer a higher association efficiency and a higher permeability of biologic membranes, due to the significantly lower particle size. The formulation of PLGA nanoparticles was optimized by factorial design. The delivery system comprising PLGA nanoparticles into guar gum films (GfNp) did not compromise the viability of the cell lines TR146 and Caco-2/HT29-MTX at tested concentrations. Moreover, GfNp promoted a slower peptide release and in vitro permeability across TR146 and Caco-2/HT29-MTX cell layers when compared with the films and nanoparticles alone or with a free peptide solution (control). However, apparent permeability was higher for GfNp when compared with remaining formulations. Results may be due to the intimate contact between the peptide and the epithelia, promoted by GfNp. It was also possible to observe that the peptide carried by GfNp presented a higher in vitro capacity to inhibit the angiotensin-converting enzyme after being subjected to the simulation of gastrointestinal tract, therefore presenting higher antihypertensive potential, when compared with the peptide carried by the nanoparticles or films alone or with the control solution (free peptide). The previously optimized system as carrier and delivery system for the antihypertensive peptide was also used to enhance the bioavailability of the relaxing peptide alpha-casozepine (sequence: YLGYLEQLLR). It was possible to conclude, through MTT assay, that none of the formulations compromised cell viability of TR146 cell line or Caco-2/HT29-MTX co-culture. Moreover, peptide permeability across in vitro buccal and intestinal epithelial models, while being subjected to simulated gastrointestinal tract, was higher and faster (higher apparent permeability) for the association of guar gum films with PLGA nanoparticles, when compared with PLGA nanoparticles or guar gum films alone or with the free peptide solution (control). Obtained results are related with the increased mucoadhesion conferred by the association of PLGA nanoparticles with guar gum films, verified through the analysis of adhesivity and work of adhesion to cow tongue. After validation of the effectivity of the formulations regarding release and permeability of caffeine and bioactive peptides, preliminary studies were performed to understand the stability of GfB and the opinion of potential future consumers of the developed products. Formulations were subjected to accelerated degradation conditions according to the International Conference of Harmonization (ICH) and it was verified, through analysis of spectra (by FTIR-ATR spectroscopy) and retention times (by HPLC-UV), that no chemical alterations of caffeine molecule carried by GfB were observed in any of the set time points (i.e. immediately after preparation of GfB and 3, 6 and 9 months after preparation, under accelerated degradation conditions – 40 ºC and 75% of relative humidity). Moreover, an increased water content was observed along the three time points. Further, a focus group and a sensory analysis study with a naïve panel allowed to understand the suitability of the flavours but also the tolerability to acidity and bitterness by the consumer. A slight tendency to the acceptance of mint flavour and some tolerance to bitterness and acidity was verified when mint was used in the formulation. Developed and optimized oral delivery systems in the scope of the present thesis induced significant improvements on the in vitro pharmacokinetic behaviour of carried bioactive molecules. Indeed, the association of oral films with micro- and nanoparticles may represent conceptually new delivery systems that offer higher effectivity and consumer/patient compliance.

AbstractBiobased polymers are of great interest due to the release of tension on non-renewable petroleum-based polymers for environmental concerns. However, biobased polymers usually have poor mechanical and barrier properties when used as the main component of coatings and films, but they can be improved by adding nanoscale reinforcing agents (nanoparticles - NPs or fillers), thus forming nanocomposites. The nano-sized components have a larger surface area that favors the filler-matrix interactions and the resulting material yield. For example, natural fibers from renewable plants could be used to improve the mechanical strength of the biobased composites. In addition to the mechanical properties, the optical, thermal and barrier properties are mainly effective on the selection of type or the ratio of biobased components. Biobased nanocomposites are one of the best alternatives to conventional polymer composites due to their low density, transparency, better surface properties and biodegradability, even with low filler contents. In addition, these biomaterials are also incorporated into composite films as nano-sized bio-fillers for the reinforcement or as carriers of some bioactive compounds. Therefore, nanostructures may provide antimicrobial properties, oxygen scavenging ability, enzyme immobilization or act as a temperature or oxygen sensor. The promising result of biobased functional polymer nanocomposites is shelf life extension of foods, and continuous improvements will face the future challenges. This chapter will focus on biobased materials used in nanocomposite polymers with their functional properties for food packaging applications.

Since implant surface is the region in contact with living tissue, corrosion resistance is an important property for biomedical implant materials. The deposition of biocompatible materials by magnetron sputtering is a viable surface modification method for improving the biocompatibility, corrosion resistance, and extends the life time of traditional biomedical implants. Titanium, niobium, and zirconium materials have excellent mechanical properties and corrosion resistance which make them potential candidates for coating implants. In this work, thin films of Ti, Nb, and Zr were deposited on Si(111) substrates in an oxygen/argon atmosphere to create models of biocompatible coatings. The chemical composition, morphology, Young’s modulus, and hardness of the three thin films were analyzed by X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectrometry (ToF-SIMS), atomic force microscopy (AFM), and nanoindentation. It was found that all thin films were completely oxidized, had nanostructured grains, bulk uniformity, and good mechanical properties, thus thin TiO2, Nb2O5, and ZrO2 films can be considered as good candidates for bioactive implant coatings.

Bioactive glasses are very attractive materials, used for tissue engineering materials, usually to fill and restore bone defects. This category of biomaterials, show considerable potential for orthopaedic surgery because they can promote bone tissue regeneration. Many trace elements have been incorporated in the glass network, an example is metallic glasses to obtain the desired properties. Because of tolerable mechanical properties, and because they are able to bond to living bone and stimulate its regeneration, this bioactive glasses have a particular interest and are in a continuous research and improvement. The chapter presents the history of bioactive glasses, classification, include a summary of common fabrication methods, applications, surface coatings, applications and future trends in relation to human bone. This review highlight new trends and areas of future research for bioactive glasses.

Regenerative therapies aim to develop novel treatments to restore tissue function. Several strategies have been investigated including the use of biomedical implants as three-dimensional artificial matrices to fill the defect side, to replace damaged tissues or for drug delivery. Bioactive implants are used to provide growth environments for tissue formation for a variety of applications including nerve, lung, skin and orthopaedic tissues. Implants can either be biodegradable or non-degradable, should be nontoxic and biocompatible, and should not trigger an immunological response. Implants can be designed to provide suitable surface area-to-volume ratios, ranges of porosities, pore interconnectivities and adequate mechanical strengths. Due to their broad range of properties, numerous biomaterials have been used for implant manufacture. To enhance an implant’s bioactivity, materials can be functionalised in several ways, including surface modification using proteins, incorporation of bioactive drugs, growth factors and/or cells. These strategies have been employed to create local bioactive microenvironments to direct cellular responses and to promote tissue regeneration and controlled drug release. This chapter provides an overview of current bioactive biomedical implants, their fabrication and applications, as well as implant materials used in drug delivery and tissue regeneration. Additionally, cell- and drug-based bioactivity, manufacturing considerations and future trends will be discussed.

Mostly, food packaging employs synthetic materials obtained from nonrenewable sources. These packaging materials are based on petrochemicals and cause substantial environmental problems by producing massive amounts of non-biodegradable solid wastes. Edible coatings and films are considered as the potential solution to these problems of non-biodegradable packaging solid wastes for maintaining food-environment interactions, retaining food quality, and extending shelf life. In addition, edible coatings and films offer prevention from microbial spoilage of packed foods by controlling moisture and gas barrier characteristics. Increasing environmental concerns and consumer demands for high-quality eco-friendly packaging have fueled the advancement of innovative packaging technologies, for instance, the development of biodegradable films from renewable agricultural and food processing industry wastes. Therefore, the current chapter presents the application of edible coatings and films as an alternative to conventional packaging, emphasizing the fundamental characterization that these biodegradable packaging should hold for specific applications such as food preservation and shelf life enhancement. The primary employed components (e.g., biopolymers, bioactive, and additives components), manufacturing processes (for edible films or coatings), and their application to specific foods have all been given special consideration in this chapter. Besides, a future vision for the use of edible films and coatings as quality indicators for perishable foods is presented.

Seaweeds are an abundant source of bioactive compounds and hydrocolloids that can be used in several industries, especially in the food industry. They can be directly consumed as a whole, used as an ingredient, or a source of several compounds that can enrich the nutrient content and sensory experience of foods. The demand for more sustainable packaging options resulted in the increase in research and development of edible seaweed-based films that ensure the safety and quality of the food products, while reducing packaging waste. This chapter will make an overview of the seaweed compounds used in the food industry, their use in the formulation of edible active packaging, and how this packaging can become widely available for consumers.<br>

In addition to being the oldest cultivated plant from past to present as a raw material source, Cannabis sativa L. is a strategically characteristic plant used for food, medicine, cosmetics, and pharmacological purposes. Molecular modeling is an important means for elucidating molecular structures and plays an important role in the design and development of new pharmaceuticals. This chapter includes the beneficial properties of the primary and secondary metabolites contained in cannabis such as foaming, emulsifying, gelling, film formation; antioxidant, antimicrobial activities and bakery products by means of their superior nutritional values, snack products, dairy products, processed meat products, in beverages such as coffees, teas, soda and functional food ingredients in many other areas of the food industry, nutritional supplements, usability in the development of new food formulations on hemp and molecular modeling studies on constituents of cannabis sativa.

Ginseng, the root of Panax species is a well-known conventional and perennial herb belonging to Araliaceae of various countries China, Korea, and Japan that is also known as the king of all herbs and famous for many years worldwide. It is a short underground rhizome that is associated with the fleshy root. Pharmacognostic details of cultivation and collection with different morphological characters are discussed. Phytocontent present is saponins glycosides, carbohydrates, polyacetylenes, phytosterols, nitrogenous substances, amino acids, peptides, vitamins, volatile oil, minerals, and enzymes details are discussed. The main focusing of the bioactive constituent of ginseng is ginsenosides are triterpenoid saponin glycosides having multifunctional pharmacological activities including anticancer, anti-inflammatory, antimicrobial, antioxidant and many more will be discussed. Ginseng is helpful in the treatment of microbial infection, inflammation, oxidative stress, diabetes, and obesity. Nanoparticles and nanocomposite film technologies had developed in it as novel drug delivery for cancer, inflammation, and neurological disorder. Multifaceted ginseng will be crucial for future development. This chapter review pharmacological, phytochemical, and pharmacognostic studies of this plant.

Plasma polymer deposition is the method of choice for the finishing of metallic implant materials like titanium with nitrogen-containing bioactive coatings. The deposited cell-adhesive plasma polymer films have to possess special properties such as homogeneity, film stability on air as well as in different media, sufficient density of functional groups and the appropriate surface charge. But also the knowledge of long-term stability is essential for the application as implant surface. Therefore, aging studies of plasma polymer coatings on titanium surfaces are important to detect the changes of surface chemistry over a longer time period. For this purpose, results of physicochemical surface diagnostics were combined with adequate tissue culture experiments. The objective of this paper was to measure surface chemical characteristics of thin plasma polymerized allylamine (PPAAm) coatings on polished titanium with the main focus on FT-IR studies over a time period of one year and to correlate these data with the adhesion of human MG-63 osteoblastic cells.

The objective of this work was to produce, through the thermoplastic extrusion process followed by blowing, manioc starch-based flexible films added with Brazilian pepper oil as an antioxidant and plasticizer agent, and verify if the bioactive compounds contained in the fresh pepper oil are present after the drying step of the thermoplastic extrusion. After analysis by gas chromatography-mass spectrometry volatiles compounds were identified in the films. Pepper oil also influenced the mechanichal properties of the films.These results suggest that the temperatures used in the process, kept some of the existing compounds in the Brazilian pepper essential oil adhered to the packages. Keywords: Termoplastic extrusion, temperature, bioactivs compounds

Objective. We report the study of feasibility to produce the thing bioactive coating from experimental dental cement using pulsed laser deposition (PLD) technique. Methods. The targets for PLD system (disks 30 mm in diameter × 5 mm thick) were sintered from micronized powder of set Alborg White Portland cement (WPC). The parameters for sintering process were chosen based thermo-gravimetric analysis and differential scanning calorimetry (TGA/DSC). The coatings were deposited by PLD on silicon substrates. The effect of laser power on coating crystallinity and morphology was evaluated by scanning electron microscope (SEM) and X-ray diffraction (XRD). The material transfer from target to substrate were evaluated by X-ray fluorescence (XRF) and X-ray energy dispersive spectroscopy (EDS). The bioactivity of deposited films was evaluated by ability produce the hydroxyapatite (HA) layer on a surface of specimen immersed in a simulated body fluid (Dulbecco’s Phosphate-Buffered Saline (DPBS). The formation of hydroxyapatite was confirmed by SEM, X-ray energy dispersive spectroscopy (EDS), XRD and micro-Raman spectroscopy. The formation of HA was evaluated after 1, 3, 7, 14, and 21 days of immersion. Results. This study demonstrated that White Portland cement can be used as a target material for manufacturing of bio-functional coatings. The films deposited on Si substrates have mainly amorphous structure; the crystallinity of the film can be achieved by increasing the laser power. The biological performance of deposited films was tested by HA forming ability in simulated body fluid. The HA layer was formed on a coated surface after first day of immersion.

In this work, we report a simple fabrication method for metal nanoparticles and nanorods on halloysite supports. Silver nanorods of 15 nm diameter were synthesized by thermal decomposition of silver acetate within halloysite lumen. Nanorods had crystalline nature with [111] axis oriented ∼ 68° from the halloysite tubule main axis. Linear arrays from gold, iron, cobalt and palladium nanoparticles on halloysite external surface were also synthesized by chemical and thermal reduction method. Samples were analyzed by high-resolution transmission electron microscopy and field-emission scanning electron microscopy. These in situ syntheses offer a simple method for large scale fabrication of metallic nanorods and core-shell ceramic nanocomposites, which can be used as antimicrobial additives in plastic composites, nanoelectronic and optical materials with biocompatibility and environmentally friendly. Antimicrobial thin films were prepared based on halloysite-silver nanocomposites and tested on E. Coli and S Aureus bacterial culture. Antibacterial performance of the nanocomposite material was superior to the other conventional antimicrobial additives (silver doped bioactive glasses and carbon nanotubes). Radiation protection coatings based on fabricated nanocomposite materials is under development.

Fermented products have gained worldwide popularity for their nutritional and health aspects. Many studies have been done on this topic, including fermented cabbage (sauerkraut). Yet little or no studies are done on evaluation of fermented cabbage juice which is considered as by-product of sauerkraut production, still rich in bioactive compounds. In order to reduce food waste, sustainable solutions are being searched for to preserve valuable fermented cabbage juice. The aim of this study was to evaluate chemical and physical composition of concentrated fermented cabbage juice and their changes after storage. The fermented cabbage juice was concentrated on falling film evaporator from 9.2 till 34.3 °Brix. Physio-chemical (moisture, pH, total soluble solids, total phenol content, antiradical activity by DPPH and ABTS+, ascorbic acid, total sugar profile, nitrates and minerals) and microbiological (lactic acid bacteria, total plate count) analyses were carried out. Concentrated fermented cabbage juice is a source of minerals and phenol compounds as well as salt substitute in food applications. After 6 months of storage there is significant degradation of ascorbic acid but total phenol content is not affected. The evaporation process did not inhibit microbiological activity; as a result, there is a decrease in lactic acid bacteria but increase in total plate count.

The objective of the study was to determine the content of vitamin C (L-ascorbic acid) in two leafy parsley (Petroselinum crispum Mill. Fuss) cultivars subjected to different post-harvest treatments. Ascorbic acid (AA), due to its instability, is one of the indicators of leafy vegetable freshness and quality. High content of this compound in vegetables is desired because of its bioactive properties. The cultivars differ in morphology of usable parts: ‘Rialto’ has flat lamina and ‘Petra’ triple-curled lamina. The plant material was obtained in 2015 and 2016 from experimental field in Warsaw-Wilanów. The leaves were washed directly after harvest: a) in tap water or b) in tap water with ozone added. Two methods of postharvest storage were applied: A) at the cold store and B) under simulated retail conditions. In the case of cold store method (A), the plants were tied in tufts and stored at the temperature of 0 °C and RH 90 % for 7, 14, 28 days in two variants: 1) in containers, where leaves petioles were immersed in water, and 2) in special bulk modified atmosphere packaging (MAP), dedicated to fresh herbs (Stepac, Israel). In the case of storage under simulated retail conditions (B), the leaves were kept for 48 hours at 10 °C, RH 30-40 % in two variants: 1) tufts wrapped in perforated PE film with petioles immersed in water, and 2) packed to retail MAPs, dedicated to leafy herbs (Stepac, Israel). Concentration of L-ascorbic acid in the leaves was determined spectrophotometrically, with the method based on reaction of Folin’s phenol reagent in low pH. Fresh and stored ‘Rialto’ leaves were characterised by a higher concentration of AA than ‘Petra’ leaves (110 and 44 mg g -1 f.w., respectively). Significant decrease of AA after ozone treatment was observed only for ‘Rialto’ directly after harvest. Storage length had significant influence on the content of AA in both cultivars. Decrease of AA content was observed during storage period. Average concentration of AA after 7 days of storage was 85 and 44 -1 f.w. for ‘Rialto’ and ‘Petra’, respectively. After 28 days of storage the concentration was 54% lower for ‘Rialto’ and 36% lower for ‘Petra’, compared to the initial content. Leaves after simulated retail conditions showed slightly lower AA content than those stored for 7 days in the cold room. Retail MAPs guaranteed significantly higher preservation of AA in both cultivars than wrapping in film with immersing in water. Washing in ozone-added water showed influence only on AA content in ‘Rialto’ leaves wrapped in film – they showed lower concentration of AA under simulated retail conditions. Our study showed that the content of AA in parsley leaves was cultivar dependent. Ozone treatment did not have negative effect on AA during the storage. Storage duration, rather than packaging methods, plays a key role in preservation of high content of AA. However, method of packaging is important under retail conditions, where temperature is higher and RH is much lower, than at a cold room.

Background. In this proposal we suggest developing a common solution for three seemingly unrelated acute problems: (1) improving sustainability of fast-growing mushroom industry producing worldwide millions of tons of underutilized leftovers; (2) alleviating the epidemic of vitamin D deficiency adversely affecting the public health in both countries and in other regions; (3) reducing spoilage of perishable fruit and vegetable products leading to food wastage. Based on our previous experience we propose utilizing appropriately processed mushroom byproducts as a source of two valuable bioactive materials: antimicrobial and wholesome polysaccharide chitosan and health-strengthening nutrient ergocalciferol⁽ᵛⁱᵗᵃᵐⁱⁿ ᴰ2⁾. ᴬᵈᵈⁱᵗⁱᵒⁿᵃˡ ᵇᵉⁿᵉᶠⁱᵗ ᵒᶠ ᵗʰᵉˢᵉ ᵐᵃᵗᵉʳⁱᵃˡˢ ⁱˢ ᵗʰᵉⁱʳ ᵒʳⁱᵍⁱⁿ ᶠʳᵒᵐ ⁿᵒⁿ⁻ᵃⁿⁱᵐᵃˡ ᶠᵒᵒᵈ⁻ᵍʳᵃᵈᵉ source. We proposed using chitosan and vitamin D as ingredients in active edible coatings on two model foods: highly perishable fresh-cut melon and less perishable health bars. Objectives and work program. The general aim of the project is improving storability, safety and health value of foods by developing and applying a novel active edible coating based on utilization of mushroom industry leftovers. The work plan includes the following tasks: (a) optimizing the UV-B treatment of mushroom leftover stalks to enrich them with vitamin D without compromising chitosan quality - Done; (b) developing effective extraction procedures to yield chitosan and vitamin D from the stalks - Done; (c) utilizing LbL approach to prepare fungal chitosan-based edible coatings with optimal properties - Done; (d) enrichment of the coating matrix with fungal vitamin D utilizing molecular encapsulation and nano-encapsulation approaches - Done, it was found that no encapsulation methods are needed to enrich chitosan matrix with vitamin D; (e) testing the performance of the coating for controlling spoilage of fresh cut melons - Done; (f) testing the performance of the coating for nutritional enhancement and quality preservation of heath bars - Done. Achievements. In this study numerous results were achieved. Mushroom waste, leftover stalks, was treated ʷⁱᵗʰ ᵁⱽ⁻ᴮ ˡⁱᵍʰᵗ ᵃⁿᵈ ᵗʳᵉᵃᵗᵐᵉⁿᵗ ⁱⁿᵈᵘᶜᵉˢ ᵃ ᵛᵉʳʸ ʰⁱᵍʰ ᵃᶜᶜᵘᵐᵘˡᵃᵗⁱᵒⁿ ᵒᶠ ᵛⁱᵗᵃᵐⁱⁿ ᴰ2, ᶠᵃʳ ᵉˣᶜᵉᵉᵈⁱⁿᵍ any other dietary vitamin D source. The straightforward vitamin D extraction procedure and ᵃ ˢⁱᵐᵖˡⁱᶠⁱᵉᵈ ᵃⁿᵃˡʸᵗⁱᶜᵃˡ ᵖʳᵒᵗᵒᶜᵒˡ ᶠᵒʳ ᵗⁱᵐᵉ⁻ᵉᶠᶠⁱᶜⁱᵉⁿᵗ ᵈᵉᵗᵉʳᵐⁱⁿᵃᵗⁱᵒⁿ ᵒᶠ ᵗʰᵉ ᵛⁱᵗᵃᵐⁱⁿ ᴰ2 ᶜᵒⁿᵗᵉⁿᵗ suitable for routine product quality control were developed. Concerning the fungal chitosan extraction, new freeze-thawing protocol was developed, tested on three different mushroom sources and compared to the classic protocol. The new protocol resulted in up to 2-fold increase in the obtained chitosan yield, up to 3-fold increase in its deacetylation degree, high whitening index and good antimicrobial activity. The fungal chitosan films enriched with Vitamin D were prepared and compared to the films based on animal origin chitosan demonstrating similar density, porosity and water vapor permeability. Layer-by-layer chitosan-alginate electrostatic deposition was used to coat fruit bars. The coatings helped to preserve the quality and increase the shelf-life of fruit bars, delaying degradation of ascorbic acid and antioxidant capacity loss as well as reducing bar softening. Microbiological analyses also showed a delay in yeast and fungal growth when compared with single layer coatings of fungal or animal chitosan or alginate. Edible coatings were also applied on fresh-cut melons and provided significant improvement of physiological quality (firmness, weight ˡᵒˢˢ⁾, ᵐⁱᶜʳᵒᵇⁱᵃˡ ˢᵃᶠᵉᵗʸ ⁽ᵇᵃᶜᵗᵉʳⁱᵃ, ᵐᵒˡᵈ, ʸᵉᵃˢᵗ⁾, ⁿᵒʳᵐᵃˡ ʳᵉˢᵖⁱʳᵃᵗⁱᵒⁿ ᵖʳᵒᶜᵉˢˢ ⁽Cᴼ2, ᴼ²⁾ ᵃⁿᵈ ᵈⁱᵈ not cause off-flavor (EtOH). It was also found that the performance of edible coating from fungal stalk leftovers does not concede to the chitosan coatings sourced from animal or good quality mushrooms. Implications. The proposal helped attaining triple benefit: valorization of mushroom industry byproducts; improving public health by fortification of food products with vitamin D from natural non-animal source; and reducing food wastage by using shelf- life-extending antimicrobial edible coatings. New observations with scientific impact were found. The program resulted in 5 research papers. Several effective and straightforward procedures that can be adopted by mushroom growers and food industries were developed. BARD Report - Project 4784