In this study, we introduce novel, truly non-ionic hydrogen-bonded layer-by-layer (LbL) coatings for cell surface engineering capable of long-term support of cell function. Utilizing the LbL technique imparts the ability to tailor membrane permeability, which is of particular importance for encapsulation of living cells as cell viability critically depends on the diffusion of nutrients through the artificial polymer membrane. Ultrathin, permeable polymer membranes are constructed on living cells without a cationic pre-layer, which is usually employed to increase the stability of LbL coatings. In the absence of the cytotoxic PEI pre-layer, viability of encapsulated cells drastically increases to 94%, as compared to 20-50% in electrostatically-bonded shells. Engineering surfaces of living cells with natural or synthetic compounds can mediate intercellular communication, render the cells less sensitive to environmental changes, and provide a protective barrier from hostile agents. Surface engineered cells show great potential for biomedical applications, including biomimetics, biosensing, enhancing biocompatibility of implantable materials, and may represent an important step toward construction of an artificial cell.

Cancer is a disease that varies dramatically from person to person due to the specifics of the individual's physiology and the source of the cancer. In most cases, the origin of the cancer can be determined but metastasis can lead to tumors anywhere and thus many cancers require treatment of the whole body. Since many of the drugs that are used to treat cancer are toxic to healthy cells as well as cancerous ones, there has been considerable interest in developing ways to convey the drug specifically to the cancer cells with minimal exposure to healthy cells. Colloid drug delivery vehicles have shown considerable progress toward this end, while also reducing degradation of the drug prior to delivery to targeted sites (particularly important for oligonucleotide and protein therapeutics), and controlling release rates. Toward the end of improved drug delivery, this thesis work investigates the encapsulation of DNA in gelatin microspheres (GMS) and the subsequent temperature controlled release of the encapsulated DNA from these GMS. DNA-loaded GMS were then used as templates for colloidal satellite assemblies and the released DNA was shown to competitively displace the original partner strands of immobilized DNA on the surface of the assemblies. To support these investigations, hybridization of DNA at colloidal surfaces was also investigated using in situ measurements and found to significantly deviate from solution behavior. DNA hybridization is of particular interest as means of controlling the functionality of colloidal structures because it is uniquely reversible and tunable as well as biocompatible. Gelatin was chosen as the encapsulation matrix for its superior biocompatibility, convenient gel to liquid phase transition at ~35 oC, and economical availability.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Osteoarthritis is a degenerative joint disease characterized by progressive erosion of the articular cartilage. Epidemiological studies have established a relationship between osteoarthritis and menopause suggesting that estrogen may be important in the development of cartilage regeneration therapies. The overall goal of this research project was to advance the field of cartilage tissue regeneration by investigating the role of 17 ß -estradiol (E2), an active estrogen metabolite, on the chondrocyte phenotype. The central hypothesis was that E2 plays an important and sex-specific role in regulating chondrogenesis. Specific Aim-1 focused on establishing and characterizing a primary human articular chondrocyte (HAC) cell source, and then examining the response of the cells in culture to E2. It was demonstrated that the response of HACs to E2 treatment was sex-specific despite both male and females cells expressing estrogen receptors. Female HACs showed changes in proliferation, matrix production, and differentiation while male cells did not. In addition, the female response was regulated through a rapid membrane signaling pathway mediated by protein kinase C. Specific Aim-2 involved establishing an ovariectomized animal model to investigate the effects of E2 on orthopaedic tissue implants. Human demineralized bone matrix (DBM) was implanted intramuscularly into female nude mice and rats. Ovariectomy was shown to reduce the ability of DBM to induce the formation of cartilage and bone tissue. Moreover, the inductive properties of DBM were reestablished with subcutaneous E2 supplementation. Specific Aim-3 entailed developing and characterizing a microencapsulation method for in vitro culture and in vivo delivery of chondrocytes to study the effects of E2 on chondrogenesis. Rat growth plate chondrocytes and HACs were microencapsulated in alginate using an extrusion method in conjunction with high electrostatic potential. Chondrocytes maintained their phenotype in alginate suspension but were unable to form cartilage tissue when implanted into our animal model. Further optimization of the system is required before the role of E2 on chondrogenesis of tissue engineered constructs can be determined. In summary, our results suggest that the successful production of tissue engineered therapies will likely depend on understanding and manipulating the actions of sex hormones in both the in vitro and in vivo environment.

Tissue repair is a complex process that is difficult to emulate. The addition of the glycosaminoglycan heparan sulfate (HS), a multi-potential regulator of numerous growth factors and cytokines endogenously expressed during the repair process, may represent a valuable tool for tissue engineering. The addition of exogenous HS into wound site has previously been shown to promote tissue repair in a number of models, however, the incorporation of HS into controlled release systems or biomaterials for tissue engineering had not been explored prior to the work presented here. Thus, this thesis explores the incorporation of HS and its analogue heparin into synthetic biodegradable polymer biomaterials with different potential applications, either as a slow releasing drug reservoir, or as a drug releasing cell scaffold. Polycaprolactone was used to make microcapsules and electrospun fibers for HS or heparin entrapment. These materials were characterized for their drug release profiles, biocompatibility and bioactivity. Microcapsules encapsulating heparin or HS were made by the oil - in - water solvent evaporation method which allowed fabrication of slow releasing drug reservoirs. Either pure water or a poly(vinyl alcohol) solution was used in the drug phase which resulted in capsules with similar size and drug loading. However the internal morphology and drug release profiles showed differences depending on the drug phase, in either case release was sustained for over 30 days. These capsules elicited no pro-inflammatory response from macrophages in vitro, and the released HS retained its bioactivity to induce the proliferation of human mesenchymal stem cells, an important cell type for bone tissue engineering. Heparin and HS were incorporated into electrospun fibers as a drug releasing scaffold for two different tissue engineering applications. Heparin fibers were studied as a drug releasing membrane that could be used in vascular repair to prevent the unwanted proliferation of vascular smooth muscle cells. Heparin release was sustained from the fibers for at least 2 weeks. The fibers did not induce a pro-inflammatory response from macrophages in vitro and the released heparin retained the ability to inhibit the proliferation in vascular smooth muscle cells. HS fibers were studied as a tissue engineering scaffold for bone repair using human mesenchymal stem cells. HS release was maintained for over 30 days which is thought to be an appropriate time for bone repair applications. The release profiles depended on the HS concentration in the spinning solution which affected the morphology of the fibers. The fibers did not elicit a pro-inflammatory response in cultured macrophages and supported the proliferation and mineralization of human mesechymal stem cells. The HS fibers were then taken through to an in vivo model to study ectopic bone formation of pre-osteoblast cells on HS releasing scaffolds. The fibers produced a chronic inflammatory response in vivo, which lead to the clearance of implanted cells and no mineralization of the scaffold. The HS and heparin materials made in this work showed sustained release over appropriate time frames for different tissue repair applications. The released HS and heparin maintained bioactivity and showed good biocompatibility in vitro, however, further in vivo studies are required to fully test their efficacy for tissue engineering.

Fortification of instant Asian noodles with folic acid has the potential to enhance dietary folate intakes. Recent studies show folate deficiency is prevalent in many countries. Furthermore, this vitamin is unstable upon exposure to light, air, heat and extreme conditions of acidity and alkalinity. Internationally, folate in foods has traditionally been analysed by a microbiological assay, however, due to the extensive time required for sample preparation and analysis, alternative procedures for analysis require consideration. The aims of the current study have been to investigate the stability of added folic acid in fortified instant fried noodles by analytical methods of capillary electrophoresis and reversed-phase HPLC. Additionally, procedures for the microencapsulation of folic acid by spray drying have been evaluated along with their significance in increasing the stability of the vitamin during processing and boiling of instant noodles. Optimisation of capillary electrophoretic conditions showed that the maximum response of folic acid relative to an internal standard was achieved using various concentrations of phosphate and borate. Analytical parameters including the effects of pH, voltage and temperature were studied along with enzymatic treatments for liberation of folic acid from the noodle matrix based. Higher recoveries were obtained using the enzymes however these exceeded 100% due to sample matrix interference. Standard addition or internal calibration were both effective in correcting for matrix interferences. Comparative investigations with reversed-phase HPLC confirmed the results obtained with the capillary electrophoresis. Using either a phosphate based buffer in conjunction with an ion-pairing agent at alkaline pH or an acidic mobile phase, the results attained were in good agreement as folic acid exhibited excellent stability under typical processing conditions. Various food approved hydrocolloids were evaluated for encapsulation of folic acid by spray drying. Incorporation of the microcapsules into formulations of instant fried noodles showed that after boiling the folic acid was chemically degraded to some extent and leaching also occurred. The microcapsules exhibited similar properties regardless of the binding agent used, with losses still occurring during the boiling stage. In order to enhance the structural integrity of the spray dried microcapsules, CaCl2 was used as a cross-linking agent for capsules prepared using alginate or pectin binding agents. Considerable increases in retention of core material were observed as the network exhibited a reduction in swelling and hydration, and subsequently a decrease in the release of folic acid. In summary, capillary electrophoresis and reversed-phase HPLC provided excellent separation and good quantitatation of added folic acid in instant Asian noodles. Excellent resolution was obtained between the sample matrix interference of instant noodles and the analysed vitamin. Folic acid displayed high stability throughout the processing of instant noodles whereas there was consistent evidence that unencapsulated folic acid was degraded during boiling. Microencapsulation of folic acid with combinations of alginate and pectin as the binding agents, proved to be effective in maintaining folic acid stability when calcium treatment was performed after spray drying. These findings provide an effective way to retain folic acid used in fortifying Asian instant noodles.

MENDES, Luana Guabiraba. Microencapsulação do corante natural de urucum: uma análise da eficiência da goma do cajueiro com material de parede. 2012. 130 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Tecnologia de Alimentos, Fortaleza-CE, 2012 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-06-14T16:13:27Z No. of bitstreams: 1 2012_dis_lgmendes.pdf: 1934768 bytes, checksum: e4667dcb8eb5ef8e88f0c80e4d29e75b (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-06-14T16:13:37Z (GMT) No. of bitstreams: 1 2012_dis_lgmendes.pdf: 1934768 bytes, checksum: e4667dcb8eb5ef8e88f0c80e4d29e75b (MD5) / Made available in DSpace on 2016-06-14T16:13:37Z (GMT). No. of bitstreams: 1 2012_dis_lgmendes.pdf: 1934768 bytes, checksum: e4667dcb8eb5ef8e88f0c80e4d29e75b (MD5) Previous issue date: 2012 / Na prática, alguns corantes naturais, como a bixina, apresentam dificuldades tecnológicas de utilização, dificultando sua aplicação na indústria de alimentos. A microencapsulação pode amenizar essa situação, aumentando sua estabilidade e tornando possível sua incorporação em sistemas alimentícios sem a perda de suas propriedades. Assim, o objetivo dessa pesquisa foi microencapsular o corante natural de urucum, utilizando a goma de cajueiro como material de parede, aplicando os processos de liofilização e atomização, caracterizar as microcápsulas obtidas através de análises morfológicas, físico-quimicas, bem como determinar a eficiência da microencapsulação pela quantificação do teor de bixina total e superficial. Goma arábica foi utilizada como material de parede de referência, para efeito comparativo. Assim, foram preparadas três formulações, a saber: corante de urucum/ goma do cajueiro (U-GC) 4:1 m/m, corante de urucum /goma arábica (U-GA) 4:1 m/m e corante de urucum/ goma do cajueiro/ e goma arábica (1:1) 4:1 m/m (U-GCA). As suspensões, previamente preparadas também foram caracterizadas por análises físico-químicas. Na avaliação colorimétrica estudou-se a estabilidade do pigmento microencapsulado, em temperatura ambiente, a 25°C± 2°C na presença e na ausência de luz, durante 40 dias de armazenamento, em duas situações, ou seja, dissolvidos em solução-tampão pH 4,0 e na forma de pó. Nas três suspensões formuladas a análise colorimétrica e o pH não apresentaram grandes diferenças, mas já na viscosidade a suspensão goma do cajueiro/ corante de urucum, resultou em menor viscosidade (0,01746±0,000) nas mesmas concentrações de sólidos. As microcápsulas produzidas por atomização mostraram também formas irregulares com predominância circular e superfície dentada e, por liofilização, mostraram formas indefinidas e com tamanhos muito variáveis. Todas as microcápsulas mostraram-se solúveis em água. As microcápsulas elaboradas com goma arábica e goma do cajuerio/goma arábica (1:1) apresentaram as melhores eficiências na microencapsulação, 43,14 e 31,21%, respectivamente, além de conferirem maior estabilidade à bixina durante a exposição à luz e na estocagem no escuro. As diferentes formulações resultaram em diferentes teores do corante de urucum microencapsulado, sendo que a goma arábica apresentou maior eficiência e a goma do cajueiro menor eficiência, considerando os processos de liofilização e atomização. Houve uma maior degradação da bixina total, em torno de 65,79% e, da bixina superficial, em torno de 60,52%, nas microcápsulas armazenadas sob incidência de luz a 25°C±2°C, evidenciando o efeito deletério da luz sobre o corante de urucum. O método de microencapsulação por liofilização foi o que levou a menor degradação da bixina encapsulada. Dentre dos parâmetros de concentração utilizados e dos e dos métodos de microencsapsulação empregados, a goma do cajueiro, individualmente, não se apresentou como um eficiente material de parede. No entanto, a sua mistura com a goma arábica forneceu resultados promissores, os quais tornaram favorável a utilização desse material (U-GCA), como na indústria de alimentos, bem como em outros segmentos tecnologicos como os cosméticos e a farmacêutica. / In practice, some natural dyes, such as bixin present technological difficulties of use, hindering their application in the food industry. Microencapsulation can improve this situation by increasing their stability and making their incorporation in food systems possible without losing their properties. The objective of this research was to microencapsulate the natural dye annatto, using cashew gum as wall material, applying the freeze-drying and spray drying processes, to characterize the microcapsules obtained according to morphological and physico-chemical properties as well as to determine the effectiveness of the microencapsulation by quantification of the total bixin and surface bixin. Gum arabic was used as wall material, the reference for comparison. Thus, three formulations were prepared as follows: annatto colorant / cashew gum (U-GC) 4:1 w / w of annatto colorant / gum arabic (GA-U) 4:1 m / m of annatto colorant / cashew gum / gum arabic and (1:1) 4:1 m / m (U-GCA). The suspensions, which had been previously prepared, were also characterized in terms physical and chemical analyses. In colorimetric evaluation the stability of the microencapsulated pigment was studied, at a room temperature of 25 ° C ± 2 ° C in the presence and absence of light for 40 days storage in two situations, that is dissolved in buffer solution pH 4.0 and in powder form. In the three formulated suspensions , there were no major differences between the pH and colorimetric analyses, but the suspension viscosity cashew gum / dye annatto, resulted in lower viscosity (0.01746 ± 0.000) at the same concentrations of solids. The microcapsules produced by atomization also showed predominantly irregularly shaped circular and toothed surfaces, and lyophilization, show indefinite shapes and sizes vary widely. All the microspheres proved soluble in water. Microcapsules prepared with gum arabic and gum cajuerio / gum arabic (1:1) showed the best efficiency in the microencapsulation, 43.14 and 31.21%, respectivelyand this confers greater stability to the bixin during exposure to light and storage in the dark. The different formulations resulted in different concentrations of microencapsulated annatto dye, with the gum arabic being the most efficient and the cashew gum the least efficient in terms of lyophilization and spray drying. There was a greater total degradation of bixin, about 65.79% and the surface bixin, about 60.52%, in the microcapsules stored under light incidence at 25 ° C ± 2°C, demonstrating the detrimental effect of light on the dye annatto. The microencapsulation method by means of lyophilization is the one which led to the lowest levels of degradation of encapsulated bixin. Among the parameters used and the concentration and methods of microencapsulation employed, cashew gum, on its own, is not presented as an effective wall material. However, its mixture with gum arabic provided promising results, which favors making use of this material (U-GCA) in the food industry and in other technological sectors such as cosmetics and pharmaceuticals.

MOREIRA, Samira Pereira. Avaliação da qualidade e segurança de melão minimamente processado revestido em matriz de quitosana adicionada de compostos bioativos microencapsulados extraídos de subprodutos de acerola. 2014. 166 f. Dissertação (Mestrado em Ciência e Tecnologia de Alimentos)-Universidade Federal do Ceará, Fortaleza, 2014. / Submitted by Vitor Campos (vitband@gmail.com) on 2016-10-03T23:19:30Z No. of bitstreams: 1 2014_dis_spmoreira.pdf: 2559693 bytes, checksum: d4f3061bd25a32ab80831f9b6ec70674 (MD5) / Approved for entry into archive by Jairo Viana (jairo@ufc.br) on 2016-10-04T23:33:53Z (GMT) No. of bitstreams: 1 2014_dis_spmoreira.pdf: 2559693 bytes, checksum: d4f3061bd25a32ab80831f9b6ec70674 (MD5) / Made available in DSpace on 2016-10-04T23:33:53Z (GMT). No. of bitstreams: 1 2014_dis_spmoreira.pdf: 2559693 bytes, checksum: d4f3061bd25a32ab80831f9b6ec70674 (MD5) Previous issue date: 2014 / The minimal processing of fruits and vegetables offers the consumer food fresh and practicality. However, these products rapidly lose quality after processing due to physical changes and their consequential biochemical reactions that can reduce the nutritional value and favours besides the growth of deteriorant and pathogenic microorganisms. Thus, this study aimed to develop a system of active packaging, consisting of an microencapsulated antimicrobial edible coating using chitosan added with bioactive compounds extracted from byproducts of processing of acerola (Malpighia emarginata D.C.) for minimally processed melon to maintain the quality and microbiological safety of this product. The parameters extraction of bioactive compounds of lyophilized byproducts (seed, peel and fibers) were optimized as the rotation speed, extraction time, extraction solution and by proportion: extraction solution. The extract showed high content of antioxidants, especially polyphenols. The coating was produced from the 1:1 mixture (v / v) solution of chitosan and 1% ethanolic extract of bioactive compounds, and chitosan itself as the encapsulating material. Two treatments (coated and uncoated melons) each 3 days during 12 days of storage at 10˚C ± 1 were evaluated as attributes of quality, bioactive compounds, total antioxidant activity, spoilage and pathogenic microorganisms (Salmonella spp and Listeria monocytogenes). The results showed that the coating samples provided the melons minor loss of firmness and weight, increased the content of yellow flavonoids and total extractable polyphenols, consequently high total antioxidant activity when compared to control cubes. Moreover, the coating showed antimicrobial activity against bacteria (mesophilic, psicotrophic and lactic), molds and yeasts in coated fruits. The novel product is according to RDC ANVISA 12˚ whose recommendation for fresh fruits must be absence of Salmonella spp in 25 g. Therefore, this coating is a viable alternative for maintain the quality of minimally processed melons , since it provided better nutrition by maintaining bioactive fruit compounds, the safety, beyond the scope of the life of the product. / O processamento mínimo de frutas e hortaliças oferece ao consumidor alimentos frescos e práticos. Contudo, estes produtos perdem rapidamente a qualidade após o processamento devido às alterações físicas e suas conseqüentes reações bioquímicas reduzindo o valor nutricional, além de favorecer o crescimento de microrganismos patogênicos. Assim, este trabalho objetivou desenvolver um sistema de embalagem ativa, consistindo de um revestimento comestível antimicrobiano para revestimento de melão minimamente processado à base de quitosana incorporado de compostos bioativos microencapsulados extraídos de subprodutos do processamento de acerola (Malpighia emarginata D.C) visando a manutenção da qualidade e segurança microbiológica do produto. Os parâmetros de extração dos compostos bioativos dos subprodutos liofilizados (semente, película e fibras) foram otimizados quanto a velocidade de rotação, tempo de extração, solução extratora e proporção subprodutos : solução extratora . O extrato apresentou alto teor de compostos antioxidantes, principalmente polifenóis. O revestimento utilizado nesse estudo foi produzido a partir da mistura na proporção 1:1 (v/v) de solução de quitosana 1% e extrato etanólico de compostos bioativos, tendo a própria quitosana como material encapsulante. Dois tratamentos (melões revestidos e não revestidos) foram avaliados a cada 3 dias durante 12 dias de armazenamento a 10˚C ± 1 quanto aos atributos de qualidade, compostos bioativos, atividade antioxidante total, microorganismos deteriorantes e patogênicos (Salmonella spp e Listeria monocytogenes). Os resultados demonstraram que o revestimento proporcionou aos melões menor perda de firmeza e de peso, além de maiores conteúdos de flavonóides amarelos e de polifenóis extraíveis totais, consequentemente elevada atividade antioxidante total quando comparado ao controle. Foi obsevado ação antimicrobiana contra bactérias (mesófilas, psicotróficas e láticas), bolores e leveduras nos frutos revestidos. O produto apresentou-se em conformidade com a RDC n˚ 12 da ANVISA cuja recomendação para frutas in natura é de ausência de Salmonela spp em 25 g. Portanto, este revestimento é uma alternativa viável para a manutenção da qualidade de melões minimamente processados, uma vez que proporcionou a melhoria nutricional, através da manutenção de compostos bioativos da fruta a segurança, além da extensão da vida útil desse produto.

Os microrganismos probióticos são considerados suplementos alimentares vivos, apresentando benefícios ao hospedeiro e melhorando o balanço intestinal. A produção de Lactobacillus com alta densidade celular vem sendo estudada e possui grande interesse por parte da indústria, bem como o estudo de novos meios de cultivo alternativos. Outros interesses são a melhora da sobrevivência dos microrganismos durante a passagem pelo trato gastrintestinal por meio da microencapsulação e a elaboração de um produto com potencial probiótico que não necessite da cadeia do frio. Dentro deste contexto o presente trabalho objetivou a produção de microrganismo potencialmente probiótico em meio de cultivo vegetal e após microencapsulado, para obtenção de um pó alimentício para ser diretamente utilizado em alimentos. Na primeira etapa deste trabalho foi realizada uma avaliação de variáveis para fixar os parâmetros de processo e o meio de cultivo em biorreator submerso, para produção de biomassa de Lactobacillus plantarum BL011. O meio de cultivo e parâmetros de processo que apresentaram os melhores resultados para a produção de biomassa e ácido láctico foram: 40 g L-1 de açúcares totais (soro ácido de soja); 15 g L-1 de extrato de levedura; velocidade de agitação de 200 rpm; 25 °C e 4,5 vvm. Os resultados obtidos permitiram uma produção de biomassa de 17,87 g L-1 e 37,59 g L-1 de ácido láctico. Em uma segunda etapa deste trabalho o microrganismo foi microencapsulado pela técnica de electrospraying, utilizando como agentes encapsulantes alginato de sódio (ALG) e uma mistura de alginato de sódio e pectina cítrica (ALG-PEC). As células microbianas livres e microencapsuladas foram submetidas ao suco gástrico simulado (SGS) e suco intestinal simulado (SIS). O microrganismo controle (células livres) demonstrou uma diminuição de 6 e 4,2 log UFC mL-1 depois de 120 min de exposição, respectivamente. No entanto, as células microencapsuladas em ALG e em ALG-PEC apresentaram resistência considerável, diminuindo 2,9 log UFC mL-1 para SGS e 2,7 log UFC mL-1 para SIS. Testes de armazenamento sob temperatura de refrigeração por 21 dias apresentaram boa sobrevivência bacteriana de 9,3 log UFC mL-1 (ALG) e 8,6 log UFC mL-1 (ALG-PEC) para células microencapsuladas, enquanto que as células livres apresentaram uma sobrevivência de apenas 1,2 log UFC mL-1 Na terceira etapa foram realizados experimentos para obtenção do pó alimentício com potencial probiótico, onde o microrganismo microencapsulado em ALG foi liofilizado e analisada a viabilidade no período de 6 meses de armazenamento a temperatura ambiente (25 °C), aqual foi mantida acima de 7 log UFC g-1 de pó alimentício, a análise microbiológica (conforme legislação brasileira) realizada antes e após o período de armazenamento não demonstrou contaminações para os patógenos avaliados. Realizou-se uma análise sensorial adicionando o pó alimentício em suco natural de laranja, obtendo aceitação sensorial elevada, maior que 88 %. O suco com adição do pó alimentício foi exposto aos SGS e SIS e apresentou, após 120 min, redução de apenas 2,4 log UFC mL-1 para SGS e 1,3 log UFC mL-1 para SIS. / Probiotic microorganisms are considered living dietary supplements showing benefic effects to hosts by improving the intestinal balance. The high cell density production of Lactobacillus has been the interest of many studies and presents great interest for industry, along with the development of new alternative culture media. Other concerns are the improvement of the survival of microorganisms during passage through the gastrointestinal tract by means of microencapsulation, and the preparation of a product with probiotic potential that would require no cold chain. In this context, this study aimed at producing potentially probiotic bacterium with alternative sources of cultivation substrates and its microencapsulation to obtain a food powder to be used directly in food. In the first step of this study a screnning of variables was carried out to set the process parameters and culture medium in the submerged bioreactor for the production of L. plantarum BL011 The optimized culture medium and processing parameters for biomass and lactic acid formation were: 40 g L-1 total sugar (liquid acid protein residue of soybean); 15 g L-1 yeast extract; stirring speed of 200 rpm; 25 °C, and 4.5 vvm. The results obtained allowed for a production of 17.87 g L-1 of biomass and 37.59 g L-1 of lactic acid. In a second step of this study L. plantarum BL011 was microencapsulated using the electrospraying technique, using as encapsulating agents sodium alginate (ALG) and a mixture of sodium alginate and citrus pectin (ALG-PEC). The free and microencapsulated cells were subjected to the simulated gastric juice (SGJ) and simulated intestinal juice (SIJ). The microorganism control (free cells) showed a decrease of 6 and 4.2 log CFU mL-1 after 120 min of exposure, respectively. However, the microencapsulated cells in ALG and in ALG-PEC showed significant resistance, decreasing by 2.9 log CFU mL-1 in SGJ, and 2.7 log CFU mL-1 in SIJ. Storage tests under refrigeration temperature for 21 days showed good bacterial survival of 9.3 log CFU mL-1 (ALG) and 8.6 log CFU mL-1 (ALG-PEC) for microencapsulated cells, whereas free cells showed a survival of only 1.2 log CFU mL-1 In the third step of the work, it was obtained a food powder with probiotic potential, where the ALG-microencapsulated bacterium was lyophilized and viability was investigated within 6 months of storage at room temperature (25 °C), keeping 7 log CFU g-1 product of its initial value. Microbiological analyses (according to Brazilian legislation) performed before and after the storage period did not show any contaminations by pathogens. The formulated orange juice containing L. plantarum BL011 obtained high sensory acceptance (> 88 %) in the sensory analysis. The juice with the addition of food powder was exposed to SGJ and SIJ and presented, after 120 min, reduction of 2.4 log CFU mL-1 for SGJ and 1.3 log CFU mL-1 for SIJ.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The benefits provided by probiotics to the human body have provided their addition to various products, spreading their consumption. However, due to various factors such as storage at low temperatures, acidity and the passage through the human gastrointestinal tract undermine the viability of these organisms. Microencapsulation is alternative for the protection of these probiotics to the human intestine. The aim of this study was to develop probiotic microcapsules Bifidobacterium BB-12 by internal ionic gelation in the wet and freeze-dried form. Moreover, it was analyzed the survival of probiotics under simulated gastrointestinal conditions, tolerance "in vitro" when inoculated at different pH solutions (4.5, 6.0 and 7.5) and viability during storage at different temperatures (-18, 7, 25 °C) at different times for 120 days. In addition to the morphology, mean diameter and physicochemical characterization of microparticles. Under the conditions of 1.5% sodium alginate, 190 ×g rotation speed and 1.5% emulsifier, the microparticles had a mean diameter of 55 μm and an encapsulation yield greater than 90%. In relation to tests simulating gastrointestinal conditions, both the moist microcapsules as lyophilized were resistant, with microbial counts of 12.93 and 11.13 log CFU g-1 respectively, and these are within the standards required by Brazilian law to occur benefits exercised by probiotics. Both moist microcapsules as lyophilized showed good protection in acidic solution (pH 4.5) and total liberation of probiotics in weakly basic solution (pH 7.5). The viability of wet microcapsules was maintained for 75 days at room temperature, and there was a reduction of 6.74 log CFU g-1 over existing storage due to metabolic activity, thus resulting in cell death and loss of cell viability. Compared to other storage temperatures, the refrigeration temperature was further reduction, which was 10.52 log CFU g-1. While, in the freezing showed the best results with a probiotic viability of 7.31 log UFC g-1 after the 120 days. Analyzing the lyophilized microcapsules at room temperature caused a probiotic viability by 60 days. However, refrigeration temperatures and freeze resulted in viable microparticles for 120 days of storage. The results of the physico-chemical characterization indicated encapsulation yield and stability of the microparticles with high efficiency, to facilitate incorporation into food products. / Os benefícios proporcionados pelos probióticos ao organismo humano têm proporcionado sua adição a diversos produtos, difundindo seu consumo. No entanto, devido a vários fatores como o armazenamento em baixas temperaturas, acidez e a passagem pelo trato gastrointestinal humano prejudicam a viabilidade destes microrganismos. A microencapsulação vem como alternativa de proteção destes probióticos até o intestino humano. O objetivo geral deste trabalho foi desenvolver microcápsulas probióticas de Bifidobacterium BB-12 por gelificação iônica interna na forma úmida e liofilizada. Além disto, foi analisado a sobrevivência dos probióticos sob condições gastrointestinais simuladas, tolerância ―in vitro‖ quando inoculados em diferentes soluções de pH (4.5, 6.0 e 7.5) e viabilidade durante armazenamento sob diferentes temperaturas (-18, 7 e 25 °C) em diferentes tempos por 120 dias. Além da morfologia, diâmetro médio e caracterização físico-química das micropartículas. Sob as condições de 1.5% de alginato de sódio, 190 ×g de velocidade de rotação e 1.5% de emulsificante, as micropartículas apresentaram um diâmetro médio de 55 μm e uma eficiência de encapsulação superior a 90%. Em relação aos testes simulando as condições gastrointestinais, tanto as microcápsulas úmidas como as liofilizadas foram resistentes, apresentando uma contagem de 12.93 e 11.13 log UFC g-1 respectivamente, estando dentro dos padrões exigidos pela legislação brasileira para que ocorram os benefícios exercidos pelos probióticos. Tanto as microcápsulas úmidas como as liofilizadas apresentaram boa proteção em solução ácida (pH 4.5) e liberação total dos probióticos em solução fracamente básica (pH 7.5). A viabilidade das microcápsulas úmidas, foi mantida durante 75 dias à temperatura ambiente, sendo que houve uma redução de 6.74 log UFC g-1 ao longo do armazenamento devido a atividade metabólica existente, resultando assim na morte de células e perda de viabilidade celular. Comparado as outras temperaturas de armazenamento, na temperatura de refrigeração houve maior redução, que foi de 10.52 log UFC g-1. Enquanto, que no congelamento apresentou os melhores resultados com uma viabilidade probiótica de 7.31 log UFC g-1 após os 120 dias. Analisando as microcápsulas liofilizadas, a temperatura ambiente ocasionou uma viabilidade probiótica até os 60 dias. Entretanto, as temperaturas de refrigeração e congelamento acarretaram em micropartículas viáveis por 120 dias de estocagem. Os resultados da caracterização físico-química e eficiência de encapsulação indicaram uma estabilidade das micropartículas com alta eficiência, facilitando a incorporação em produtos alimentícios.