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Design and Preparation of Gelatin-Based Carriers for Imaging Probes to Visualize Cell Functions / 細胞機能を可視化するイメージングプローブのためのゼラチンからなるキャリアのデザインと作製Murata, Yuki 23 March 2021 (has links)
京都大学 / 新制・課程博士 / 博士(工学) / 甲第23161号 / 工博第4805号 / 新制||工||1751(附属図書館) / 京都大学大学院工学研究科高分子化学専攻 / (主査)教授 田畑 泰彦, 教授 秋吉 一成, 教授 沼田 圭司 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM
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Obtenção e caracterização de membrana de gelatina e membrana de gelatina com prata para uso em regeneração tecidual guiada / Preparation and characterization of membrane gelatin and membrane gelatin silver for guided tissue regenerationSousa, Lorena Oliveira de 26 August 2013 (has links)
O estudo de biomateriais aplicados à cicatrização levou à modalidade do tratamento chamada de regeneração tecidual guiada (RTG). A RTG reconstitui novos tecidos usando membranas como barreira de proteção da área defeituosa, impedindo a invasão de outros tecidos, especialmente dos tecidos conjuntivos fibrosos. Em outras palavras, a RTG é uma forma de isolar o tecido ósseo do tecido mole, resultando dessa forma em uma ótima condição para a formação óssea. Atualmente, mais de 20 tipos diferentes de polímeros são usados em aplicações biomédicas, e entre eles está o colágeno, polímero ao qual, pela sua hidrólise parcial pode-se obter a gelatina. A gelatina é um polipeptídeo biodegradável, biocompatível, que apresenta anti-genicidade, plasticidade e elasticidade. A prata tem seu efeito antimicrobiano bem conhecido, com utilidade em diferentes campos da medicina, odontologia e biomateriais. Nesse contexto, este trabalho desenvolve e estuda o comportamento das membranas de gelatina e membranas de gelatina com prata para aplicações em RTG. As Membranas de gelatina foram obtidas com concentração de 4% em massa e as membranas de gelatina com prata foram obtidas nas concentrações de 0,01%, 0,05% e 0,1% da solução. Na etapa de reticulação, as membranas secas foram imersas em solução tampão de fosfato de sódio com concentrações de glutaraldeído (GTA) a 0,5%, 1% e 2%, 24 horas. O ensaio de intumescimento foi realizado em membranas reticuladas e não reticuladas, pesadas e imersas em solução tampão de fosfato pH 7,4 por períodos de 5 minutos, 15 minutos, 30 minutos, 1 hora, 3 horas, 22 horas e 24 horas e durante 8 semanas. Obteve-se membranas de gelatina com prata mediante a metodologia de adição da solução de Ag compravada pelas técnicas de DRX, FTIR, MEV e pelo teste de halo de inibição. As membranas reticuladas apresentaram menor valor de porcentagem de intumescimento quanto maior foi a concentração do agente reticulante, com exceção das amostras com concentração de 0,1M de prata. Além da atuação do GTA a prata também interfere no processo de intumescimento, aspecto e comportamento das membranas de gelatina. O efeito bactericida foi avaliado utilizando ensaio de cultura de bactérias - Teste de halo de inibição, onde as membranas de gelatina com Ag apresentaram efeito bacteriostático contra bactéria Gram positiva e Gram negativa. / The study of biomaterials applied to healing led to the treatment modality called guided tissue regeneration (GTR). The RTG reconstruct new tissue using the membranes as a barrier to protect the defective area, preventing the invasion of other tissues, especially connective tissue fibers. In other words, the RTG is a way of isolating the bone tissue of the soft tissue, thereby resulting in an excellent condition for bone formation. Currently, more than 20 different types of polymers are used in biomedical applications, and among them is the collagen polymer to which, by its partial hydrolysis can get the gelatin. Gelatin is a polypeptide biodegradable, which has antigenicity, plasticity and elasticity. Silver has antimicrobial effect well known, are useful in different fields of medicine, dentistry and biomaterials. In this context, this paper develops and studies the behavior of membranes of gelatin and gelatin silver membranes for applications in RTG. Membranes were obtained with gelatin concentration of 4% by weight and gelatin silver membranes were obtained at concentrations of 0.01%, 0.05% and 0.1% of the solution. In the crosslinking step, the dried membranes were immersed in sodium phosphate buffer to concentrations of glutaraldehyde (GTA) at 0.5%, 1% and 2% 24 hours. The swelling test was performed on crosslinked and uncrosslinked membranes, weighed and immersed in phosphate buffer pH 7.4 for a period of 5 minutes, 15 minutes, 30 minutes, 1 hour, 3 hours 22 hours and 24 hours, and for 8 weeks. Obtained gelatin membranes with silver by the method of adding the solution of Ag proven by XRD techniques, FTIR, SEM and the zone of inhibition test. The crosslinked membranes had lower percentage of swelling the higher the concentration of crosslinking agent, except for the samples with a concentration of 0.1 M silver. In addition to operating the GTA silver also interferes with the process of swelling, appearance and behavior of gelatin membranes. The bactericidal effect was evaluated by bacterial culture test - Test of inhibition zone, where the gelatin membrane with Ag Ag exhibited bacteriostatic effect exhibited against Gram positive and Gram negative.
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Preparação e caracterização de hidrogéis neutros de colágeno aniônico:gelatina:extrato de semente de uva / Preparation and characterization of anionic collagen:gelatin:grape seed extract neutral hydrogelsLancelotti, Cindia 09 June 2014 (has links)
O desenvolvimento de uma matriz de colágeno associado à gelatina tem potencial como biomaterial devido sua alta biocompatibilidade, capacidade de alterar suas propriedades físico-químicas e estruturais por modificações químicas e a habilidade de formar géis estáveis. Porém, um ponto negativo de sua aplicação está na biodegradabilidade. Desta forma, para reduzir esta degradação, agentes de reticulação, como a proantocianidina (PA), que age formando ligações de hidrogênio as quais estabilizam o complexo proteína-PA, podem ser empregados. Este trabalho teve como objetivo a obtenção de hidrogéis neutros de colágeno aniônico:gelatina:extrato de semente de uva. Para tanto, foram utilizadas duas proporções de extrato, 0,25 e 0,50% e três diferentes tempos de hidrólise alcalina do colágeno, 24, 72 e 120 horas, gerando nove diferentes biomateriais, incluindo os hidrogéis sem tratamento com o extrato. O colágeno foi extraído de tendão bovino e o agente reticulante foi o extrato de semente de uva cujo componente majoritário é a proantocianidina. A caracterização foi feita por termogravimetria (TG), calorimetria exploratória diferencial (DSC), espectroscopia de absorção no infravermelho (FTIR), microscopia eletrônica de varredura (MEV), cinética de absorção de água e ensaios de citotoxicidade in vitro, pelos métodos de difusão em ágar e difusão de extrato em solução (MTT). Estudos de TG mostraram a perda de água em um processo único, representando em média 95% do hidrogel. As curvas DSC mostraram que quanto maior a concentração de extrato, maior é a temperatura de desnaturação, aumentando em média 6,8°C com a adição de 0,50% de extrato, o que indica a eficácia da proantocianidina na reticulação do colágeno. Além disso, notaram-se temperaturas menores para maiores tempos de hidrólise alcalina do colágeno, sendo de 58,4°C, 49,7°C e 46,5°C para hidrogéis preparados com colágeno 24, 72 e 120 horas, respectivamente. A presença do extrato não causou alterações significativas nos espectros FTIR, apenas surgimento das bandas em 1118 e 1288 cm-1 referentes ao anel aromático da proantocianidina, mas gerou mudanças nas estruturas internas dos hidrogéis, visualizadas por MEV, como aumento do número de poros e interconectividade entre eles. A cinética de absorção de água mostrou que o equilíbrio é atingido em aproximadamente 10 minutos, indicando vantagem para a aplicação do hidrogel, já que é obtido rapidamente a partir de sua forma liofilizada, adequada para o armazenamento. Também foi observado que quanto menor é o tempo de hidrólise, maiores são as absorções, variando de 540 até 1360%. Com os ensaios de citotoxicidade foi possível concluir que o hidrogel C24GE50 mostrou-se mais adequado para uma aplicação como biomaterial, com um índice de 92,7% de sobrevivência celular. / The development of a collagen matrix associated with gelatin has potential as biomaterial due to its high compatibility, ability to change its physical-chemical and structural properties by chemical modifications and also the ability to form stable gels. However, a negative point of its application is the biodegradability. Thus, to reduce this degradation, crosslinking agents, such as proanthocyanidin (PA), which acts forming hydrogen bonds which stabilize the PA protein complex may be employed. This project aimed to obtain neutral hydrogels prepared by mixture of anionic collagen: gelatin: grape seed extract. It was used two extract proportions (0.25 and 0.50%) and three different periods of time for collagen alkaline hydrolysis (24, 72, 120 hours), giving nine different biomaterials, including hydrogels without treatment with the extract. The collagen was extracted from bovine tendon and the crosslinking agent was grape seed extract whose major component is the proanthocyanidin. The characterization was done by thermogravimetry (TG), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy, scanning electron microscopy (SEM), water absorption kinetics and in vitro cytotoxicity assays, by agar diffusion method and diffusion of extract in solution (MTT). TG results showed loss of water in a single process, representing about 95% of the hydrogel.DSC curves showed that the higher the concentration of the extract, the higher the melting temperature, increasing on average 6.8 °C with the addition of 0.50% extract, which indicates the effectiveness of the proanthocyanidin crosslinking of the collagen. Moreover, lower temperatures were observed for longer periods of alkaline hydrolysis of collagen, being 58.4°C, 49.7°C and 46.5°C for preparations with 24, 72 and 120 hours, respectively. The presence of the extract did not cause significant changes in the FTIR spectra, only appearance of bands at 1118 and 1288 cm-1 related to the aromatic ring of proanthocyanidin but led to changes in internal structures of the hydrogels viewed by SEM, showing increased number of pores and interconnectivity between them. The water absorption kinetics showed that the equilibrium is achieved in approximately 10 minutes, indicating advantages in using this hydrogel, which is quickly obtained from its lyophilized form, a suitable form for storage. It was also observed that the shorter the time of hydrolysis, the greater the absorptions, ranging from 540 to 1360%. It was possible to conclude from the cytotoxicity analysis that the C24GE50 is more suitable for application as a biomaterial, with an index of 92.7% cell survival.
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Preparação e caracterização de hidrogéis neutros de colágeno aniônico:gelatina:extrato de semente de uva / Preparation and characterization of anionic collagen:gelatin:grape seed extract neutral hydrogelsCindia Lancelotti 09 June 2014 (has links)
O desenvolvimento de uma matriz de colágeno associado à gelatina tem potencial como biomaterial devido sua alta biocompatibilidade, capacidade de alterar suas propriedades físico-químicas e estruturais por modificações químicas e a habilidade de formar géis estáveis. Porém, um ponto negativo de sua aplicação está na biodegradabilidade. Desta forma, para reduzir esta degradação, agentes de reticulação, como a proantocianidina (PA), que age formando ligações de hidrogênio as quais estabilizam o complexo proteína-PA, podem ser empregados. Este trabalho teve como objetivo a obtenção de hidrogéis neutros de colágeno aniônico:gelatina:extrato de semente de uva. Para tanto, foram utilizadas duas proporções de extrato, 0,25 e 0,50% e três diferentes tempos de hidrólise alcalina do colágeno, 24, 72 e 120 horas, gerando nove diferentes biomateriais, incluindo os hidrogéis sem tratamento com o extrato. O colágeno foi extraído de tendão bovino e o agente reticulante foi o extrato de semente de uva cujo componente majoritário é a proantocianidina. A caracterização foi feita por termogravimetria (TG), calorimetria exploratória diferencial (DSC), espectroscopia de absorção no infravermelho (FTIR), microscopia eletrônica de varredura (MEV), cinética de absorção de água e ensaios de citotoxicidade in vitro, pelos métodos de difusão em ágar e difusão de extrato em solução (MTT). Estudos de TG mostraram a perda de água em um processo único, representando em média 95% do hidrogel. As curvas DSC mostraram que quanto maior a concentração de extrato, maior é a temperatura de desnaturação, aumentando em média 6,8°C com a adição de 0,50% de extrato, o que indica a eficácia da proantocianidina na reticulação do colágeno. Além disso, notaram-se temperaturas menores para maiores tempos de hidrólise alcalina do colágeno, sendo de 58,4°C, 49,7°C e 46,5°C para hidrogéis preparados com colágeno 24, 72 e 120 horas, respectivamente. A presença do extrato não causou alterações significativas nos espectros FTIR, apenas surgimento das bandas em 1118 e 1288 cm-1 referentes ao anel aromático da proantocianidina, mas gerou mudanças nas estruturas internas dos hidrogéis, visualizadas por MEV, como aumento do número de poros e interconectividade entre eles. A cinética de absorção de água mostrou que o equilíbrio é atingido em aproximadamente 10 minutos, indicando vantagem para a aplicação do hidrogel, já que é obtido rapidamente a partir de sua forma liofilizada, adequada para o armazenamento. Também foi observado que quanto menor é o tempo de hidrólise, maiores são as absorções, variando de 540 até 1360%. Com os ensaios de citotoxicidade foi possível concluir que o hidrogel C24GE50 mostrou-se mais adequado para uma aplicação como biomaterial, com um índice de 92,7% de sobrevivência celular. / The development of a collagen matrix associated with gelatin has potential as biomaterial due to its high compatibility, ability to change its physical-chemical and structural properties by chemical modifications and also the ability to form stable gels. However, a negative point of its application is the biodegradability. Thus, to reduce this degradation, crosslinking agents, such as proanthocyanidin (PA), which acts forming hydrogen bonds which stabilize the PA protein complex may be employed. This project aimed to obtain neutral hydrogels prepared by mixture of anionic collagen: gelatin: grape seed extract. It was used two extract proportions (0.25 and 0.50%) and three different periods of time for collagen alkaline hydrolysis (24, 72, 120 hours), giving nine different biomaterials, including hydrogels without treatment with the extract. The collagen was extracted from bovine tendon and the crosslinking agent was grape seed extract whose major component is the proanthocyanidin. The characterization was done by thermogravimetry (TG), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy, scanning electron microscopy (SEM), water absorption kinetics and in vitro cytotoxicity assays, by agar diffusion method and diffusion of extract in solution (MTT). TG results showed loss of water in a single process, representing about 95% of the hydrogel.DSC curves showed that the higher the concentration of the extract, the higher the melting temperature, increasing on average 6.8 °C with the addition of 0.50% extract, which indicates the effectiveness of the proanthocyanidin crosslinking of the collagen. Moreover, lower temperatures were observed for longer periods of alkaline hydrolysis of collagen, being 58.4°C, 49.7°C and 46.5°C for preparations with 24, 72 and 120 hours, respectively. The presence of the extract did not cause significant changes in the FTIR spectra, only appearance of bands at 1118 and 1288 cm-1 related to the aromatic ring of proanthocyanidin but led to changes in internal structures of the hydrogels viewed by SEM, showing increased number of pores and interconnectivity between them. The water absorption kinetics showed that the equilibrium is achieved in approximately 10 minutes, indicating advantages in using this hydrogel, which is quickly obtained from its lyophilized form, a suitable form for storage. It was also observed that the shorter the time of hydrolysis, the greater the absorptions, ranging from 540 to 1360%. It was possible to conclude from the cytotoxicity analysis that the C24GE50 is more suitable for application as a biomaterial, with an index of 92.7% cell survival.
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Characterisation and extrusion of Metroxylon sago starchAnsharullah, University of Western Sydney, Hawkesbury, Faculty of Environmental Management and Agriculture, School of Food Science January 1997 (has links)
The study presented here was firstly to investigate the physiochemical properties of native sago starch (obtained from Metroxylon sp. and designated as sago INA), in comparison with those of Metroxylon sago starch obtained from a different source, sago starch derived from Arenga sp. palms, wheat, corn, and tapioca starches. The properties analysed were chemical composition, total starch content, apparent amylose content, pasting properties, endothermic thermal behaviour, starch paste clarity, freeze-thaw stability, hardness of gel, and microscopic structure of the granules. The results obtained indicated that sago INA starch sample contained less fat and protein, compared to cereal starches. The sago starch sample had larger sized granules and had a more transparent paste. The gels of the starch were harder, and showed a relatively better stability to freeze-thaw treatment. The other part of the study was extrusion of sago INA starch both in the absence and presence of enzyme by utilising a response surface design. In the absence of the enzyme, the experiment was conducted to establish the extrusion process conditions including moisture contents, melt temperature, and screw speed. The extruded products were then analysed for degree of molecular degradation, light microscopic structure, reducing sugars of the water soluble materials, water absorption index, water solubility index, enzyme susceptibility, and gelatinisation endothermic energy. Increased mechanical and thermal energy input received by the products in the extruder resulted in a significant degradation of the molecular weight of the macromolecules. Light photomicrographs also suggested that the granule structures of the extrudates have been reshaped. All extrudate samples had a very low gelatinisation endothermic energy compared to its native starch. The specific mechanical energy received by the products in the extruder was calculated and related to the process variables. The possibility of using the products in food application was also discussed. / Doctor of Philosophy (PhD)
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Encapsulation of Protein Microfiber Networks Supporting Pancreatic IsletsSTEELE, JOSEPH ALLAN MCKINNON 24 August 2011 (has links)
A method was developed to produce and incorporate a network of discrete, genipin-crosslinked
gelatin microfibers around a pancreatic islet within a barium alginate microcapsule. This
technique allows for the encapsulation of a porous fibrous matrix without the geometrical
restrictions required for cellular aggregate seeding. Microfibers were produced from a novel
vortex-drawn extrusion system with an alginate support matrix. Optimization culminated in a
hydrated fiber diameter of 22.3 ± 0.4 μm, a 98% reduction in cross sectional area, while making
the process more reliable and less labour intensive. The optimized microfibers were encapsulated
at 40 vol% within 294 ± 4 μm 1.6% barium alginate microparticles by an electrostatic-mediated
dropwise extrusion system. Pancreatic islets extracted from Sprague Dawley rats were
encapsulated within the microparticles, and analyzed over a 21-day preliminary in vitro study.
Acridine orange and propidium iodide fluorescent viability staining and light microscopy
indicated a significant increase in viability for the fiber-laden particles relative to fiber-free
control particles at days 7, 14, and 21. The fiber-laden system also reduced the incidence of
disrupted islet cohesion from 31% to 8% at day 21, and showed evidence of islet-fiber adhesion.
Preliminary investigations into insulin secretion and metabolic activity showed no significant
difference between test and control groups. Further investigation into benefits of islet
encapsulation within an extracellular matrix fiber network will be the subject of future studies
with this body of work serving as a foundation.
The system developed in this investigation could be developed into a modular scaffold system for
tissue engineering beyond the field of islet research. / Thesis (Master, Chemical Engineering) -- Queen's University, 2011-08-18 15:05:50.917
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New Automated Industrial Technologies for Improving Chemical Penetration of Bovine Pieces in the Raw Material Processing and Conditioning Areas of Gelatine ManufactureWittich, William John January 2005 (has links)
The production of gelatine at Gelita N.Z. Ltd. is a time consuming process. The time limiting step in the process is the pre-treatment of the collagen tissue of the raw material in a lime/sodium sulfide solution. The liming solution breaks down the collagen in the tissue to gelatine. This is a necessary step prior to the extraction of gelatine from the hide pieces. The current liming process takes nearly 50 days to complete. Methods were investigated to increase the rate of penetration of the chemicals into the bovine hide raw material. An increase in the penetration of the liming solutions would lead to shorter processing times for this step in the process. The methods that were investigated were temperature controlled mixing, fluidization of the hide pieces and the use of ultrasound. Of all the methods tested, the fluidization of the hide pieces gave the best results. The pretreatment time of the hide pieces was reduced 9 days with this technique. Methods were also investigated to monitor the levels of conditioning in the raw material An accurate technique to measure hide conditioning was important to pilot plant trials. This helped determine how well any of the trail methods increased the penetration of chemicals into the hide pieces. The use of an ultraviolet dye proved an effective method of measuring conditioning for all the pilot plant trials. The level of chemical penetration was monitored by assessing the penetration of the UV dye. The penetration of the UV dye could be quantified by using imaging software. A possible method of monitoring conditioning in full-scale production was tested. It was determined that the glycosaminoglycans and soluble collagen released into the liming solution could be accurately measured, and related to the overall conditioning of the raw material.
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The combining weight of gelatin as an acid; The mobility of the gelatinate ion; The transference numbers and activities of sodium hydroxide in aqueous solution ...Schluchter, Alfred William, Ferguson, Alfred Lynn, January 1926 (has links)
Thesis (PH. D.)--University of Michigan, 1926. / "By A.L. Ferguson and A.W. Schluchter." Also issued in print.
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A new competitive inhibition technique for determination of plasma fibronectin activity and plasma fibronectin in B-thalassemia/HbE ; The role of plasma fibronectin as a non-immunological opsonin to promote monocyte phagocytic function in B-thalassemia HbE /Chawalit Kritpetchrarut, Sophark Rojanasthien, January 1986 (has links) (PDF)
Thesis (M.Sc. (Clinical Pathology))--Mahidol University, 1986.
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Diffusion potential studies: their application to the systems hydrochloric acid-sodium chloride, hydrochloric acid-gelatin and a new method for the determination of the equivalent weight of gelatin ...Bacon, Egbert King, Ferguson, Alfred Lynn, January 1927 (has links)
Thesis (Ph. D.)--University of Michigan, 1926. / Reprinted from two articles by Alfred L. Ferguson and Egbert K. Bacon, published in the Journal of the American Chemical Society, v. 49, August, 1927.
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