Yes / The synthesis of nanoparticles of conjugates of caffeic acid-modified chitosan with ruthenium arene complexes is described. The chemical structure and physical properties of the nanoparticles were characterised by electronic absorption spectroscopy (UV-vis), Fourier transform infrared spectroscopy (FT-IR), 1H NMR spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), and circular dichroism (CD) analysis. The multi-spectral results revealed that caffeic acid is covalently bound to chitosan and chelates to {Ru(p-cymene)Cl}+. The DLS studies indicated that the Ru–caffeic acid modified chitosan nanoparticles are well-defined and of nanometre size. Such well-defined nanocomposites of chitosan and metal complexes might find a range of applications, for example in drug delivery. / We thank the National Natural Science Foundation of China (Project No. 21571154), the Jiangsu Overseas Research & Training Program for University Prominent Young & Middle-aged Teachers and Presidents, Leverhulme Trust (Early Career Fellowship No. ECF-2013-414 to NPEB), the ERC (Grant No. 247450 to PJS), EPSRC (EP/F034210/1 to PJS) and Science City (AWM/ERDF) for support, and EU COST Action CM1105 for stimulating discussions.

Over the past several decades, gene therapy technologies have been developed for a diverse number of applications ranging from DNA-based vaccines to gene silencing with RNAi. While all are powerful tools, a common limitation for these technologies is the need for effective and safe delivery to target sites within the body. Such delivery vectors are necessary for retention of bioactivity and stability, while also providing a method of cellular and tissue uptake and distribution, which may require endosomal escape. Although, viral and lipid-based technologies have shown promise as nucleic acid delivery vectors, both have inherent issues such as cytoxicity, oncogenicity, and immunogenicity. Thus, the development of polymer-based non-viral vectors has been an area of great focus over the past decade. While many polymeric vectors have been developed for plasmid DNA (pDNA) delivery, very few have shown effective delivery of short interfering RNA (siRNA), a powerful tool for gene silencing via the RNA interference mechanism. Furthermore, very few prospective delivery vectors have shown versatility for the administration of siRNA through multiple routes of administration. The overall goal of this research was to develop a biocompatible non-viral delivery system for the delivery of plasmid DNA, siRNA, and contrast agents through the modification of the natural biopolymer chitosan. We have synthesized an imidazole modified chitosan (chitosan-IAA) by conjugation of imidazole acetic acid to chitosan. Extensive evaluation and characterization of the modified polymer demonstrates enhanced solubility and buffering capacity within the physiological and endosomal pHs, thus providing enhanced endosomal escape by exploiting the "proton sponge" effect. We have demonstrated effective in vitro gene expression and gene silencing with chitosan-IAA mediated delivery of pDNA and siRNA, respectively. Furthermore, we have demonstrated in vivo gene silencing by delivery of siRNA through both intranasal and intravenous routes of delivery with chitosan-IAA/siRNA nanocomplexes. We have also demonstrated delivery of contrast agents up to 45 nm in size through mucosal tissue following treatment with chitosan and no contrast agent modification in both human and animal tissue. In conclusion, we have successfully developed a versatile and highly effective delivery vector for both nucleic acids and contrast agents. / text

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-09-06T12:43:49Z No. of bitstreams: 1 arquivototal.pdf: 4989490 bytes, checksum: e588d94dcdcc484ca970d62d2e581dd7 (MD5) / Made available in DSpace on 2017-09-06T12:43:49Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4989490 bytes, checksum: e588d94dcdcc484ca970d62d2e581dd7 (MD5) Previous issue date: 2016-08-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The aim of this study was to evaluate the inclusion of chitosan and modified chitosan (chitosan-glucose) in appropriate concentration to meet the health claim in sausages with reduced fat, on the technological functionality and shelf life of these sausages. For this, an initial study was conducted, in which chitosan (2% w/w) was added to sausages prepared with pork and three levels of fat (5%, 12.5% and 20% w/w). The sausages were submitted to cold storage at 4 ± 1 °C for 15 days and evaluated in terms of microbiological, physic-chemical and sensory parameters. Subsequently, considering the absence of studies on incorporation of chitosan in meat goat products, carried out a second study, similar to the above to evaluate the effect of incorporation of chitosan (2% w/w) in sausage made with goat meat, and similar levels of fat in the microbiological and physico-chemical quality during cold storage (4 ± 1 °C) for 15 days. Finally, in the third study was evaluated the effect of the derivative of chitosan (2% w/w) in comparison with chitosan base (2% w/w) in sausage made with goat meat and reduced fat content (10% w/w). The sausages were stored at 4 ± 1 ° C for 21 days and evaluated by microbiological, physic-chemical and sensory parameters. In all three studies a formulation without the addition of polymers (chitosan and modified chitosan) was used as control. The results indicated that the use of 2% (w / w) chitosan and its derivative (corresponding to 1 g of chitosan per sausage of 50g each in a portion of 150g) to ensure ingestion of 3 g of chitosan per day is technologically feasible for use in the preparation of sausage with pork and goat meat. In addition to the functional potential, chitosan and chitosan derivative provided increased stability of the treated sausages since it caused the reduction of microbial growth when compared to the control sample. Additionally, they provided an improvement in red color, resulting in positive effects on the appearance, a more stable emulsion for the greater ability to bind water and fat, and a firmer texture to increase the compression forces, without negatively affecting the sensory properties of sausages. The chitosan and modified chitosan showed similar behavior, but the use of chitosan resulted in higher antioxidant activity than its derivative, considering further reduction of TBARS values and increased content of unsaturated fatty acids and poly-unsaturated. However, the chitosan derivative promoted a greater red color (a *) and lower hardness than chitosan. Therefore, the results of the use of chitosan in the manufacture of goat sausages, which was studied for the first time, indicated that the chitosan and modified chitosan showed potential interest to be used in the preparation of meat products, since they can provide higher quality and shelf life, adding value to the raw material of low commercial value, besides allow to obtain of a functional product with the health claim and fat reduction. / O objetivo deste estudo foi avaliar a inclusão da quitosana e quitosana modificada (quitosana-glicose), em concentração adequada para atender a alegação de saúde, em salsichas com reduzido teor de gordura, sobre a funcionalidade tecnológica e a vida de prateleira destas salsichas. Para isso, um primeiro estudo foi realizado, no qual a quitosana (2 % m/m) foi adicionada em salsichas elaboradas com carne suína e três níveis de gordura (5%, 12,5% e 20%, m/m). As salsichas foram submetidas ao armazenamento refrigerado a 4 ± 1 ºC por 15 dias e avaliadas em relação aos parâmetros microbiológicos, físico-químicos e sensoriais. Posteriormente, considerando a inexistência de estudos sobre a incorporação de quitosana em produtos cárneos caprino, realizou-se um segundo estudo, semelhante ao anterior, para avaliar o efeito da incorporação da quitosana (2 % m/m) em salsicha elaborada com carne caprina e similares níveis de gordura na qualidade microbiológica e físico-química durante o armazenamento refrigerado (4 ± 1 ºC) por 15 dias. Por fim, no terceiro estudo foi avaliado o efeito do derivado da quitosana (2% m/m), comparando com a quitosana base (2% m/m), em salsicha elaborada com carne caprina e reduzido teor de gordura (10% m/m). As salsichas foram armazenadas a 4 ± 1 ºC por 21 dias, e avaliadas nos parâmetros microbiológicos, físico-químicos e sensoriais. Nos três estudos uma formulação sem adição dos polímeros (quitosana e quitosana modificada) foi usada como controle. Os resultados indicaram que o uso de 2% (m / m) de quitosana e seu derivado (correspondendo a 1 g de quitosana por salsicha de 50g cada em uma porção de 150g) para garantir a ingestão de 3 g de quitosana por dia é tecnologicamente viável para uso na elaboração de salsicha com carne de porco e cabra. Além do potencial funcional, a quitosana e o derivado da quitosana proporcionaram aumento da estabilidade das salsichas tratadas, uma vez que provocaram a redução do crescimento microbiológico quando comparadas a amostra controle. Adicionalmente, proporcionaram uma melhoria na cor vermelha, resultando em efeitos positivos na aparência, uma emulsão mais estável, pela maior capacidade de ligar água e gordura, e uma textura mais firme por aumentar as forças de compressão, sem afetar negativamente as propriedades sensoriais das salsichas. A quitosana e a quitosana modificada mostraram comportamento semelhante, mas o uso da quitosana resultou em maior atividade antioxidante do que o seu derivado, considerando a maior redução dos valores de TBARS e maior teor de ácidos graxos insaturados e poli-insaturados. Entretanto, o derivado da quitosana promoveu maior cor vermelha (a*) e menor dureza que a quitosana. Portanto, os resultados da utilização da quitosana na fabricação de salsichas caprina, a qual foi estudada pela primeira vez, indicaram que a quitosana e a quitosana modificada apresentaram potencial interessante para serem utilizados na elaboração de produtos cárneos, uma vez que podem proporcionar maior qualidade e vida de prateleira, agregando valor à matéria-prima de baixo valor comercial, além de permitir a obtenção de um produto funcional com alegação de saúde e redução de gordura.