Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:13Z No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T22:23:31Z (GMT) No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) / Made available in DSpace on 2016-04-28T22:23:31Z (GMT). No. of bitstreams: 1 MariliaMedeirosFernandesDeNegreiros_DISSERT.pdf: 2997173 bytes, checksum: 85466eb8c67b6b55509f4861197c4d35 (MD5) Previous issue date: 2015-08-07 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / A produ??o de nanoparticulas de prata com extratos naturais tem sido apontada como uma excelente alternativa para potencializar ou fornecer novas aplicabilidades aos extratos. Extratos de polissacar?deos sulfatados de algas (ASP) apresentam propriedades farmacol?gicas, por?m h? poucos relatos da produ??o de nanopart?culas de prata com extratos ricos em polissacar?deos sulfatados (SPN). Assim, neste trabalho sintetizou-se SPN de algas encontradas no Brasil: Spatoglossum schr?ederi, Dictyopteris justii, Sargassum filipendula e Dictyota mertensii. A obten??o dos extratos ricos em polissacar?deos ocorreu por prote?lise seguida por precipita??o com metanol. A s?ntese das diferentes nanopart?culas ocorreu com a adi??o de solu??es de prata 1 mM em solu??es dos diferentes polissacar?deos e mantidos em repouso. Posteriormente as amostras foram centrifugadas e liofilizadas. A forma??o SPN foi confirmada por espectroscopia UV/vis?vel, microscopia eletr?nica de varredura e microscopia de for?a at?mica. O tamanho das SPN foi de 108 ? 2 nm; 82 ? 1nm; 288 ? 52 nm; 104 ? 2 nm para S. schr?ederi; D. justii; S. filipendula; D. mertensii, respectivamente e se manteve est?vel por catorze meses, os potenciais zeta foram negativos e as formas das SPN foram esf?ricas. Os resultados de diversos testes in vitro mostraram que as SPN potencializam as atividades antioxidantes de ASP. As SPN tamb?m foram biocompat?veis com c?lulas normais 3T3 (fibroblastos murinicos). Por outro lado, SPN de S schr?ederi e de D. mertensii tiveram atividade citot?xica (~60%) frente as c?lulas de melanoma mur?nico (B16F10) e acredita-se que a maior atividade citot?xica destas SPN ocorram devido aos seus pequenos tamanhos. SPN tamb?m tiveram grande capacidade antibacteriana. Nanopart?culas de D. justii e S. filipendula tiveram os melhores resultados, sendo necess?rio somente 50 ?g/mL para a morte da bact?ria E. coli e 100 ?g/mL para a morte de S. aureus. Hipotet?za-se que esta atividade ocorra por libera??o de prata pelas SPN. SPN tamb?m foram capazes de induzir a produ??o de ?xido n?trico e citocinas com perfil semelhante a os seus respectivos ASP, com exce??o para SPN de S. schr?ederi. Em geral, os resultados revelaram que a s?ntese de SPN a partir das ASP potencializa efeitos antioxidantes, citot?xico e antibacteriano dos ASP, al?m de apresentar o mesmo efeito imunomodulador de seus respectivos ASP. Os dados obtidos levam a propor que a s?ntese de SPN constituiu-se como um poss?vel mecanismo potencializador de atividades biol?gicas. / The production of silver nanoparticle containing natural extracts has been identified as an excellent alternative to enhance or provide new applicability for these extracts. Sulfated polysaccharides-rich extracts (ASP) from seaweeds showed several pharmacological properties, but there are few reports of the production of silver nanoparticle with sulfated polysaccharides (SPN). Therefore, this paper we synthesized SPN from seaweeds founded in Brazil: Spatoglossum schr?ederi, Dictyopteris justii, Sargassum filipendula e Dictyota mertensii. The obtainment of polysaccharides rich extracts it occurred by proteolysis followed by methanol precipitation. The synthesis of various nanoparticles occurred with the addition of silver solution 1 mM on different polysaccharides solutions and kept at rest. Subsequently the samples were centrifuge and lyophilized. The SPN formation was confirmed by UV/ visible spectroscopy, scanning electron microscopy and atomic force microscopy. The size of SPN was 108 ? 2 nm; 82 ? 1nm; 288 ? 52 nm; 104 ? 2 nm for S. schr?ederi; D. justii; S. filipendula; D. mertensii, respectively. The nanoparticules remained stable for fourteen months, the Zeta potential were negative and forms of SPN were rounded. Data from various antioxidant in vitro tests showed that the SPN were more effective than ASP. The SPN were also biocompatible with the normal cells 3T3 (murine fibroblast). Moreover, SPN of S. schr?ederi and of D. mertensii had cytotoxic activity (~60%) against melanoma cells (B16F10). We believe that the increased of cytotoxic activity of these SPN occur due to their small size. SPN also had great antibacterial capacity. D. justii and S. filipendula nanoparticles had the best results, requiring only 50 mg/mL and 100 mg/mL for induces E. coli and S. aureus death. We hypothesized that this activity occurs by release of SPN?s silver. SPN also increase the nitric oxide and cytokines release from macrophages as similar profile to the respective ASP, except for S. schr?ederi SPN. In general, the results showed that SPN were more effective as antioxidant, antibacterial and cytotoxic agents than their respective ASP, as well as, SPN have shown the same immunomodulation effect of the respective ASP. The results lead to propose that the SPN synthesis constituted as a possible potentiating mechanism of biological activities of ASPs.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-06-02T22:15:27Z No. of bitstreams: 1 LarisseAraujoDantas_DISSERT.pdf: 2039205 bytes, checksum: c6567bc4fed86a3072ac9d28162203b1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-06-08T19:00:32Z (GMT) No. of bitstreams: 1 LarisseAraujoDantas_DISSERT.pdf: 2039205 bytes, checksum: c6567bc4fed86a3072ac9d28162203b1 (MD5) / Made available in DSpace on 2017-06-08T19:00:32Z (GMT). No. of bitstreams: 1 LarisseAraujoDantas_DISSERT.pdf: 2039205 bytes, checksum: c6567bc4fed86a3072ac9d28162203b1 (MD5) Previous issue date: 2017-04-17 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Nanopart?culas de prata (AgNPs) possuem diversas aplica??es biom?dicas, dentre elas destaca-se o potencial antiproliferativo. In?meros tipos de s?ntese de nanopart?culas de prata encontram-se descritos na literatura, no entanto, um tipo de s?ntese menos agressiva e mais economicamente vi?vel, a s?ntese verde, tem ganhado import?ncia dentre as demais. Neste processo de s?ntese ? necess?rio o uso de um agente redutor e estabilizante que n?o sejam t?xicos para as c?lulas animais e para o meio ambiente. Nesse contexto, a associa??o de biomol?culas de origem marinha com atividades biol?gicas j? descritas, como os polissacar?deos ?cidos de algas, ?s AgNPs ganham import?ncia. Muitos polissacar?deos ?cidos (PA) extra?dos de algas marinhas destacam-se por exibir diversas atividades, como exemplo as fucanas da alga Spatoglossum schr?ederi. Estas fucanas apresentam atividade antioxidante, antiangiog?nica, antitumoral, dentre outras. Devido aos poucos relatos de s?ntese de AgNPs envolvendo polissacar?deos de algas, este trabalho teve por objetivo sintetizar AgNPs com uma fra??o rica em ?cido alg?nico e fucana A da alga S. schr?ederi por um processo de s?ntese verde, e testar as AgNPs obtidas frente ? linhagem de melanoma B16F10. Os PA da alga S. schr?ederi foram extra?dos por um processo de digest?o proteol?tica e fracionados com acetona. A fra??o de maior rendimento e rica em ?cido alg?nico e fucana A, a F0.5 v, foi empregada para a s?ntese das AgNPs. A caracteriza??o das AgNPs obtidas contemplou a an?lise de absor??o de luz UV, composi??o qu?mica, di?metro m?dio, polidispers?o, potencial zeta, espectroscopia de infravermelho (FTIR) e a espectroscopia de energia dispersiva (EDS), microscopia eletr?nica de varredura (MEV) e microscopia de for?a at?mica (MFA). As an?lises por espectroscopia de UV-vis?vel na faixa entre 400 e 440 nm confirmaram a forma??o das AgNPs, estas apresentaram di?metro de 196 ? 13 nm, valores de polidispers?o abaixo de 0,4 e potencial zeta negativo. As dosagens qu?micas, o FTIR e o EDS revelaram que a maior parte das AgNPs ? composta por polissacar?deos ?cidos. As imagens obtidas por MEV e MFA indicaram um formato esf?rico para as AgNPs. A atividade antiproliferativa foi avaliada pelo m?todo do MTT frente ?s linhagens celulares B16F10 e fibroblastos 3T3, e por citometria de fluxo foi investigado sua a??o na morte e no ciclo celular. As AgNPs afetaram a capacidade de redu??o do MTT das linhagens 3T3 e B16F10, sendo esta mais evidenciada para a B16F10, pois as AgNPs (0,5 mg/mL) conseguiram reduz?-la em torno de 50%, enquanto a prata i?nica e a F0.5 v em nenhuma das concentra??es influenciaram na redu??o do MTT. As an?lises por citometria de fluxo apontaram uma elevada taxa de marca??o para anexina V e iodeto de prop?dio para as c?lulas B16F10 expostas ?s AgNPs, al?m de um aumento na porcentagem das c?lulas em Sub-G1 em detrimento ?s outras fases. Estudos adicionais s?o necess?rios para elucidar completamente o mecanismo de a??o antiproliferativo das AgNPs e descobrir outras propriedades da fra??o que s?o intensificadas pela s?ntese de nanopart?culas. Entretanto, pode-se concluir que ? poss?vel sintetizar AgNPs com polissacar?deos ?cidos de S. schr?ederi usando um m?todo verde e que essas AgNPs apresentaram atividade antiproliferativa mais pronunciada que seus percussores. / Silver nanoparticles (AgNPs) have several biomedical applications, among them the antiproliferative potential. Numerous types of synthesis of silver nanoparticles are found in the literature, however, a type of synthesis less aggressive and more economically viable, a green synthesisl, has gained value among the others. In this synthesis process, it is necessary to use a reducing agent and stabilizer which are non-toxic to animal cells and to the environment. In this context, an association of biomolecules of marine origin with biological activities already described, such as the polysaccharide acids of algae, to the AgNPs gain importance. Many acidic polysaccharides (PA) extracted from marine algae are distinguished by their different activities, such as the algae Spatoglossum schr?ederi. These fucans have antioxidant, antiangiogenic, antitumor activity, among others. Due to the few reports of AgNPs synthesis involving algae polysaccharides, the objective of this work was to synthesize AgNPs with a rich fraction of alginic acid and fucan A from the S. schr?ederi algae by a green synthesis process and to test the AgNPs obtained on the lineage of melanoma B16F10. The PA of the S. schr?ederi algae was extracted by a proteolytic digestion process and fractionated with acetone. The higher yield fraction rich in alginic acid and fucan A, F0.5 v, was used for synthesis of the AgNPs. The characterization of the AgNPs obtained contemplated an analysis of UV light absorption, chemical composition, average diameter, polydispersity, zeta potential, infrared spectroscopy (FTIR), dispersive energy spectroscopy (EDS), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The analyzes by UV-visible spectroscopy in the range between 400 and 440 nm confirmed the formation of AgNPs, they presented 196 ? 13 nm diameter, polydispersion values below 0.4 and negative zeta potential. The chemical dosages, FTIR and EDS revealed that most AgNPs are composed of acidic polysaccharides. The images obtained by SEM and AFM indicated a spherical format for AgNPs. The antiproliferative activity was evaluated by the MTT method against B16F10 cell lines and 3T3 fibroblasts, and by flow cytometry, its action on death and cell cycle was investigated. The AgNPs affected the ability to reduce MTT from the 3T3 and B16F10 lineages, but these were evidenced for a B16F10, the AgNPs (0.5 mg/mL) were able to reduce by around 50%, while a silver ion and F0.5 v at any of the concentrations influenced the reduction of MTT. Flow cytometry analyzes indicated a high labeling rate for annexin V and propidium iodide for the B16F10 cells exposed to AgNPs, in addition to an increase in the percentage of cells in SubG1 in detriment to the other phases. Additional studies are needed to fully elucidate the mechanism of antiproliferative action of AgNPs and to discover other properties of the fraction that are enhanced by the synthesis of nanoparticles. However, it can be concluded that it is possible to synthesize AgNPs with acidic polysaccharides from S. schr?ederi using a green method and that these AgNPs showed more pronounced antiproliferative activity than their precursors.