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Submitted by Mariana Rillo Sato (mari_sato_@hotmail.com) on 2020-10-27T19:07:55Z No. of bitstreams: 2 TESE PARCIAL FINAL ENTREGUE - MARIANA RILLO SATO.pdf: 2732250 bytes, checksum: 4cc1b40ef5936cf487c551ede38422c7 (MD5) TESE FINAL INTEGRAL ENTREGUE - MARIANA RILLO SATO.pdf: 8151565 bytes, checksum: ecdb4916a9cf97a305039bf15df9fb1e (MD5) Approved for entry into archive by Maria Irani Coito (irani@fcfar.unesp.br) on 2020-10-27T19:48:05Z (GMT) No. of bitstreams: 1 sato_mr_dr_arafcf_par.pdf: 2777263 bytes, checksum: e8a4a568713d4295f5b6dd2039aba4f9 (MD5) Made available in DSpace on 2020-10-27T19:48:05Z (GMT). No. of bitstreams: 1 sato_mr_dr_arafcf_par.pdf: 2777263 bytes, checksum: e8a4a568713d4295f5b6dd2039aba4f9 (MD5) Previous issue date: 2020-01-30 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) A candidíase vulvovaginal (CVV) é uma infecção fúngica causada tipicamente por Candida albicans, a qual apresenta resistência aos fármacos utilizados no tratamento. Carreadores lipídicos nanoestruturados (CLN) com hipericina (HIP) encapsulada dispersos em hidrogéis (HG) de geleificação in situ demonstram ser uma opção interessante para viabilizar o tratamento da CVV, associada a métodos alternativos, como a terapia fotodinâmica (TFD). Este trabalho visa avaliar o potencial dos CLN com e sem HIP encapsulada dispersos em HG, associados à TFD para administração vaginal. Os CLN sem (F6.5) e com HIP encapsulada (F6.5_0,05, F6.5_0,25 e F6.5_0,5) foram obtidos por sonicação a 35% de amplitude por 5min, contendo poloxamer 407 e na fase oleosa, estearato de polioxietileno, triglicérides do ácido cáprico/caprílico, óleo de rícino hidrogenado, etoxilado e/ou HIP. Os HG foram constituídos de poloxamer 407 e quitosana, sendo acrescidos de CLN com e sem HIP encapsulada em diferentes proporções de CLN:HG dando origem às formulações CLN_HIP-HG1, CLN_HIP-HG2, CLN_HIP-HG3, CLN_HIP-HG4 e CLN_HIP-HG5. Os sistemas F6.5, F6.5_0,05, F6.5_0,25 e F6.5_0,5 caracterizados por espalhamento de luz dinâmico, apresentaram diâmetros nanométricos com baixa polidispersão, e revelaram excelente estabilidade física. A formulação F6.5 apresentou na análise de rastreamento de nanopartículas distribuição unimodal com predominância de partículas de 151 nm. Na microscopia eletrônica de transmissão foram observadas partículas de morfologias praticamente esféricas e em escalas nanométricas. A eficiência de encapsulação da HIP foi de 89,3%. A microscopia eletrônica de varredura dos HG acrescidos de CLN com HIP encapsulada obtidos em diferentes proporções, revelou valores médios de rugosidade superficial sem diferença estatística entre os sistemas. A reologia contínua mostrou que praticamente todos os sistemas tiveram um comportamento Newtoniano; enquanto na reologia oscilatória a maioria dos sistemas apresentou valores de G’ superiores a G’’, com exceção de CLN_HIP-HG1. O HG livre e os sistemas CLN_HIP-HG1 e CLN_HIP-HG2 apresentaram satisfatórias propriedades mucoadesivas, além de valores de seringabilidade que comparados a relatos da literatura podem conferir a facilidade de estes serem aplicados na via vaginal. O ensaio de calorimetria exploratória diferencial evidenciou eventos endotérmicos com picos similares e característicos a componentes presentes nas formulações. Os dois métodos analíticos validados para quantificar a HIP (HIP solubilizada em metanol e HIP na solução receptora composta de meio tampão fosfato com 3% de Tween 80) mostraram-se condizentes aos valores preconizados pela ANVISA. O perfil de liberação in vitro da HIP liberada a partir de CLN_HIP-HG1, indicou comportamento de liberação sustentada do composto ativo durante as 12h de análise, e que os principais mecanismos de liberação seguem os modelos de Peppas e de Higuchi. Tais sistemas não apresentaram permeação na mucosa vaginal suína, além de baixos valores de retenção. Os CLN com e sem HIP encapsulada dispersos ou não em HG revelaram potente ação antifúngica frente à cepa de C. albicans ATCC 10231 e comportamento fungistático frente à levedura. Os CLN com e sem HIP encapsulada dispersos ou não em HG, tratados com TFD, reduziram significativamente as culturas planctônicas de C. albicans variando de aproximadamente 1 a 2 log10 (UFC/mL) comparados ao YNB; enquanto a TFD mediada pelos sistemas F6.5_0,05 e CLN_HIP-HG1 apresentaram diminuições na viabilidade do biofilme de C. albicans de 2 e 1,2 log10 (UFC/mL), respectivamente, em relação ao PBS. Na análise de SOSG, a solução de HIP livre, bem como os CLN com HIP encapsulada dispersos em HG foram capazes de fornecer efetivamente oxigênio à solução de metanol. Na análise antifúngica in vivo associada ou não à TFD, o antifúngico e os grupos tratados com TFD apresentaram uma predominância de células fúngicas de diferentes tamanhos, além disso, CLN_HIP-HG1/TFD se destacou entre os grupos tratados com TFD obtendo uma melhora significativa do perfil terapêutico comparado ao grupo infectado. Assim, os resultados evidenciaram o potencial antifúngico dos sistemas in situ para a administração vaginal da HIP demonstrando serem capazes de contribuir no tratamento da CVV. Vulvovaginal candidiasis (VVC) is a fungal infection typically caused by Candida albicans, which is resistant to the drugs used in the treatment. The hypericin (HYP)- loaded nanostructured lipid carrier (NLC) dispersed in situ hydrogels (HG) proves to be an interesting option for the treatment of VVC, associated to alternative methods such as photodynamic therapy (PDT). This work aims to evaluate the potential of HYPloaded and empty NLC or NLC dispersed in the HG, associated to PDT for vaginal administration. The empty NLC (F6.5) and HYP-loaded NLC (F6.5_0.05, F6.5_0.25 and F6.5_0.5) and were obtained by sonication at 35% amplitude for 5min, containing poloxamer 407 the aqueous phase and in the oil phase, polyoxyethylene stearate, capric/caprylic acid triglycerides, ethoxylated hydrogenated castor oil and/or HYP. The HG were composed of poloxamer 407 and chitosan, being incorporated into HYPloaded and empty NLC in different proportions of NLC:HG giving rise to formulations NLC_HYP-HG1, NLC_HYP-HG2, NLC_HYP-HG3, NLC_HYP-HG4 and NLC_HYPHG5. The F6.5, F6.5_0.05, F6.5_0.25 and F6.5_0.5 systems, characterized by dynamic light scattering, presented nanometric diameters with low polydispersity and showed excellent physical stability. Formulation F6.5 showed unimodal distribution in the nanoparticle screening with predominance of 151 nm particles. Transmission electron microscopy showed particles of practically spherical morphologies and nanometric scales. The HYP encapsulation efficiency was 89.3%. Scanning electron microscopy of the HG and HYP-loaded NLC in different proportions of NLC:HG, revealed average values of surface roughness without statistical difference between the systems. Continuous rheology showed that all systems had Newtonian behavior; while in oscillatory rheology most systems showed G’ values higher than G’’, except of NLC_HYP-HG1. The HG and the NLC_HYP-HG1 and NLC_HYP-HG2 systems showed satisfactory mucoadhesive properties, in addition to syringability values that, compared to reports in the literature, showed their ease of application in the vaginal route. Differential scanning calorimetry showed endothermic events with similar peaks and characteristics to components present in the formulations. Two analytical methods were validated to quantify the HYP (HYP solubilized in methanol and HYP in the receiving solution of phosphate buffered medium with 3% Tween 80), being consistent with the values recommended by ANVISA. The in vitro release profile of HYP released from NLC_HYP-HG1, indicated a sustained drug release behavior during the 12h of analysis, and that the main release mechanisms follow the Peppas and Higuchi models. Such systems showed no permeation in the porcine vaginal mucosa, in addition to low retention values. The NLC with encapsulated HYP dispersed or not in HG, showed potent antifungal action against the strain of C. albicans ATCC 10231 and fungistatic behavior against yeast. The HG incorporated into HYP-loaded and empty NLC in different proportions, treated with PDT, significantly reduced the planktonic cultures of C. albicans ranging from approximately 1 to 2 log10 (CFU/mL) compared to YNB; while the PDT mediated by the F6.5_0.05 and NLC_HYP-HG1 systems showed decreases in the viability of the C. albicans biofilm of 2 and 1.2 log10 (CFU/mL), respectively, in relation to the PBS. In the SOSG analysis, the free HYP solution, as well as the HYP-loaded NLC dispersed in HG were able to effectively supply oxygen to the methanol solution. In the in vivo antifungal analysis associated or not with PDT, the antifungal and groups treated with PDT showed a predominance of fungal cells of different sizes, in addition, NLC_HYP-HG1/PDT stood out among the groups treated with PDT, obtaining a significant improvement in the therapeutic profile compared to the infected group. Thus, the results showed the antifungal potential of in situ systems for the vaginal administration of HYP, demonstrating that they are capable of contributing to the treatment of VVC. FAPESP: 2016/11198-4

Submitted by MARIANA RILLO SATO null (marillosato@gmail.com) on 2016-02-22T18:19:25Z No. of bitstreams: 1 Dissertação FINAL - COM FICHA.pdf: 2624639 bytes, checksum: 73f8b5a016f99a538e050ac383d48e8e (MD5) Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-02-24T14:12:59Z (GMT) No. of bitstreams: 1 sato_mr_me_arafcf_par.pdf: 1019774 bytes, checksum: ef4be326766e21e326047c976e98d864 (MD5) Made available in DSpace on 2016-02-24T14:12:59Z (GMT). No. of bitstreams: 1 sato_mr_me_arafcf_par.pdf: 1019774 bytes, checksum: ef4be326766e21e326047c976e98d864 (MD5) Previous issue date: 2016-02-03 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Tuberculosis (TB) is known worldwide as the leading infectious disease of chronic evolution in which the abandonment of treatment before the recommended period tends to cause the development of more severe disease, such as the appearance of resistant strains. Copper interacts with biologically active ligands developing copper(II) complex with activities antimicrobial. Nanostructured lipid carriers (NLCs) have gained prominence in the pharmaceutical field, since they have the ability to compartmentalize efficiently lipophilic drugs and modify their properties and behavior in the biological environment. The aims of this study was to evaluate the potential of copper(II) complexes-loaded NLCs applicable in the study against Mycobacterium tuberculosis. Copper(II) complexes containing isoniazid binder were obtained and they were entitled C1, C2, and C3 where they have co-binders anionic chloride, cyanate and thiocyanate, respectively. NLCs were obtained by fusion-emulsification technique, formulations were obtained by Ultra turrax at 5000 rpm for 10 minutes followed by sonicator at 8% amplitude for 20 minutes to obtain stable NLCs, those were incorporated of 3.50% and 0.50% to surfactant poloxamer 407 and cetrimonium bromide, respectively, and 2.07%, 2.50% and 0.88% of polyoxyethylene stearate lipids, triglycerides of capric acid/caprylic and ethoxylated hydrogenated castor oil 40 OE (F32) or castor oil (F38), respectively. Copper(II) complexes were incorporated into NLCs (F32.OE.C1, F32.OE.C2, F32.OE.C3 and F38.OR.C1, F38.OR.C2, F38.OR.C3) and characterized by diameter (Dmn) , polydispersivity index (PDI) and zeta potential (ZP) for 1, 15, 30, 45 and 60 days, which revealed homogeneous dispersion and excellent physical stability over the same period; the atomic force microscopy confirmed that developed NLCs are spherical particles dispersions at nanometer scales; differential scanning calorimetry revealed that the constituents used in the formulations interacted in the systems and confirmed that copper(II) complexes were incorporated in the NLCs. The antimicrobial activity of the complexes against M. tuberculosis H37Rv was determined by the resazurin microdilution assay demonstrated potential anti-TB when complexes were incorporated into NLCs. Cytotoxicity assay in murine macrophage-like cells J774A.1 (ATCC TIB-67) e MRC-5 (ATCC-CCL-171) for 24 h and acute toxicity of the complex against Artemia salina L (Artemiidae) showed the formulations have toxicity in all tested concentrations; although published reports indicated the safety of the components used of the formulations, studies should be conducted in animal models to assess the acute toxicity. These results suggest that NLCs have potential application in the study against Mycobacterium tuberculosis. A Tuberculose (TB) é mundialmente conhecida como a principal doença infecto-contagiosa de evolução crônica, em que o abandono do tratamento antes do período recomendado tende a ocasionar o desenvolvimento de casos mais graves da doença, gerando o aparecimento de cepas resistentes. O cobre interage com ligantes biologicamente ativos desenvolvendo complexos de cobre(II) com ações antimicrobiana. Carreadores lipídicos nanoestruturados (CLNs) vêm se destacando na área farmacêutica, pois apresentam a capacidade de compartimentalizar, de maneira eficiente fármacos lipofílicos e de modificar suas propriedades e comportamento em meio biológico. O objetivo deste trabalho foi avaliar o potencial de CLNs acrescidos de complexos de cobre(II) aplicáveis no estudo frente ao Mycobacterium tuberculosis. Foram obtidos 3 complexos de cobre(II), intitulados C1, C2 e C3 que contem o ligante isoniazida e os co-ligantes aniônicos cloreto, tiocianato e cianato, respectivamente. Os CLNs foram obtidos pela técnica de fusão-emulsificação, testando-se diversas formulações por Ultra turrax a 5000 rpm por 10 minutos e por sonicador com amplitude 8% por 20 minutos até a obtenção de CLNs estáveis, que foram constituídos de 3,50% e 0,50 % de poloxamer 407 e brometo de cetrimônio, respectivamente, e de 2,07%, 2,50% e 0,88% de estearato de polioxietileno, triglicérides do ácido cáprico/caprílico e óleo de rícino hidrogenado e etoxilado 40 (F32) ou óleo de rícino (F38), respectivamente. Os complexos de cobre(II) foram incorporados nos CLNs (F32.OE.C1, F32.OE.C2, F32.OE.C3, e F38.OR.C1, F38.OR.C2, F38.OR.C3), e caracterizados por diâmetro hidrodinâmico médio (Dnm), índice de polidispersidade (IPD) e potencial zeta (PZ) por 1, 15, 30, 45 e 60 dias, revelando homogeneidade da dispersão e excelente estabilidade física; a microscopia de força atômica confirmou que os CLNs tratam-se de dispersões de partículas esféricas em escalas nanométricas; a calorimetria exploratória diferencial revelou que os componentes utilizados nas formulações interagiram para formação do sistema e que os complexos de cobre(II) foram incorporados nos CLNs. A atividade antimicrobiana dos complexos, frente ao M. tuberculosis H37Rv determinada pelo ensaio de microdiluição com Resazurina evidenciaram potencialização da ação anti-TB pela incorporação dos complexos nos CLNs. Os ensaios de citotoxicidade frente à linhagem de macrófagos J774A.1 (ATCC TIB-67) e de MRC-5 (ATCC-CCL-171) em 24 h e a toxicidade aguda dos complexos frente à Artemia salina L., mostraram que as formulações apresentaram toxicidade em todas as concentrações testadas; apesar de relatos da literatura indicarem a segurança dos componentes das formulações, estudos devem ser realizados em modelos animais para avaliar a toxicidade aguda. Os resultados sugerem que os CLNs apresentam aplicação potencial no estudo frente ao Mycobacterium tuberculosis.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2022-02-22T12:44:18Z No. of bitstreams: 1 PDF - Karen Loraine Macena Santos.pdf: 1188485 bytes, checksum: fd5d6da33c9f4bc8a3c89aa3affbe57e (MD5) Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2022-02-22T14:02:13Z (GMT) No. of bitstreams: 1 PDF - Karen Loraine Macena Santos.pdf: 1188485 bytes, checksum: fd5d6da33c9f4bc8a3c89aa3affbe57e (MD5) Made available in DSpace on 2022-02-22T14:03:44Z (GMT). No. of bitstreams: 1 PDF - Karen Loraine Macena Santos.pdf: 1188485 bytes, checksum: fd5d6da33c9f4bc8a3c89aa3affbe57e (MD5) Previous issue date: 2020-09-28 Photodynamic therapy (PDT) is a promising, minimally invasive therapeutic modality that has low toxicity in the biological system. PDT is based on the local or systemic application of a photosensitizer (PS), which accumulates in pathological tissues. The FS molecules absorb light from an appropriate wavelength, initiating photochemical reaction processes that result in the formation of singlet oxygen (1O2) and reactive oxygen species (ROS). Currently, there are several PSs, among them, we have Phthalocyanine (PC), a second generation FS. However, one of its limitations is related to its hydrophobic nature, which can induce aggregation in biological systems with reduced photodynamic activity. The incorporation of PC in nanostructured lipid carriers (NLC) can be a promising alternative to overcome this problem. Thus, the objective of this work was to develop and characterize PC-loaded NLCs for later application in photodynamic therapy. For the development of the formulations, the fusion-emulsification technique followed by sonication was applied. The NLC and NLC-PC were characterized in terms of mean diameter (D.mn), polydispersion index (PdI), zeta potential (Pz), encapsulation efficiency (% EE), transmission electron microscopy (TEM), differential scanning calorimetry (DSC), photobleaching and singlet oxygen generation in cell systems (SOSG). In addition, in vitro release tests were performed using the beaker method, using dialysis membranes. The average size of the NLC and NLC-PC formulations were less than 200 nm, PdI

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2022-03-21T12:28:27Z No. of bitstreams: 1 PDF - Larissa Pereira Alves.pdf: 1340796 bytes, checksum: e21a15af70424df69defcee4c056571c (MD5) Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2022-03-22T10:47:05Z (GMT) No. of bitstreams: 1 PDF - Larissa Pereira Alves.pdf: 1340796 bytes, checksum: e21a15af70424df69defcee4c056571c (MD5) Made available in DSpace on 2022-03-22T11:26:04Z (GMT). No. of bitstreams: 1 PDF - Larissa Pereira Alves.pdf: 1340796 bytes, checksum: e21a15af70424df69defcee4c056571c (MD5) Previous issue date: 2020-02-17 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Hypercholesterolemia represents one of the risk factors for the development and progression of atherosclerosis and consequently cardiovascular diseases, which are one of the main causes of morbidity and mortality in the world. Simvastatin, responsible for reducing the level of cholesterol in the blood, has low oral bioavailability (