Desenvolvimento de nanopartículas poliméricas pelo método nano spray dryer para encapsulação de corantes fotossensíveis
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Data
2019-08-29
Autores
Gobo, Graciely Gomides
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Neste trabalho de mestrado profissional foi possível obter um sistema nanoestruturado polimérico contendo Quinizarina (QZ), um corante fotossensibilizador que emerge como nova alternativa para tratamento antitumoral, utilizando a terapia fotodinâmica (TFD). As nanopartículas foram produzidas em escala, utilizando um método inovador de Nano Spray Dryer as quais foram caracterizadas por diferentes métodos, tais como: Espectroscopia por Absorção e Emissao de Fluorescência na região do Ultra-Violeta Visível (UV-Vis), Espalhamento de Luz dinâmica (DLS) e Microscopia Eletrônica de Varredura (MEV). Através das caracterizações observou-se nanopartículas monodispersas com diâmetro hidrodinâmico reduzido, verificou-se também que as nanopartículas foram preparadas de forma eficiente, o qual obteve-se a encapsulação das partículas QZ camada por camada (layer by layer). As nanopartículas foram preparadas utilizando uma rota de baixo custo e com altissimo rendimento (>70%). Após a completa caracterização do sistema, as nanopartículas foram aplicadas em testes biológicos in vitro utilizando dois tipos de linhagens celulares: a fibroblastóide (NIH-3T3) e cancerígena de origem melanocítica (B16-F10). Realizou-se estudos de biocompatibilidade, internalização utilizando QZ livre e encapsulada (uptake) e microscopia confocal em ambas as linhagens utilizadas e testes de fototoxicidade na linhagem melanocítica. Observou-se através dos estudos de uptake que houve uma maior internalização da quinizarina nanoestruturada, devido à sua alta biocompatibilidade, em relação a quinizarina livre. O estudo de internalização através da microscopia confocal demonstrou que o sistema nanoestruturado QZ funciona como um potente corante (fármaco fotossensibilizante), devido à sua capacidade de marcação nuclear, o que ainda não é encontrado na literatura utilizando a linhagem melanocítica. Os estudos in vitro foram realizados na ausência e presença de luz (LED), sendo que na ausência de luz (escuro), observou-se viabilidade compatível, não apresentando significativa morte celular, já os estudos na presença de luz o efeito fotodinâmico foi expressivo, onde houve grande morte celular. Dentro deste contexto as nanopartículas poliméricas contendo Quinizarina (QZ) é um potencial fármaco fotossensibilizante, podendo ser aplicado em terapia fotodinâmica para o câncer de pele melanômico.
In this professional master's work it was possible to obtain a polymeric nanostructured system containing Quinizarin (QZ), a photosensitizing dye that appears as a new alternative for antitumor treatment, using a photodynamic therapy (PDT). The nanoparticles were produced in scale using an innovative Nano Spray Dryer method, and characterized by different methods such as: Absorption Spectroscopy and Fluorescence Emission in the Ultra-Violet and Visible Region (UV-Vis), Dynamic Light Scattering (DLS) and Scanning Electron Microscopy (SEM). Through caracterizations, was observed that the nanoparticles produced where monodispered and small hydrodynamic diameter, the particles where prepared in a efficient way, which encapsulated the QZ particles layer by layer. The nanoparticles were prepared using a low cost, high yield (> 70%) route. After a complete characterization of the system, nanoparticles were applied in in vitro biological tests using two types of cell line: a fibroblast (NIH-3T3) and a melanocytic cancer (B16-F10). Biocompatibility, uptake, using free and encapsulated QZ and confocal microscopy studies were performed in both cell lines, and phototoxicity tests were performed in melanocytic cell line. It was observed through uptake studies that there was a greater internalization of nanostructured quinizarin, due to its high biocompatibility, compared to free quinizarin. The internalization study by confocal microscopy demonstrates that the QZ nanostructured system functions as a potent dye (photosensitizing drug) due to its nuclear labeling capability, or that has not yet been found in the literature using a melanocytic line. In vitro studies were performed in the absence and presence of light (LED), and the absence of light (dark), prevented the viability of use, showed no significant cell death, while studies in the presence of light or photodynamic effect were expressive. where there was great cell death. Within this context, as polymeric nanoparticles with Quinizarin (QZ), is a potential photosensitizing drug and can be applied in photodynamic therapy for melanomic skin cancer.
In this professional master's work it was possible to obtain a polymeric nanostructured system containing Quinizarin (QZ), a photosensitizing dye that appears as a new alternative for antitumor treatment, using a photodynamic therapy (PDT). The nanoparticles were produced in scale using an innovative Nano Spray Dryer method, and characterized by different methods such as: Absorption Spectroscopy and Fluorescence Emission in the Ultra-Violet and Visible Region (UV-Vis), Dynamic Light Scattering (DLS) and Scanning Electron Microscopy (SEM). Through caracterizations, was observed that the nanoparticles produced where monodispered and small hydrodynamic diameter, the particles where prepared in a efficient way, which encapsulated the QZ particles layer by layer. The nanoparticles were prepared using a low cost, high yield (> 70%) route. After a complete characterization of the system, nanoparticles were applied in in vitro biological tests using two types of cell line: a fibroblast (NIH-3T3) and a melanocytic cancer (B16-F10). Biocompatibility, uptake, using free and encapsulated QZ and confocal microscopy studies were performed in both cell lines, and phototoxicity tests were performed in melanocytic cell line. It was observed through uptake studies that there was a greater internalization of nanostructured quinizarin, due to its high biocompatibility, compared to free quinizarin. The internalization study by confocal microscopy demonstrates that the QZ nanostructured system functions as a potent dye (photosensitizing drug) due to its nuclear labeling capability, or that has not yet been found in the literature using a melanocytic line. In vitro studies were performed in the absence and presence of light (LED), and the absence of light (dark), prevented the viability of use, showed no significant cell death, while studies in the presence of light or photodynamic effect were expressive. where there was great cell death. Within this context, as polymeric nanoparticles with Quinizarin (QZ), is a potential photosensitizing drug and can be applied in photodynamic therapy for melanomic skin cancer.
Descrição
Palavras-chave
Nanopartículas, Nano Spray Dryer, Quinizarina, Terapia fotodinâmica, Tratamento de câncer, Nanoparticles, Quinizarin, Photodynamic therapy, Cancer treatment