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Mônica Cristina de Oliveirahttp://lattes.cnpq.br/0383215992450256Sávia Caldeira de Araújo LopesLucas Antônio Miranda FerreiraFrederic Jean Georges FrezardIzabella Thaís da Silvahttp://lattes.cnpq.br/5967785123600047Eliza Rocha Gomes2022-11-21T19:02:38Z2022-11-21T19:02:38Z2022-08-18http://hdl.handle.net/1843/47356A doxorrubicina (DOX) apresenta uma potente ação antineoplásica e vem sendo utilizada no tratamento de diversos tumores. Entretanto, graves efeitos tóxicos têm limitado seu uso, principalmente a cardiotoxicidade. Recentemente, nanocarreadores híbridos obtidos pela fusão das membranas lipídicas de exossomas e lipossomas vêm sendo estudados, a fim de aumentar a eficácia antitumoral, minimizar os efeitos adversos e contornar os problemas relacionados a resistência de quimioterápicos. A proposta desse estudo consistiu na fusão de exossomas derivados de células de mama tumorais com lipossomas pH-sensíveis de circulação prolongada contendo DOX para tratamento de câncer de mama (ExoSpHL-DOX). O diâmetro médio das vesículas foi de 100,8 ± 7,8 nm, o índice de polidispersão igual a 0,122 ± 0,004 e o teor de DOX encapsulado foi de 83,5 ± 2,5%. A fusão de exossomas com lipossomas pH-sensíveis de circulação prolongada foi confirmada por espectroscopia de infravermelho com transformada de Fourier, espectroscopia Raman e nanocitometria de fluxo. A avaliação da estabilidade de armazenamento dos ExoSpHL-DOX a 4°C comprovou a manutenção das características físico-químicas da formulação por 60 dias. O estudo de liberação de DOX a partir de ExoSpHL-DOX em meios de diluição apresentando diferentes valores de pH, comprovou a característica de pH-sensibilidade do nanossistema. A formulação mostrou-se citotóxica para células tumorais 4T1 de mama murina. A toxicidade aguda foi determinada pela avaliação da mortalidade e morbidade dos animais, análises hematológicas, bioquímicas e histopatológicas, após uma única administração intravenosa de ExoSpHL-DOX. O intervalo de dose letal mediana (LD50) encontrado após o tratamento com ExoSpHL-DOX (17,5 - 20 mg/kg) é maior do que o encontrado com DOX livre (12,5 - 15 mg/kg). Além disso, ExoSpHL-DOX não apresentou sinais de nefrotoxicidade mesmo na dose mais elevada de DOX, indicando que o nanocarreador híbrido pode alterar a distribuição de DOX e reduzir o dano renal. Em relação à atividade antitumoral, ExoSpHL-DOX inibiu em aproximadamente 50% o crescimento do tumor comparado ao grupo controle. Além disso, o nanocarreador híbrido de exossomas-lipossomas reduziu o número de focos metastáticos nos pulmões. Esses resultados indicam que ExoSpHL-DOX pode ser um nanocarreador promissor para o tratamento do câncer de mama, reduzindo a toxicidade e inibindo a metástase, principalmente nos pulmões.Doxorubicin (DOX) has a potent antineoplastic action and has been used in the treatment of several tumors. However, serious toxic effects have limited its use, especially cardiotoxicity. Recently, hybrid nanocarriers obtained by fusion of lipid membranes of exosomes and liposomes have been studied in order to increase antitumor efficacy, minimize adverse effects and overcome problems related to chemotherapy resistance. Thus, the purpose of this study was the fusion of exosomes derived from breast tumor cells with long-circulating and pH-sensitive liposomes containing DOX (ExoSpHL-DOX) for the treatment of breast cancer. The mean diameter of ExoSpHL-DOX was 100.8 ± 7.8 nm, the polydispersity index was 0.122 ± 0.004, and the encapsulated DOX content was equal to 83.5 ± 2.5%. The fusion of exosomes with long-circulating and pH-sensitive liposomes was confirmed by Fourier transform infrared spectroscopy, Raman spectroscopy, and nano-flow cytometry. The physicochemical characteristics of ExoSpHL-DOX were maintained for 60 days, at 4°C. The study of the release of DOX from ExoSpHL-DOX in dilution media with different pH values showed the pH-sensitivity of the nanosystem. The cytotoxic study against the 4T1 breast cancer cell line demonstrated that ExoSpHL-DOX treatment significantly reduced the cancer cell viability. Acute toxicity was determined by evaluating the mortality and morbidity of the animals, hematological, biochemical, and histopathological analyses, after a single intravenous administration of ExoSpHL-DOX. The results of the study indicated that the median lethal dose range (LD50) of the ExoSpHL-DOX treatment (17.5 - 20 mg/kg) is higher than that found for treatment with free DOX (12.5 - 15 mg/kg). In addition, ExoSpHL-DOX treatment showed no signs of nephrotoxicity even at the highest dose of DOX, indicating that the presence of hybrid nanocarrier may alter the distribution of DOX and reduce kidney damage. Regarding to the antitumor activity, ExoSpHL-DOX treatment inhibited close to 50% the tumor growth compared to control group. Furthermore, the hybrid nanocarrier of tumor-derived exosomes fused with long-circulating and pH-sensitive liposomes reduced the number of metastatic foci in the lungs. These results indicate that ExoSpHL-DOX may be a promising nanocarrier for the treatment of breast cancer, reducing toxicity and inhibiting metastasis, mainly in the lungs.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Ciências FarmacêuticasUFMGBrasilFARMACIA - FACULDADE DE FARMACIACâncer de mamaLipossomasExossomasDoxorrubicinaMetástaseExossomas fundidos com lipossomas pH-sensíveis de circulação prolongada contendo doxorrubicina: preparo, caracterização, avaliação da toxicidade aguda e da atividade antitumoral.Exosomes fused with long-circulating and pH-sensitive liposomes loaded with doxorubicin: preparation, characterization, investigation of the acute toxicity and antitumor activityinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALTese_PPGCF_Eliza.pdfTese_PPGCF_Eliza.pdfapplication/pdf2772156https://repositorio.ufmg.br/bitstream/1843/47356/3/Tese_PPGCF_Eliza.pdf0c956315221ac0e19e3596101c377f51MD53LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/47356/4/license.txtcda590c95a0b51b4d15f60c9642ca272MD541843/473562022-11-21 16:02:39.063oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2022-11-21T19:02:39Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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