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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2023.tde-08122023-165236
Document
Author
Full name
Gabriella Sales Calaço Roque
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2023
Supervisor
Committee
Souza, Ana Olívia de (President)
Boas, Silas Granato Villas
Coelho, Maria Gabriela Berzoti
Maria, Durvanei Augusto
Rodrigues, Fabio
Title in Portuguese
Estudo da ação antitumoral de nanopartículas de prata biogênicas
Keywords in Portuguese
AgNPs
Aspergillus tubingensis
Citotoxicidade
Fungos
MDA-MB-231.
Abstract in Portuguese
Dados da literatura evidenciam promissoras propriedades antitumorais de nanopartículas de prata (AgNPs) de origem biológica que representam uma nova alternativa para o tratamento do câncer. Com esta perspectiva, neste estudo, sete AgNPs micogênicas foram obtidas utilizando as espécies de fungos Aspergillus tubingensis (AgNP-AT), Aspergillus spp. (AgNP-Asp), Bionectria ochroleuca (AgNP-BO), Cladosporium pini-ponderosae (AgNP-CPP), Fusarium proliferatum (AgNP-FP), Epicoccum nigrum (AgNP-EN) e Exserohilum rostratum (AgNP-ER). As AgNPs foram caracterizadas por espalhamento dinâmico de luz (DLS), índice de polidispersão (PDI), potencial Zeta (PZ), microscopia eletrônica de transmissão (MET) e espectroscopia no infravermelho por transformada de Fourier (FTIR), e apresentaram forma esférica e tamanho de 21,8 a 35,8 nm. A citotoxicidade das AgNPs foi avaliada em células de câncer de mama (MCF-7 e MDA-MB-231), células de câncer de pancreas (MIA PaCa-2), e células normais de fibroblastos humanos (FN1) pelo ensaio de MTT ((3-(4,5-dimetiltiazol-2 il)-2,5-difenil brometo de tetrazólio) nos tempos de 24, 48 e 72 h. Os valores das concentrações efetivas em inibir 50% da proliferação celular em células tumorais foram definidos como correspondentes ao IC50 e em células normais como CC50. Em 48 h, as AgNP-BO, AgNP-AT, e AgNP-Asp demonstraram alta citotoxicidade para as células MDA-MB-231, com IC50 de 0,0016; 0,0025; e 0,0253 mM, respectivamente, enquanto apresentaram baixa citotoxicidade para as células FN1, com CC50 de 0,0982; 0,2130, e 1,3190 mM. Os valores de IC50 e CC50 foram utilizados para calcular o índice de seletividade das AgNPs (IS = CC50/IC50) em 48 h, sendo 85,20; 61,38 e 52,13 para AgNP-AT, AgNP-BO e AgNP-Asp, indicando a especificidade das AgNPs pelas células tumorais. Considerando a alta especificidade das AgNP-AT pelas células MDA-MB-231, a ação dessas nanopartículas foi também avaliada nas fases do ciclo celular, na proliferação celular, na indução de apoptose e na mutagenicidade. A 0,03 mM as AgNP-AT induziram parada do ciclo celular na fase S, aumento de apoptose em 1,7 vezes e ausência de danos aos cromossomos de células V79-4. Os dados comprovam a citotoxicidade e especificidade das AgNP-AT para células tumorais MDA-MB-231, com IS de 85,20, indicando baixa toxicidade para células normais.
Title in English
Study of the antitumor action of biogenic silver nanoparticles
Keywords in English
AgNPs
Aspergillus tubingensis
Cytotoxicity
Fungi
MDA-MB-231.
Abstract in English
Data from the literature show promising antitumor properties of silver nanoparticles (AgNPs) of biological origin that represent a new alternative for the treatment of cancer. With this perspective, in this study, seven mycogenic AgNPs were obtained using the fungal species Aspergillus tubingensis (AgNP-AT), Aspergillus spp. (AgNP-Asp), Bionectria ochroleuca (AgNP-BO), Cladosporium pini-ponderosae (AgNP-CPP), Fusarium proliferatum (AgNP-FP), Epicoccum nigrum (AgNP-EN) e Exserohilum rostratum (AgNP-ER). The AgNPs were characterized by dynamic light scattering (DLS), polydispersion index (PDI), Zeta potential (PZ), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) and showed spherical shape and size from 21.8 to 35.8 nm. Cytotoxicity of AgNPs was evaluated in breast cancer cells (MCF-7 and MDA-MB-231), pancreas cancer cells (MIA PaCa-2), and normal human fibroblast cells (FN1) by MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay at times of 24, 48, and 72 h. The values of concentrations effective in inhibiting 50% of cell proliferation in tumor cells were defined as corresponding to IC50 and in normal cells as CC50. In 48 h, AgNP-BO, AgNP-AT, and AgNP-Asp demonstrated high cytotoxicity for MDA-MB-231 cells, with IC50 of 0.0016; 0.0025; and 0.0253 mM, respectively, while showing low cytotoxicity for FN1 cells, with a CC50 of 0.0982; 0.2130, and 1.3190. IC50 and CC50 values were used to calculate the AgNPs selectivity index (SI = CC50/IC50) in 48 h, being 85.20; 61.38 and 52.13 for AgNP-AT, AgNP-BO and AgNP-Asp, indicating the specificity of AgNPs for tumor cells. Considering the high specificity of AgNP-AT for MDA-MB-231 cells, its action was also evaluated in the cell cycle phases, cell proliferation, induction of apoptosis and mutagenicity. At 0.03 mM, AgNP-AT induced cell cycle arrest in the S phase, a 1.7-fold increase in apoptosis and no damage to the chromosomes of V79-4 cells. The data confirm the cytotoxicity and specificity of AgNP-AT for MDA-MB-231 tumor cells, with an IS of 85.20, indicating low toxicity for normal cells.
 
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Publishing Date
2023-12-14
 
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