RESUMO
Silver nanoparticles (AgNPs) hold promise for cancer therapy. This study aimed to evaluate their impact on tumor and non-tumor cell number, viability, and morphology. Antitumor activity was tested on U-87MG (glioblastoma) and DU-145 (prostate cancer) cell lines. Treatment with AgNPs notably reached a reduction of U-87MG and DU-145 cell growth by 89.30% and 79.74%, respectively, resulting in slower growth rates. AgNPs induced DNA damage, evidenced by reduced nuclear area and DNA content via fluorescent image-based analyses. Conversely, HFF-1 non-tumor cells displayed no significant changes post-AgNPs exposure. Viability assays revealed substantial reductions in U-87MG and DU-145 cells (79% and 63% in MTT assays, 30% and 52.2% in high-content analyses), while HFF-1 cells exhibited lower sensitivity. Tumor cells had notably lower IC50 values than non-tumor cells, indicating selective susceptibility. Transmission electron microscopy (TEM) showed morphological changes post-AgNPs administration, including increased vacuoles, myelin figures, membrane ghosts, cellular extravasation, and membrane projections. The findings suggest the potential of AgNPs against glioblastoma and prostate cancer, necessitating further exploration across other cancer cell lines.
Assuntos
Antineoplásicos , Sobrevivência Celular , Glioblastoma , Nanopartículas Metálicas , Neoplasias da Próstata , Prata , Humanos , Nanopartículas Metálicas/toxicidade , Masculino , Prata/toxicidade , Glioblastoma/tratamento farmacológico , Glioblastoma/patologia , Linhagem Celular Tumoral , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/patologia , Sobrevivência Celular/efeitos dos fármacos , Antineoplásicos/farmacologia , Contagem de Células , Dano ao DNA/efeitos dos fármacosRESUMO
The aim of this work was to develop and test the in vitro biological activity of nanocapsules loaded with a doxorubicin (DOX) free base dissolved in a core of castor oil shelled by poly(methyl vinyl ether-co-maleic anhydride) conjugated to n-octadecylamine residues. This system was stable and monodisperse, with a hydrodynamic diameter of about 300 nm. These nanocapsules changed the intracellular distribution of DOX, from the nuclei to the cytoplasm, and exhibited higher toxicity towards cancer cells - 4T1 and MCF-7 - and significantly lower toxicity towards normal cells - NIH-3T3 and MCF-10A - in vitro. In conclusion, these nanocapsules are suitable DOX carriers, which remain to be studied in in vivo tumor models.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/metabolismo , Portadores de Fármacos/química , Nanocápsulas/química , Animais , Neoplasias da Mama/patologia , Óleo de Rícino , Linhagem Celular , Linhagem Celular Tumoral , Núcleo Celular , Citoplasma , Doxorrubicina/toxicidade , Portadores de Fármacos/normas , Humanos , Células MCF-7 , Camundongos , Células NIH 3T3RESUMO
Magnetic nanoparticles can be used in different areas of biology. It is therefore important to know the effects of such nanomaterials on germline cells as they may traverse the blood-testis barrier. This work aimed to evaluate the response of bull sperm after exposure to a magnetic fluid containing DMSA-coated maghemite nanoparticles (MNP-DMSA) in order to determine nanotoxicity. Bull sperm was incubated with MNP-DMSA at final concentrations of 0.06, 0.03 or 0.015 mg Fe/mL. Sperm kinetics, plasma membrane integrity and acrosome reaction were evaluated over a 4 h incubation period. The sperm cells were also evaluated by transmission electron microscopy. Exposure of bull sperm to MNP-DMSA did not affect sperm kinetics or integrity. Neither ultrastructural damage of sperm cells nor uptake of nanoparticles by the spermatozoa was observed. In conclusion, MNP-DMSA does not affect sperm function or structure under the conditions tested.
Assuntos
Compostos Férricos/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Espermatozoides/efeitos dos fármacos , Succímero/administração & dosagem , Animais , Bovinos , Membrana Celular/efeitos dos fármacos , Compostos Férricos/química , Masculino , Nanopartículas Metálicas/química , Microscopia Eletrônica de Transmissão , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Espermatozoides/ultraestrutura , Succímero/químicaRESUMO
The ultrastructural analysis of oocytes and ovarian follicles has been used to evaluate the effects of assisted reproductive techniques, such as cryopreservation or in vitro oocyte maturation. It also benefits the understanding of such complex mechanisms that occur during folliculogenesis. From the beginning of primordial follicles growth until oocyte maturation in preovulatory follicles oocyte cytoplasmic organelles undergo dynamic alterations that reflect physiological changes and development. This review aims to make a retrospective survey of the relevant features of follicles and oocytes ultrastructure, highlighting the differences between mammalian species, specially the domestic ones.
Assuntos
Oócitos/citologia , Oócitos/ultraestrutura , Oogênese/fisiologia , Animais , Animais Domésticos , Diferenciação Celular , Feminino , Células da Granulosa/citologia , Células da Granulosa/ultraestrutura , Mamíferos , Folículo Ovariano/citologia , Folículo Ovariano/crescimento & desenvolvimentoRESUMO
The purpose of this study was to assess the cytotoxic effect of the non-fractionated aqueous extract of A. sylvaticus mushroom in cultures of non-tumor cells (NIH3T3) and tumor cells (OSCC-3). The cells were maintained in DMEN cell culture medium added of 10% of fetal bovine serum and 1% antibiotic. For the cytotoxicity test we prepared the aqueous mushroom extract at concentrations of 0.01 mg.ml⁻¹, 0.02 mg.ml⁻¹, 0.04 mg.ml⁻¹, 0.08 mg.ml⁻¹, 0.16 mg.ml⁻¹, and 0.32 mg.ml⁻¹. For the culture, 2 x 10⁵ cells/ml was deposited in 96-well microplates during 24 hour incubation with subsequent exchange of medium by another containing the mushroom concentrations. After 24 hour incubation the medium was discarded and 100 ml of tetrazolium blue (MTT) was added at a concentration of 5 mg.ml⁻¹. The microplates were incubated for 2 h at 37° C. Spectrophotometric analysis was performed using 570 nm wavelength. From the values of the optical densities we determined the drug concentration capable of reducing cell viability by 50%. Therefore, the mushroom A. sylvaticus, at all concentrations tested, did not show cytotoxic effects, once the inhibitory concentration (IC₅₀) obtained for tumor cells OSCC-3 was 0.06194 mg.ml⁻¹, and the IC₅₀ checked for non-tumor cells NIH3T3 was 0,06468 mg.ml⁻¹. This test made it possible to determine that A. sylvaticus mushroom has no cytotoxic effects, suggesting its use safe for human consumption (AU)
El objetivo de este estudio fue evaluar el efecto citotóxico de un extracto acuoso no fraccionado de la seta A. sylvaticus en cultivos de células no tumorales (NIH3T3) y tumorales (OSCC-3). Se mantuvo a las células en un medio de cultivo celular DMEN al que se añadió suero de ternera fetal al 10% y antibiótico al 1%. Para la prueba de toxicidad, se preparó el extracto acuoso del hongo a concentraciones 0,01 mg.ml-1, 0,02 mg.ml-1, 0,04 mg.ml-1, 0,08 mg.ml-1, 0,16 mg.ml-1 y 0,32 mg.ml-1. Para el cultivo, 2 x 105 células/ml se depositaron en microplacas de 96 pocillos durante 24 horas de incubación con el subsiguiente recambio del medio por otro que contenía las concentraciones del hongo. Tras 24 horas de incubación, se desechó el medio y se añadieron 100 ml de azul de tetrazolio (MTT) a una concentración de 5 mg.ml-1. Se incubaron las microplacas durante 2 h a 37° C. Se realizó un análisis espectrofotométrico con una longitud de onda de 570 nm. A partir de los valores de las densidades ópticas, se determinó la concentración de la droga capaz de reducir la viabilidad celular en un 50 %. La seta A. sylvaticus, a todas las concentraciones probadas, no mostró efectos citotóxicos una vez que la concentración inhibitoria (IC50) obtenida para las células tumorales OSCC-3 fue de 0,06194 mg.ml-1 y la IC50 comprobada para las células no tumorales NIH3T3 fue de 0,06468 mg.ml-1. Esta prueba consiguió determinar que la seta A. sylvaticus no posee efectos citotóxicos lo que sugiere un uso seguro para el consumo humano (AU)
Assuntos
Humanos , Citotoxinas/farmacocinética , Agaricus/patogenicidade , Medidas de Toxicidade , Contaminação de Alimentos/análise , Sais de Tetrazólio , Testes de Toxicidade/métodosRESUMO
The purpose of this study was to assess the cytotoxic effect of the non-fractionated aqueous extract of A. sylvaticus mushroom in cultures of non-tumor cells (NIH3T3) and tumor cells (OSCC-3). The cells were maintained in DMEN cell culture medium added of 10% of fetal bovine serum and 1% antibiotic. For the cytotoxicity test we prepared the aqueous mushroom extract at concentrations of 0.01 mg.ml⻹, 0.02 mg.ml⻹, 0.04 mg.ml⻹, 0.08 mg.ml⻹, 0.16 mg.ml⻹, and 0.32 mg.ml⻹. For the culture, 2 x 105 cells/ml was deposited in 96-well microplates during 24 hour incubation with subsequent exchange of medium by another containing the mushroom concentrations. After 24 hour incubation the medium was discarded and 100 ml of tetrazolium blue (MTT) was added at a concentration of 5 mg.ml⻹. The microplates were incubated for 2 h at 37° C. Spectrophotometric analysis was performed using 570 nm wavelength. From the values of the optical densities we determined the drug concentration capable of reducing cell viability by 50%. Therefore, the mushroom A. sylvaticus, at all concentrations tested, did not show cytotoxic effects, once the inhibitory concentration (IC50) obtained for tumor cells OSCC-3 was 0.06194 mg.ml⻹, and the IC50 checked for non-tumor cells NIH3T3 was 0,06468 mg.ml⻹. This test made it possible to determine that A. sylvaticus mushroom has no cytotoxic effects, suggesting its use safe for human consumption.
El objetivo de este estudio fue evaluar el efecto citotóxico de un extracto acuoso no fraccionado de la seta A. sylvaticus en cultivos de células no tumorales (NIH3T3) y tumorales (OSCC-3). Se mantuvo a las células en un medio de cultivo celular DMEN al que se añadió suero de ternera fetal al 10% y antibiótico al 1%. Para la prueba de toxicidad, se preparó el extracto acuoso del hongo a concentraciones 0,01 mg.ml-1, 0,02 mg.ml-1, 0,04 mg.ml-1, 0,08 mg.ml-1, 0,16 mg.ml-1 y 0,32 mg.ml-1. Para el cultivo, 2 x 105 células/ml se depositaron en microplacas de 96 pocillos durante 24 horas de incubación con el subsiguiente recambio del medio por otro que contenía las concentraciones del hongo. Tras 24 horas de incubación, se desechó el medio y se añadieron 100 ml de azul de tetrazolio (MTT) a una concentración de 5 mg.ml-1. Se incubaron las microplacas durante 2 h a 37° C. Se realizó un análisis espectrofotométrico con una longitud de onda de 570 nm. A partir de los valores de las densidades ópticas, se determinó la concentración de la droga capaz de reducir la viabilidad celular en un 50 %. La seta A. sylvaticus, a todas las concentraciones probadas, no mostró efectos citotóxicos una vez que la concentración inhibitoria (IC50) obtenida para las células tumorales OSCC-3 fue de 0,06194 mg.ml-1 y la IC50 comprobada para las células no tumorales NIH3T3 fue de 0,06468 mg.ml-1. Esta prueba consiguió determinar que la seta A. sylvaticus no posee efectos citotóxicos lo que sugiere un uso seguro para el consumo humano.
Assuntos
Agaricus/química , Sobrevivência Celular/efeitos dos fármacos , Intoxicação Alimentar por Cogumelos , Células 3T3 , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Corantes , Meios de Cultura , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Camundongos , Espectrofotometria Ultravioleta , Sais de Tetrazólio , TiazóisRESUMO
This work aimed to develop solid lipid nanoparticles (SLNs) loaded with doxorubicin evaluating the influence of docosahexaenoic acid (DHA), a polyunsaturated fatty acid that enhances the activity of anticancer drugs, on drug encapsulation efficiency (EE). The SLN were characterized for size, zeta potential, entrapment efficiency (EE) and drug release. Studies of in vitro antitumor activity and cellular uptake were also conducted. The reduction in particle size (from 127 ± 14 to 94 ± 1 nm) and negative charges were obtained for SLN loaded with DHA and triethanolamine (TEA), amine used to increase the solubility of doxorubicin in melted lipid. The EE was significantly improved from 36 ± 4% to 99 ± 2% for SLN without and with DHA at 0.4%, respectively. The doxorubicin release in a slightly acid medium (pH 5.0) was higher than that observed at physiological pH. The in vitro studies clearly showed the higher cytotoxicity of doxorubicin-DHA-loaded SLN than free doxorubicin+DHA on human lung tumor cell line (A549) and the improved cellular uptake achieved with the drug encapsulation can be an explanation. These findings suggest that DHA-doxorubicin-loaded SLN is a promising alternative for the treatment of cancer.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Portadores de Fármacos/administração & dosagem , Nanopartículas/administração & dosagem , 1-Octanol/química , Antibióticos Antineoplásicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/química , Doxorrubicina/química , Portadores de Fármacos/química , Humanos , Microscopia Eletrônica de Transmissão , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da Partícula , Água/químicaRESUMO
The aim of this study was to evaluate the effects of pituitary (pFSH) or recombinant (rFSH) FSH on the survival and growth of caprine preantral follicles. Caprine ovarian tissues were in vitro cultured for one or seven days in Minimum Essential Medium (MEM) alone or containing 10, 50, 100 and 1000 ng/ml of pFSH or rFSH. Control tissues (non-cultured) and those cultured were processed for histological and ultrastructural studies. In addition, follicular and oocyte diameter were analysed. After seven days of culture, only 50ng/ml of rFSH maintained the percentage of normal follicles similar to control. Moreover, 10 ng/ml of pFSH and all the concentrations of rFSH promoted primordial follicles activation. In addition, the presence of 50 ng/ml of rFSH promoted the highest follicular diameter at day seven of culture. In conclusion, 50 ng/ml of rFSH maintained the ultrastructural integrity of caprine preantral follicles, promoted primordial follicles activation and further growth of cultured follicles.
O objetivo deste estudo foi avaliar os efeitos do FSH pituitário (pFSH)ou recombinante (rFSH) sobre a sobrevivência e o crescimento de folículos pré-antrais caprinos. O tecido ovariano foi cultivado in vitro por um ou sete dias em Meio Essencial Mínimo (MEM) sozinho, ou contendo 10, 50, 100 e 1000 ng/ml de pFSH ou rFSH. O grupo controle (não cultivado) e aqueles cultivados foram processados para análises histológica e ultra-estrutural. Além disso, os diâmetros folicular e oocitário foram avaliados. Após sete dias de cultivo, apenas 50 ng/ml de rFSH manteve o percentual de folículos normais semelhante ao controle. Além disso, 10 ng/ml de pFSH e todas as concentrações de rFSH promoveram ativação de folículos primordiais. A presença de50 ng/ml de rFSH promoveu o maior diâmetro folicular após sete dias de cultivo. Em conclusão, 50 ng/ml de rFSH manteve a integridadede folículos pré-antrais caprinos e promoveu a ativação e o crescimento dos folículos cultivados.