Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/20518 |
Resumo: | Cancer is one of the diseases with the highest mortality in Brazil and worldwide. In addition, the conventional therapies present low efficacy and high toxicity. In this context, the diphenyl diselenide [(PhSe)2], an organoselenium compound, has been investigated in view of its promising pharmacological properties, including the anti-tumoral effect. However, such molecule has some physicochemical (lipophilic) and toxicological issues, which limit its clinical application. The development of a nanocapsules formulation could be an interesting approach to overcome such restrictions and potentiates its pharmacological effects. Considering the aforementioned context, the current study aimed to develop a (PhSe)2-loaded poly-Ɛ-caprolactone nanocapsules (NC (PhSe)2) (5 mg/mL) formulation as well as to investigate the potential cytotoxicity against melanoma and glioma cell lines, which are both highly aggressive and prevalent cancers. The formulations were prepared by the interfacial deposition of preformed polymer method. A detailed physicochemical characterization was performed using the following parameters: average particle size, polydispersity index (PDI), zeta potential, pH, compound content, encapsulation efficiency, morphology, thermal behavior (thermogravimetric analysis and differential scanning calorimetry), X-ray diffractrometry and fourier-transform infrared spectroscopy. Stability studies were also carried out using a photodegradation assay and monitoring the formulations over a 30-day storage period. The obtained nanocapsules suspension had suitable physicochemical properties as well as adequate compatibility with the constituents and increased the compound stability. Different in vitro techniques (MTT assay, propidium iodide uptake and nitric oxide content) were applied to investigate the cytotoxic potential of the NC (PhSe)2 against human melanoma cells lines (A375 and SK-Mel-103), rat glioma cells lines (C6) and healthy cells (human red blood cells and keratinocytes and rat astrocytes). Overall, the results demonstrated that the nanoencapsulation provided selective and superior cytotoxic effect against the tumoral cells in comparison to the free compound, which was also toxic to keratinocytes. The NC (PhSe)2 caused an increase in the nitrite content to both SK-Mel-103 and C6 cells, suggesting that the oxide nitric mediated pathways could be associated with the cytotoxic potential of the formulation. Following, in order to develop an alternative adjuvant therapy to melanoma treatment, a hydrogel formulation was developed. A mortar and pestle were used to prepare the hydrogels by thickening the nanocapsules suspension or a solution of free compound, both at a final (PhSe)2 concentration 5mg/g. Such formulations were characterized concerning average particle size, polydispersity index (PDI), pH, compound content, physicochemical stability, spreadability assessment, rheological behavior and cutaneous permeation profile (human skin). Both formulations had compatible characteristics intending cutaneous application and the nano-based hydrogel provided superior permeation of (PhSe)2 to dermis layer in comparison to the free compound formulation. Regarding the toxicological potential of the nanostructured formulation, two different protocols were carried out to better understand such characteristic. The fist protocol evaluated the nanocapsules formulation effect in a Zebra-fish model by assessing many behavioral parameters; the second one was performed using a repeated administration schedule by the intragastric route in male adult Wistar rats (5 mg/Kg – 1 mL/Kg, once a day, during 15 days) in which theplasma biochemical markers of liver and kidney function as well as the oxidative status of different organs (brain, liver and kidney) were evaluated. The exposure of Zebra-fish to NC (PhSe)2 did not trigger behavioral impairments or oxidative stress-related injuries in the samples of rats that received the repeated treatment. Finally, a preclinical model of glioblastoma multiforme was carried out. Male adult Wistar rats received an injection of C6 cells in the right striatum (1 x 106 cells/3 μL) for tumor developing. Five days after glioma implantation the animals were treated with free compound or NC (PhSe)2 for 15 days (1 mg/Kg- 1 mL/Kg/once a day) by the intragastric route. At the end of the experiment, the groups that received the compound, free or nanoencapsulated, had a reduction in tumor size in comparison to the vehicle-treated groups. Therefore, as a conclusion, the results obtained herein showed the great biological potential of nanocapsule formulations containing (PhSe)2 intending selective anti-tumoral effect. |
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Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do gliomaNanotechnological based-systems containing diphenyl diselenide intending melanoma and glioma treatmentSelênioNanopartículasCâncerHidrogelToxicidadePermeação cutâneaCNPQ::CIENCIAS DA SAUDE::FARMACIACancer is one of the diseases with the highest mortality in Brazil and worldwide. In addition, the conventional therapies present low efficacy and high toxicity. In this context, the diphenyl diselenide [(PhSe)2], an organoselenium compound, has been investigated in view of its promising pharmacological properties, including the anti-tumoral effect. However, such molecule has some physicochemical (lipophilic) and toxicological issues, which limit its clinical application. The development of a nanocapsules formulation could be an interesting approach to overcome such restrictions and potentiates its pharmacological effects. Considering the aforementioned context, the current study aimed to develop a (PhSe)2-loaded poly-Ɛ-caprolactone nanocapsules (NC (PhSe)2) (5 mg/mL) formulation as well as to investigate the potential cytotoxicity against melanoma and glioma cell lines, which are both highly aggressive and prevalent cancers. The formulations were prepared by the interfacial deposition of preformed polymer method. A detailed physicochemical characterization was performed using the following parameters: average particle size, polydispersity index (PDI), zeta potential, pH, compound content, encapsulation efficiency, morphology, thermal behavior (thermogravimetric analysis and differential scanning calorimetry), X-ray diffractrometry and fourier-transform infrared spectroscopy. Stability studies were also carried out using a photodegradation assay and monitoring the formulations over a 30-day storage period. The obtained nanocapsules suspension had suitable physicochemical properties as well as adequate compatibility with the constituents and increased the compound stability. Different in vitro techniques (MTT assay, propidium iodide uptake and nitric oxide content) were applied to investigate the cytotoxic potential of the NC (PhSe)2 against human melanoma cells lines (A375 and SK-Mel-103), rat glioma cells lines (C6) and healthy cells (human red blood cells and keratinocytes and rat astrocytes). Overall, the results demonstrated that the nanoencapsulation provided selective and superior cytotoxic effect against the tumoral cells in comparison to the free compound, which was also toxic to keratinocytes. The NC (PhSe)2 caused an increase in the nitrite content to both SK-Mel-103 and C6 cells, suggesting that the oxide nitric mediated pathways could be associated with the cytotoxic potential of the formulation. Following, in order to develop an alternative adjuvant therapy to melanoma treatment, a hydrogel formulation was developed. A mortar and pestle were used to prepare the hydrogels by thickening the nanocapsules suspension or a solution of free compound, both at a final (PhSe)2 concentration 5mg/g. Such formulations were characterized concerning average particle size, polydispersity index (PDI), pH, compound content, physicochemical stability, spreadability assessment, rheological behavior and cutaneous permeation profile (human skin). Both formulations had compatible characteristics intending cutaneous application and the nano-based hydrogel provided superior permeation of (PhSe)2 to dermis layer in comparison to the free compound formulation. Regarding the toxicological potential of the nanostructured formulation, two different protocols were carried out to better understand such characteristic. The fist protocol evaluated the nanocapsules formulation effect in a Zebra-fish model by assessing many behavioral parameters; the second one was performed using a repeated administration schedule by the intragastric route in male adult Wistar rats (5 mg/Kg – 1 mL/Kg, once a day, during 15 days) in which theplasma biochemical markers of liver and kidney function as well as the oxidative status of different organs (brain, liver and kidney) were evaluated. The exposure of Zebra-fish to NC (PhSe)2 did not trigger behavioral impairments or oxidative stress-related injuries in the samples of rats that received the repeated treatment. Finally, a preclinical model of glioblastoma multiforme was carried out. Male adult Wistar rats received an injection of C6 cells in the right striatum (1 x 106 cells/3 μL) for tumor developing. Five days after glioma implantation the animals were treated with free compound or NC (PhSe)2 for 15 days (1 mg/Kg- 1 mL/Kg/once a day) by the intragastric route. At the end of the experiment, the groups that received the compound, free or nanoencapsulated, had a reduction in tumor size in comparison to the vehicle-treated groups. Therefore, as a conclusion, the results obtained herein showed the great biological potential of nanocapsule formulations containing (PhSe)2 intending selective anti-tumoral effect.O câncer é atualmente uma das doenças com maior grau de mortalidade no Brasil e no mundo. As terapias convencionais apresentam na maioria dos casos uma baixa eficácia e elevada toxicidade. Sendo assim, o disseneleto de difenila [(PhSe)2], um composto orgânico de selênio, vem sendo estudado por suas promissoras atividades farmacológicas, incluindo antitumoral. Porém, este composto apresenta elevada lipofilia e efeitos tóxicos, o que dificulta seu uso clínico. Nesse sentido, as nanocápsulas poliméricas tornam-se uma alternativa interessante para contornar as limitações do (PhSe)2. Diante disso, este estudo teve como objetivo desenvolver nanocápsulas poliméricas para a incorporação do (PhSe)2 e avaliar, de forma inédita, a sua atividade antitumoral contra o melanoma e o glioma, dois tipos de câncer de elevada agressividade. As suspensões de nanocápsulas de poli-Ɛ-caprolactona contendo (PhSe)2 (NC (PhSe)2) (5mg/mL) foram preparadas pelo método de deposição interfacial do polímero pré-formado e caracterizadas em relação ao tamanho médio de partícula, índice de polidispersão, potencial zeta, pH, teor do composto, eficiência de encapsulamento (EE), morfologia, estabilidade físico-química durante 30 dias e por análises térmicas (calorimetria diferencial exploratória e termogavimetria), interações entre os constituintes (raio-X e infravermelho) e fotoestabilidade. Foram observadas características compatíveis com sistemas nanoestruturados para liberação de fármacos, elevada estabilidade físico-química sem incompatibilidades entre os componentes e a formulação foi capaz de aumentar a fotoestabilidade do composto. Análises in vitro empregando os testes do MTT, iodeto de propidio e níveis de óxido nítrico foram utilizadas para avaliar o potencial citotóxico frente a células de melanoma humano A375 e SK-Mel-103 e células de glioma de rato C6. Além disso, o teste do MTT foi usado para determinar a citotoxicidade em células sanguíneas humanas, queratinócitos e astrócitos. Tanto o composto livre quanto o nanoencapsulado foram capazes de reduzir a viabilidade das células tumorais, sendo que as NC (PhSe)2 tiveram efeito superior nas células SK-Mel-103 e C6. Além disso, foi observado que as suspensões de nanocápsulas causaram aumento nos níveis de óxido nítrico, sugerindo que a modulação desta via possivelmente esteja relacionada com o efeito antitumoral da formulação Em relação à citotoxicidade, as NC (PhSe)2 não causaram toxicidade em nenhuma linhagem de células normais, enquanto que o composto livre foi citotóxico para os queratinócitos. A fim de propor um hidrogel como terapia adjuvante para o tratamento do melanoma, a suspensão de nanocápsula foi espessada com goma xantana. Os hidrogéis foram preparados utilizando pistilo e gral de vidro para espessar a suspensão de NC (PhSe)2 e uma solução do composto livre (5 mg/g) e estas foram caracterizadas em relação ao tamanho médio de partículas, índice de polidispersão, pH, teor do composto, estabilidade físico-química, espalhabilidade, comportamento reológico e perfil de permeação in vitro. Ambas as formulações apresentaram características compatíveis com a aplicação tópica e o perfil de permeação cutânea demonstrou que as nanocápsulas favoreceram a permeação do composto até a derme em uma maior quantidade que a formulação contendo o composto livre. A fim de estudar a toxicidade da formulação nanoestruturada, dois protocolos foram conduzidos: o primeiro foi realizado empregando peixe-zebra como modelo animal e o teste do tanque novo para avaliar os efeitos da formulação (0,1 – 2 μM); o segundo protocolo utilizou ratos como modelo animal e análises bioquímicas plasmáticas e teciduais foram realizadas para determinar a toxicidade (5 mg/Kg – 1 mL/kg, durante 15 dias, uma vez ao dia, pela via intragástrica). No primeiro protocolo, as NC (PhSe)2 não provocaram alterações comportamentais ou bioquímicas nos peixes e no segundo protocolo, tanto o composto livre quanto o nanoencapsulado, não causaram nenhuma alteração nos parâmetros bioquímicos avaliados. Por fim, um modelo pré-clínico de glioblastoma foi conduzido para determinar a atividade antitumoral do (PhSe)2 e NC (PhSe)2. Para isso, uma alíquota das células C6 foi injetada no cérebro dos ratos para o desenvolvimento do tumor. Após 5 dias, os animais foram tratados durante 15 dias (1 mg/Kg – 1 mL/kg, uma vez ao dia, pela via intragástrica). Foi observado que o composto, independente de estar na forma livre ou incorporado às nanocápsulas, foi capaz de reduzir o tamanho do tumor. Desta forma, o estudo realizado mostrou que nanocápsulas poliméricas, tanto em suspensão aquosa quanto em hidrogel, apresentam grande potencial como nanocarreadores do (PhSe)2 em sistemas biológicos visando efeito antitumoral.Universidade Federal de Santa MariaBrasilAnálises Clínicas e ToxicológicasUFSMPrograma de Pós-Graduação em Ciências FarmacêuticasCentro de Ciências da SaúdeCruz, Letíciahttp://lattes.cnpq.br/3095970241017527Nogueira, Cristina WayneSilva, Cristiane de Bona daPrigol, MarinaWilhelm, Ethel AntunesCodevilla, Cristiane FrancoFerreira, Luana Mota2021-04-09T13:24:50Z2021-04-09T13:24:50Z2018-08-31info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/20518porAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-08-03T20:44:00Zoai:repositorio.ufsm.br:1/20518Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-08-03T20:44Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma Nanotechnological based-systems containing diphenyl diselenide intending melanoma and glioma treatment |
title |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
spellingShingle |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma Ferreira, Luana Mota Selênio Nanopartículas Câncer Hidrogel Toxicidade Permeação cutânea CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
title_full |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
title_fullStr |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
title_full_unstemmed |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
title_sort |
Sistemas de base nanotecnológica contendo disseleneto de difenila para o tratamento do melanoma e do glioma |
author |
Ferreira, Luana Mota |
author_facet |
Ferreira, Luana Mota |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cruz, Letícia http://lattes.cnpq.br/3095970241017527 Nogueira, Cristina Wayne Silva, Cristiane de Bona da Prigol, Marina Wilhelm, Ethel Antunes Codevilla, Cristiane Franco |
dc.contributor.author.fl_str_mv |
Ferreira, Luana Mota |
dc.subject.por.fl_str_mv |
Selênio Nanopartículas Câncer Hidrogel Toxicidade Permeação cutânea CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Selênio Nanopartículas Câncer Hidrogel Toxicidade Permeação cutânea CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Cancer is one of the diseases with the highest mortality in Brazil and worldwide. In addition, the conventional therapies present low efficacy and high toxicity. In this context, the diphenyl diselenide [(PhSe)2], an organoselenium compound, has been investigated in view of its promising pharmacological properties, including the anti-tumoral effect. However, such molecule has some physicochemical (lipophilic) and toxicological issues, which limit its clinical application. The development of a nanocapsules formulation could be an interesting approach to overcome such restrictions and potentiates its pharmacological effects. Considering the aforementioned context, the current study aimed to develop a (PhSe)2-loaded poly-Ɛ-caprolactone nanocapsules (NC (PhSe)2) (5 mg/mL) formulation as well as to investigate the potential cytotoxicity against melanoma and glioma cell lines, which are both highly aggressive and prevalent cancers. The formulations were prepared by the interfacial deposition of preformed polymer method. A detailed physicochemical characterization was performed using the following parameters: average particle size, polydispersity index (PDI), zeta potential, pH, compound content, encapsulation efficiency, morphology, thermal behavior (thermogravimetric analysis and differential scanning calorimetry), X-ray diffractrometry and fourier-transform infrared spectroscopy. Stability studies were also carried out using a photodegradation assay and monitoring the formulations over a 30-day storage period. The obtained nanocapsules suspension had suitable physicochemical properties as well as adequate compatibility with the constituents and increased the compound stability. Different in vitro techniques (MTT assay, propidium iodide uptake and nitric oxide content) were applied to investigate the cytotoxic potential of the NC (PhSe)2 against human melanoma cells lines (A375 and SK-Mel-103), rat glioma cells lines (C6) and healthy cells (human red blood cells and keratinocytes and rat astrocytes). Overall, the results demonstrated that the nanoencapsulation provided selective and superior cytotoxic effect against the tumoral cells in comparison to the free compound, which was also toxic to keratinocytes. The NC (PhSe)2 caused an increase in the nitrite content to both SK-Mel-103 and C6 cells, suggesting that the oxide nitric mediated pathways could be associated with the cytotoxic potential of the formulation. Following, in order to develop an alternative adjuvant therapy to melanoma treatment, a hydrogel formulation was developed. A mortar and pestle were used to prepare the hydrogels by thickening the nanocapsules suspension or a solution of free compound, both at a final (PhSe)2 concentration 5mg/g. Such formulations were characterized concerning average particle size, polydispersity index (PDI), pH, compound content, physicochemical stability, spreadability assessment, rheological behavior and cutaneous permeation profile (human skin). Both formulations had compatible characteristics intending cutaneous application and the nano-based hydrogel provided superior permeation of (PhSe)2 to dermis layer in comparison to the free compound formulation. Regarding the toxicological potential of the nanostructured formulation, two different protocols were carried out to better understand such characteristic. The fist protocol evaluated the nanocapsules formulation effect in a Zebra-fish model by assessing many behavioral parameters; the second one was performed using a repeated administration schedule by the intragastric route in male adult Wistar rats (5 mg/Kg – 1 mL/Kg, once a day, during 15 days) in which theplasma biochemical markers of liver and kidney function as well as the oxidative status of different organs (brain, liver and kidney) were evaluated. The exposure of Zebra-fish to NC (PhSe)2 did not trigger behavioral impairments or oxidative stress-related injuries in the samples of rats that received the repeated treatment. Finally, a preclinical model of glioblastoma multiforme was carried out. Male adult Wistar rats received an injection of C6 cells in the right striatum (1 x 106 cells/3 μL) for tumor developing. Five days after glioma implantation the animals were treated with free compound or NC (PhSe)2 for 15 days (1 mg/Kg- 1 mL/Kg/once a day) by the intragastric route. At the end of the experiment, the groups that received the compound, free or nanoencapsulated, had a reduction in tumor size in comparison to the vehicle-treated groups. Therefore, as a conclusion, the results obtained herein showed the great biological potential of nanocapsule formulations containing (PhSe)2 intending selective anti-tumoral effect. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-08-31 2021-04-09T13:24:50Z 2021-04-09T13:24:50Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/20518 |
url |
http://repositorio.ufsm.br/handle/1/20518 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Análises Clínicas e Toxicológicas UFSM Programa de Pós-Graduação em Ciências Farmacêuticas Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Brasil Análises Clínicas e Toxicológicas UFSM Programa de Pós-Graduação em Ciências Farmacêuticas Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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1805922074988904448 |