Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo
Ano de defesa: | 2022 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de São Carlos
Câmpus São Carlos |
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv
|
Departamento: |
Não Informado pela instituição
|
País: |
Não Informado pela instituição
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | https://repositorio.ufscar.br/handle/ufscar/17593 |
Resumo: | Known for their biocompatible potential and magnetic property, iron oxide nanoparticles are the target of several studies for different applications. However, the synthesis of new nanoparticles with different physicochemical characteristics leads to unknown biological interactions that may pose health risks to their manipulators. In this study, the mixed iron oxide nanoparticle (Fe3O4 NP) with sodium (SO3)-Na+) sulfonate ligands, of interest in the energy sector, was evaluated through nanotoxicological aspects. Initially, morphological, chemical, and physical characterization of the Fe3O4 NP was performed using SEM-FEG (High-Resolution Scanning Electron Microscopy), DLS (Dynamic Light Scattering), Zeta Potential, UV-Vis (Ultraviolet-Visible Absorption Spectroscopy) and ATR-FTIR (Fourier Transform Attenuated Total Reflection Infrared Spectroscopy) techniques. For the in vitro experiments, different concentrations of Fe3O4 NP (250; 100 and 50 μg/mL) were analyzed against murine fibroblasts (LA-9) in periods of 24, 48 and 72h. Thus, its cytotoxic potential was established through analysis of mitochondrial function (MTT), morphological changes and cell internalization (Prussian Blue), production of Reactive Oxygen Species (ROS), Nitric Oxide (NO) (Griess reaction), cytokines (IL-6 and TNF), determination of cell death (flow cytometry) and analysis of clonogenic potential on the 7th day after exposure. For the in vivo experiments, mice (BALB/c) were divided into 5 experimental groups (100; 50; 25 and 10 mg/Kg of Fe3O4 NP) and were intraperitoneally exposed to 4 doses over 14 days, and the toxicity of repeated doses was evaluated. Indicative signs of toxicity, hematological parameters (overall and differential leukocyte count), liver function analyses (TGO and TGP), quantification of cytokines (IFN-ˠ; TNF; IL-6 and IL-10) and histological analysis of the organs, heart, lung, liver, spleen and kidneys were evaluated. The results showed that Fe3O4 NP has a circular shape, with heterogeneous particle distribution, compacted and agglomerate formation. There was the internalization of Fe3O4 NP for all concentrations tested in vitro (250, 50 and 100 μg/mL) in a time/dependent manner. However, the increase in ROS occurred only for the 250 μg/mL concentration. Moreover, it is suggested that this same concentration triggered a senescence process in fibroblasts (LA-9), due to the low cellular metabolism and consequent reduction in colony formation within seven days. All concentrations tested during in vivo experimentation (100; 50; 25 and 10 mg/Kg) promoted damage to mouse organs, especially to the liver. There was activation of the immune system through the production of leukocytes and pro-inflammatory cytokines. Our results contribute to the knowledge about the toxicology of iron oxide nanoparticles, demonstrating the importance of analyses in longer periods after exposure. Keywords: Nanotoxicology; Inflammation; BALB/c; LA-9; Histology; Senescence. |
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Feitosa, Karina AlvesAnibal, Fernanda de Freitashttp://lattes.cnpq.br/4918261968772806http://lattes.cnpq.br/17110119670646662023-04-04T17:27:01Z2023-04-04T17:27:01Z2022-10-25FEITOSA, Karina Alves. Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo. 2022. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17593.https://repositorio.ufscar.br/handle/ufscar/17593Known for their biocompatible potential and magnetic property, iron oxide nanoparticles are the target of several studies for different applications. However, the synthesis of new nanoparticles with different physicochemical characteristics leads to unknown biological interactions that may pose health risks to their manipulators. In this study, the mixed iron oxide nanoparticle (Fe3O4 NP) with sodium (SO3)-Na+) sulfonate ligands, of interest in the energy sector, was evaluated through nanotoxicological aspects. Initially, morphological, chemical, and physical characterization of the Fe3O4 NP was performed using SEM-FEG (High-Resolution Scanning Electron Microscopy), DLS (Dynamic Light Scattering), Zeta Potential, UV-Vis (Ultraviolet-Visible Absorption Spectroscopy) and ATR-FTIR (Fourier Transform Attenuated Total Reflection Infrared Spectroscopy) techniques. For the in vitro experiments, different concentrations of Fe3O4 NP (250; 100 and 50 μg/mL) were analyzed against murine fibroblasts (LA-9) in periods of 24, 48 and 72h. Thus, its cytotoxic potential was established through analysis of mitochondrial function (MTT), morphological changes and cell internalization (Prussian Blue), production of Reactive Oxygen Species (ROS), Nitric Oxide (NO) (Griess reaction), cytokines (IL-6 and TNF), determination of cell death (flow cytometry) and analysis of clonogenic potential on the 7th day after exposure. For the in vivo experiments, mice (BALB/c) were divided into 5 experimental groups (100; 50; 25 and 10 mg/Kg of Fe3O4 NP) and were intraperitoneally exposed to 4 doses over 14 days, and the toxicity of repeated doses was evaluated. Indicative signs of toxicity, hematological parameters (overall and differential leukocyte count), liver function analyses (TGO and TGP), quantification of cytokines (IFN-ˠ; TNF; IL-6 and IL-10) and histological analysis of the organs, heart, lung, liver, spleen and kidneys were evaluated. The results showed that Fe3O4 NP has a circular shape, with heterogeneous particle distribution, compacted and agglomerate formation. There was the internalization of Fe3O4 NP for all concentrations tested in vitro (250, 50 and 100 μg/mL) in a time/dependent manner. However, the increase in ROS occurred only for the 250 μg/mL concentration. Moreover, it is suggested that this same concentration triggered a senescence process in fibroblasts (LA-9), due to the low cellular metabolism and consequent reduction in colony formation within seven days. All concentrations tested during in vivo experimentation (100; 50; 25 and 10 mg/Kg) promoted damage to mouse organs, especially to the liver. There was activation of the immune system through the production of leukocytes and pro-inflammatory cytokines. Our results contribute to the knowledge about the toxicology of iron oxide nanoparticles, demonstrating the importance of analyses in longer periods after exposure. Keywords: Nanotoxicology; Inflammation; BALB/c; LA-9; Histology; Senescence.Conhecida pelo seu potencial biocompatível e sua propriedade magnética, as nanopartículas de óxido de ferro são alvos de diversos estudos para diferentes aplicações. Entretanto, a síntese de novas nanopartículas com diferentes características físico-químicas acarretam em desconhecidas interações biológicas que podem trazer riscos à saúde de seus manipuladores. Nesse estudo, a nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3) -Na+), e de interesse no setor energético, foi avaliada por meio de aspectos nanotoxicológicos. Inicialmente foi realizada caracterização morfológica, química e física da NP Fe3O4, através das técnicas MEV-FEG (Microscopia Eletrônica de Varredura de Alta Resolução), DLS (Espalhamento de Luz Dinâmico), Potencial Zeta, UV-Vis (Espectroscopia de Absorção Ultravioleta Visível) e ATR-FTIR (Espectroscopia Infravermelho de Reflexão Total Atenuada por Transformada de Fourier). Para os experimentos in vitro, foram analisadas diferentes concentrações de NP Fe3O4 (250; 100 e 50 μg/mL) frente aos fibroblastos murinos (LA-9) em períodos de 24, 48 e 72h. Dessa forma foi estabelecido seu potencial citotóxico através de análise de função mitocondrial (MTT), alterações morfológicas e internalização celular (Azul de Prússia), produção de Espécies Reativas de Oxigênio (EROs), Óxido nítrico (NO) (Reação de Griess), citocinas (IL-6 e TNF), determinação da morte celular (citometria de fluxo) e análise do potencial clonogênico no 7º dia após exposição. Para os experimentos in vivo, camundongos (BALB/c) foram divididos em 5 grupos experimentais (100; 50; 25 e 10 mg/Kg de NP Fe3O4), e foram expostos intraperitonealmente a 4 doses no período de 14 dias, sendo avaliado a toxicidade de doses repetidas. Foram avaliados sinais indicativos de toxicidade, parâmetros hematológicos (contagem global e diferencial de leucócitos), análises da função hepática (TGO e TGP), quantificação de citocinas (IFN-ˠ; TNF; IL-6 e IL-10) e análise histológica dos órgãos, coração, pulmão, fígado, baço e rins. Os resultados demonstraram que a NP Fe3O4 possui formato circular, com distribuição das partículas de forma heterogênea, compactada e com formação de aglomerados. Houve internalização de NP Fe3O4 para todas as concentrações testadas in vitro (250, 50 e 100 μg/mL) de forma tempo/dependente. Porém o aumento de EROs ocorreu somente para a concentração de 250 μg/mL. Além do mais, sugere-se que essa mesma concentração, desencadeou um processo de senescência nos fibroblastos (LA-9), devido ao baixo metabolismo celular e consequente redução na formação de colônias no período de sete dias. Todas as concentrações testadas durante experimentação in vivo (100; 50; 25 e 10 mg/Kg), promoveram danos aos órgãos dos camundongos, sobretudo para o fígado. Houve ativação do sistema imune mediante produção de leucócitos e citocinas pró-inflamatórias. Nossos resultados contribuem para o conhecimento acerca da toxicologia de nanopartículas de óxido de ferro, demonstrando a importância de análises em períodos maiores após exposição. Palavras-chave: Nanotoxicologia; Inflamação; BALB/c; LA-9; Histologia; Senescência.OutraCentro de Pesquisas, Desenvolvimento e Inovação Leopoldo Américo Miguez de Mello - CENPES/Petrobras/Projeto: Proc. Nº 2017/00010-7.porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEvUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessNanotoxicologiaInflamaçãoBALB/cLA-9HistologiaSenescênciaNanotoxicologyInflammationHistologySenescenceCIENCIAS BIOLOGICAS::BIOQUIMICACIENCIAS BIOLOGICASAnálise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivoToxicological analysis of mixed iron oxide nanoparticle (NP Fe3O4) with sodium sulfonate (SO3)-Na+ ligands: in vitro and in vivoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTESE- Karina Alves Feitosa - 03-04-23.pdfTESE- Karina Alves Feitosa - 03-04-23.pdfTESE - KARINA ALVES FEITOSA - 03-04-23application/pdf2748102https://repositorio.ufscar.br/bitstream/ufscar/17593/1/TESE-%20Karina%20Alves%20Feitosa%20-%2003-04-23.pdfab106d562de23a5c7b729014e368a14bMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8810https://repositorio.ufscar.br/bitstream/ufscar/17593/2/license_rdff337d95da1fce0a22c77480e5e9a7aecMD52TEXTTESE- Karina Alves Feitosa - 03-04-23.pdf.txtTESE- Karina Alves Feitosa - 03-04-23.pdf.txtExtracted texttext/plain194705https://repositorio.ufscar.br/bitstream/ufscar/17593/3/TESE-%20Karina%20Alves%20Feitosa%20-%2003-04-23.pdf.txtf8d841a49413843fb63581bead074187MD53THUMBNAILTESE- Karina Alves Feitosa - 03-04-23.pdf.jpgTESE- Karina Alves Feitosa - 03-04-23.pdf.jpgIM Thumbnailimage/jpeg7238https://repositorio.ufscar.br/bitstream/ufscar/17593/4/TESE-%20Karina%20Alves%20Feitosa%20-%2003-04-23.pdf.jpg4b2e67a70538f49cc42dacc9dc5be0e9MD54ufscar/175932023-04-05 03:21:44.675oai:repositorio.ufscar.br:ufscar/17593Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-05-25T13:05:41.108389Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
dc.title.alternative.eng.fl_str_mv |
Toxicological analysis of mixed iron oxide nanoparticle (NP Fe3O4) with sodium sulfonate (SO3)-Na+ ligands: in vitro and in vivo |
title |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
spellingShingle |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo Feitosa, Karina Alves Nanotoxicologia Inflamação BALB/c LA-9 Histologia Senescência Nanotoxicology Inflammation Histology Senescence CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS |
title_short |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
title_full |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
title_fullStr |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
title_full_unstemmed |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
title_sort |
Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo |
author |
Feitosa, Karina Alves |
author_facet |
Feitosa, Karina Alves |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/1711011967064666 |
dc.contributor.author.fl_str_mv |
Feitosa, Karina Alves |
dc.contributor.advisor1.fl_str_mv |
Anibal, Fernanda de Freitas |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4918261968772806 |
contributor_str_mv |
Anibal, Fernanda de Freitas |
dc.subject.por.fl_str_mv |
Nanotoxicologia Inflamação BALB/c LA-9 Histologia Senescência |
topic |
Nanotoxicologia Inflamação BALB/c LA-9 Histologia Senescência Nanotoxicology Inflammation Histology Senescence CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS |
dc.subject.eng.fl_str_mv |
Nanotoxicology Inflammation Histology Senescence |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS |
description |
Known for their biocompatible potential and magnetic property, iron oxide nanoparticles are the target of several studies for different applications. However, the synthesis of new nanoparticles with different physicochemical characteristics leads to unknown biological interactions that may pose health risks to their manipulators. In this study, the mixed iron oxide nanoparticle (Fe3O4 NP) with sodium (SO3)-Na+) sulfonate ligands, of interest in the energy sector, was evaluated through nanotoxicological aspects. Initially, morphological, chemical, and physical characterization of the Fe3O4 NP was performed using SEM-FEG (High-Resolution Scanning Electron Microscopy), DLS (Dynamic Light Scattering), Zeta Potential, UV-Vis (Ultraviolet-Visible Absorption Spectroscopy) and ATR-FTIR (Fourier Transform Attenuated Total Reflection Infrared Spectroscopy) techniques. For the in vitro experiments, different concentrations of Fe3O4 NP (250; 100 and 50 μg/mL) were analyzed against murine fibroblasts (LA-9) in periods of 24, 48 and 72h. Thus, its cytotoxic potential was established through analysis of mitochondrial function (MTT), morphological changes and cell internalization (Prussian Blue), production of Reactive Oxygen Species (ROS), Nitric Oxide (NO) (Griess reaction), cytokines (IL-6 and TNF), determination of cell death (flow cytometry) and analysis of clonogenic potential on the 7th day after exposure. For the in vivo experiments, mice (BALB/c) were divided into 5 experimental groups (100; 50; 25 and 10 mg/Kg of Fe3O4 NP) and were intraperitoneally exposed to 4 doses over 14 days, and the toxicity of repeated doses was evaluated. Indicative signs of toxicity, hematological parameters (overall and differential leukocyte count), liver function analyses (TGO and TGP), quantification of cytokines (IFN-ˠ; TNF; IL-6 and IL-10) and histological analysis of the organs, heart, lung, liver, spleen and kidneys were evaluated. The results showed that Fe3O4 NP has a circular shape, with heterogeneous particle distribution, compacted and agglomerate formation. There was the internalization of Fe3O4 NP for all concentrations tested in vitro (250, 50 and 100 μg/mL) in a time/dependent manner. However, the increase in ROS occurred only for the 250 μg/mL concentration. Moreover, it is suggested that this same concentration triggered a senescence process in fibroblasts (LA-9), due to the low cellular metabolism and consequent reduction in colony formation within seven days. All concentrations tested during in vivo experimentation (100; 50; 25 and 10 mg/Kg) promoted damage to mouse organs, especially to the liver. There was activation of the immune system through the production of leukocytes and pro-inflammatory cytokines. Our results contribute to the knowledge about the toxicology of iron oxide nanoparticles, demonstrating the importance of analyses in longer periods after exposure. Keywords: Nanotoxicology; Inflammation; BALB/c; LA-9; Histology; Senescence. |
publishDate |
2022 |
dc.date.issued.fl_str_mv |
2022-10-25 |
dc.date.accessioned.fl_str_mv |
2023-04-04T17:27:01Z |
dc.date.available.fl_str_mv |
2023-04-04T17:27:01Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
FEITOSA, Karina Alves. Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo. 2022. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17593. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/17593 |
identifier_str_mv |
FEITOSA, Karina Alves. Análise toxicológica da nanopartícula de óxido de ferro misto (NP Fe3O4) com ligantes de sulfonato de sódio (SO3)-Na+): in vitro e in vivo. 2022. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/17593. |
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https://repositorio.ufscar.br/handle/ufscar/17593 |
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por |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
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openAccess |
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Universidade Federal de São Carlos Câmpus São Carlos |
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Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv |
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UFSCar |
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Universidade Federal de São Carlos Câmpus São Carlos |
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