Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Sanches, Priscila Laviola
Orientador(a): Ribeiro, Ana Rosa Lopes Pereira
Banca de defesa: Takamori, Esther Rieko, Sant’Anna Filho, Celso Barbosa de, Granjeiro, José Mauro
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade do Grande Rio
Programa de Pós-Graduação: Programa de Pós-Graduação em Ensino das Ciências
Departamento: Unigranrio::Ensino das Ciências
País: Brasil
Palavras-chave em Português:
Área do conhecimento CNPq:
Link de acesso: http://localhost:8080/tede/handle/tede/369
Resumo: The development of the nanotechnology area, in particular in whatregards the production of nanomaterials, is growing exponentially. Human exposure to nanoparticles is high and originated from a variety of sources, such as chemical processes, medical devices (implants, prostheses, controlled drug delivery systems), food, pharmaceutical and cosmetic industries. However, the study of the cytotoxicity of these nanomaterials is contradictory. Therefore, it is extremely important to develop research that analyzes the impact of nanoparticles on human health, as well as generating regulations that up to now is insufficient. Currently, the sunscreens have in their composition nanoparticles such as those of titanium dioxide (TiO2).The use of TiO2 in sunscreen formulation is due to its ability to reflect and spread UVB (290-320 nm) and UVA (320-400 nm) rays ensuring good protection against solar radiation. Therefore, this work aimed to evaluate the potential cytotoxicity of TiO2 nanoparticles in the rutile crystalline structure in human skin cells. Due to the agglomeration of nanoparticles, a nanoparticle dispersion protocol was optimized, first in water and then in biological medium using an indirect sonicator. The dispersion was then characterized by dynamic light scattering (DLS) and electronic transmission microscopy (MET). A study of ions and proteins adsorption was carried out by energy dispersivex-ray spectroscopy (EDS) and mass spectrometry, respectively. We performed an evaluation of the nanoparticles cytotoxic profile in primary cultures of fibroblasts and keratinocytes by the neutral red assay and by flow cytometry. MET allowed the study of the nanoparticles internalization inside cells. The results demonstrate the formation of a bio-complex around nanoparticles surfaces due to the selective adsorption of ions and proteins from the culture medium, such as calcium and phosphorus ions and proteins as albumin, fibronectin and thrombospondin. We observed that human keratinocytes and fibroblasts internalized the nanoparticles, which located in microvesicles that migrate preferentially to the perinuclear region. However, after nanoparticle internalization, there was a significant increase in cell death due to apoptosis and a significant reduction in cell viability in the fibroblasts, as well as increased apoptosis death and necrosis in keratinocytes. We suggest that the bio-complex formation can mask the nanoparticles and modulate their penetration into human skin cells.
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spelling Ribeiro, Ana Rosa Lopes PereiraBenchimol, MarleneTakamori, Esther RiekoSant’Anna Filho, Celso Barbosa deGranjeiro, José MauroSanches, Priscila Laviola2020-01-07T18:10:33Z2017-05-24SANCHES, Priscila Laviola. Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos. 2017. 98 f. Dissertação (Mestrado em Biomedicina Translacional) - Universidade do Grande Rio, Duque de Caxias, 2017.http://localhost:8080/tede/handle/tede/369The development of the nanotechnology area, in particular in whatregards the production of nanomaterials, is growing exponentially. Human exposure to nanoparticles is high and originated from a variety of sources, such as chemical processes, medical devices (implants, prostheses, controlled drug delivery systems), food, pharmaceutical and cosmetic industries. However, the study of the cytotoxicity of these nanomaterials is contradictory. Therefore, it is extremely important to develop research that analyzes the impact of nanoparticles on human health, as well as generating regulations that up to now is insufficient. Currently, the sunscreens have in their composition nanoparticles such as those of titanium dioxide (TiO2).The use of TiO2 in sunscreen formulation is due to its ability to reflect and spread UVB (290-320 nm) and UVA (320-400 nm) rays ensuring good protection against solar radiation. Therefore, this work aimed to evaluate the potential cytotoxicity of TiO2 nanoparticles in the rutile crystalline structure in human skin cells. Due to the agglomeration of nanoparticles, a nanoparticle dispersion protocol was optimized, first in water and then in biological medium using an indirect sonicator. The dispersion was then characterized by dynamic light scattering (DLS) and electronic transmission microscopy (MET). A study of ions and proteins adsorption was carried out by energy dispersivex-ray spectroscopy (EDS) and mass spectrometry, respectively. We performed an evaluation of the nanoparticles cytotoxic profile in primary cultures of fibroblasts and keratinocytes by the neutral red assay and by flow cytometry. MET allowed the study of the nanoparticles internalization inside cells. The results demonstrate the formation of a bio-complex around nanoparticles surfaces due to the selective adsorption of ions and proteins from the culture medium, such as calcium and phosphorus ions and proteins as albumin, fibronectin and thrombospondin. We observed that human keratinocytes and fibroblasts internalized the nanoparticles, which located in microvesicles that migrate preferentially to the perinuclear region. However, after nanoparticle internalization, there was a significant increase in cell death due to apoptosis and a significant reduction in cell viability in the fibroblasts, as well as increased apoptosis death and necrosis in keratinocytes. We suggest that the bio-complex formation can mask the nanoparticles and modulate their penetration into human skin cells.O desenvolvimento da área de nanotecnologia, em particular no que diz respeito à produção de nanomateriais, encontra-se em crescimento exponencial. A exposição humana a nanopartículas é elevada e originada de diversas fontes, tais como processos químicos, dispositivos médicos (implantes, próteses, sistemas de liberação controlada de fármacos), indústria alimentar, farmacêutica e de cosméticos. Entretanto, os resultados dos estudos da citotoxicidade destes nanomateriais é muitas das vezes contraditório. Desta forma, é de extrema importância o desenvolvimento de pesquisas que analisem o impacto de nanopartículas na saúde humana, assim como gerar regulamentação que até ao momento é insuficiente. Atualmente, os protetores solares apresentam na sua composição nanopartículas como as de dióxido de titânio (TiO2). A utilização do TiO2 em fórmulas de filtros solares deve-se à sua capacidade de refletir e espalhar raios UVB (290–320 nm) e UVA (320–400 nm) garantindo uma boa proteção contra a radiação solar. Portanto, este trabalho teve como objetivo avaliar a potencial citotoxicidade de nanopartículas de TiO2 na estrutura cristalina de rutilo em células de pele humana. Devido à aglomeração das nanopartículas, foi otimizado um protocolo de dispersão das nanopartículas, primeiro em água e depois em meio biológico, utilizando um sonicador no modo indireto. A dispersão foi, em seguida, caracterizada por espalhamento de luz dinâmico (DLS) e microscopia eletrônica de transmissão (MET). Um estudo de adsorção de íons e proteínas foi realizado pela técnica de espetroscopia por energia dispersiva (EDS) e por espectrometria de massas, respectivamente. A avaliação do perfil citotóxico das nanopartículas em culturas primárias de fibroblastos e queratinócitos foi realizada pelo ensaio de vermelho neutro e por citometria de fluxo. Os estudos da internalização das nanopartículas no interior de células foram realizados por MET. Os resultados demonstram que houve formação de um bio-complexo nas superfícies das nanopartículas devido à adsorção seletiva de íons e proteínas do meio de cultivo, como por exemplo, os íons de cálcio e fósforo e proteínas como a albumina, fibronectina e trombospondina. Foi observado também que as nanopartículas foram internalizadas por queratinócitos e fibroblastos primários da pele humana e migram preferencialmente para a região perinuclear. Após a internalização das nanopartículas, houve um aumento significativo da morte celular por apoptose nos fibroblastos e redução significativa da viabilidade celular, assim como aumento de morte por apoptose e necrose nos queratinócitos, para as concentrações mais altas de nanopartículas. Supõe-se que a formação do bio-complexo pode mascarar as nanopartículas e modular a penetração das mesmas nas células da pele humana.Submitted by Janser dos Santos Nascimento (janser.nascimento@unigranrio.com.br) on 2020-01-07T18:10:33Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Priscila Laviola Sanches.pdf: 2756259 bytes, checksum: 161c38ecb5ded6dd78b024bc8a61037f (MD5)Made available in DSpace on 2020-01-07T18:10:33Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Priscila Laviola Sanches.pdf: 2756259 bytes, checksum: 161c38ecb5ded6dd78b024bc8a61037f (MD5) Previous issue date: 2017-05-24CNPqapplication/pdfporUniversidade do Grande RioPrograma de Pós-Graduação em Ensino das CiênciasUNIGRANRIOBrasilUnigranrio::Ensino das Ciênciashttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessBiomedicinaNanopartículasDióxido de titânioCIÊNCIAS DA SAÚDEEstudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisreponame:Biblioteca Digital de Teses e Dissertações da UNIGRANRIOinstname:Universidade do Grande Rio (UNIGRANRIO)instacron:UNIGRANRIOLICENSElicense.txtlicense.txttext/plain; charset=utf-81982http://localhost:8080/tede/bitstream/tede/369/1/license.txt4a50535e8405f611398e6e2d408dbd1bMD51CC-LICENSElicense_urllicense_urltext/plain; charset=utf-849http://localhost:8080/tede/bitstream/tede/369/2/license_url4afdbb8c545fd630ea7db775da747b2fMD52license_textlicense_texttext/html; charset=utf-80http://localhost:8080/tede/bitstream/tede/369/3/license_textd41d8cd98f00b204e9800998ecf8427eMD53license_rdflicense_rdfapplication/rdf+xml; charset=utf-80http://localhost:8080/tede/bitstream/tede/369/4/license_rdfd41d8cd98f00b204e9800998ecf8427eMD54ORIGINALPriscila Laviola Sanches.pdfPriscila Laviola Sanches.pdfapplication/pdf2756259http://localhost:8080/tede/bitstream/tede/369/5/Priscila+Laviola+Sanches.pdf161c38ecb5ded6dd78b024bc8a61037fMD55tede/3692020-01-07 16:10:33.078oai:localhost: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede.unigranrio.edu.br/PUBhttp://tede.unigranrio.edu.br/oai/requestrepositorio@instituicao.br||repositorio@instituicao.bropendoar:2020-01-07T18:10:33Biblioteca Digital de Teses e Dissertações da UNIGRANRIO - Universidade do Grande Rio (UNIGRANRIO)false
dc.title.por.fl_str_mv Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
title Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
spellingShingle Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
Sanches, Priscila Laviola
Biomedicina
Nanopartículas
Dióxido de titânio
CIÊNCIAS DA SAÚDE
title_short Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
title_full Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
title_fullStr Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
title_full_unstemmed Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
title_sort Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos
author Sanches, Priscila Laviola
author_facet Sanches, Priscila Laviola
author_role author
dc.contributor.advisor1.fl_str_mv Ribeiro, Ana Rosa Lopes Pereira
dc.contributor.advisor-co1.fl_str_mv Benchimol, Marlene
dc.contributor.referee1.fl_str_mv Takamori, Esther Rieko
dc.contributor.referee2.fl_str_mv Sant’Anna Filho, Celso Barbosa de
dc.contributor.referee3.fl_str_mv Granjeiro, José Mauro
dc.contributor.author.fl_str_mv Sanches, Priscila Laviola
contributor_str_mv Ribeiro, Ana Rosa Lopes Pereira
Benchimol, Marlene
Takamori, Esther Rieko
Sant’Anna Filho, Celso Barbosa de
Granjeiro, José Mauro
dc.subject.por.fl_str_mv Biomedicina
Nanopartículas
Dióxido de titânio
topic Biomedicina
Nanopartículas
Dióxido de titânio
CIÊNCIAS DA SAÚDE
dc.subject.cnpq.fl_str_mv CIÊNCIAS DA SAÚDE
description The development of the nanotechnology area, in particular in whatregards the production of nanomaterials, is growing exponentially. Human exposure to nanoparticles is high and originated from a variety of sources, such as chemical processes, medical devices (implants, prostheses, controlled drug delivery systems), food, pharmaceutical and cosmetic industries. However, the study of the cytotoxicity of these nanomaterials is contradictory. Therefore, it is extremely important to develop research that analyzes the impact of nanoparticles on human health, as well as generating regulations that up to now is insufficient. Currently, the sunscreens have in their composition nanoparticles such as those of titanium dioxide (TiO2).The use of TiO2 in sunscreen formulation is due to its ability to reflect and spread UVB (290-320 nm) and UVA (320-400 nm) rays ensuring good protection against solar radiation. Therefore, this work aimed to evaluate the potential cytotoxicity of TiO2 nanoparticles in the rutile crystalline structure in human skin cells. Due to the agglomeration of nanoparticles, a nanoparticle dispersion protocol was optimized, first in water and then in biological medium using an indirect sonicator. The dispersion was then characterized by dynamic light scattering (DLS) and electronic transmission microscopy (MET). A study of ions and proteins adsorption was carried out by energy dispersivex-ray spectroscopy (EDS) and mass spectrometry, respectively. We performed an evaluation of the nanoparticles cytotoxic profile in primary cultures of fibroblasts and keratinocytes by the neutral red assay and by flow cytometry. MET allowed the study of the nanoparticles internalization inside cells. The results demonstrate the formation of a bio-complex around nanoparticles surfaces due to the selective adsorption of ions and proteins from the culture medium, such as calcium and phosphorus ions and proteins as albumin, fibronectin and thrombospondin. We observed that human keratinocytes and fibroblasts internalized the nanoparticles, which located in microvesicles that migrate preferentially to the perinuclear region. However, after nanoparticle internalization, there was a significant increase in cell death due to apoptosis and a significant reduction in cell viability in the fibroblasts, as well as increased apoptosis death and necrosis in keratinocytes. We suggest that the bio-complex formation can mask the nanoparticles and modulate their penetration into human skin cells.
publishDate 2017
dc.date.issued.fl_str_mv 2017-05-24
dc.date.accessioned.fl_str_mv 2020-01-07T18:10:33Z
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dc.identifier.citation.fl_str_mv SANCHES, Priscila Laviola. Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos. 2017. 98 f. Dissertação (Mestrado em Biomedicina Translacional) - Universidade do Grande Rio, Duque de Caxias, 2017.
dc.identifier.uri.fl_str_mv http://localhost:8080/tede/handle/tede/369
identifier_str_mv SANCHES, Priscila Laviola. Estudo da citotoxicidade e biointerface de nanopartículas de dióxido de titânio utilizadas nas indústrias de cosméticos. 2017. 98 f. Dissertação (Mestrado em Biomedicina Translacional) - Universidade do Grande Rio, Duque de Caxias, 2017.
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