A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/16556 |
Resumo: | In vivo studies have reported that reduced dimensions bioactive ceramic coating overcomes the advantages of the PHSA. The aims of this study were to determine the effect of a uniform bioceramic coating in the nanothickness range onto a moderately rough surface on human harvested bone cells cultivated isolated or in coculture (osteogenic cells and PBMC). The cells were plated over titanium as-machined (S1) surfaces, alumina-blasted/acid etched surfaces (S2), and alumina-blasted/acid-etched + 300-500 nm thickness amorphous Ca e P based coating obtained by ion beam assisted deposition (S3) and evaluated regarding viability, adhesion, morphology, cytokines quantification (IL1, TGF1, IL17, IL10) and immunolocalization of osteopontin (OPN) and alkaline phosphatase (ALP). The results showed that the surface treatment did not interfere on cell viability of PBMC cell culture or osteogenic cells (p>0.005), though the adhesion of osteogenic cells have demonstrated a different pattern for each surface. At 1 day, S2 and S3 showed the betters results, showing greater adhesion than the S1surface (p<0.001). At 7 days the S1 surface showed a significantly higher adhesion than the S3 surface (p<0.005). The cell morphology evaluated on PBMC cultures showed spread morphology in S1 group. The osteogenic cells and the coculture showed a similar pattern for the tree surfaces in the evaluated times. However, when compared both cell cultures, coculture looked like less spread and smaller than osteogenic cells. The % of osteogenic cells showing intracellular label for OPN at day 1 was significantly higher for the S3 surface compared to the S1 surface (p<0.03), not showing differences in coculture. The % of ALP intracellular labeling at 7 days was significantly higher for the S2 surface compared to the S1 surface (p<0.0065), the same characteristic was observed on coculture (p<0,05). At 14 days, no differences in % ALP labeling was observed in osteogenic cells between the surfaces (p>0.09). However in coculture % ALP labeling was significantly higher for the S3 group compared to the S2 group (p<0.005). Regarding the cytokine levels were not observed a pattern for osteogenic cells, monocytes or coculture. The cytokine levels ranged among the three groups (S1, S2, S3) for each cell culture. Probably, cell interactions and cell surface contact have had an important effect on cytokine release. These results indicate that surface composition and texture influence the early events related to in vitro osteogenesis and that the cytokine production probably was modulated by cell interactions. |
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A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitroImplanteTitânioOsteoblastoNanotecnologiaImunidade celularMateriais biomédicosImplantTitaniumOsteoblastNanotechnologyCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADAIn vivo studies have reported that reduced dimensions bioactive ceramic coating overcomes the advantages of the PHSA. The aims of this study were to determine the effect of a uniform bioceramic coating in the nanothickness range onto a moderately rough surface on human harvested bone cells cultivated isolated or in coculture (osteogenic cells and PBMC). The cells were plated over titanium as-machined (S1) surfaces, alumina-blasted/acid etched surfaces (S2), and alumina-blasted/acid-etched + 300-500 nm thickness amorphous Ca e P based coating obtained by ion beam assisted deposition (S3) and evaluated regarding viability, adhesion, morphology, cytokines quantification (IL1, TGF1, IL17, IL10) and immunolocalization of osteopontin (OPN) and alkaline phosphatase (ALP). The results showed that the surface treatment did not interfere on cell viability of PBMC cell culture or osteogenic cells (p>0.005), though the adhesion of osteogenic cells have demonstrated a different pattern for each surface. At 1 day, S2 and S3 showed the betters results, showing greater adhesion than the S1surface (p<0.001). At 7 days the S1 surface showed a significantly higher adhesion than the S3 surface (p<0.005). The cell morphology evaluated on PBMC cultures showed spread morphology in S1 group. The osteogenic cells and the coculture showed a similar pattern for the tree surfaces in the evaluated times. However, when compared both cell cultures, coculture looked like less spread and smaller than osteogenic cells. The % of osteogenic cells showing intracellular label for OPN at day 1 was significantly higher for the S3 surface compared to the S1 surface (p<0.03), not showing differences in coculture. The % of ALP intracellular labeling at 7 days was significantly higher for the S2 surface compared to the S1 surface (p<0.0065), the same characteristic was observed on coculture (p<0,05). At 14 days, no differences in % ALP labeling was observed in osteogenic cells between the surfaces (p>0.09). However in coculture % ALP labeling was significantly higher for the S3 group compared to the S2 group (p<0.005). Regarding the cytokine levels were not observed a pattern for osteogenic cells, monocytes or coculture. The cytokine levels ranged among the three groups (S1, S2, S3) for each cell culture. Probably, cell interactions and cell surface contact have had an important effect on cytokine release. These results indicate that surface composition and texture influence the early events related to in vitro osteogenesis and that the cytokine production probably was modulated by cell interactions.Fundação de Amparo a Pesquisa do Estado de Minas GeraisDoutor em Imunologia e Parasitologia AplicadasEstudos in vivo têm demonstrado resultados favoráveis para os implantes de titânio com coberturas de cerâmica bioativa de reduzidas dimensões (nanométrica), quando comparada ao método tradicional de deposição por meio de plasma spray de hidroxiapatita (PHSA). O objetivo deste estudo foi avaliar o efeito de uma cobertura uniforme de biocerâmica, em escala nanométrica, gerando superfícies moderadamente rugosas, sobre células osteogênicas e mononucleares (PBMC) humanas cultivadas isoladas ou em cocultura. Para isso, as células foram plaqueadas sobre três superfícies: usinada (grupo S1), jateada com óxido de alumínio seguido por subtração ácida (grupo S2), e jateada/subtraída seguida por deposição assistida por feixe iônico de Ca e P amorfo na ordem de 300-500 nm (grupo S3). Nas culturas de mononucleares (PBMC) e células osteogênicas foram realizadas avaliações da adesão, viabilidade celular e morfologia. Também foram realizadas imunolocalização das proteínas osteopontina (OPN) e fosfatase alcalina (ALP) na cultura de células osteogências e nas coculturas, e dosagem de citocinas (IL1, TGF1, IL17, IL10). Os resultados mostraram que o tratamento de superfície não interfere na viabilidade dos PBMC e das células osteogências (p>0,05), embora a adesão de células osteogênicas tenha variado em função da superfície e do tempo. Em 24 horas, os grupos S2 e S3 apresentaram os melhores resultados, com maiores taxas de adesão que o grupo S1 (p<0,001). Aos 7 dias, o grupo S1 apresentou um maior número de células aderidas que o grupo S3 (p< 0,05). Em relação à morfologia, os PBMC mostraram-se mais espraiados no grupo S1, enquanto as células osteogênicas e da cocultura apresentaram morfologia similar nas três superfícies nos diferentes tempos experimentais. No entanto, ao comparar as células osteogênicas e cocultura, estas apresentaram se geralmente menores e menos espraiadas. A porcentagem de células osteogênicas mostrando marcação intracelular para OPN foi significativamente maior em S3 quando comparado à S1 (p<0.03), não apresentando diferenças significativas na cocultura. A porcentagem de células mostrando marcação intracelular para ALP aos 7 dias foi significativamente maior no grupo S2 comparado ao grupo S1, tanto na cultura de células osteogênicas como na cocultura (p<0,05). Aos 14 dias, na cultura de células osteogênicas, não foram observadas diferenças na % de marcação intracelular para ALP entre os grupos, enquanto na cocultura foram detectadas diferenças entre os grupos S2 e S3 (p>0.09). Os níveis das citocinas variaram para cada cultura em função do tempo e da superfície, não apresentando um padrão de secreção. Os resultados do presente estudo indicam que composição e a textura da superfície influenciam os eventos iniciais da osteogênese in vitro e que a produção das citocinas é modulada pelas interações celulares sobre essas superfícies.Universidade Federal de UberlândiaBRPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasCiências BiológicasUFUDechichi, Paulahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4704960P7Souza, Maria Aparecida dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762379E9Beletti, Marcelo Emíliohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703947D5Fernandes, Liliana Bastos FreitasPeixoto, Pablohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703659Y7Moura, Camilla Christian Gomes2016-06-22T18:46:17Z2009-08-182016-06-22T18:46:17Z2009-07-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfapplication/pdfMOURA, Camilla Christian Gomes. A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro. 2009. 71 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2009.https://repositorio.ufu.br/handle/123456789/16556porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2020-01-24T06:01:05Zoai:repositorio.ufu.br:123456789/16556Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2020-01-24T06:01:05Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
title |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
spellingShingle |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro Moura, Camilla Christian Gomes Implante Titânio Osteoblasto Nanotecnologia Imunidade celular Materiais biomédicos Implant Titanium Osteoblast Nanotechnology CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
title_short |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
title_full |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
title_fullStr |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
title_full_unstemmed |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
title_sort |
A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro |
author |
Moura, Camilla Christian Gomes |
author_facet |
Moura, Camilla Christian Gomes |
author_role |
author |
dc.contributor.none.fl_str_mv |
Dechichi, Paula http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4704960P7 Souza, Maria Aparecida de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4762379E9 Beletti, Marcelo Emílio http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703947D5 Fernandes, Liliana Bastos Freitas Peixoto, Pablo http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4703659Y7 |
dc.contributor.author.fl_str_mv |
Moura, Camilla Christian Gomes |
dc.subject.por.fl_str_mv |
Implante Titânio Osteoblasto Nanotecnologia Imunidade celular Materiais biomédicos Implant Titanium Osteoblast Nanotechnology CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
topic |
Implante Titânio Osteoblasto Nanotecnologia Imunidade celular Materiais biomédicos Implant Titanium Osteoblast Nanotechnology CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
description |
In vivo studies have reported that reduced dimensions bioactive ceramic coating overcomes the advantages of the PHSA. The aims of this study were to determine the effect of a uniform bioceramic coating in the nanothickness range onto a moderately rough surface on human harvested bone cells cultivated isolated or in coculture (osteogenic cells and PBMC). The cells were plated over titanium as-machined (S1) surfaces, alumina-blasted/acid etched surfaces (S2), and alumina-blasted/acid-etched + 300-500 nm thickness amorphous Ca e P based coating obtained by ion beam assisted deposition (S3) and evaluated regarding viability, adhesion, morphology, cytokines quantification (IL1, TGF1, IL17, IL10) and immunolocalization of osteopontin (OPN) and alkaline phosphatase (ALP). The results showed that the surface treatment did not interfere on cell viability of PBMC cell culture or osteogenic cells (p>0.005), though the adhesion of osteogenic cells have demonstrated a different pattern for each surface. At 1 day, S2 and S3 showed the betters results, showing greater adhesion than the S1surface (p<0.001). At 7 days the S1 surface showed a significantly higher adhesion than the S3 surface (p<0.005). The cell morphology evaluated on PBMC cultures showed spread morphology in S1 group. The osteogenic cells and the coculture showed a similar pattern for the tree surfaces in the evaluated times. However, when compared both cell cultures, coculture looked like less spread and smaller than osteogenic cells. The % of osteogenic cells showing intracellular label for OPN at day 1 was significantly higher for the S3 surface compared to the S1 surface (p<0.03), not showing differences in coculture. The % of ALP intracellular labeling at 7 days was significantly higher for the S2 surface compared to the S1 surface (p<0.0065), the same characteristic was observed on coculture (p<0,05). At 14 days, no differences in % ALP labeling was observed in osteogenic cells between the surfaces (p>0.09). However in coculture % ALP labeling was significantly higher for the S3 group compared to the S2 group (p<0.005). Regarding the cytokine levels were not observed a pattern for osteogenic cells, monocytes or coculture. The cytokine levels ranged among the three groups (S1, S2, S3) for each cell culture. Probably, cell interactions and cell surface contact have had an important effect on cytokine release. These results indicate that surface composition and texture influence the early events related to in vitro osteogenesis and that the cytokine production probably was modulated by cell interactions. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-08-18 2009-07-17 2016-06-22T18:46:17Z 2016-06-22T18:46:17Z |
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 |
MOURA, Camilla Christian Gomes. A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro. 2009. 71 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2009. https://repositorio.ufu.br/handle/123456789/16556 |
identifier_str_mv |
MOURA, Camilla Christian Gomes. A influência do nano-recobrimento de cálcio e fósforo no processo de osteogênese in vitro. 2009. 71 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2009. |
url |
https://repositorio.ufu.br/handle/123456789/16556 |
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por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia BR Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas Ciências Biológicas UFU |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia BR Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas Ciências Biológicas UFU |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
instname_str |
Universidade Federal de Uberlândia (UFU) |
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UFU |
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UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU) |
repository.mail.fl_str_mv |
diinf@dirbi.ufu.br |
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1813711490498166784 |