Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10348/1466 |
Resumo: | Recent studies suggest that bone marrow stromal cells are a potential source of osteoblasts and chondrocytes and can be used to regenerate damaged tissues using a tissue-engineering (TE) approach. However, these strategies require the use of an appropriate scaffold architecture that can support the formation de novo of either bone and cartilage tissue, or both, as in the case of osteochondral defects. The later has been attracting a great deal of attention since it is considered a difficult goal to achieve. This work consisted on developing novel hydroxyapatite/chitosan (HA/CS) bilayered scaffold by combining a sintering and a freeze-drying technique, and aims to show the potential of such type of scaffolds for being used in TE of osteochondral defects. The developed HA/CS bilayered scaffolds were characterized by Fourier transform infra-red spectroscopy, X-ray diffraction analysis, micro-computed tomography, and scanning electron microscopy (SEM). Additionally, the mechanical properties of HA/CS bilayered scaffolds were assessed under compression. In vitro tests were also carried out, in order to study the water-uptake and weight loss profile of the HA/CS bilayered scaffolds. This was done by means of soaking the scaffolds into a phosphate buffered saline for 1 up to 30 days. The intrinsic cytotoxicity of the HA scaffolds and HA/CS bilayered scaffolds extract fluids was investigated by carrying out a cellular viability assay (MTS test) using Mouse fibroblastic-like cells. Results have shown that materials do not exert any cytotoxic effect. Complementarily, in vitro (phase I) cell culture studies were carried out to evaluate the capacity of HA and CS layers to separately, support the growth and differentiation of goat marrow stromal cells (GBMCs) into osteoblasts and chondrocytes, respectively. Cell adhesion and morphology were analysed by SEM while the cell viability and proliferation were assessed by MTS test and DNA quantification. The chondrogenic differentiation of GBMCs was evaluated measuring the glucosaminoglycans synthesis. Data showed that GBMCs were able to adhere, proliferate and osteogenic differentiation was evaluated by alkaline phosphatase activity and immunocytochemistry assays after 14 days in osteogenic medium and into chondrocytes after 21 days in culture with chondrogenic medium. The obtained results concerning the physicochemical and biological properties of the developed HA/CS bilayered scaffolds, show that these constructs exhibit great potential for their use in TE strategies leading to the formation of adequate tissue substitutes for the regeneration of osteochondral defects. |
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Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cellsHydroxyapatiteChitosanBilayered scaffoldOsteochondral tissue engineeringStromal cellsAutologous modelRecent studies suggest that bone marrow stromal cells are a potential source of osteoblasts and chondrocytes and can be used to regenerate damaged tissues using a tissue-engineering (TE) approach. However, these strategies require the use of an appropriate scaffold architecture that can support the formation de novo of either bone and cartilage tissue, or both, as in the case of osteochondral defects. The later has been attracting a great deal of attention since it is considered a difficult goal to achieve. This work consisted on developing novel hydroxyapatite/chitosan (HA/CS) bilayered scaffold by combining a sintering and a freeze-drying technique, and aims to show the potential of such type of scaffolds for being used in TE of osteochondral defects. The developed HA/CS bilayered scaffolds were characterized by Fourier transform infra-red spectroscopy, X-ray diffraction analysis, micro-computed tomography, and scanning electron microscopy (SEM). Additionally, the mechanical properties of HA/CS bilayered scaffolds were assessed under compression. In vitro tests were also carried out, in order to study the water-uptake and weight loss profile of the HA/CS bilayered scaffolds. This was done by means of soaking the scaffolds into a phosphate buffered saline for 1 up to 30 days. The intrinsic cytotoxicity of the HA scaffolds and HA/CS bilayered scaffolds extract fluids was investigated by carrying out a cellular viability assay (MTS test) using Mouse fibroblastic-like cells. Results have shown that materials do not exert any cytotoxic effect. Complementarily, in vitro (phase I) cell culture studies were carried out to evaluate the capacity of HA and CS layers to separately, support the growth and differentiation of goat marrow stromal cells (GBMCs) into osteoblasts and chondrocytes, respectively. Cell adhesion and morphology were analysed by SEM while the cell viability and proliferation were assessed by MTS test and DNA quantification. The chondrogenic differentiation of GBMCs was evaluated measuring the glucosaminoglycans synthesis. Data showed that GBMCs were able to adhere, proliferate and osteogenic differentiation was evaluated by alkaline phosphatase activity and immunocytochemistry assays after 14 days in osteogenic medium and into chondrocytes after 21 days in culture with chondrogenic medium. The obtained results concerning the physicochemical and biological properties of the developed HA/CS bilayered scaffolds, show that these constructs exhibit great potential for their use in TE strategies leading to the formation of adequate tissue substitutes for the regeneration of osteochondral defects.Elsevier2011-12-28T10:16:09Z2006-01-01T00:00:00Z2006info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10348/1466engmetadata only accessinfo:eu-repo/semantics/openAccessOliveira, Joaquim M.Rodrigues, Márcia T.Silva, Simone S.Malafaya, Patrícia B.Gomes, Manuela E.Viegas, Carlos A.Azevedo, Jorge Manuel Teixeira deDias, Maria Isabel R.Mano, João F.Reis, Rui L.reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-02T13:01:14Zoai:repositorio.utad.pt:10348/1466Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:07:20.692564Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
title |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
spellingShingle |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells Oliveira, Joaquim M. Hydroxyapatite Chitosan Bilayered scaffold Osteochondral tissue engineering Stromal cells Autologous model |
title_short |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
title_full |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
title_fullStr |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
title_full_unstemmed |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
title_sort |
Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications : scaffold design and its performance when seeded with goat bone marrow stromal cells |
author |
Oliveira, Joaquim M. |
author_facet |
Oliveira, Joaquim M. Rodrigues, Márcia T. Silva, Simone S. Malafaya, Patrícia B. Gomes, Manuela E. Viegas, Carlos A. Azevedo, Jorge Manuel Teixeira de Dias, Maria Isabel R. Mano, João F. Reis, Rui L. |
author_role |
author |
author2 |
Rodrigues, Márcia T. Silva, Simone S. Malafaya, Patrícia B. Gomes, Manuela E. Viegas, Carlos A. Azevedo, Jorge Manuel Teixeira de Dias, Maria Isabel R. Mano, João F. Reis, Rui L. |
author2_role |
author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Oliveira, Joaquim M. Rodrigues, Márcia T. Silva, Simone S. Malafaya, Patrícia B. Gomes, Manuela E. Viegas, Carlos A. Azevedo, Jorge Manuel Teixeira de Dias, Maria Isabel R. Mano, João F. Reis, Rui L. |
dc.subject.por.fl_str_mv |
Hydroxyapatite Chitosan Bilayered scaffold Osteochondral tissue engineering Stromal cells Autologous model |
topic |
Hydroxyapatite Chitosan Bilayered scaffold Osteochondral tissue engineering Stromal cells Autologous model |
description |
Recent studies suggest that bone marrow stromal cells are a potential source of osteoblasts and chondrocytes and can be used to regenerate damaged tissues using a tissue-engineering (TE) approach. However, these strategies require the use of an appropriate scaffold architecture that can support the formation de novo of either bone and cartilage tissue, or both, as in the case of osteochondral defects. The later has been attracting a great deal of attention since it is considered a difficult goal to achieve. This work consisted on developing novel hydroxyapatite/chitosan (HA/CS) bilayered scaffold by combining a sintering and a freeze-drying technique, and aims to show the potential of such type of scaffolds for being used in TE of osteochondral defects. The developed HA/CS bilayered scaffolds were characterized by Fourier transform infra-red spectroscopy, X-ray diffraction analysis, micro-computed tomography, and scanning electron microscopy (SEM). Additionally, the mechanical properties of HA/CS bilayered scaffolds were assessed under compression. In vitro tests were also carried out, in order to study the water-uptake and weight loss profile of the HA/CS bilayered scaffolds. This was done by means of soaking the scaffolds into a phosphate buffered saline for 1 up to 30 days. The intrinsic cytotoxicity of the HA scaffolds and HA/CS bilayered scaffolds extract fluids was investigated by carrying out a cellular viability assay (MTS test) using Mouse fibroblastic-like cells. Results have shown that materials do not exert any cytotoxic effect. Complementarily, in vitro (phase I) cell culture studies were carried out to evaluate the capacity of HA and CS layers to separately, support the growth and differentiation of goat marrow stromal cells (GBMCs) into osteoblasts and chondrocytes, respectively. Cell adhesion and morphology were analysed by SEM while the cell viability and proliferation were assessed by MTS test and DNA quantification. The chondrogenic differentiation of GBMCs was evaluated measuring the glucosaminoglycans synthesis. Data showed that GBMCs were able to adhere, proliferate and osteogenic differentiation was evaluated by alkaline phosphatase activity and immunocytochemistry assays after 14 days in osteogenic medium and into chondrocytes after 21 days in culture with chondrogenic medium. The obtained results concerning the physicochemical and biological properties of the developed HA/CS bilayered scaffolds, show that these constructs exhibit great potential for their use in TE strategies leading to the formation of adequate tissue substitutes for the regeneration of osteochondral defects. |
publishDate |
2006 |
dc.date.none.fl_str_mv |
2006-01-01T00:00:00Z 2006 2011-12-28T10:16:09Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10348/1466 |
url |
http://hdl.handle.net/10348/1466 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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metadata only access info:eu-repo/semantics/openAccess |
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metadata only access |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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