Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Journal of applied oral science (Online) |
Texto Completo: | https://www.revistas.usp.br/jaos/article/view/100990 |
Resumo: | Objective : To investigate the influence of a three-dimensional cell culture model on the expression of osteoblastic phenotype in human periodontal ligament fibroblast (hPDLF) cultures. Material and Methods : hPDLF were seeded on bi-dimensional (2D) and three-dimensional (3D) collagen type I (experimental groups) and and on a plastic coverslip (control) for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity were performed. Also, cell morphology and immunolabeling for alkaline phosphatase (ALP) and osteopontin (OPN) were assessed by epifluorescence and confocal microscopy. The expression of osteogenic markers, including alkaline phosphatase, osteopontin, osteocalcin (OC), collagen I (COL I) and runt-related transcription factor 2 (RUNX2), were analyzed using real-time polymerase chain reaction (RT-PCR). Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Results : Experimental cultures produced an increase in cell proliferation. Immunolabeling for OPN and ALP in hPDLF were increased and ALP activity was inhibited by three-dimensional conditions. OPN and RUNX2 gene expression was significantly higher on 3D culture when compared with control surface. Moreover, ALP and COL I gene expression were significantly higher in three-dimensional collagen than in 2D cultures at 7 days. However, at 14 days, 3D cultures exhibited ALP and COL I gene expression significantly lower than the control, and the COL I gene expression was also significantly lower in 3D than in 2D cultures. Significant calcium mineralization was detected and quantified by alizarin red assay, and calcified nodule formation was not affected by tridimensionality. Conclusion : This study suggests that the 3D cultures are able to support hPDLF proliferation and favor the differentiation and mineralized matrix formation, which may be a potential periodontal regenerative therapy. |
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oai:revistas.usp.br:article/100990 |
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USP-17 |
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Journal of applied oral science (Online) |
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Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel Objective : To investigate the influence of a three-dimensional cell culture model on the expression of osteoblastic phenotype in human periodontal ligament fibroblast (hPDLF) cultures. Material and Methods : hPDLF were seeded on bi-dimensional (2D) and three-dimensional (3D) collagen type I (experimental groups) and and on a plastic coverslip (control) for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity were performed. Also, cell morphology and immunolabeling for alkaline phosphatase (ALP) and osteopontin (OPN) were assessed by epifluorescence and confocal microscopy. The expression of osteogenic markers, including alkaline phosphatase, osteopontin, osteocalcin (OC), collagen I (COL I) and runt-related transcription factor 2 (RUNX2), were analyzed using real-time polymerase chain reaction (RT-PCR). Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Results : Experimental cultures produced an increase in cell proliferation. Immunolabeling for OPN and ALP in hPDLF were increased and ALP activity was inhibited by three-dimensional conditions. OPN and RUNX2 gene expression was significantly higher on 3D culture when compared with control surface. Moreover, ALP and COL I gene expression were significantly higher in three-dimensional collagen than in 2D cultures at 7 days. However, at 14 days, 3D cultures exhibited ALP and COL I gene expression significantly lower than the control, and the COL I gene expression was also significantly lower in 3D than in 2D cultures. Significant calcium mineralization was detected and quantified by alizarin red assay, and calcified nodule formation was not affected by tridimensionality. Conclusion : This study suggests that the 3D cultures are able to support hPDLF proliferation and favor the differentiation and mineralized matrix formation, which may be a potential periodontal regenerative therapy. Universidade de São Paulo. Faculdade de Odontologia de Bauru2015-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/jaos/article/view/10099010.1590/1678-775720140462Journal of Applied Oral Science; Vol. 23 No. 2 (2015); 206-214Journal of Applied Oral Science; Vol. 23 Núm. 2 (2015); 206-214Journal of Applied Oral Science; v. 23 n. 2 (2015); 206-2141678-77651678-7757reponame:Journal of applied oral science (Online)instname:Universidade de São Paulo (USP)instacron:USPenghttps://www.revistas.usp.br/jaos/article/view/100990/99654Copyright (c) 2015 Journal of Applied Oral Scienceinfo:eu-repo/semantics/openAccessALVES, Luciana Bastos MARIGUELA, Viviane Casagrande GRISI, Márcio Fernando de Moraes SOUZA, Sérgio Luiz Scaombatti de NOVAES JUNIOR, Arthur Belém TABA JUNIOR, Mário OLIVEIRA, Paulo Tambasco de PALIOTO, Daniela Bazan 2015-07-28T17:13:11Zoai:revistas.usp.br:article/100990Revistahttp://www.scielo.br/jaosPUBhttps://www.revistas.usp.br/jaos/oai||jaos@usp.br1678-77651678-7757opendoar:2015-07-28T17:13:11Journal of applied oral science (Online) - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
title |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
spellingShingle |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel ALVES, Luciana Bastos |
title_short |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
title_full |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
title_fullStr |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
title_full_unstemmed |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
title_sort |
Expression of osteoblastic phenotype in periodontal ligament fibroblasts cultured in three-dimensional collagen gel |
author |
ALVES, Luciana Bastos |
author_facet |
ALVES, Luciana Bastos MARIGUELA, Viviane Casagrande GRISI, Márcio Fernando de Moraes SOUZA, Sérgio Luiz Scaombatti de NOVAES JUNIOR, Arthur Belém TABA JUNIOR, Mário OLIVEIRA, Paulo Tambasco de PALIOTO, Daniela Bazan |
author_role |
author |
author2 |
MARIGUELA, Viviane Casagrande GRISI, Márcio Fernando de Moraes SOUZA, Sérgio Luiz Scaombatti de NOVAES JUNIOR, Arthur Belém TABA JUNIOR, Mário OLIVEIRA, Paulo Tambasco de PALIOTO, Daniela Bazan |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
ALVES, Luciana Bastos MARIGUELA, Viviane Casagrande GRISI, Márcio Fernando de Moraes SOUZA, Sérgio Luiz Scaombatti de NOVAES JUNIOR, Arthur Belém TABA JUNIOR, Mário OLIVEIRA, Paulo Tambasco de PALIOTO, Daniela Bazan |
description |
Objective : To investigate the influence of a three-dimensional cell culture model on the expression of osteoblastic phenotype in human periodontal ligament fibroblast (hPDLF) cultures. Material and Methods : hPDLF were seeded on bi-dimensional (2D) and three-dimensional (3D) collagen type I (experimental groups) and and on a plastic coverslip (control) for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity were performed. Also, cell morphology and immunolabeling for alkaline phosphatase (ALP) and osteopontin (OPN) were assessed by epifluorescence and confocal microscopy. The expression of osteogenic markers, including alkaline phosphatase, osteopontin, osteocalcin (OC), collagen I (COL I) and runt-related transcription factor 2 (RUNX2), were analyzed using real-time polymerase chain reaction (RT-PCR). Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Results : Experimental cultures produced an increase in cell proliferation. Immunolabeling for OPN and ALP in hPDLF were increased and ALP activity was inhibited by three-dimensional conditions. OPN and RUNX2 gene expression was significantly higher on 3D culture when compared with control surface. Moreover, ALP and COL I gene expression were significantly higher in three-dimensional collagen than in 2D cultures at 7 days. However, at 14 days, 3D cultures exhibited ALP and COL I gene expression significantly lower than the control, and the COL I gene expression was also significantly lower in 3D than in 2D cultures. Significant calcium mineralization was detected and quantified by alizarin red assay, and calcified nodule formation was not affected by tridimensionality. Conclusion : This study suggests that the 3D cultures are able to support hPDLF proliferation and favor the differentiation and mineralized matrix formation, which may be a potential periodontal regenerative therapy. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-04-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/jaos/article/view/100990 10.1590/1678-775720140462 |
url |
https://www.revistas.usp.br/jaos/article/view/100990 |
identifier_str_mv |
10.1590/1678-775720140462 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/jaos/article/view/100990/99654 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2015 Journal of Applied Oral Science info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2015 Journal of Applied Oral Science |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Odontologia de Bauru |
publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Odontologia de Bauru |
dc.source.none.fl_str_mv |
Journal of Applied Oral Science; Vol. 23 No. 2 (2015); 206-214 Journal of Applied Oral Science; Vol. 23 Núm. 2 (2015); 206-214 Journal of Applied Oral Science; v. 23 n. 2 (2015); 206-214 1678-7765 1678-7757 reponame:Journal of applied oral science (Online) instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Journal of applied oral science (Online) |
collection |
Journal of applied oral science (Online) |
repository.name.fl_str_mv |
Journal of applied oral science (Online) - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
||jaos@usp.br |
_version_ |
1800221678758789120 |