Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.mimet.2018.07.004 http://hdl.handle.net/11449/176641 |
Resumo: | The aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorporating specific numbers of keratinocytes (NOK-si) onto cellularised, collagen gel scaffolds containing human gingival fibroblasts incubated in KGM media and cultured for 14 days. The detection of the appropriate oral mucosa/epithelial structure was evaluated by histology (hematoxylin and eosin (HE), periodic acid–Schiff (P.A.S.) and Picrosirius red), and immunocytochemistry (cytokeratin 13, cytokeratin 14, Ki-67 and collagen IV) compared to a normal human gingiva. The morphology of the reconstituted tissue was analyzed by Transmission Electron Microscopy. To further quantitate tissue damage, lactate dehydrogenase (LDH) was measured in the tissue supernatant. NOK-si grown upon a gingival scaffold provided an organotypic model in an in vitro setting and exhibited structural characteristics typically associated with normal oral mucosa. Immunocytochemistry revealed the detection of epithelial cytokeratins 13 and 14, Col IV and Ki-67 in the reconstituted oral mucosa model. Infection was detected after 8 h and 16 h. This study presents an in vitro cellularised, organotypic model of reconstituted oral mucosa, which enables close control and characterization of its structure and differentiation over a mid-length period of time in culture. |
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Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactionsCandida albicansHost-pathogenInfectionSecreted factorsStaphylococcus aureusTissue engineeringThe aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorporating specific numbers of keratinocytes (NOK-si) onto cellularised, collagen gel scaffolds containing human gingival fibroblasts incubated in KGM media and cultured for 14 days. The detection of the appropriate oral mucosa/epithelial structure was evaluated by histology (hematoxylin and eosin (HE), periodic acid–Schiff (P.A.S.) and Picrosirius red), and immunocytochemistry (cytokeratin 13, cytokeratin 14, Ki-67 and collagen IV) compared to a normal human gingiva. The morphology of the reconstituted tissue was analyzed by Transmission Electron Microscopy. To further quantitate tissue damage, lactate dehydrogenase (LDH) was measured in the tissue supernatant. NOK-si grown upon a gingival scaffold provided an organotypic model in an in vitro setting and exhibited structural characteristics typically associated with normal oral mucosa. Immunocytochemistry revealed the detection of epithelial cytokeratins 13 and 14, Col IV and Ki-67 in the reconstituted oral mucosa model. Infection was detected after 8 h and 16 h. This study presents an in vitro cellularised, organotypic model of reconstituted oral mucosa, which enables close control and characterization of its structure and differentiation over a mid-length period of time in culture.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Department of Dental Materials and Prosthodontics Oral Rehabilitation Program Araraquara School of Dentistry UNESP Univ. Estadual Paulista, CentroDepartment of Morphology Laboratory of Histology and Embryology Araraquara School of Dentistry UNESP Univ. Estadual PaulistaPlymouth University Peninsula Schools of Medicine and DentistryDepartment of Dental Materials and Prosthodontics Oral Rehabilitation Program Araraquara School of Dentistry UNESP Univ. Estadual Paulista, CentroDepartment of Morphology Laboratory of Histology and Embryology Araraquara School of Dentistry UNESP Univ. Estadual PaulistaCNPq: 163551/2012-0CNPq: 400658/2012-7CAPES: 99999.007120/2015-00Universidade Estadual Paulista (Unesp)Peninsula Schools of Medicine and Dentistryde Carvalho Dias, Kássia [UNESP]de Sousa, Denise Lins [UNESP]Barbugli, Paula Aboud [UNESP]Cerri, Paulo Sérgio [UNESP]Salih, Vehid MaxVergani, Carlos Eduardo [UNESP]2018-12-11T17:21:52Z2018-12-11T17:21:52Z2018-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article52-60application/pdfhttp://dx.doi.org/10.1016/j.mimet.2018.07.004Journal of Microbiological Methods, v. 152, p. 52-60.1872-83590167-7012http://hdl.handle.net/11449/17664110.1016/j.mimet.2018.07.0042-s2.0-850505536952-s2.0-85050553695.pdf32784959112078820000-0001-5756-5828Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Microbiological Methods0,696info:eu-repo/semantics/openAccess2023-10-27T06:09:03Zoai:repositorio.unesp.br:11449/176641Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T16:09:00.903392Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
title |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
spellingShingle |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions de Carvalho Dias, Kássia [UNESP] Candida albicans Host-pathogen Infection Secreted factors Staphylococcus aureus Tissue engineering |
title_short |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
title_full |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
title_fullStr |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
title_full_unstemmed |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
title_sort |
Development and characterization of a 3D oral mucosa model as a tool for host-pathogen interactions |
author |
de Carvalho Dias, Kássia [UNESP] |
author_facet |
de Carvalho Dias, Kássia [UNESP] de Sousa, Denise Lins [UNESP] Barbugli, Paula Aboud [UNESP] Cerri, Paulo Sérgio [UNESP] Salih, Vehid Max Vergani, Carlos Eduardo [UNESP] |
author_role |
author |
author2 |
de Sousa, Denise Lins [UNESP] Barbugli, Paula Aboud [UNESP] Cerri, Paulo Sérgio [UNESP] Salih, Vehid Max Vergani, Carlos Eduardo [UNESP] |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Peninsula Schools of Medicine and Dentistry |
dc.contributor.author.fl_str_mv |
de Carvalho Dias, Kássia [UNESP] de Sousa, Denise Lins [UNESP] Barbugli, Paula Aboud [UNESP] Cerri, Paulo Sérgio [UNESP] Salih, Vehid Max Vergani, Carlos Eduardo [UNESP] |
dc.subject.por.fl_str_mv |
Candida albicans Host-pathogen Infection Secreted factors Staphylococcus aureus Tissue engineering |
topic |
Candida albicans Host-pathogen Infection Secreted factors Staphylococcus aureus Tissue engineering |
description |
The aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorporating specific numbers of keratinocytes (NOK-si) onto cellularised, collagen gel scaffolds containing human gingival fibroblasts incubated in KGM media and cultured for 14 days. The detection of the appropriate oral mucosa/epithelial structure was evaluated by histology (hematoxylin and eosin (HE), periodic acid–Schiff (P.A.S.) and Picrosirius red), and immunocytochemistry (cytokeratin 13, cytokeratin 14, Ki-67 and collagen IV) compared to a normal human gingiva. The morphology of the reconstituted tissue was analyzed by Transmission Electron Microscopy. To further quantitate tissue damage, lactate dehydrogenase (LDH) was measured in the tissue supernatant. NOK-si grown upon a gingival scaffold provided an organotypic model in an in vitro setting and exhibited structural characteristics typically associated with normal oral mucosa. Immunocytochemistry revealed the detection of epithelial cytokeratins 13 and 14, Col IV and Ki-67 in the reconstituted oral mucosa model. Infection was detected after 8 h and 16 h. This study presents an in vitro cellularised, organotypic model of reconstituted oral mucosa, which enables close control and characterization of its structure and differentiation over a mid-length period of time in culture. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-11T17:21:52Z 2018-12-11T17:21:52Z 2018-09-01 |
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://dx.doi.org/10.1016/j.mimet.2018.07.004 Journal of Microbiological Methods, v. 152, p. 52-60. 1872-8359 0167-7012 http://hdl.handle.net/11449/176641 10.1016/j.mimet.2018.07.004 2-s2.0-85050553695 2-s2.0-85050553695.pdf 3278495911207882 0000-0001-5756-5828 |
url |
http://dx.doi.org/10.1016/j.mimet.2018.07.004 http://hdl.handle.net/11449/176641 |
identifier_str_mv |
Journal of Microbiological Methods, v. 152, p. 52-60. 1872-8359 0167-7012 10.1016/j.mimet.2018.07.004 2-s2.0-85050553695 2-s2.0-85050553695.pdf 3278495911207882 0000-0001-5756-5828 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Microbiological Methods 0,696 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
52-60 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128612100997120 |