Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Dental Journal |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402020000300244 |
Resumo: | Abstract This in vitro study evaluated cell viability and metabolism, nitric oxide release and production of two chemokines and one cytokine by cultured human dental pulp fibroblasts (HDPF) in contact with two glass ionomer cements (Ketac Molar-KM and Vitrebond-VB), Single Bond (SB) and calcium hydroxide (Dycal-DY). Cultures of HDPF were established by means of an explant technique. The specimens were prepared under sterile conditions and in disks measuring 5 mm x 2 mm obtained from a prefabricated mold and placed on a permeable membrane to avoid direct contact with the cells. Cytotoxicity was assessed by Trypan Blue exclusion method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide release in cell supernatant was detected by the Griess Method whereas stromal derived factor-1 alpha (SDF-1α or CXCL12), chemokine (C-X-C motif) ligand 8 [Interleukin 8 (IL-8 or CXCL8)] and interleukin-6 (IL-6) were detected by ELISA. RT-qPCR was employed for gene expression analysis. Statistical analyses were performed by One-way ANOVA followed by Tukey’s post hoc test for materials independent of the time, and Two-way ANOVA followed by Bonferroni correction test for the comparisons between materials and experimental time (p<0.05). Cytotoxic tests showed significant differences only for DY. Protein levels and mRNA expression were significantly increased for IL-8 for both periods of time. IL-6 production increased when fibroblasts were stimulated by KM. SDF-1α protein production and mRNA expression were not affected by any of the materials. There was a decrease in nitrate/nitrite levels only for KM. Although DY caused intense cell death and did not stimulate the production of the inflammatory mediators evaluated in this work, it is known that this event seems to be fundamental for the process of repair of the pulp tissue and formation of mineralized barrier. KM and VB increased production of proteins related to the inflammatory process, thus favoring tissue repair. Therefore, although these glass ionomer cements did not lead to large cell death, they should be used with caution. |
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Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materialscytotoxicitydental materialschemokinescytokinesfibroblastsAbstract This in vitro study evaluated cell viability and metabolism, nitric oxide release and production of two chemokines and one cytokine by cultured human dental pulp fibroblasts (HDPF) in contact with two glass ionomer cements (Ketac Molar-KM and Vitrebond-VB), Single Bond (SB) and calcium hydroxide (Dycal-DY). Cultures of HDPF were established by means of an explant technique. The specimens were prepared under sterile conditions and in disks measuring 5 mm x 2 mm obtained from a prefabricated mold and placed on a permeable membrane to avoid direct contact with the cells. Cytotoxicity was assessed by Trypan Blue exclusion method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide release in cell supernatant was detected by the Griess Method whereas stromal derived factor-1 alpha (SDF-1α or CXCL12), chemokine (C-X-C motif) ligand 8 [Interleukin 8 (IL-8 or CXCL8)] and interleukin-6 (IL-6) were detected by ELISA. RT-qPCR was employed for gene expression analysis. Statistical analyses were performed by One-way ANOVA followed by Tukey’s post hoc test for materials independent of the time, and Two-way ANOVA followed by Bonferroni correction test for the comparisons between materials and experimental time (p<0.05). Cytotoxic tests showed significant differences only for DY. Protein levels and mRNA expression were significantly increased for IL-8 for both periods of time. IL-6 production increased when fibroblasts were stimulated by KM. SDF-1α protein production and mRNA expression were not affected by any of the materials. There was a decrease in nitrate/nitrite levels only for KM. Although DY caused intense cell death and did not stimulate the production of the inflammatory mediators evaluated in this work, it is known that this event seems to be fundamental for the process of repair of the pulp tissue and formation of mineralized barrier. KM and VB increased production of proteins related to the inflammatory process, thus favoring tissue repair. Therefore, although these glass ionomer cements did not lead to large cell death, they should be used with caution.Fundação Odontológica de Ribeirão Preto2020-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402020000300244Brazilian Dental Journal v.31 n.3 2020reponame:Brazilian Dental Journalinstname:Fundação Odontológica de Ribeirão Preto (FUNORP)instacron:FUNORP10.1590/0103-6440202003523info:eu-repo/semantics/openAccessModena,Karin Cristina da SilvaCalvo,Adriana MariaSipert,Carla RenataColombini-Ishikiriama,Bella LunaDionísio,Thiago JoséNavarro,Maria Fidela de LimaAtta,Maria TeresaSantos,Carlos Ferreiraeng2020-07-09T00:00:00Zoai:scielo:S0103-64402020000300244Revistahttps://www.scielo.br/j/bdj/https://old.scielo.br/oai/scielo-oai.phpbdj@forp.usp.br||sergio@fosjc.unesp.br1806-47600103-6440opendoar:2020-07-09T00:00Brazilian Dental Journal - Fundação Odontológica de Ribeirão Preto (FUNORP)false |
dc.title.none.fl_str_mv |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
title |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
spellingShingle |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials Modena,Karin Cristina da Silva cytotoxicity dental materials chemokines cytokines fibroblasts |
title_short |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
title_full |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
title_fullStr |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
title_full_unstemmed |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
title_sort |
Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials |
author |
Modena,Karin Cristina da Silva |
author_facet |
Modena,Karin Cristina da Silva Calvo,Adriana Maria Sipert,Carla Renata Colombini-Ishikiriama,Bella Luna Dionísio,Thiago José Navarro,Maria Fidela de Lima Atta,Maria Teresa Santos,Carlos Ferreira |
author_role |
author |
author2 |
Calvo,Adriana Maria Sipert,Carla Renata Colombini-Ishikiriama,Bella Luna Dionísio,Thiago José Navarro,Maria Fidela de Lima Atta,Maria Teresa Santos,Carlos Ferreira |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Modena,Karin Cristina da Silva Calvo,Adriana Maria Sipert,Carla Renata Colombini-Ishikiriama,Bella Luna Dionísio,Thiago José Navarro,Maria Fidela de Lima Atta,Maria Teresa Santos,Carlos Ferreira |
dc.subject.por.fl_str_mv |
cytotoxicity dental materials chemokines cytokines fibroblasts |
topic |
cytotoxicity dental materials chemokines cytokines fibroblasts |
description |
Abstract This in vitro study evaluated cell viability and metabolism, nitric oxide release and production of two chemokines and one cytokine by cultured human dental pulp fibroblasts (HDPF) in contact with two glass ionomer cements (Ketac Molar-KM and Vitrebond-VB), Single Bond (SB) and calcium hydroxide (Dycal-DY). Cultures of HDPF were established by means of an explant technique. The specimens were prepared under sterile conditions and in disks measuring 5 mm x 2 mm obtained from a prefabricated mold and placed on a permeable membrane to avoid direct contact with the cells. Cytotoxicity was assessed by Trypan Blue exclusion method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide release in cell supernatant was detected by the Griess Method whereas stromal derived factor-1 alpha (SDF-1α or CXCL12), chemokine (C-X-C motif) ligand 8 [Interleukin 8 (IL-8 or CXCL8)] and interleukin-6 (IL-6) were detected by ELISA. RT-qPCR was employed for gene expression analysis. Statistical analyses were performed by One-way ANOVA followed by Tukey’s post hoc test for materials independent of the time, and Two-way ANOVA followed by Bonferroni correction test for the comparisons between materials and experimental time (p<0.05). Cytotoxic tests showed significant differences only for DY. Protein levels and mRNA expression were significantly increased for IL-8 for both periods of time. IL-6 production increased when fibroblasts were stimulated by KM. SDF-1α protein production and mRNA expression were not affected by any of the materials. There was a decrease in nitrate/nitrite levels only for KM. Although DY caused intense cell death and did not stimulate the production of the inflammatory mediators evaluated in this work, it is known that this event seems to be fundamental for the process of repair of the pulp tissue and formation of mineralized barrier. KM and VB increased production of proteins related to the inflammatory process, thus favoring tissue repair. Therefore, although these glass ionomer cements did not lead to large cell death, they should be used with caution. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-06-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402020000300244 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-64402020000300244 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/0103-6440202003523 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Fundação Odontológica de Ribeirão Preto |
publisher.none.fl_str_mv |
Fundação Odontológica de Ribeirão Preto |
dc.source.none.fl_str_mv |
Brazilian Dental Journal v.31 n.3 2020 reponame:Brazilian Dental Journal instname:Fundação Odontológica de Ribeirão Preto (FUNORP) instacron:FUNORP |
instname_str |
Fundação Odontológica de Ribeirão Preto (FUNORP) |
instacron_str |
FUNORP |
institution |
FUNORP |
reponame_str |
Brazilian Dental Journal |
collection |
Brazilian Dental Journal |
repository.name.fl_str_mv |
Brazilian Dental Journal - Fundação Odontológica de Ribeirão Preto (FUNORP) |
repository.mail.fl_str_mv |
bdj@forp.usp.br||sergio@fosjc.unesp.br |
_version_ |
1754204095820857344 |