Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts

Detalhes bibliográficos
Autor(a) principal: Basso, Fernanda Goncalves
Data de Publicação: 2013
Outros Autores: Pansani, Taisa Nogueira, Oliveira, Camila Favero De, Turrioni, Ana Paula Silveira, Soares, Diana Gabriela, Hebling, Josimeri, Costa, Carlos Alberto De Souza
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/0103-6440201302229
http://hdl.handle.net/11449/126179
Resumo: Bisphosphonate-induced osteonecrosis has been related to the cytotoxicity of these drugs on oral mucosa cells. A previous study showed that 5 µM of zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is the highest concentration of this drug found in the oral cavity of patients under treatment. Therefore, in order to simulate an osteonecrosis clinical condition, the aim of this study was to evaluate the highest concentration of ZA applied on human epithelial cells (HaCaT) and gingival fibroblasts. For this purpose, cells (3×104 cells/cm2) were seeded in wells for 48 h using complete culture medium (cDMEM). After 48 h incubation, the cDMEM was replaced by fresh serum-free culture medium (DMEM-FBS) in which the cells were maintained for additional 24 h. Then, 5 µM ZA were added to the DMEM–FBS and the cells incubated in contact with the drug for 48 h. After this period, the number of viable cells (trypan blue), cell viability (MTT assay), total protein (TP) production and cell morphology (SEM analysis) were assessed. Data were analyzed statistically by Mann-Whitney, ANOVA and Tukey's test (α=0.05). ZA caused a significant reduction in the number of viable cells and decreased the metabolic activity of both cell lines. However, decrease of TP production occurred only in the epithelial cell cultures. Morphological alterations were observed in both cell types treated with ZA. In conclusion, ZA (5 µM) was cytotoxic to human epithelial cells and gingival fibroblast cultures, which could be associated, clinically, with the development of bisphosphonate-induced osteonecrosis.
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spelling Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblastsbisphosphonatescytotoxicityepithelial cellsfibroblastsBisphosphonate-induced osteonecrosis has been related to the cytotoxicity of these drugs on oral mucosa cells. A previous study showed that 5 µM of zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is the highest concentration of this drug found in the oral cavity of patients under treatment. Therefore, in order to simulate an osteonecrosis clinical condition, the aim of this study was to evaluate the highest concentration of ZA applied on human epithelial cells (HaCaT) and gingival fibroblasts. For this purpose, cells (3×104 cells/cm2) were seeded in wells for 48 h using complete culture medium (cDMEM). After 48 h incubation, the cDMEM was replaced by fresh serum-free culture medium (DMEM-FBS) in which the cells were maintained for additional 24 h. Then, 5 µM ZA were added to the DMEM–FBS and the cells incubated in contact with the drug for 48 h. After this period, the number of viable cells (trypan blue), cell viability (MTT assay), total protein (TP) production and cell morphology (SEM analysis) were assessed. Data were analyzed statistically by Mann-Whitney, ANOVA and Tukey's test (α=0.05). ZA caused a significant reduction in the number of viable cells and decreased the metabolic activity of both cell lines. However, decrease of TP production occurred only in the epithelial cell cultures. Morphological alterations were observed in both cell types treated with ZA. In conclusion, ZA (5 µM) was cytotoxic to human epithelial cells and gingival fibroblast cultures, which could be associated, clinically, with the development of bisphosphonate-induced osteonecrosis.A osteonecrose induzida por bisfosfonatos tem sido associada a um efeito citotóxico destes medicamentos sobre as células da mucosa oral. Um estudo recente demonstrou que 5 µM de ácido zoledrônico (AZ), um potente bisfosfonato nitrogenado, foi a maior concentração encontrada na cavidade oral da pacientes em tratamento com este medicamento. Portanto, para simular esta condição in vivo, o objetivo deste estudo foi avaliar o efeito da aplicação desta concentração de AZ sobre células epiteliais (HaCaT) e fibroblasto de gengiva. As células foram semeadas (3×104 células/cm2) e incubadas por 48 h em placas de 24 compartimentos, utilizando meio de cultura completo (cDMEM). Após permanecer por 24 h em DMEM sem soro fetal bovino (DMEM-SFB), 5 µM do AZ foram adicionados a este meio de cultura, o qual foi incubado em contato com as células por 48 h. Após este período, foram avaliados o número de células viáveis (trypan blue), viabilidade celular (teste de MTT), produção de proteína total e a morfologia celular (MEV). Os dados obtidos foram submetidos aos testes estatísticos de Mann-Whitney e ANOVA complementada por testes de Tukey (p>0,05). Foi demonstrado que o AZ causou diminuição significativa no número de células viáveis, além de redução do metabolismo celular para ambos os tipos celulares avaliados. Porém, redução na produção de proteína total ocorreu apenas para as células epiteliais. Alterações morfológicas foram observadas em ambos os tipos celulares tratados com AZ. Estes dados científicos indicam que a concentração de AZ avaliada neste estudo (5 µM) apresenta ação citotóxica sobre células epiteliais e fibroblastos de gengiva, o que poderia estar associado, clinicamente, ao desenvolvimento da osteonecrose induzida por bisfosfonatos.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Piracicaba School of Dentistry UNICAMP University of CampinasAraraquara School of Dentistry UNESP - Univ Estadual PaulistaButantan Institute/Departament of Morphology of UNIFESP - Federal University of Sao Paulo Laboratory of GeneticsAraraquara School of Dentistry UNESP - Univ Estadual PaulistaFundação Odontológica de Ribeirão PretoUniversidade Estadual de Campinas (UNICAMP)Universidade Estadual Paulista (Unesp)Universidade de São Paulo (USP)Basso, Fernanda GoncalvesPansani, Taisa NogueiraOliveira, Camila Favero DeTurrioni, Ana Paula SilveiraSoares, Diana GabrielaHebling, JosimeriCosta, Carlos Alberto De Souza2015-08-06T16:14:44Z2015-08-06T16:14:44Z2013-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article551-558application/pdfhttp://dx.doi.org/10.1590/0103-6440201302229Brazilian Dental Journal. Fundação Odontológica de Ribeirão Preto, v. 24, n. 6, p. 551-558, 2013.0103-6440http://hdl.handle.net/11449/12617910.1590/0103-6440201302229S0103-64402013000600551S0103-64402013000600551.pdfSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBrazilian Dental Journal0,476info:eu-repo/semantics/openAccess2023-12-24T06:19:23Zoai:repositorio.unesp.br:11449/126179Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:10:52.070287Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
title Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
spellingShingle Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
Basso, Fernanda Goncalves
bisphosphonates
cytotoxicity
epithelial cells
fibroblasts
title_short Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
title_full Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
title_fullStr Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
title_full_unstemmed Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
title_sort Cytotoxic Effects of Zoledronic Acid on Human Epithelial Cells and Gingival Fibroblasts
author Basso, Fernanda Goncalves
author_facet Basso, Fernanda Goncalves
Pansani, Taisa Nogueira
Oliveira, Camila Favero De
Turrioni, Ana Paula Silveira
Soares, Diana Gabriela
Hebling, Josimeri
Costa, Carlos Alberto De Souza
author_role author
author2 Pansani, Taisa Nogueira
Oliveira, Camila Favero De
Turrioni, Ana Paula Silveira
Soares, Diana Gabriela
Hebling, Josimeri
Costa, Carlos Alberto De Souza
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual de Campinas (UNICAMP)
Universidade Estadual Paulista (Unesp)
Universidade de São Paulo (USP)
dc.contributor.author.fl_str_mv Basso, Fernanda Goncalves
Pansani, Taisa Nogueira
Oliveira, Camila Favero De
Turrioni, Ana Paula Silveira
Soares, Diana Gabriela
Hebling, Josimeri
Costa, Carlos Alberto De Souza
dc.subject.por.fl_str_mv bisphosphonates
cytotoxicity
epithelial cells
fibroblasts
topic bisphosphonates
cytotoxicity
epithelial cells
fibroblasts
description Bisphosphonate-induced osteonecrosis has been related to the cytotoxicity of these drugs on oral mucosa cells. A previous study showed that 5 µM of zoledronic acid (ZA), a nitrogen-containing bisphosphonate, is the highest concentration of this drug found in the oral cavity of patients under treatment. Therefore, in order to simulate an osteonecrosis clinical condition, the aim of this study was to evaluate the highest concentration of ZA applied on human epithelial cells (HaCaT) and gingival fibroblasts. For this purpose, cells (3×104 cells/cm2) were seeded in wells for 48 h using complete culture medium (cDMEM). After 48 h incubation, the cDMEM was replaced by fresh serum-free culture medium (DMEM-FBS) in which the cells were maintained for additional 24 h. Then, 5 µM ZA were added to the DMEM–FBS and the cells incubated in contact with the drug for 48 h. After this period, the number of viable cells (trypan blue), cell viability (MTT assay), total protein (TP) production and cell morphology (SEM analysis) were assessed. Data were analyzed statistically by Mann-Whitney, ANOVA and Tukey's test (α=0.05). ZA caused a significant reduction in the number of viable cells and decreased the metabolic activity of both cell lines. However, decrease of TP production occurred only in the epithelial cell cultures. Morphological alterations were observed in both cell types treated with ZA. In conclusion, ZA (5 µM) was cytotoxic to human epithelial cells and gingival fibroblast cultures, which could be associated, clinically, with the development of bisphosphonate-induced osteonecrosis.
publishDate 2013
dc.date.none.fl_str_mv 2013-12-01
2015-08-06T16:14:44Z
2015-08-06T16:14:44Z
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/10.1590/0103-6440201302229
Brazilian Dental Journal. Fundação Odontológica de Ribeirão Preto, v. 24, n. 6, p. 551-558, 2013.
0103-6440
http://hdl.handle.net/11449/126179
10.1590/0103-6440201302229
S0103-64402013000600551
S0103-64402013000600551.pdf
url http://dx.doi.org/10.1590/0103-6440201302229
http://hdl.handle.net/11449/126179
identifier_str_mv Brazilian Dental Journal. Fundação Odontológica de Ribeirão Preto, v. 24, n. 6, p. 551-558, 2013.
0103-6440
10.1590/0103-6440201302229
S0103-64402013000600551
S0103-64402013000600551.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Brazilian Dental Journal
0,476
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 551-558
application/pdf
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 SciELO
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
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