Isolation and characterization of gingival fibroblasts positive for alkaline phosphatase in patients with chronic periodontitis and drug-induced gingival hyperplasia

Isolamento e caracterização de fibroblastos gengivais positivos para fosfatase alcalina de pacientes portadores de periodontite crônica e hiperplasia gengival medicamentosa

Vagner Rodrigues Santos^I; Rafael Tomaz Gomes^II; Maurício Resende^III; Oslei Paes de Almeida^IV; Ricardo Della Colleta^IV

^IDepartamento de Clínica, Patologia e Cirurgia Odontológicas, Faculdade de Odontologia, UFMG, Belo Horizonte, MG, Brazil
^IICurso de Medicina, Faculdade de Medicina, UFMG, Belo Horizonte, MG, Brazil ]]> ^IIIDepartamento de Microbiologia, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG, Brazil
^IVDepartamento de Diagnóstico Oral, Faculdade de Odontologia de Piracicaba, UNICAMP, Piracicaba, SP, Brazil

Correspondence

ABSTRACT

PURPOSE: To investigate the expression of alkaline phosphatase (ALP) activity in gingival fibroblasts from individuals with chronic periodontitis (CP) and drug-induced gingival hyperplasia (DGH) induced by diphenylhydantoin.
METHODS: Gingival fragments were obtained from 13 patients (8 women and 5 men, from 22 to 74 years of age), with 4 fragments from clinically normal gingiva (NG), 5 from biopsy of periodontal pockets with CP, and 4 from DGH induced by diphenylhydantoin. Using an enzymatic digestion procedure, gingival cell suspensions containing ALP-positive fibroblasts were prepared without affecting ALP activity. Cytochemistry and histochemistry analyses were performed.
RESULTS: Fibroblasts from NG presented low levels of ALP when compared to CP and DGH, which showed elevated and intermediate levels of ALP, respectively. Little cell proliferation was observed for fibroblasts from CP and DGH as compared to NG. However, the quantity of cells recovered from the subcultures was similar to the quantity recovered from the initial cell culture for the three sources.
CONCLUSION: The expression of ALP is increased in CP and DGH, and fibroblasts in CP and DGH show low proliferation. This suggests that periodontal inflammation and diphenylhydantoin may influence ALP expression and human gingival fibroblast expansion. Other studies are necessary to better assess the importance of ALP in the development and progression of CP and DGH.

RESUMO

OBJETIVO: Investigar a expressão da atividade de fosfatase alcalina (ALP) em fibroblastos gengivais de sujeitos com periodontite crônica (PC) e hiperplasia gengival medicamentosa (HGM) induzida por difenilhidantoína.
METODOLOGIA: Os fragmentos gengivais foram obtidos de 13 pacientes (8 mulheres e 5 homens, 22 a 74 anos), sendo 4 fragmentos procedentes de gengiva clinicamente normal (GN), 5 de biópsias de bolsas periodontais com PC e 4 de HGM induzida pela difenilhidantoína. Através de digestão enzimática, suspensões de células gengivais foram preparadas contendo fibroblastos ALP-positivos sem afetar a atividade da enzima ALP. Análises citoquímica e histoquímica foram realizadas.
RESULTADOS: Fibroblastos de GN apresentaram fraca positividade para ALP quando comparados com PC e HGM. PC e HGM exibiram níveis elevados e intermediários, respectivamente, de ALP. Observou-se menor proliferação celular na cultura de células em PC e HGM que em GN. Entretanto, a quantidade de células recuperadas das sub-culturas foi similar à quantidade recuperada na cultura inicial para as três linhagens.
CONCLUSÕES: A expressão de ALP na PC e na HGM encontra-se aumentada e ocorre baixa proliferação de fibroblastos na PC e na HGM. Isto sugere que a inflamação periodontal e a difenilhidantoína podem influenciar a expressão de ALP e a multiplicação de fibroblastos gengivais humanos. Outros estudos são necessários para melhor avaliar a importância de ALP no desenvolvimento e na progressão de PC e HGM.

Palavras-chave: Fibroblastos; fosfatase alcalina; doença periodontal; hiperplasia gengival medicamentosa

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Acknowledgments

This study was financially supported by FAPEMIG. We thank Prof. Ricardo Della Colleta (FOP-UNICAMP) and Prof. Maurício Resende (ICB-UFMG) for their collaboration.

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Correspondence:
Vagner Rodrigues Santos
Faculdade de Odontologia UFMG
Av. Antônio Carlos 6627 - Campus Pampulha
Belo Horizonte, MG - Brasil 31270-901
E-mail: vegneer2003@yahoo.com.br

Received: December 22, 2008 ]]> Accepted: May 21, 2009