ORIGINAL ARTICLE

 

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

 

 

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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.

Key words: Fibroblasts; alkaline phosphatase; periodontal disease; drug-induced gingival hyperplasia

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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.

 

References

1. Lisanti MP, Rodriguez-Boulan E, Saltiel AR. Emerging functional roles for the glycosyl-phosphatidylinositol membrane proteine anchor. J Membr Biol 1990;117:1-10.         [ Links ]

2. Weiss MJ, Ray K, Henthorn PS, Lamb B, Kadesh T, Harris H. Structure of the human liver/bone/kidney alkaline phosphatase gene. J Biol Chem 1988;263:12002-10.         [ Links ]

3. Harris H. The human alkaline phosphatases: What we know and what we don't know. Clin Chim Acta 1990;186:133-50.         [ Links ]

4. Somerman MJ, Archer SY, Imm GR, Foster RA. A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. J Dent Res 1988;67:66-70.         [ Links ]

5. Piche JE, Carnes DL Jr, Graves DT. Initial characterization of cells derived from humam periodontia. J Dent Res 1989;68:761-7.         [ Links ]

6. Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999;4;1-6.         [ Links ]

7. Colleta RD, Almeida OP, Graner E, Page RC, Bozzo L. Differential proliferation of fibroblasts cultured from hereditary gingival fibromatosis and normal gingiva. J Periodontal Res 1998;33:469-75.         [ Links ]

8. Abe T, Akamine A, Hara Y, Maeda K. Expression of membrane alkaline phosphatase activity on gingival fibroblasts in chronic inflammatory periodontal disease. J Periodontal Res 1994;29:259-65.         [ Links ]

9. Low MG. The glicosyl-phosphatidylinositol anchor of membrane proteins. Biochim Biophys Acta 1989;988:427-54.         [ Links ]

10. Noda M, Yoon K, Rodan GA, Koppel DE. High lateral mobility of endogenous and transfected alkaline phosphatase: a phosphatidylinositol-anchored membrane protein. J Cell Biol 1987;105:1671-7.         [ Links ]

11. Abe T, Hara Y, Abe Y, Aida Y, Maeda K. Isolation of alkaline phosphatase-positive gingival fibroblasts from patients with chronic inflammatory periodontal disease. J Periodontal Res 1996;31:285-93.         [ Links ]

12. Bartold PM, Page RC. Proteoglycans synthesized by cultured fibroblasts derived from normal and inflamed human gingiva. In Vitro Cell Dev Biol 1986;22:407-17.         [ Links ]

13. Larjava H, Heino J, Kähäri VM, Krusius T, Vuorio E. Characterization of one phenotype of human periodontal granulation-tissue fibroblasts. J Dent Res 1989;68:20-5.         [ Links ]

14. Huang JS, Ho KY, Chen CC, Wu YM, Wang CC, Ho YP, Liu CS. Collagen synthesis in idiopatic and dilantin-induced gingival fibromatosis. Kaohsiung J Med Sci 1997;13:141-8.         [ Links ]

15. Mornet E, Stura E, Lia-Baldini AS, Stigbrand T, Ménez A, Le Du MH. Structural evidence for a functional role of human tissue nonspecific alkaline phosphatase in bone mineralization. J Biol Chem 2001;276:31171-8.         [ Links ]

16. Komuro T. Re-evaluation of fibroblasts and fibroblast-like cells. Anat Embryol (Berl.) 1990;182:103-12.         [ Links ]

17. Schmitt-Gräff A, Desmoulière A, Gabbiani G. Heterogeneity of myofibroblast phenotypic features: an example of fibroblastic cell plasticity. Virchows Arch 1994;425:3-24.         [ Links ]

 

 

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