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Isolation of alkaline phosphatase‐positive gingival fibroblasts from patients with chronic inflammatory periodontal disease
Author(s) -
Abe Tatsuya,
Hara Yoshitaka,
Abe Yukiko,
Aida Yoshitomi,
Maeda Katsumasa
Publication year - 1996
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.1996.tb00495.x
Subject(s) - alkaline phosphatase , fibroblast , subculture (biology) , periodontitis , microbiology and biotechnology , chronic periodontitis , biology , cell , in vitro , pathology , immunology , chemistry , enzyme , medicine , biochemistry
We have reported recently that increased expression of membrane alkaline phosphatase (ALP) activity is a phenotypical characteristic of gingival fibroblasts located in chronic inflammatory periodontal lesions. To understand the cellular properties of these cells, we isolated ALP‐positive gingival fibroblasts from patients with adult periodontitis and evaluated their proliferative potential. Using an enzymatic digestion procedure, we prepared gingival cell suspensions containing ALP‐positive fibroblasts without affecting their ALP activities. These cell suspensions were then subjected to 1 g sedimentation, followed by allowing cells to adhere to substrata. Using this procedure, 71.9% of isolated cells were ALP‐positive. Dissociation of ALP‐positive fibroblasts and contamination by non‐fibroblastic cells were examined by cytochemical and immunocytochemical analyses. The proliferative capacity of ALP‐positive fibroblasts in culture was assessed by monitoring the proportion of ALP‐positive cells after repeated subculture passages and by labelling DNA‐synthesizing cells with bromodeoxyuridine (BrdU). The proportion of ALP‐positive fibroblasts decreased during cell culture passages without an apparent change in the ALP‐positive phenotype. The percentage of BrdU‐positive cells was significantly lower among ALP‐positive than among ALP‐negative fibroblasts. These results indicate that ALP‐positive fibroblasts in chronic inflammatory periodontal lesions have low growth potential. We suggest that their reduced capacity to grow in vitro reflects a more differentiated state induced under inflammatory conditions in vivo .