Isolation of alkaline phosphatase-positive gingival fibroblasts from patients with chronic inflammatory periodontal disease.
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.
Abe
T
T
Department of Periodontology and Endodontology, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
Hara
Y
Y
Abe
Y
Y
Aida
Y
Y
Maeda
K
K
D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis*; D001973:Bromodeoxyuridine; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D002469:Cell Separation; D002478:Cells, Cultured; D002908:Chronic Disease; D004247:DNA / Q000096:biosynthesis; D005260:Female; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology*; D005881:Gingiva / Q000201:enzymology / Q000473:pathology*; D006651:Histocytochemistry; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008565:Membrane Proteins / Q000032:analysis; D008875:Middle Aged; D010518:Periodontitis / Q000201:enzymology / Q000473:pathology*; D010641:Phenotype
D008565:Membrane Proteins; D004247:DNA; D000469:Alkaline Phosphatase; D001973:Bromodeoxyuridine