1989 Fiscal Year Final Research Report Summary
ULTRASTRUCTURAL AND CYTOCHEMICAL STUDIES OF CLONAL CELL LINES IN FIBROBLASTS DERIVED FROM HUMAN PERIODONTAL LIGAMENTS
| Project/Area Number |
62440071
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Morphological basic dentistry
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| Research Institution | Tokyo Dental College |
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Principal Investigator |
MIAKE Kiyoshi TOKYO DENT. COLL. SCH. OF DENT. PROFESSOR, 歯学部, 教授 (30085691)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAYAMA Akihiko TOKYO DENT. COLL. SCH. OF DENT. ASSISTANT, 歯学部, 助手 (30130131)
YAMADA Marie TOKYO DENT. COLL. SCH. OF DENT. ASSOCIATE PROFESSOR, 歯学部, 助教授 (70115088)
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| Project Period (FY) |
1987 – 1989
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| Keywords | HUMAN PERIODONTAL LIGAMENT / FIBROBLASTS / SINGLE CELL CLONE / STRETCHING FORCE / ALKALINE PHOSPHATASE / ENZYME HISTOCHEMISTRY / ULTRASTRUCTURE |
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| Research Abstract |
It has recently been suggested that fibroblasts in periodontal ligaments have cytochemical characteristics similar to those of osteoblasts and different from those other in vivo connective tissues. The authors isolated clonal cell lines of fibroblast-like cells from human periodontal ligaments in order to clarify their nature.
Digestion with collagenase was used to isolated the cells form a human periodontal ligament. From the 768 cells cultured from human periodontal ligament, 7 clonal cell lines were isolated in vitro. Cultures of these clonal cell lines resulted in typical, spindle-shaped, fibroblast-like cells, all of which were homogeneous. Very high ALPase activity was observed in 4 of the 7 cell lines. Enzyme reaction products occurred mainly along cell membranes.
These clonal cell lines established from human periodontal ligaments were used in in vitro experiments to ascertain periodontal ligament responses, on the cellular level, to mechanical stretching. A procedure developed b
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y Hasegawa et al.(1985) was used in applying mechanical stretching. Unstretched cultures were used as controls. All cultures were processed for investigations of fine structures, histochemical and cytochemical detection of ALPase activity, and localization of alpha-tubulin. Cultured cells to which mechanical stretching was intermittently applied showed little change in overall appearance, cell shape and arrangement, and distribution of alpha-tubulin. Although fine structural characteristics remained unchanged in both stretched and unstretched cultures, mechanical stretching force clearly increased the intensity of ALPase activity. Histochemical and cytochemical examinations indicated that the added intensity resulted from increases in numbers of cells demonstrating enzymatic activity.
Finally, we tried to develop chemically-defined and reproducible methods for this cell culture study, and cultured the clonal cells in media containing different concentrations of serum. The results indicated that the appearance of high ALPase activity depended upon the supplementation of serum. Less
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