1987 Fiscal Year Final Research Report Summary
Osteoblastic Characterization of Human Periodontal Ligament Fibroblast-like Cells
| Project/Area Number |
61570900
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Kanagawa Dental College |
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Principal Investigator |
TOSHIO Kawase Kanagawa Dental College, Assistant Professor, 歯学部, 助教授 (30084784)
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| Co-Investigator(Kenkyū-buntansha) |
TAMOTSU Nakano Kanagawa Dental College, Research Associate, 歯学部, 助手 (80164250)
KIYOSHI Imai Kanagawa Dental College, Research Associate, 歯学部, 助手 (10151656)
KATSUHIRO Nishiyama Kanagawa Dental College, Research Associate, 歯学部, 助手 (20084783)
SHIGERU Satio Kanagawa Dental College, Rpofessor, 歯学部, 教授 (80084713)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Human periodontal Ligament Fibroblast-like Cells / Human Alveolasr Bone Cells / Alkaline Phosphatase |
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| Research Abstract |
In this study, we have devised a simple procedure for culturing cells from human periodontaol ligmaent(HPL) and determined the activity and phenotype of alkaline phosphatase(ALPase) as the first step of the chartacterization of HPL fibroblasts(HPLF).
premolars extracted for orthodontic reasons were obtained fresh. Small pieces of HPL were removed from the middle one-third of the root and placed between the bottom of a 2cm well and 15mm thermanox coverslip in D-MEM containing 2 mg FCP/ml, 50ug ascorbic acis/ml and penichillin/streptomycin. HPLF were grown out of this explant for 7-10 days, and then subcultured after trypsinization. During the groeth pshase, these cultures were treacted with or without 5x10^<-9> M 1,25(OH)_2D_3.
HPLF has a high ALPase activity of approximately 17 nmole p-nitrophenol produced/min/1x10^5 cells which is similar than that of the freshly isolated rat calvatial osteoblast-like cells(23 nmole/mip/1x10^5 cells) and cultured human alveolar bone cells(HABC: 20 nmole/min/1x10^5cells).
The Km of ALPase at pH 10.15 was 0.43mM which remained through at least several passeges. The ALPase fo HPLF and HABC was heat labile. It was inhibited remarkedly by 1-homoarginine and only slightly by 1-phenylalanine and 1-phenylalanylglycylglycine.
During the proliferrative phase, ALPase of HPLF was not present. But at confluence, HPLF possesed ALPase which was stimulated by 1,25(OH)_2D_3.
ALPase of HPLF can bind to the non-collagenous extract which could be hyaluronic acid, but does not associate with collagenous protein. The ALPase in the cultured HPL fibroblast-like cells might be located not only on a plasma membrane but also in an extracelluar matrix.
Therefore, we conclude that the ALPase of HPLF is similar to that present osteoblasts. HPLF could be termed osteoblastic fibroblasts.
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