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doi: 10.1242/10.1242/jcs.00098

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A cell line with characteristics of the periodontal ligament fibroblasts is negatively regulated for mineralization and Runx2/Cbfa1/Osf2 activity, part of which can be overcome by bone morphogenetic protein-2

Yoshinori Saito^1,2,*, Tatsuya Yoshizawa^1,*, Fumio Takizawa^1,2, Mika Ikegame^1,3, Osamu Ishibashi¹, Kazuhiro Okuda², Kohji Hara², Kotaro Ishibashi⁴, Masuo Obinata⁵ and Hiroyuki Kawashima^1,

¹ Divisions of Cell Biology and Molecular Pharmacology, Niigata University Graduate School of Medical and Dental Sciences, 5274 2-Bancho, Gakkocho-dori, Niigata-city, Niigata 951-8514, Japan
² Division of Periodontology, Niigata University Graduate School of Medical and Dental Sciences, 5274 2-Bancho, Gakkocho-dori, Niigata-city, Niigata 951-8514, Japan
³ Divisions of Anatomy and Cell Biology of the Hard Tissue, Niigata University Graduate School of Medical and Dental Sciences, 5274 2-Bancho, Gakkocho-dori, Niigata-city, Niigata 951-8514, Japan
⁴ Daiichi Pharmaceutical Co. Ltd., 1-16-13 Kita-kasai, Edogawa-ku, Tokyo 134-8630, Japan
⁵ Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-cho, Aoba-ku, Sendai, Miyagi 980-8575, Japan

Author for correspondence (e-mail: kawashim{at}dent.niigata-u.ac.jp)

Accepted 13 August 2002

The periodontal ligament (PDL) is a connective tissue located between the cementum of teeth and the alveolar bone of the mandibula. It plays an integral role in the maintenance and regeneration of periodontal tissue. The cells responsible for maintaining this tissue are thought to be fibroblasts, which can be either multipotent or composed of heterogenous cell populations. However, as no established cell lines from the PDL are available, it is difficult to assess what type of cell promotes all of these functions. As a first step to circumvent this problem, we have cloned and characterized cell lines from the PDL from mice harboring a temperature-sensitive SV 40 large T-antigen gene. RT-PCR and in situ hybridization studies demonstrated that a cell line, designated PDL-L2, mimics the gene expression of the PDL in vivo: it expresses genes such as alkaline phosphatase, type I collagen, periostin, runt-related transcription factor-2 (Runx2) and EGF receptor, but does not express genes such as bone sialoprotein and osteocalcin. Unlike osteoblastic cells and a mixed cell population from the PDL, PDL-L2 cells do not produce mineralized nodules in the minearlization medium. When PDL-L2 cells were incubated in the presence of recombinant human bone morphogenetic protein-2 alkaline phosphatase activity increased and mineralized nodules were eventually produced, although the extent of mineralization is much less than that in osteoblastic MC3T3-E1 cells. Furthermore, PDL-L2 cells appeared to have a regulatory mechanism by which the function of Runx2 is normally suppressed.

Key words: Periodontal ligament, Cell line, Runx2/Cbfa1/Osf-2, Mineralization, BMP-2

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