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      • KCI등재후보

        BMP Expression by Human Cementum-Derived Cells in vitro

        Ko, Hyun-Jung,,Grzesik, Wojciech J Korean Academy of Oral Biology and the UCLA Dental 2005 International Journal of Oral Biology Vol.30 No.3

        Bone morphogenetic proteins (BMPs), members of a large group of TGF-beta family, are important molecular regulators of morphogenesis of numerous tissues and organs, including bones and teeth. Most BMPs are capable of inducing bone formation in vivo and therefore are of considerable clinical interest for regenerating mineralized tissues. Recently, we have developed a method to culture cells from human cementum (human cementum-derived cells, HCDCs). HCDCs, when attached to synthetic hydroxyapatite/tricalcium phosphate (HA/TCP) ceramic and transplanted into immunodeficient mice, formed histologically identifiable cementum-like tissue. Since it is unclear to what extent BMPs are involved in cementogenesis, the aim of this study was to establish which BMPs are expressed by cementogenic HCDCs and whether the expression of BMPs is related to the degree of cellular differentiation in vitro HCDCs were maintained in growth medium (DMEM/F12 supplemented with 10% FBS) until confluent (proliferation stage). Upon reaching confluence, cells were incubated in the differentiation medium (DMEM/F12 medium containing 10% FBS and 50㎎/㎖ ascorbic acid) for 14 days (differentiation stage). Next, HCDCs were incubated in mineralization medium (DMEM/F12, 50 ㎎/㎖ ascorbic acid, 2.5 ㎎/㎖ of ITS (insulin-transferrin-selenium), 5 mM beta-glycerophosphate and10^(-8)M dexamethasone) for another 14 days (mineralization stage). At the end of each differentiation stage, total RNA wan isolated and evaluated for BMPs (2 through 8) expression by employing real time RT-PCR. HCDCs expressed most of BMPs examined except BMP-7 and BMP-8. Furthermore, on average, the highest levels of BMPs were expressed at the earlier differentiation stage, prior to the initiation of mineralization in vitro. These results indicate that several BMPs are expressed during cementoblastic differentiation and suggest that BMPs may be involved in the homeostasis of human cementum.

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        Inhibition of osteoclast formation by putative human cementoblasts

        Kim, Mi Ri,Yang, Kyung Yang,Grzesik, Wojciech,Ko, Hyun Jung KOREAN ACADAMY OF ORAL BIOLOGY 2008 International Journal of Oral Biology Vol.33 No.3

        Cementum is the mineralized tissue of the tooth. It is similar to bone in several aspects but it differs from bone. Human bone marrow stromal cells (BMSC and human cementum derived cells (HCDC)(10,000 cells/cm²) were plated in 6 well plates as feeder cells. The next day, mouse bone marrow cells (1.5 million cells/cm²) were added. One group of these plates were incubated in serum-free conditioned medium (SFCM) generated from BMSC or HCDC supplemented with 2% FBS, parathyroid hormone (PTH), 1, 25 dihydroxyvitamin D₃(Vit, D₃) and dexamethasone, or plain medium with the same supplements. Another group of plates were cocultured with BMSC or HCDC in plain medium supplemented with 2% FBS, PTH, Vit, D₃and dexamethasone. Plates grown without SFCM or coculture were used as controls. After 10 days, the cells were stained for tartrate-resistant acid phosphatase(TRAP). BMSC were found to support osteoclast formation under normal conditions. This was inhibited however by both SFCM generated from HCDC and also by coculture with HCDC. In addition, HCDC themselves did not support osteoclast formation under any conditions. Our results thus indicate that HCDC do not support osteoclast formation in vitro and that soluble factor (s) from HCDC may inhibit this process. In addition, we show that this inhibition also involves an active mechanism that is independent of osteoprotegerin, a feature that may distinguish cementoblasts from other cells present in periodontium.

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