| 2603 Multi-layered Cell Sheet Engineering to Achieve Periodontal Tissue Regeneration | ||
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M. GOMEZ FLORES1, M. HASEGAWA2, M. YAMATO3, T. OKANO3, and I. ISHIKAWA4, 1Tokyo Medical & Dental University, Institute of Advance Biomedical Engineering and Science, Tokyo Women's Medical University, Bunkyo-ku Tokyo, Japan, 2Tokyo Medical & Dental University, Bunkyo-ku Tokyo, Japan, 3Institute of Advance Biomedical Engineering and Science, Tokyo Women's Medical University, Japan, 4Tokyo Medical & Dental University -, Bunkyo-ku Tokyo, Japan Objectives: The aim of periodontal therapy is to regenerate the different components of the periodontium. It might be possible to engineer periodontal tissue structure, maintaining its architecture and organization features, by laminating cell sheets. The aim of this study is to evaluate if multi-layered human periodontal ligament cells could induce mineralized tissue. Methods: Periodontal ligament cells were isolated. Then plated in culture dishes grafted with a temperature-responsive polymer to allow cell detachment as a continuos cell sheet. In Control group, PDL cells were cultured with D-MEM with 10% FBS and 10% antibiotics. In Experimental group, PDL cells were cultured with D-MEM and osteoblastic differentiation medium containing; 10nM Dexamethasone, 50μg/ml Ascorbic acid and 10mMβ-glycerophosphate. After 3 weeks Real Time-PCR for type I collagen, Scanning electron Microscope, alkaline phosphatase (ALP) and Von Kossa staining were performed. PDL cell sheets were layered and placed onto dentin blocks. Three groups were transplanted subcutaneously into the back of immunodeficient rats. Periods of 3 and 6 weeks after transplantation, the animals were sacrificed. Demineralized tissue sections were stained for H&E, Azan, and analyzed. Results: After 3 weeks of culture with osteoblastic medium, PDL cells produced mineral-like nodules. These also showed positive staining for ALP and Von Kossa. Higher level of mRNA expression of type I collagen. These were not observed in control group. After 6 weeks of transplantation with PDL cells induced with osteoblastic medium, we observed that the dentin surface showed a newly immature cementum-like tissue formation, and collagen fibers with perpendicular orientation and adherence to the newly deposited cementum-like tissue. Conclusions: We propose a novel strategy for periodontal regeneration to construct transplantable periodontal tissues by layering HPDL cell sheets in vitro. It might be useful for intended periodontal regeneration to apply the multi-layer PDL cell sheets. | ||
| Seq #217 - Periodontal Therapy 10:15 AM-12:15 PM, Saturday, 1 July 2006 Brisbane Convention & Exhibition Centre P4 | ||
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