2004 Fiscal Year Final Research Report Summary
THE EVALUATION OF OSTEOGEMC POTENTIAL OF CULTURED PERIOSTEUM-DERIVED CELLS
| Project/Area Number |
15592109
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
UENO Takaaki OKAYAMA UNIVEERSITY, GRADUATE SCHOOL OF MEDICINE AND DENITSTRY, ORAL AND MAXILLOFACIAL RECONSTRCUIIVE SURGERY, ASSISTANT PROFESSOR, 大学院・医歯学総合研究科, 助手 (60252996)
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| Co-Investigator(Kenkyū-buntansha) |
KAGAWA Toshimasa OKAYAMAUNIVIRSITY MEDICALAND DENTAL HOSPITAL, ORAL AND MAXILLOFACIAL, RECONSTRCUTIVE SURGERY, ASSISTANT PROFESSOR, 医学部・歯学部附属病院, 助手 (80346444)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Periosteal graft / Cultured periosteum derived cell / Bone morphogenetic protein-4 / Bone repair / エレクトロポレーション / 遺伝子導入 / BMP-4遺伝子 |
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| Research Abstract |
Periosteum has osteogenic potential and considerable attention has been focused on its use as a grafting material for the repair of bone and joint defects. Recently, the osteogenic potential of cultured periosteal cells has been reported. We present findings of bone formation induced from cultured periosteum-derived cells of rats tibiae. Periosteum was placed in culture medium with 10% Fetal Bovine Serum for 14 days. After reaching confluence, periosteal cells were resuspended with 025% trypsin/EDTA and then re-cultured three dimensionally on a collagen sponge for 7 days. Periosteal cell/collagen complex was grafted into rat submandibular muscle and A solution of plasmid DNA containing mouse BMP-4 (pMiw-BMP-4, kindly provided by Dr.Brigid I.M. Hogan and Dr.Kishimoto, Tohoku Univ.) was injected into grfated tissue and electric pulses were applied through paired-needle electrodes. As a control plasmid, Normal saline solution was injected. Immunosuppressant (FK506, 1.0 mg/kg/day) was administered intramuscularly. At 14, 21 and 35 days post grafting, grafted tissue was extirpated and compared histologically and radiographically with control groups. In the experimental group, periosteal cells had proliferated and differentiated into osteogenic cells by 14 days post grafting. Cell proliferation marker PCNA positive cells significantly increased compared with control groups. At 21 days post grafting, new bone formation was evident. At this stage, VEGF and BMP-4 positive cells significantly increased compared with control groups. By 35 days post grafting, bone growth was observed and new calcification was detected radiographically in the defect. New bone formation in experimental group developed and total calcified area was larger than control group. Cultured periosteum-derived cells treated with BMP-4 gene showed higher osteogenic potential in the early stage of periosteal cell graft model of rat.
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