KURATANI Tsuyoshi Okayama University Graduate School of medicine and dentistry, Research assistant, 大学院・医歯学総合研究科, 助手 (90311802)
KAMIOKA Hiroshi Okayama University Dental Hospital, Assistant Professor, 歯学部附属病院, 講師 (80253219)
TAKANO Teruko (YAMAMOTO,TERUKO) Okayama University Graduate School of Medicine and Dentistry, Professor, 大学院・医歯学総合研究科, 教授 (00127250)
|Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,400,000 (Direct Cost: ¥2,400,000)
The purpose of the present study was to investigate the CTGF gene delivery for enhancing the repair of fractured condylar and mandibular ramus. Using six-week-old wistar rat, 12 hours, 1, 3, 5, 7, and 14 days after the fracture, mandible including fractured condylar and ramus were dissected and decalcified with EDTA. After embedded in paraffin, 5 μ, β horizontal and sagittal sections were made. RNA probes of CTGF, collagen type I, II, III, X, Cbfal and TGF 0 were used for in situ hybridization to examine the expression of these factors during fracture healing. CTGF were expressed not only hypertrophic but hyperplastic chondrocyte in the healing site and this expression was changed in the time course. The present findings suggest that CTGF may play an important roll for repair and healing in condylar fracture and this model is useful as a model of fracture healing.
Following experiments were performed in vitro and in vivo, to examine the optimum conditions of HVJ liposome method for the condylar fracture model. (1) Luciferase gene was delivered to mesenchymal cell, isolated from the bone marrow in mouse femur, by using HVJ-liposome method in vitro. Luciferase reporter assay was performed to evaluate delivery rate. (2) After the fracture, β-galactosidase gene was delivered by using HVJ-liposome method. Three days after gene introduction, X-gal staining was performed to evaluate the delivery rate. These pilot studies indicated that the gene delivery by using HVJ-liposome was efficient to the condylar fracture model.
CTGF gene delivered by using HVJ-liposome in rat is performed and both in situ hybridization and immuno-histochemistry were used to analyze the expression of CTGF during fracture healing. We are preparing the paper about these observations to submit to the international journals, and some of them have been accepted.