| Project/Area Number |
14370463
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Kobe University |
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Principal Investigator |
KUROSAKA Masahiro Kobe University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (70170115)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIOKA Hiroyuki Kobe University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 助手 (10252777)
YAMAMOTO Tetsuji Kobe University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 講師 (80220482)
YOSHIYA Shinnichi Kobe University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (00201070)
KOKUBU Takeshi Kobe University, Graduate School of Medicine, Physician, 医学部附属病院, 医員
MATSUI Nobuzou Kobe University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 助手 (60304094)
土井田 稔 神戸大学, 医学部附属病院, 講師 (60237170)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥8,500,000 (Direct Cost: ¥8,500,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2002: ¥5,800,000 (Direct Cost: ¥5,800,000)
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| Keywords | Anterior Cruciate Ligament / Low Intensity Pulsed Ultrasound / Tenomodulin / GDF-5 / bFGF / GDF-5 |
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| Research Abstract |
Muscle-derived cells (MDC) have been reported to differentiate into osteogenic and adipogenic cells. We investigated that low intensity pulsed ultrasound (LIPUS) acting as mechanical stress make MDCs differentiate into ligament cell lineage. Muscle-derived cells were isolated from the muscle of beagle dogs. Minced muscle was digested with collagenase and trypsinized. The cells were cultivated on T25 flask. Then, the attached cells were passaged for cell expansion. MDCs were plated in 6-well culture plates at a density of 2x10^5 and after incubation for 24 hrs, the medium was replaced. Following this the MDCs were exposed to LIPUS, consisting of 200 microseconds burst sine waves of 1.5 MHz repeating at 1.0 kHz, for 20 minutes. The incident intensity was 30 mW/cm^2 At 0 (immediately after 20 minutes LIPUS exposure) and one hour after LIPUS exposure, total RNA was extracted for the following RT-PCR. We semi-quantified mRNA of growth factors (TGFb1, bFGF, PDGF, GDF-5). We also investigated mRNA of type I and III collagen, which is major components of ligament fiber, and tenomodulin, as a differentiation marker for ligament. The up-regulations of bFGF was observed after the 20 minutes of LIPUS exposure. Type I and III collagen mRNA was expressed constitutively in both the MDCs exposed to LIPUS and the unexposed cells but that tenomodulin mRNA was induced in the MDCs exposed to LIPUS. An up-regulation of tenomodulin mRNA was observed in the MDCs exposed to LIPUS at immediately after the 20 minutes of LIPUS exposure. MDCs differentiated into another tissue lineage like tendon or ligament as a result of LIPUS exposure. This study can contribute to a strategy to successfully culture replacement human ligament tissues in vitro.
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