| Project/Area Number |
13671540
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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 |
Orthopaedic surgery
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| Research Institution | Keio University |
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Principal Investigator |
CHIBA Kazuhiro Keio University, School of Medicine, Assistant Professor, 医学部, 講師 (80179952)
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| Co-Investigator(Kenkyū-buntansha) |
IMABAYASHI Hideaki Keio University, School of Medicine, Orthopedic Surgery, Resident, 医学部, 助手 (40296629)
TSUJI Takashi Keio University, School of Medicine, Orthopedic Surgery, Resident (2001-2001), 医学部, 助手13年度 (60296639)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | intervertebral disc / nucleus pulposus / annulus fibrosus / tissue culture / matrix metabolism / proteoglycan / collagen / alginate / 線維輪 / 細胞培養 / 細胞外基質 / 基質代謝 |
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| Research Abstract |
Purpose of the current study was to establish a reliable, cost effective and reproducible in vitro tissue culture system for the intervertebral disc to investigate the metabolism of the intervertebral disc as a whole organ, thereby obtaining information which may lead to the elucidation of possible mechanisms of intervertebral disc degeneration
Rabbit intervertebral discs were extirpated and embedded in alginate gel and were cultured for up to 4 weeks. At weekly intervals, the contents of DNA, sulfated proteoglycans, antigenic keratan sulfate,hyaluronic acid and collagen in the cultured disc tissues were measured to evaluate the tissue integrity, Incorporation rates of 3SS sulfate were also measured at each time point to investigate proteoglycan synthesis and newly synthesized radiolabelled proteoglycans were characterized by separation with sepharose CL2B column sieve chromatography. Intervetebral disc cells cultured in medium alone were used as controls throughout the experiments
The annulus fibrosus and ucleus pulposus of the intervertebral disc cultured in alginate gel maintained a higher content of various extracellular matrix molecules and sustained a higher rate of proteoglycan synthesis than those cultured in medium alone at all time points. Newly synthesized proteoglycans in the discs embedded in alginate gel also maintained their molecular sizes close to intact discs
Incorporation of whole disc tissues in alginate gel proved useful to maintain the integrity of intervertebral disc and may be useful for studying the metabolism of intervertebral disc as a whole organ
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