2007 Fiscal Year Final Research Report Summary
Regeneration of cartilage by laser irradiation
| Project/Area Number |
18591671
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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 | Kyoto Prefectural University of Medicine |
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Principal Investigator |
HOJO Tatsuya Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Assistant (40298740)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Kenji Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Assistant (30347447)
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| Project Period (FY) |
2007 – 2008
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| Keywords | chondrocyte / svnovial fibroblast / laser / heat |
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| Research Abstract |
Heat stimulation was applied to the knee joints of Japanese white rabbits for 20 min using a microwave (MW) applicator (2.45-GHz, 0-80 W) . Alter 8-72 hrs, the articular cartilage was removed from the knee joints and proteins and total RNA were extracted. As controls, the knee joints without heat stimulation were analyzed. Expression of heat shock protein (HSP) 70 was confirmed by real-time PCR and Western blotting. Expression of proteoglycan core protein (PG) and type II collagen (Col II) were quantified using real-time PCR to assess cartilage matrix metabolism Compared to controls, HSP70 expression was higher with more than 40 W of heat stimulation. The expression of PG and Col II mRNA were higher with more than 20 W of heat stimulation and peaked with 40 W The highest expression of HSP70, PG, and Col II mRNA were observed 8hrs, 48hrs, and 72hrs after heat stimulation, respectively. On the other hand, the highest HSP70 protein expression was observed 24 h afar heat stimulation. When
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quercetin was used to inhibit the induction of HSP70 expression, PG mRNAexpression did not increase.
The present study demonstrated that appropriate heat stimulation increased the expressions of extracellular matrix gene, which might be partially mediated by HSP70. These findings showed that controlling HSP70 expression in the cartilage using thermotherapy might offer a novel strategy for suppressing cartilage degeneration in osteoarthritis.
Low-energy laser irradiation (LELI) as been hind to modulate various biological effects.,especially, cell proliferation of various type of cell. We reveal the mechanisms involved in cell proliferation effects induced by LET T Synovial fibroblasts were irradiated with a 660nm laser for 30 seconds or 2 minutes.After 48 hours LELI both for 30 seconds or 2 minutes promoted cell proliferation significantly by 13%. BrdU incorporation was determined in time course of 6, 12, 18, 24, and 48 hours after laser irradiation. The ratio of S phase on irradiation was significantly higher than non-irradiated groups at 24 hours. Cells treated with culture medium containing 0.2%FBS had positive expression of p15 in the nulear, otherwise cells treated with LELI had positive expression of p15 in the cytoplasm. In this study, we found that 660nm LELI stimulated synovial fibroblasts proliferation by modulating p15 nuclear transition. Less
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