Regeneration of cartilage by laser irradiation

用微波(2.45 GHz，0~80W)对日本大耳白兔膝关节进行热刺激20min。8-72小时后，取膝关节软骨，提取蛋白质和总RNA。作为对照，对无热刺激的膝关节进行分析。实时荧光定量聚合酶链式反应和Western blotting检测热休克蛋白70的表达。实时定量聚合酶链式反应检测软骨基质中蛋白多糖核心蛋白(PG)和II型胶原(Col II)的表达，热刺激40W以上时HSP70的表达高于对照组。热刺激20W以上，PG和Col II基因表达水平较高，40W时达到高峰，热刺激后8h、48h和72hHSP70、PG和Col II基因表达最高。热刺激后24 h，HSP70蛋白表达最高。当…适当的热刺激可促进细胞外基质基因的表达，这种作用可能部分是由HSP70介导的。这些结果表明，利用热疗控制HSP70在软骨中的表达可能为抑制骨关节炎软骨退变提供一种新的策略。低能量激光照射(LELI)可以调节多种生物学效应，特别是各种细胞的增殖。用660 nm激光照射T滑膜成纤维细胞30秒或2分钟，48小时后，30秒或2分钟的LELI均能显着促进细胞增殖，促进细胞增殖的作用达13%。分别于激光照射后6、12、18、24、48h测定BrdU掺入。照射后24小时S时相比例明显高于未照射组。在含0.2%胎牛血清的培养液中，p15在胞核呈阳性表达，而经Leli处理的细胞在胞浆中呈阳性表达。在本研究中，我们发现660 nm Leli通过调节p15核转变刺激滑膜成纤维细胞的增殖。较少

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 … More 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