Regeneration of cartilage by laser irradiation

采用2.45 ghz， 0-80 W微波照射器对大白兔膝关节进行热刺激20 min。8 ~ 72小时，取膝关节关节软骨，提取蛋白和总RNA。作为对照，分析无热刺激的膝关节。通过实时荧光定量PCR和Western blotting检测热休克蛋白（HSP） 70的表达。采用实时荧光定量PCR法定量测定蛋白多糖核心蛋白（PG）和II型胶原蛋白（Col II）的表达，评估软骨基质代谢。与对照组相比，热刺激超过40 W时，HSP70的表达更高。PG和Col II mRNA的表达量在热刺激20 W以上时最高，在热刺激40 W时达到峰值，HSP70、PG和Col II mRNA的表达量分别在热刺激后8小时、48小时和72小时达到最高。另一方面，热刺激24 h时HSP70蛋白表达量最高。当槲皮素抑制HSP70的表达时，PG mrna的表达不增加。本研究表明，适当的热刺激增加了细胞外基质基因的表达，这可能是由HSP70介导的。这些发现表明，利用热疗控制软骨中HSP70的表达可能为抑制骨关节炎软骨退变提供了一种新的策略。低能量激光照射（LELI）已被用于调节各种生物效应。特别是各种类型细胞的增殖。我们揭示了LET - T诱导滑膜成纤维细胞增殖效应的机制，用660nm激光照射30秒或2分钟。48 h后，LELI作用30秒或2分钟均能显著促进细胞增殖13%。分别在激光照射后6、12、18、24、48小时测定BrdU的掺入情况。24 h时辐照组S相比例显著高于未辐照组。含有0.2%FBS的培养液处理的细胞核中p15阳性表达，而LELI处理的细胞细胞质中p15阳性表达。在本研究中，我们发现660nm LELI通过调节p15核跃迁刺激滑膜成纤维细胞增殖。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