Ogita Mayumi, Tsuchida Sachio, Aoki Akira, Satoh Mamoru, Kado Sayaka, Sawabe Masanori, Nanbara Hiromi, Kobayashi Hiroaki, Takeuchi Yasuo, Mizutani Koji, Sasaki Yoshiyuki, Nomura Fumio, Izumi Yuichi
Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan.
Lasers Med Sci. 2015 Sep;30(7):1855-66. doi: 10.1007/s10103-014-1691-4. Epub 2014 Nov 28.
Erbium-doped yttrium aluminum garnet (Er:YAG) laser treatment has demonstrated favorable wound healing effect after periodontal therapy. One of the reasons may be the positive biological effect of the low-level laser on the irradiated tissues, although the mechanism remains unclear. The aim of this study was to investigate the effect of low-level Er:YAG laser irradiation on cell proliferation and laser-induced differential expression of proteins in human gingival fibroblasts (HGFs) by proteomic analysis. In the first experiment, HGFs were exposed to low-level Er:YAG laser irradiation and the laser-induced cell proliferation and damage were evaluated on day 3. In the second experiment, proteomic analysis was performed on day 1 after irradiation. The peptides prepared from HGFs were analyzed by a hybrid ion trap-Fourier transform mass spectrometer, Mascot search engine, and UniProtKB database. A significant increase in cell proliferation without cell damage after irradiation was observed. Among the total identified 377 proteins, 59 proteins, including galectin-7, which was associated with the process of wound healing, were upregulated and 15 proteins were downregulated in laser-treated HGFs. In the third experiment, the increase in messenger RNA (mRNA) and protein expression of galectin-7 in the irradiated HGFs was validated by various analytical techniques. In addition, the effect of recombinant human galectin-7 on the modulation of HGFs proliferation was confirmed. The results indicate that low-level Er:YAG laser irradiation can promote HGF proliferation and induce a significant change in protein expression and the upregulation of galectin-7 expression may partly contribute to the increase in cell proliferation.
掺铒钇铝石榴石(Er:YAG)激光治疗在牙周治疗后已显示出良好的伤口愈合效果。尽管其机制尚不清楚,但原因之一可能是低强度激光对受照射组织具有积极的生物学效应。本研究的目的是通过蛋白质组学分析,研究低强度Er:YAG激光照射对人牙龈成纤维细胞(HGFs)增殖及激光诱导的蛋白质差异表达的影响。在第一个实验中,将HGFs暴露于低强度Er:YAG激光照射下,并在第3天评估激光诱导的细胞增殖和损伤情况。在第二个实验中,在照射后第1天进行蛋白质组学分析。从HGFs制备的肽段通过混合离子阱-傅里叶变换质谱仪、Mascot搜索引擎和UniProtKB数据库进行分析。照射后观察到细胞增殖显著增加且无细胞损伤。在总共鉴定出的377种蛋白质中,59种蛋白质上调,包括与伤口愈合过程相关的半乳糖凝集素-7,15种蛋白质在激光处理的HGFs中下调。在第三个实验中,通过各种分析技术验证了照射后HGFs中半乳糖凝集素-7信使核糖核酸(mRNA)和蛋白质表达的增加。此外,证实了重组人半乳糖凝集素-7对HGFs增殖调节的作用。结果表明,低强度Er:YAG激光照射可促进HGFs增殖,并诱导蛋白质表达发生显著变化,半乳糖凝集素-7表达上调可能部分促成了细胞增殖的增加。