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高频近红外二极管激光照射对小鼠颅骨成骨细胞增殖和迁移的影响。

Effects of high-frequency near-infrared diode laser irradiation on the proliferation and migration of mouse calvarial osteoblasts.

作者信息

Kunimatsu Ryo, Gunji Hidemi, Tsuka Yuji, Yoshimi Yuki, Awada Tetsuya, Sumi Keisuke, Nakajima Kengo, Kimura Aya, Hiraki Tomoka, Abe Takaharu, Naoto Hirose, Yanoshita Makoto, Tanimoto Kotaro

机构信息

Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan.

出版信息

Lasers Med Sci. 2018 Jul;33(5):959-966. doi: 10.1007/s10103-017-2426-0. Epub 2018 Jan 4.

Abstract

Laser irradiation activates a range of cellular processes and can promote tissue repair. Here, we examined the effects of high-frequency near-infrared (NIR) diode laser irradiation on the proliferation and migration of mouse calvarial osteoblastic cells (MC3T3-E1). MC3T3-E1 cells were cultured and exposed to high-frequency (30 kHz) 910-nm diode laser irradiation at a dose of 0, 1.42, 2.85, 5.7, or 17.1 J/cm. Cell proliferation was evaluated with BrdU and ATP concentration assays. Cell migration was analyzed by quantitative assessment of wound healing using the Incucyt ZOOM system. In addition, phosphorylation of mitogen-activated protein kinase (MAPK) family members including p38 mitogen-activated protein kinase (p38), stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK), and extracellular signal-regulated protein kinase (ERK)1/2) after laser irradiation was examined with western blotting. Compared to the control, cell proliferation was significantly increased by laser irradiation at a dose of 2.85, 5.7, or 17.1 J/cm. Laser irradiation at a dose of 2.85 J/cm induced MC3T3-E1 cells to migrate more rapidly than non-irradiated control cells. Irradiation with the high-frequency 910-nm diode laser at a dose of 2.85 J/cm induced phosphorylation of MAPK/ERK1/2 15 and 30 min later. However, phosphorylation of p38 MAPK and SAPK/JNK was not changed by NIR diode laser irradiation at a dose of 2.85 J/cm. Irradiation with a high-frequency NIR diode laser increased cell division and migration of MT3T3-E1 cells, possibly via MAPK/ERK signaling. These observations may be important for enhancing proliferation and migration of osteoblasts to improve regeneration of bone tissues.

摘要

激光照射可激活一系列细胞过程,并能促进组织修复。在此,我们研究了高频近红外(NIR)二极管激光照射对小鼠颅骨成骨细胞(MC3T3-E1)增殖和迁移的影响。培养MC3T3-E1细胞,并使其分别接受剂量为0、1.42、2.85、5.7或17.1 J/cm的高频(30 kHz)910 nm二极管激光照射。通过BrdU和ATP浓度测定评估细胞增殖。使用Incucyt ZOOM系统通过定量评估伤口愈合情况来分析细胞迁移。此外,通过蛋白质印迹法检测激光照射后丝裂原活化蛋白激酶(MAPK)家族成员包括p38丝裂原活化蛋白激酶(p38)、应激激活蛋白激酶/ Jun氨基末端激酶(SAPK/JNK)和细胞外信号调节蛋白激酶(ERK)1/2的磷酸化情况。与对照组相比,剂量为2.85、5.7或17.1 J/cm的激光照射可显著提高细胞增殖。剂量为2.85 J/cm的激光照射诱导MC3T3-E1细胞比未照射的对照细胞迁移得更快。剂量为2.85 J/cm的高频910 nm二极管激光照射在15和30分钟后诱导MAPK/ERK1/2磷酸化。然而,剂量为2.85 J/cm的近红外二极管激光照射并未改变p38 MAPK和SAPK/JNK的磷酸化。高频近红外二极管激光照射可能通过MAPK/ERK信号通路增加MT3T3-E1细胞的分裂和迁移。这些观察结果对于增强成骨细胞的增殖和迁移以改善骨组织再生可能具有重要意义。

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