Fujimoto Keiko, Kiyosaki Takeshi, Mitsui Narihiro, Mayahara Kotoe, Omasa Saori, Suzuki Naoto, Shimizu Noriyoshi
Department of Orthodontics, Nihon University School of Dentistry, 1-8-13, Kanda Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
Lasers Surg Med. 2010 Aug;42(6):519-26. doi: 10.1002/lsm.20880.
Previously, we reported that low-intensity laser irradiation accelerated bone formation, and that this mechanism deeply involved insulin-like growth factor I expression. However, as bone formation is supported by many local factors, the mechanism involved in laser irradiation remains incompletely understood. Therefore, the purpose of this study was to determine the effects of laser irradiation on the osteogenic response in vitro.
Mouse osteoblast-like cells, MC3T3-E1, were cultured and were irradiated for 5-20 minutes (0.96-3.82 J/cm(2)) at the subconfluent stage using a low-intensity Ga-Al-As diode laser apparatus. After laser irradiation, expression of bone morphogenetic proteins (BMPs), transcription factors (Runx2, Osterix, Dlx5, Msx2), and phosphorylation of Smad1 were determined, and calcium content of cell cultures was also determined.
Irradiation at 1.91 J/cm(2) significantly increased the expression of BMPs and Runx2, Osterix, Dlx5, Msx2, and the phosphorylation of Smad1. Noggin, a BMP receptor blocker, inhibited the laser-induced Runx2 expression and phosphorylation of Smad1. Moreover, laser irradiation significantly increased the calcium content of cell cultures, and noggin inhibited this increase.
These results suggest that low-intensity laser irradiation stimulates in vitro mineralization via increased expression of BMPs and transcription factors associated with osteoblast differentiation.
此前,我们报道低强度激光照射可加速骨形成,且该机制与胰岛素样生长因子I的表达密切相关。然而,由于骨形成受多种局部因素支持,激光照射所涉及的机制仍未完全明确。因此,本研究旨在确定激光照射对体外成骨反应的影响。
培养小鼠成骨样细胞MC3T3-E1,并在亚汇合阶段使用低强度Ga-Al-As二极管激光装置照射5-20分钟(0.96-3.82 J/cm²)。激光照射后,测定骨形态发生蛋白(BMPs)、转录因子(Runx2、Osterix、Dlx5、Msx2)的表达以及Smad1的磷酸化水平,同时测定细胞培养物中的钙含量。
1.91 J/cm²的照射显著增加了BMPs、Runx2、Osterix、Dlx5、Msx2的表达以及Smad1的磷酸化水平。BMP受体阻滞剂Noggin可抑制激光诱导的Runx2表达和Smad1的磷酸化。此外,激光照射显著增加了细胞培养物中的钙含量,而Noggin可抑制这种增加。
这些结果表明,低强度激光照射通过增加与成骨细胞分化相关的BMPs和转录因子的表达来刺激体外矿化。