Bone formation stimulation by semiconductor laser via Cbfa1/Pebp2αA

半导体激光通过 Cbfa1/Pebp2αA 刺激骨形成

• 批准号: 12672018
• 负责人: SHIMIZU Noriyoshi
• 金额: $ 1.73万
• 依托单位: Nihon-University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672018/
• 关键词: low-power laser; bone formation; osteoblast; IGF-I; calvaria; レーザー; bone nodule

Bone formation acceleration following low-intensity laser treatment has been demonstrated in both animal and cell culture models, however, the action mechanisms of irradiation on bone are unclear. We previously reported that low-intensity laser irradiation stimulated bone nodule formation through enhanced cellular proliferation and differentiation, thus, in the present study, we attempted to determine whether insulin-like growth factor (IGF)-I is responsible for the action observed. Osteoblast-like cells were isolated from fetal rat calvariae and cultured with rat recombinant (r) IGF-I, IGF-I-antibody (Ab), and/or the cells were irradiated once (3.75 J/cm^2) with a low- intensity Ga-Al-As laser (830 nm) at the subconfluent stage. The cells were cultured for 24 days and stained using the von-Kossa technique. Treatment with recombinant IGF-I for 6 days significantly stimulated the number and area of bone nodules as compared to the control (1.4-1.6-fold), and these effects were quite similar to those by laser irradiation. Moreover, the stimulatory effect by laser irradiation was abrogated dose-dependently by treatment with IGF-I-Ab for 6 days. To confirm the mechanisms responsible for the action of laser irradiation IGF-I protein levels and gene expression in rat calvarial cells were also examined. IGF-I production in the medium was significantly increased at 3 and 6 days after laser irradiation as compared to the control, and the cell mass center expressed IGF-I. Gene expression, examined by semi-quantitative RT-PCR, was also found to be significantly stimulated at 1 and 3 days after irradiation, and returned to the control level thereafter. These results indicate that the stimulatory effect of bone nodule formation by low-intensity laser irradiation may be at least partly mediated by IGF-I expression. Gene expression of Cbfa1/Pebp2αA by laser irradiation is in progress.

低强度激光治疗促进骨形成在动物和细胞培养模型中都得到了证实，然而，照射对骨的作用机制尚不清楚。我们以前报道过低强度激光照射通过促进细胞增殖和分化来刺激骨结节的形成，因此，在本研究中，我们试图确定胰岛素样生长因子-I是否与所观察到的作用有关。从胎鼠颅骨分离出成骨样细胞，与大鼠重组IGF-I、IGF-I抗体(Ab)共同培养，并在亚融合阶段用低强度Ga-Al-As激光(830 Nm)照射1次(3.75J/cm2)。细胞培养24天，用von-Kossa技术染色。重组IGF-I处理6d后，骨结节的数量和面积明显增加，是对照组的1.4-1.6倍，与激光照射的效果非常相似。IGF-I-Ab作用6d后，对激光的刺激作用呈剂量依赖性地减弱。为了证实激光照射的作用机制，我们还检测了大鼠颅骨细胞中IGF-I的蛋白水平和基因表达。激光照射后3d和6d，细胞培养上清液中IGF-I的产生明显高于对照组，细胞质心表达IGF-I。半定量RT-PCR检测也发现，照射后1天和3天，基因表达显著增强，此后恢复到对照水平。这些结果表明，低强度激光对骨结节形成的促进作用可能至少部分是通过IGF-I的表达来实现的。激光照射下Cbfa1/Pebp2αA的基因表达正在进行中。

项目成果

Shimizu N: "Periodontal tissue regeneration utilizing a low-power laser (Japanese)"In : Okuda K, Ed. Front line of periodontology. Nihonsikahyouron, Tokyo. 332-340 (2000)

Shimizu N：“利用低功率激光进行牙周组织再生（日语）”，见：Okuda K，Ed。

Ueda Y, Shimizu N.: "Pulse irradiation of low-power laser stimulates bone nodule formation"J Oral Science. 43. 55-60 (2001)

Ueda Y，Shimizu N.：“低功率激光的脉冲照射刺激骨结节形成”J Oral Science。

Shimizu N: "Laser application in orthodontics (Japanese)"In : Fujita K, Ed. Toward new health creativity in dental science. Ishiyakusyuppan, Tokyo. 210-211 (2001)

Shimizu N：“激光在牙齿矫正中的应用（日语）”见：Fujita K，Ed。

清水典佳: "レーザの矯正治療への応用低出力レーザ照射による矯正治療期間の短縮と疼痛軽減の可能性"光アライアンス. 11巻12号. 38-41 (2000)

Norika Shimizu：“激光在正畸治疗中的应用：通过低功率激光照射缩短正畸治疗周期和减轻疼痛的可能性”Hikari Alliance，第 11 卷，第 12 期。38-41 (2000)。

Ueda Y., Shimizu N.: "Pulse irradiation low-power laser stimulates bone nodule formation"Jof Oral Science. 43. 55-60 (2001)

Ueda Y.、Shimizu N.：“脉冲照射低功率激光刺激骨结节形成”口腔科学杂志。