Low Pulse Energy Nd:YAG Laser Irradiation Exerts a Biostimulative Effect on Different Cells of the Oral Microenvironment: "An In Vitro Study"
Low Pulse Energy Nd:YAG Laser Irradiation Exerts a Biostimulative Effect on Different Cells of the Oral Microenvironment: "An In Vitro Study"
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DOI：
10.1002/lsm.20861
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发表时间：
2010-08-01
影响因子：
2.4
通讯作者：
Giannelli, Marco
中科院分区：
文献类型：
--
作者：
Chellini, Flaminia;Sassoli, Chiara;Giannelli, Marco
背景和目的：牙科激光是治疗牙周及种植体疾病的一种很有前途的治疗工具，但其临床应用仍然有限。在这里，我们研究了低脉冲能量的脉冲激光对口腔微环境中不同细胞的潜在生物刺激作用，并阐明了其潜在的分子机制。材料和方法：用低脉冲能量(20mJ)、高重复频率(50~70 Hz)的脉冲激光照射SAOS-2成骨细胞、H端内皮细胞和NIH/3T3成纤维细胞，分别用光敏剂亚甲蓝(MB)处理或不处理，用共聚焦免疫荧光和实时RT-PCR方法检测细胞活力、增殖和特异性分化标志物的表达。结果：激光照射对所有受试细胞的存活率均无明显影响，但能有效刺激非致敏成骨细胞的生长，并能显著诱导骨桥蛋白的表达。亚甲基蓝预处理对未刺激和激光刺激的细胞分化有负面影响，照射后的成骨细胞中有碱性磷酸酶和Runx2的表达，成纤维细胞中有I型胶原的表达，内皮细胞中有纽蛋白的表达。值得注意的是，激光照射还通过激活TRPC1离子通道引起成骨细胞内钙离子的增加。结论：低脉冲能量、高重复频率的脉冲激光照射对口腔微环境中的不同细胞，尤其是成骨细胞可能具有生物刺激作用。在照射前用亚甲蓝预处理阻碍了这种效果，并限制了光敏染料在牙科实践激光外科42：527-539,2010(C)2010 Wiley-Liss，Inc.中的潜在临床应用。
Background and Objectives: Dental lasers represent a promising therapeutic tool in the treatment of periodontal and pen-implant diseases However, their clinical application remains still limited. Here, we Investigated the potential biostimulatory effect of low pulse energy neodymium:yttrium aluminum garnet (Nd.YAG) laser irradiation on different cells representative of the oral microenvironment and elucidated the underlying molecular mechanisms.Materials and Methods: Saos-2 osteoblasts, H-end endothelial cells, and NIH/3T3 fibroblasts pre-treated or not with photosensitizing dye methylene blue (MB), were irradiated with low pulse energy (20 mJ) and high repetition rate (50-70 Hz) Nd YAG laser, and evaluated for cell viability and proliferation as well as for the expression of specific differentiation markers by confocal immunofluorescence and real-time RT-PCR. Changes in intracellular Ca2+ levels after laser exposure were also evaluated in living osteoblasts.Results: Nd:YAG laser irradiation did not affect cell viability in all the tested cell types, even when combined with pre-treatment with MB, and efficiently stimulated cell growth in the non-sensitized osteoblasts Moreover, a significant induction in the expression of osteopontin. ALP, and Runx2 in osteoblasts, type I collagen in fibroblasts, and vinculin in endothelial cells could be observed in the irradiated cells Pre-treatment with MB negatively affected cell differentiation in the unstimulated and laser-stimulated cells. Notably, laser irradiation also caused an increase in the intracellular Ca2+ in osteoblasts through the activation of TRPC1 ion channels. Moreover, the pharmacologic or genetic inhibition of these channels strongly attenuated laser-induced osteopontin expression, suggesting a role for the laser-mediated Ca2+ influx in regulating osteoblast differentiationConclusion: Low pulse energy and high repetition rate Nd.YAG laser irradiation may exert a biostimulative effect on different cells representative of the oral microenvironment, particularly osteoblasts. Pre-treatment with MB prior to irradiation hampers this effect and limits the potential clinical application of photosensitizing dyes in dental practice Lasers Surg. Med 42:527-539, 2010 (C) 2010 Wiley-Liss, Inc.