Effects of Low-Level Er:YAG Laser Irradiation on Proliferation and Calcification of Primary Osteoblast-Like Cells Isolated From Rat Calvaria
Effects of Low-Level Er:YAG Laser Irradiation on Proliferation and Calcification of Primary Osteoblast-Like Cells Isolated From Rat Calvaria
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DOI：
10.3389/fcell.2020.00459
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发表时间：
2020-06-23
影响因子：
5.5
通讯作者：
Aoki, Akira
中科院分区：
文献类型：
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作者：
Niimi, Hiromi;Ohsugi, Yujin;Aoki, Akira
一些报告表明，各种激光对细胞的光生物调节具有良好的生物效应。然而，低水平Er:YAG激光照射对成骨细胞的影响仍不清楚。本研究的目的是评估低水平Er:YAG激光照射对从3-5日龄Wistar大鼠颅骨中分离的原代成骨细胞样细胞增殖和成骨分化的影响。用 Er:YAG 激光分别以 2.2、3.3 和 4.3 J/cm(2) 的能量注量照射细胞。照射后，测量细胞表面温度并通过流式细胞术和CCK-8评估细胞增殖。在骨诱导培养基中培养7、14和21天后，通过测量茜素红S染色面积来评估钙化。在照射后 3、6 和 12 小时以及 1、3、7 和 14 天时通过 qPCR 评估未照射和激光照射细胞中的基因表达。照射后6小时进行微阵列分析，综合评价3.3 J/cm(2)下未照射和照射细胞的基因表达。辐射没有引起细胞表面温度的明显升高。辐射不影响成骨细胞样细胞的增殖。 3.3 J/cm(2) Er:YAG激光照射后7天，成骨细胞样细胞钙化显着增加。3.3 J/cm(2)照射后6小时，Bglap表达在细胞中显着增加。微阵列分析表明，3.3 J/cm(2) 的辐射引起炎症相关基因的上调和Wisp2 的下调。基因集富集分析还阐明了炎症相关和 Notch 信号基因集的富集。总之，3.3 J/cm(2) 的低水平 Er:YAG 激光照射通过增强的 Bglap 表达和丰富的 Notch 信号传导增强了原代成骨细胞样细胞的钙化。
Several reports have shown that the photo-bio-modulation of cells by various lasers has favorable biological effects. However, the effects of low-level Er:YAG laser irradiation on osteoblasts remain unclear. The purpose of this study was to evaluate the effects of low-level Er:YAG laser irradiation on proliferation and osteogenic differentiation of primary osteoblast-like cells isolated from the calvariae of 3-5-day-old Wistar rats. Cells were irradiated by Er:YAG laser at energy fluences of 2.2, 3.3, and 4.3 J/cm(2), respectively. After irradiation, cell surface temperatures were measured and cell proliferation was evaluated by flow cytometry and CCK-8. Calcification was evaluated by measuring areas of Alizarin red S staining after 7, 14, and 21 days culture in osteoinductive medium. Gene expression in non-irradiated and laser-irradiated cells was evaluated by qPCR at 3, 6, and 12 h, as well as 1, 3, 7, and 14 days after irradiation. Microarray analysis was performed to comprehensively evaluate the gene expression of non-irradiated and irradiated cells at 3.3 J/cm(2)at 6 h after irradiation. No pronounced increase of cell surface temperature was induced by irradiation. Irradiation did not affect osteoblast-like cell proliferation. Osteoblast-like cell calcification was significantly increased 7 days after Er:YAG laser irradiation at 3.3 J/cm(2).Bglapexpression was significantly increased in cells irradiated at 3.3 J/cm(2)6 h post-irradiation. Microarray analysis showed that irradiation at 3.3 J/cm(2)caused an upregulation of inflammation-related genes and downregulation ofWisp2. Gene set enrichment analysis also clarified enrichment of inflammation-related and Notch signaling gene sets. In conclusion, low-level Er:YAG laser irradiation at 3.3 J/cm(2)enhanced calcification of primary osteoblast-like cells via enhancedBglapexpression and enriched Notch signaling.