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660nm 的光生物调节反应不同于 980nm,且更持久。

Photobiomodulation Response From 660 nm is Different and More Durable Than That From 980 nm.

机构信息

Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, Massachusetts, 02114.

Department of Dermatology, Harvard Medical School, Boston, Massachusetts, 02115.

出版信息

Lasers Surg Med. 2021 Nov;53(9):1279-1293. doi: 10.1002/lsm.23419. Epub 2021 May 16.

Abstract

BACKGROUND AND OBJECTIVES

Photobiomodulation (PBM) therapy uses light at various wavelengths to stimulate wound healing, grow hair, relieve pain, and more-but there is no consensus about optimal wavelengths or dosimetry. PBM therapy works through putative, wavelength-dependent mechanisms including direct stimulation of mitochondrial respiration, and/or activation of transmembrane signaling channels by changes in water activity. A common wavelength used in the visible red spectrum is ~660 nm, whereas recently ~980 nm is being explored and both have been proposed to work via different mechanisms. We aimed to gain more insight into identifying treatment parameters and the putative mechanisms involved.

STUDY DESIGN/MATERIALS AND METHODS: Fluence-response curves were measured in cultured keratinocytes and fibroblasts exposed to 660 or 980 nm from LED sources. Metabolic activity was assessed using the MTT assay for reductases. ATP production, a major event triggered by PBM therapy, was assessed using a luminescence assay. To measure the role of mitochondria, we used an ELISA to measure COX-1 and SDH-A protein levels. The respective contributions of cytochrome c oxidase and ATP synthase to the PBM effects were gauged using specific inhibitors.

RESULTS

Keratinocytes and fibroblasts responded differently to exposures at 660 nm (red) and 980 nm (NIR). Although 980 nm required much lower fluence for cell stimulation, the resulting increase in ATP levels was short-term, whereas 660 nm stimulation elevated ATP levels for at least 24 hours. COX-1 protein levels were increased following 660 nm treatment but were unaffected by 980 nm. In fibroblasts, SDH-A levels were affected by both wavelengths, whereas in keratinocytes only 660 nm light impacted SDH-A levels. Inhibition of ATP synthase nearly completely abolished the effects of both wavelengths on ATP synthesis. Interestingly, inhibiting cytochrome c oxidase did not prevent the rise in ATP levels in response to PBM treatment.

CONCLUSION

To the best of our knowledge, this is the first demonstration of differing kinetics in response to PBM therapy at red versus NIR wavelength. We also found cell-type-specific differences in PBM therapy response to the two wavelengths studied. These findings confirm that different response pathways are involved after 660 and 980 nm exposures and suggest that 660 nm causes a more durable response. © 2021 Wiley Periodicals LLC.

摘要

背景与目的

光生物调节(PBM)疗法使用各种波长的光来刺激伤口愈合、头发生长、缓解疼痛等,但对于最佳波长或剂量学尚无共识。PBM 疗法通过推测的、波长依赖性机制发挥作用,包括直接刺激线粒体呼吸,以及/或通过水活度的变化激活跨膜信号通道。在可见红光谱中常用的波长为660nm,而最近正在探索980nm,并且两者都被提议通过不同的机制发挥作用。我们旨在更深入地了解治疗参数和涉及的推测机制。

研究设计/材料和方法:将培养的角质形成细胞和成纤维细胞暴露于 LED 源的 660 或 980nm 下,测量其光通量-反应曲线。使用 MTT 测定法评估还原酶的代谢活性。使用发光测定法评估 PBM 治疗引发的主要事件——三磷酸腺苷(ATP)的产生。为了测量线粒体的作用,我们使用 ELISA 测量细胞色素 c 氧化酶(COX-1)和琥珀酸脱氢酶 A(SDH-A)蛋白水平。使用特定抑制剂来衡量细胞色素 c 氧化酶和 ATP 合酶对 PBM 作用的各自贡献。

结果

角质形成细胞和成纤维细胞对 660nm(红色)和 980nm(近红外)的暴露反应不同。虽然 980nm 照射细胞所需的光通量要低得多,但导致的 ATP 水平升高是短期的,而 660nm 刺激则至少持续 24 小时增加 ATP 水平。660nm 处理后 COX-1 蛋白水平增加,但 980nm 处理则不受影响。在成纤维细胞中,两种波长均影响 SDH-A 水平,而在角质形成细胞中仅 660nm 光会影响 SDH-A 水平。抑制 ATP 合酶几乎完全消除了两种波长对 ATP 合成的影响。有趣的是,抑制细胞色素 c 氧化酶并不能阻止 PBM 治疗后 ATP 水平的升高。

结论

据我们所知,这是首次证明在红色与近红外波长的 PBM 治疗中存在不同的动力学反应。我们还发现,两种研究波长的 PBM 治疗对细胞类型的反应存在差异。这些发现证实,660nm 和 980nm 照射后涉及不同的反应途径,并表明 660nm 会引起更持久的反应。©2021Wiley Periodicals LLC.

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