Differential response of human dermal fibroblast subpopulations to visible and near-infrared light: Potential of photobiomodulation for addressing cutaneous conditions
Differential response of human dermal fibroblast subpopulations to visible and near-infrared light: Potential of photobiomodulation for addressing cutaneous conditions
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DOI：
10.1002/lsm.22823
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发表时间：
2018-10-01
影响因子：
2.4
通讯作者：
Tobin, Desmond J.
中科院分区：
文献类型：
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作者：
Mignon, Charles;Uzunbajakava, Natallia E.;Tobin, Desmond J.
背景与目的在过去的十年里，光生物调节技术(PBM)得到了迅速发展，在皮肤病的治疗方面取得了令人鼓舞的成果。然而，对这种方法的信心不仅因为缺乏对光触发的分子级联的理解，而且因为发表的实验结果、研究设计和应用的光学参数的显著不一致而受到损害。本研究旨在研究人真皮成纤维细胞亚群对可见光和近红外线(NIR)光的反应，以确定具有较高潜力的光学治疗参数，以解决老化皮肤和不可愈合的慢性创面的缺陷。材料和方法从手术后剩余的人面部皮肤中分离出原代人网状和乳头状真皮成纤维细胞(DF)。一种内部开发的基于LED的设备被用来使用六个离散波长(450、490、550、590、650和850 nm)照射细胞培养物。在所有六个波长的标准氧浓度(20%)下，根据细胞代谢活动来评估光的剂量-反应。然后在蛋白质水平上分析转录组和I型前胶原蛋白的产生，其中细胞培养在更接近体内的条件下，环境氧气和2%血清。此外，使用实时荧光共聚焦显微镜成像来获取活性氧物种(ROS)的产生。结果在代谢活性方面，短波长光(530 Nm)连续照射对DF有抑制作用，而长波(~gt；=590 nm)基本上是中性作用。450 nm处理后的细胞行为是两相的，有两种不同的状态：低到中剂量水平(30J/cm(2))的抑制。细胞对蓝光的反应伴随着剂量依赖的ROS释放，ROS定位于靠近线粒体的核周区域，这种释放被抗氧化剂减弱。总体而言，网状DFS在基因表达水平上表现出比乳头状DFS更高的对光处理的敏感性，有更多的基因显著上调或下调。在细胞内信号通路水平上，只有网状DF在30J/cm(2)蓝光照射后才观察到重要通路的上调或下调。在细胞水平上，短可见波长对网状DF有较大的抑制作用。参与转化生长因子信号通路的几个基因也受到影响。此外，I型前胶原的产生也受到抑制。相比之下，850 nm的近红外(NIR)光(20J/cm(2))对这些细胞有促进代谢的作用，没有检测到细胞内ROS的形成。同样，网状DF比乳头状DF反应更快。这种刺激作用只有在类似活体的低氧条件下才能观察到，相当于正常的皮肤组织氧水平(约2%)。结论本研究强调了光对人皮肤细胞的不同影响，包括近红外光上调新陈代谢活性，以及蓝光对前胶原生成和增殖的抑制。这些发现为开发针对不同皮肤状况的治疗方法(例如，治疗瘢痕疙瘩和纤维化)或在伤口愈合的不同阶段进行区别治疗开辟了新的途径。激光外科医生。2018年50：859-882。(C)2018年威利期刊公司。
Background ObjectivesThe past decade has witnessed a rapid expansion of photobiomodulation (PBM), demonstrating encouraging results for the treatment of cutaneous disorders. Confidence in this approach, however, is impaired not only by a lack of understanding of the light-triggered molecular cascades but also by the significant inconsistency in published experimental outcomes, design of the studies and applied optical parameters. This study aimed at characterizing the response of human dermal fibroblast subpopulations to visible and near-infrared (NIR) light in an attempt to identify the optical treatment parameters with high potential to address deficits in aging skin and non-healing chronic wounds.Materials and MethodsPrimary human reticular and papillary dermal fibroblasts (DF) were isolated from the surplus of post-surgery human facial skin. An in-house developed LED-based device was used to irradiate cell cultures using six discrete wavelengths (450, 490, 550, 590, 650, and 850nm). Light dose-response at a standard oxygen concentration (20%) at all six wavelengths was evaluated in terms of cell metabolic activity. This was followed by an analysis of the transcriptome and procollagen I production at a protein level, where cells were cultured in conditions closer to in vivo at 2% environmental oxygen and 2% serum. Furthermore, the production of reactive oxygen species (ROS) was accessed using real-time fluorescence confocal microscopy imaging. Here, production of ROS in the presence or absence of antioxidants, as well as the cellular localization of ROS, was evaluated.ResultsIn terms of metabolic activity, consecutive irradiation with short-wavelength light (530nm) exerted an inhibitory effect on DF, while longer wavelengths (>=590nm) had essentially a neutral effect. Cell behavior following treatment with 450nm was biphasic with two distinct states: inhibitory at low- to mid- dose levels (30J/cm(2)). Cell response to blue light was accompanied by a dose-dependent release of ROS that was localized in the perinuclear area close to mitochondria, which was attenuated by an antioxidant. Overall, reticular DFs exhibited a greater sensitivity to light treatment at the level of gene expression than did papillary DFs, with more genes significantly up- or down- regulated. At the intra-cellular signaling pathway level, the up- or down- regulation of vital pathways was observed only for reticular DF, after treatment with 30J/cm(2) of blue light. At the cellular level, short visible wavelengths exerted a greater inhibitory effect on reticular DF. Several genes involved in the TGF- signaling pathway were also affected. In addition, procollagen I production was inhibited. By contrast, 850nm near-infrared (NIR) light (20J/cm(2)) exerted a stimulatory metabolic effect in these cells, with no detectable intracellular ROS formation. Here too, reticular DF were more responsive than papillary DF. This stimulatory effect was only observed under in vivo-like low oxygen conditions, corresponding to normal dermal tissue oxygen levels (approximately 2%).ConclusionThis study highlights a differential impact of light on human skin cells with upregulation of metabolic activity with NIR light, and inhibition of pro-collagen production and proliferation in response to blue light. These findings open-up new avenues for developing therapies for different cutaneous conditions (e.g., treatment of keloids and fibrosis) or differential therapy at distinct stages of wound healing. Lasers Surg. Med. 50:859-882, 2018. (c) 2018 Wiley Periodicals, Inc.