Ludwig Boltzmann institute for experimental and clinical traumatology, Donaueschingenstrasse 13, 1200 Vienna, Austria.
Ludwig Boltzmann institute for experimental and clinical traumatology, Donaueschingenstrasse 13, 1200 Vienna, Austria.
J Photochem Photobiol B. 2020 Aug;209:111935. doi: 10.1016/j.jphotobiol.2020.111935. Epub 2020 Jun 24.
Photobiomodulation (PBM), especially in the red wavelength range, has been demonstrated to be an effective treatment option for superficial and chronic wounds. However, ischemia and subsequent reperfusion can further challenge wound healing. Therefore, we investigated the effect of pulsed red LED light at 635 nm on cellular function in an in-vitro model of hypoxia/reoxygenation (H/R) challenge. Mouse myoblasts and fibroblasts were incubated in oxygen-deprived starvation medium (hypoxia) for 3 h after which the media was changed to oxygenated, fully supplemented media to simulate reperfusion. Cells were then treated with pulsed red LED light at a wavelength of 635 nm at 40 mW/cm. Mitochondrial respiratory activity, ATP production and ROS levels were analysed immediately post-illumination. The effects on cellular metabolic activity and proliferation were measured at 6 h and 24 h and apoptosis/necrosis was measured at 24 h post-illumination. Our results show that both cell types reacted differently to H/R challenge and PBM. PBM of H/R-challenged cells enhanced mitochondrial activity and rescued decreased ATP levels, with significant effects in fibroblasts. This was associated with increased cell proliferation rates in both cell types. The increase was again more pronounced in fibroblasts. Our study concluded that PBM with red LED light significantly restored ATP levels during H/R and effectively promoted cell growth under both normoxic and H/R conditions. In clinical applications, PBM has been repeatedly reported to resolve difficult clinical situations in which ischemia/reperfusion injuries are a major issue. Our study confirms the beneficial effects of PBM especially in H/R-challenged cells.
光生物调节（PBM），尤其是在红光波长范围内，已被证明是治疗浅表性和慢性伤口的有效方法。然而，缺血和随后的再灌注会进一步挑战伤口愈合。因此，我们研究了在缺氧/复氧（H/R）挑战的体外模型中，635nm 脉冲红光对细胞功能的影响。将小鼠成肌细胞和成纤维细胞在缺氧饥饿培养基（缺氧）中孵育 3 小时，然后将培养基更换为富含氧气的完全补充培养基，以模拟再灌注。然后，用波长为 635nm、功率密度为 40mW/cm 的脉冲红光照射细胞。立即在照射后分析线粒体呼吸活性、ATP 产生和 ROS 水平。在 6 小时和 24 小时测量细胞代谢活性和增殖的影响，并在照射后 24 小时测量细胞凋亡/坏死。我们的结果表明，两种细胞类型对 H/R 挑战和 PBM 的反应不同。PBM 对 H/R 挑战细胞增强了线粒体活性并挽救了降低的 ATP 水平，在成纤维细胞中具有显著效果。这与两种细胞类型的细胞增殖率增加有关。在成纤维细胞中，增加的幅度再次更为明显。我们的研究得出结论，PBM 用红色 LED 光在 H/R 期间显著恢复了 ATP 水平，并在正常氧和 H/R 条件下有效促进了细胞生长。在临床应用中，PBM 已多次被报道可以解决缺血/再灌注损伤是主要问题的困难临床情况。我们的研究证实了 PBM 的有益效果，特别是在 H/R 挑战的细胞中。
