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www.nature.com/scientificreports/ Figure 2. Effects of pulsed LED light on proliferation (A) and cell metabolic activity (B) of HUVEC cultured in a 2D cell culture model. Stimulation of HUVEC on day 0 with green and red light had no significant effect on metabolic activity while cell proliferation was significantly increased. In contrast, blue light significantly decreased metabolic activity and showed no effect on proliferation. *P < 0.05, **P < 0.01. Figure 3. Effects of pulsed LED light on 2D- and 3D-migration in vitro. (A) Stimulation with light enhanced wound closure in scratch assays which reached significance in the green LED group. (B) In a 3D model of HUVEC-coated Cytodex 3 beads in fibrin matrices red and green light resulted in an increased migration of cells towards the fibrin gel while blue light was not effective. (C) Effects of pulsed LED light on ROS formation. Only blue light induced a significant rise in ROS. *P < 0.05, **P < 0.01, ***P < 0.001. faster reduction of the wound scratch area determined at 6 h after LLLT compared control (Fig. 3A). While in the untreated control group wound area was reduced within 6 h by only 28.7 ± 11.1%, there was a reduction of 32.0 ± 9.6% in the blue light group, 36.6 ± 6.3% in the red light group and 40.1 ± 8.0% in the green light treated group which was statistically significant compared to control (P = 0.046). Additionally, migration was also investigated in a 3D cell culture model. The quantification of cells migrating away from Cytodex beads into the surrounding fibrin gel matrix showed that both red and green light had signif- icant influence on 3D migration after 4 days (Fig. 3B). While in the control group 19.8 ± 16.2 cells were migrating SCientifiC REpORTS | 7: 10700 | DOI:10.1038/s41598-017-11061-y 3PDF Image | impact of wavelengths of LED light-therapy on endothelial cells
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