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Irradiation at 830 nm stimulates nitric oxide production and inhibits pro‐inflammatory cytokines in diabetic wounded fibroblast cells

2010; Wiley; Volume: 42; Issue: 6 Linguagem: Inglês

10.1002/lsm.20812

1096-9101

Nicolette N. Houreld, Palesa Sekhejane, Heidi Abrahamse,

Diabetic Foot Ulcer Assessment and Management

Abstract Background and Objective Wound healing in diabetic patients remains a chief problem in the clinical setting and there is a strong need for the development of new, safe, reliable therapies. This study aimed to establish the effect of irradiating diabetic wounded fibroblast cells (WS1) in vitro on pro‐inflammatory cytokines and the production of nitric oxide (NO). Materials and Methods Normal, wounded and diabetic wounded WS1 cells were exposed to an 830 nm laser with 5 J/cm 2 and incubated for a pre‐determined amount of time. Changes in cellular viability, proliferation and apoptosis were evaluated by the Trypan blue assay, VisionBlue™ fluorescence assay and caspase 3/7 activity respectively. Changes in cytokines (interleukin—IL‐6, IL‐1β and tumour necrosis factor‐alpha, TNF‐α) were determined by ELISA. NO was determined spectrophotometrically and reactive oxygen species (ROS) was evaluated by immunofluorescent staining. Results Diabetic wounded WS1 cells showed no significant change in viability, a significant increase in proliferation at 24 and 48 hours ( P <0.001 and P <0.01 respectively) and a decrease in apoptosis 24 hours post‐irradiation ( P <0.01). TNF‐α levels were significantly decreased at both 1 and 24 hours ( P <0.05), while IL‐1β was only decreased at 24 hours ( P <0.05). There was no significant change in IL‐6. There was an increase in ROS and NO ( P <0.01) 15 minutes post‐irradiation. Conclusion Results show that irradiation of diabetic wounded fibroblast cells at 830 nm with 5 J/cm 2 has a positive effect on wound healing in vitro. There was a decrease in pro‐inflammatory cytokines (IL‐1β and TNF‐α) and irradiation stimulated the release of ROS and NO due to what appears to be direct photochemical processes. Lasers Surg. Med. 42:494–502, 2010. © 2010 Wiley–Liss, Inc.