PUBLICATION

The Effect of Fluence on Macrophage Kinetics, Oxidative Stress, and Wound Closure Using Real-Time In Vivo Imaging

Authors
Paredes, A.D., Benavidez, D., Cheng, J., Mangos, S., Patil, R., Donoghue, M., Benedetti, E., Bartholomew, A.
ID
ZDB-PUB-190507-28
Date
2019
Source
Photobiomodulation, photomedicine, and laser surgery   37: 45-52 (Journal)
Registered Authors
Mangos, Steve
Keywords
He:Ne laser, macrophage, oxidative stress, time-lapse imaging, zebrafish
MeSH Terms
  • Animals
  • Cell Movement/radiation effects
  • Kinetics
  • Low-Level Light Therapy/methods*
  • Macrophages/radiation effects*
  • Microscopy, Fluorescence
  • Oxidative Stress/radiation effects*
  • Wound Healing/radiation effects*
  • Zebrafish
PubMed
31050943 Full text @ Photobiomodul Photomed Laser Surg
Abstract
Objective: The aim of our study was to quantify the effect of doses delivered by a He:Ne laser on individual macrophage kinetics, tissue oxidative stress, and wound closure using real-time in vivo imaging. Background: Photobiomodulation has been reported to reduce tissue inflammation and accelerate wound closure; however, precise parameters of laser settings to optimize macrophage behavior have not been established. We hypothesized that quantitative and real-time in vivo imaging could identify optimal fluence for macrophage migration, reduction of reactive oxygen species, and acceleration of wound closure. Methods: Larval zebrafish Tg(mpeg-dendra2) were loaded with dihydroethidium for oxidative stress detection. Fish were caudal fin injured, treated with 635 nm continuous 5 mW He:Ne laser irradiation at 3, 9, or 18 J/cm2 and time-lapsed imaged within the first 120 min postinjury. Images taken 1 and 24-h postinjury were compared for percentage wound closure. Results: A fluence of 3 J/cm2 demonstrated significant increases in macrophage migration speed, fewer stops along the way, and greatest directed migration toward the wound. These findings were associated with a significant reduction in wound content reactive oxygen species when compared with control wounded fins. Both 3 and 9 J/cm2 significantly accelerated wound closure when compared with nonirradiated control fish. Conclusions: Wound macrophage activity could be manipulated by applied fluence, leading to reduced levels of wound reactive oxygen species and accelerated wound closure. The zebrafish model provides a means to quantitatively compare wound macrophage behavior in response to a variety of laser treatment parameters in real time.
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