Comparison of the neuroinflammatory responses to selective retina therapy and continuous-wave laser photocoagulation in mouse eyes

Graefes Arch Clin Exp Ophthalmol. 2018 Feb;256(2):341-353. doi: 10.1007/s00417-017-3883-7. Epub 2018 Jan 10.

Abstract

Purpose: This study investigated microglia and inflammatory cell responses after selective retina therapy (SRT) with microsecond-pulsed laser in comparison to continuous-wave laser photocoagulation (cwPC).

Methods: Healthy C57BL/6 J mice were treated with either a train of short pulses (SRT; 527-nm, Q-switched, 1.7-μs pulse) or a conventional thermal continuous-wave (532-nm, 100-ms pulse duration) laser. The mice were sacrificed and their eyes were enucleated 1, 3, 7, and 14 days after both laser treatments. Pattern of cell death on retinal section was evaluated by TUNEL assay, and the distribution of activated inflammatory cells and glial cells were observed under immunohistochemistry. Consecutive changes for the expression of cytokines such as IL-1β, TNF-α, and TGF-β were also examined using immunohistochemistry, and compared among each period after quantification by Western blotting.

Results: The numbers of TUNEL-positive cells in the retinal pigment epithelium (RPE) layer did not differ in SRT and cwPC lesions, but TUNEL-positive cells in neural retinas were significantly less on SRT. Vague glial cell activation was observed in SRT-treated lesions. The population of inflammatory cells was also significantly decreased after SRT, and the cells were located in the RPE layer and subretinal space. Proinflammatory cytokines, including IL-1β and TNF-α, showed significantly lower levels after SRT; conversely, the level of TGF-β was similar to the cwPC-treated lesion.

Conclusions: SRT resulted in selective RPE damage without collateral thermal injury to the neural retina, and apparently produced negligible glial activation. In addition, SRT showed a markedly less inflammatory response than cwPC, which may have important therapeutic implications for several macular diseases.

Keywords: CD11b; Continuous-wave laser photocoagulation; F4/80; IL-1β; Iba-1; Inflammatory response; Selective retina therapy; TGF-β; TNF-α.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Apoptosis
  • Blotting, Western
  • Cell Count
  • Cytokines / biosynthesis*
  • Disease Models, Animal
  • Fluorescein Angiography / methods
  • Fundus Oculi
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Laser Coagulation / methods*
  • Lasers, Solid-State / therapeutic use*
  • Mice
  • Mice, Inbred C57BL
  • Neuroglia / metabolism
  • Neuroglia / pathology*
  • Retinal Diseases / diagnosis
  • Retinal Diseases / metabolism
  • Retinal Diseases / surgery*
  • Retinal Pigment Epithelium / metabolism
  • Retinal Pigment Epithelium / pathology*
  • Tomography, Optical Coherence / methods

Substances

  • Cytokines