Release of Different Cell Mediators During Retinal Pigment Epithelium Regeneration Following Selective Retina Therapy

Invest Ophthalmol Vis Sci. 2018 Mar 1;59(3):1323-1331. doi: 10.1167/iovs.17-23163.

Abstract

Purpose: To investigate the effect of selective retina therapy (SRT) on the release of AMD-relevant cell mediators, such as matrix metalloproteinases (MMPs), VEGF, and pigment epithelium derived factor (PEDF) using different laser spot sizes and densities.

Methods: Porcine RPE-choroid explants were treated with a pulsed 532 nm Nd:YAG laser using (1) large spot sizes, (2) small spot sizes with a high-density (hd) treatment, and (3) small spot sizes with a low-density (ld) treatment. Explants were cultivated in modified Ussing chambers. RPE regeneration and RPE cell death were investigated by calcein-AM staining and immunofluorescence. The MMP release was examined via zymography and immunofluorescence. VEGF and PEDF secretion was analyzed by ELISA.

Results: During pigment epithelium regeneration (PER), mitosis and RPE cell migration were observed. Four days after SRT (large spot size) the content of active MMP2 increased significantly (P < 0.01). Hd treatment with small spot sizes resulted also in an increase of active MMP2 (P < 0.05). In immunofluorescence explants showed a localized expression of MMP2 within the healing lesions after irradiation. The PEDF level increased significantly (P = 0.01) after SRT with large spot sizes. VEGF secretion decreased significantly (P < 0.05) following SRT with large spot sizes and with hd treatment of small spot sizes.

Conclusions: SRT induces a cytokine profile, which may improve the flux across Bruch's membrane, slows down progression of early AMD by RPE regeneration, and inhibits the formation of choroidal neovascularization. The cytokine release depends on the size and density of applied laser spots.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Death
  • Cell Movement
  • Choroid / metabolism
  • Cytokines / metabolism*
  • Disease Models, Animal
  • Eye Proteins / metabolism
  • Laser Therapy* / methods
  • Lasers, Solid-State
  • Macular Degeneration / therapy
  • Matrix Metalloproteinases / metabolism
  • Mitosis
  • Nerve Growth Factors / metabolism
  • Regeneration
  • Retinal Pigment Epithelium / metabolism*
  • Retinal Pigment Epithelium / physiology
  • Serpins / metabolism
  • Swine
  • Vascular Endothelial Growth Factor A / metabolism
  • Wound Healing / physiology*

Substances

  • Cytokines
  • Eye Proteins
  • Nerve Growth Factors
  • Serpins
  • Vascular Endothelial Growth Factor A
  • pigment epithelium-derived factor
  • Matrix Metalloproteinases