Combined effects of interleukin-1β and cyclic stretching on metalloproteinase expression in corneal fibroblasts in vitro

Biomed Eng Online. 2016 Jun 10;15(1):63. doi: 10.1186/s12938-016-0198-6.

Abstract

Background: Corneal tensile strain increases if the cornea becomes thin or if intraocular pressure increases. However, the effects of mechanical stress on extracellular matrix (ECM) remodelling in the corneal repair process and the corneal anomalies are unknown.

Methods: In this study, the combined effects of interleukin-1β (IL-1β) on matrix metalloproteinases (MMPs) in corneal fibroblasts under cyclic stretching were investigated in vitro. Cultured rabbit corneal fibroblasts were subjected to 5, 10 or 15 % cyclic equibiaxial stretching at 0.1 Hz for 36 h in the presence of IL-1β. Conditioned medium was harvested for the analysis of MMP2 and MMP9 protein production using the gelatin zymography and western blot techniques.

Results and conclusions: Cyclic equibiaxial stretching changed the cell morphology by increasing the contractility of F-actin fibres. IL-1β alone induced the expression of MMP9 and increased the production of MMP2, and 5 % stretching alone decreased the production of MMP2, which indicates that a low stretching magnitude can reduce ECM degradation. In the presence of IL-1β, 5 and 10 % stretching increased the production of MMP2, whereas 15 % stretching increased the production of MMP9. These results indicate that MMP expression is enhanced by cyclic mechanical stimulation in the presence of IL-1β, which is expected to contribute to corneal ECM degradation, leading to the development of post-refractive surgery keratectasia.

Keywords: Corneal fibroblasts; Cyclic stretching; Interleukin-1β; Matrix metalloproteinases.

MeSH terms

  • Animals
  • Biomechanical Phenomena
  • Cornea / cytology*
  • Enzyme Precursors / biosynthesis
  • Enzyme Precursors / metabolism
  • Extracellular Matrix / drug effects
  • Extracellular Matrix / metabolism
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Gene Expression Regulation, Enzymologic / drug effects*
  • Humans
  • Interleukin-1beta / pharmacology*
  • Matrix Metalloproteinase 2 / biosynthesis
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 9 / biosynthesis
  • Matrix Metalloproteinase 9 / metabolism*
  • Mechanical Phenomena*
  • Rabbits

Substances

  • Enzyme Precursors
  • Interleukin-1beta
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9