Type I collagen accelerates the spreading of lens epithelial cells through the expression and activation of matrix metalloproteinases

Curr Eye Res. 2014 May;39(5):460-71. doi: 10.3109/02713683.2013.853194. Epub 2014 Jan 8.

Abstract

Purpose: Matrix metalloproteinases (MMPs) are involved in posterior capsule opacification (PCO), but the mechanisms that promote MMP expression are yet to be determined. In this study, we investigated whether type I collagen, which is only detected in aged or cataractous lens capsules, affects the expression and activation of MMPs in primary-cultured chicken lens epithelial cells (LECs).

Materials and methods: Chicken LECs were isolated from chicken embryos and cultured in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (FBS) on type I collagen-coated dishes. The activity of secreted MMPs was examined using gelatin zymography, and cell spreading was determined as the average area of randomly distributed cells. For some experiments, LECs were cultured in the presence of the broad-spectrum MMP inhibitor, GM6001. LECs cultured on uncoated dishes were used as controls. To examine the involvement of MMP in cell migration, a wound-healing assay was performed in the presence of the MMP inhibitor.

Results: Chicken LECs constitutively express the pro-form of MMP-2. When LECs were cultured on type I collagen-coated dishes, they expressed the active form of MMP-2 and the pro-form of MMP-9. This expression and activation by type I collagen was also observed in the human LEC line SRA-01/04, but not the human Müller glial cell line, MIO-M1. Type I collagen enhanced cell spreading, which was suppressed by the MMP inhibitor. Type I collagen also accelerated α-smooth muscle actin expression. In addition, LEC migration was inhibited by the MMP inhibitor in a dose-dependent manner in the wound-healing assay.

Conclusion: Type I collagen promotes the expression and activation of MMPs in a LEC-specific manner. These results suggest that type I collagen may play a role in PCO development.

MeSH terms

  • Animals
  • Cataract / metabolism*
  • Cataract / pathology
  • Cell Movement / drug effects
  • Cell Movement / physiology
  • Cell Proliferation / drug effects
  • Cell Proliferation / physiology
  • Cells, Cultured
  • Chick Embryo
  • Chickens
  • Collagen Type I / metabolism*
  • Collagen Type I / pharmacology
  • Enzyme Activation / drug effects
  • Enzyme Activation / physiology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • Lens, Crystalline / drug effects
  • Lens, Crystalline / metabolism*
  • Lens, Crystalline / pathology
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 9 / metabolism*
  • Posterior Capsule of the Lens / drug effects
  • Posterior Capsule of the Lens / metabolism
  • Posterior Capsule of the Lens / pathology
  • Wound Healing / drug effects
  • Wound Healing / physiology

Substances

  • Collagen Type I
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9