Effects of amniotic membrane extract on primary human corneal epithelial and limbal cells

Clin Exp Ophthalmol. 2015 Jul;43(5):443-8. doi: 10.1111/ceo.12480. Epub 2015 Jan 15.

Abstract

Background: To assess the effects of amniotic membrane extract (AMX) on cellular activity of primary human corneal epithelial (HCE) cells under mechanical and oxidative stress, and on human limbal cells under oxidative stress.

Methods: Corneal mechanical stress was simulated with a linear scratch in confluent HCE cell plates, then incubated with 0.1% AMX for 48 and 72 h. Subjecting HCE cultures to 0.5 mmol/L tertiary-butylhydroperoxide for 1 h simulated an oxidative stress. 0.1% AMX-treated cultures were compared with controls at 24 and 48 h using cellular viability assay, along with 12-h AMX pretreatment and human limbal cell comparisons.

Results: Mechanical stress on HCE cultures revealed a statistically significant distance ratio at 48 and 72 h in favour of 0.1% AMX-treated cultures (P = 0.021 and 0.035, respectively). Oxidative stress did not reveal any significant difference in cellular viability of AMX-treated versus control cultures. Twelve hour AMX pre-treatment prior to oxidative stress revealed a significant difference after 24 h from oxidative injury (73.3% AMX vs. 66.0% control, P = 0.035), but not after 48 h. Human limbal cells demonstrated significantly improved oxidative viability compared with HCE cells, with (91.0% vs. 82.0% control, P = 0.017) and without 0.1% AMX pre-treatment (91.2% vs. 83.7% control, P = 0.019).

Conclusions: HCE cells treated with AMX healed faster after mechanical insult, suggesting a potential benefit in acute corneal injuries. Under oxidative stress, human limbal cells, a more proliferative cell type, showed superior viability compared with HCE cells.

Keywords: Amniotic membrane; Cornea; Corneal wound healing.

MeSH terms

  • Amnion / chemistry*
  • Cell Proliferation / drug effects
  • Cell Survival
  • Epithelium, Corneal / cytology
  • Epithelium, Corneal / drug effects*
  • Humans
  • Limbus Corneae / cytology
  • Limbus Corneae / drug effects*
  • Oxidative Stress
  • Stress, Mechanical
  • Tissue Extracts / pharmacology*
  • Wound Healing / physiology
  • tert-Butylhydroperoxide / toxicity

Substances

  • Tissue Extracts
  • tert-Butylhydroperoxide