Effect of Amniotic Membrane Extract on the Tenon's Capsule Fibroblasts

Heo, Hyun; Kim, Jae Woo

Abstract

PURPOSE: To investigate the effect of amniotic membrane extract (AME) on the survival, nitric oxide production by cultured human Tenon's capsule fibroblasts (HTCF) and contraction of collagen gels.
METHODS
After exposure to 5~20 g/ml AME for 5 days, the survival and nitrite production of the primarily cultured HTCF were assessed with MTT and Griess assays respectively. Acridine orange/Hoechst 33342 double staining and flow cytometric analysis using FITC-Annexin/propidium iodide double staining were done to evaluate AME-induced apoptosis of HTCF. The effect of AME on the contraction of collagen gels cultured with fibroblast was also evaluated.
RESULTS
AME decreased cellular survival in a dose-dependent manner and did not affect the production of nitric oxide. AME caused apoptotic death of HTCF in a dose-dependent manner and caused necrosis at high concentration (20 g/ml). AME did not affect on the contraction of collagen gel.
CONCLUSIONS
AME decreased cellular survival of HTCF by apoptosis and did not affect the contraction of collagen gels. Thus AME could modulate conjunctival wound healing by suppressing the cellular survival and activity of HTCF.