Abstract
Abstract: :
Purpose: To understand whether the amniotic membrane can modulate the anti-angiogenic activity expressed by limbo-corneal epithelial cells Methods: Human limbo-corneal epithelial cells (HLE) were cultivated on 35 mm plastic dish, on intact or on denuded cryo-preserved human amniotic membrane (IAM and DAM) with SHEM-5 % FBS medium. When reaching at least 80 % confluence, the explants were cocultured for 3 days with type I collagen gels containing human umbilical endothelial cells (HUVECs) which were cultured by EGM medium (Clonetics). Conditioned media from HLE cultured under different conditions were used for HUVEC proliferation assay in 24 well for 6 days and migration assay by razor wounding method overnight. Fresh AM, cryo-preserved AM, and AM kept in 37 C incubator for 3 weeks with regular changing of culture medium (old AM) were used as controls. Results: Fresh AM had small inhibitory effect on the differentiation, migration, and proliferation of HUVECs. Such inhibitory effect was most prominent when HLE were cultivated on DAM, followed by IAM, and on plastic dish. Cryo-preserved and old AM, either intact or denuded, expressed only minimal inhibitory effect. Conclusions: Our data suggested that in addition to previously reported anti-inflammatory effect after AMT, AMT may also directly augment the anti-angiogenic activity of HLE, possibly through modulating major angiogenic/antiangiogenic factors production.
Keywords: cornea: epithelium • neovascularization • vascular cells