Corneal Neovascularization Suppressed by TIMP2 Released from Human Amniotic Membranes

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Purpose: To investigate the effects of culture medium of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice. Methods: Culture medium of amniotic membrane was prepared by cultivating AM (with epithelium side up) in EGM basic medium for 3 days, and was collected separately to three groups, e.g. control (EGM only), AM with epithelium (AM) and AM without epithelium (De-AM). Corneal neovascularization was induced in mice by using micropocket assay with Hydron polymer pellets containing 100 ng bFGF. Migration and proliferation of human umbilical cord vein endothelial cells (HUVEC) were performed in Boyden chambers and by using the CyQUANT fluorescence binding assay respectively. The levels of tissue inhibitors of metalloproteinase 1 and 2 (TIMP1, TIMP2) in culture medium were determined by ELISA assay. Results: CNV induced by bFGF was significantly suppressed by culture medium of amniotic membrane. When the medium was applied as an eyedrop Methods: To investigate the effects of culture of human amniotic membrane (AM) on corneal neovascularization (CNV) induced by basic fibroblast growth factor (bFGF) in mice. Methods: Culture medium of amniotic membrane was prepared by cultivating AM (with epithelium side up in EGM basic medium for 3 days, and were collected separately to three groups, eg control (EGM only), AM with epithelium (AM) and AM without epithelium (De- AM). Corneal neovascularization was induced in mice by using micropocket Migration and proliferation of human umbilical cord vein endothelial cells (HUVEC) were performed in Boyden chambers and by using the CyQUANT fluorescence binding assay. The levels of tissue inhibitors of metalloproteinase 1 and 2 (TIMP1 , TIMP2) in culture medium were determined by ELISA assay. Results: CNV induced by bFGF was significantly suppressed by culture medium of amniotic membrane. When the medium was app lied as an eyedrop
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