title: In vivo and in vitro inhibitory effect of amniotic extraction on neovascularization.
abstract: OBJECTIVE
  To prepare amniotic extraction (AE) and to test its antiangiogenic effect in vivo and in vitro.
  
  
  METHODS
  AE was prepared and diluted to 50, 100, and 200 microg/mL concentrations. Alkali burn-induced corneal neovascularization (NV) was produced and topically treated with different concentrations of AE or 0.1% dexamethasone for 7 days. Normal saline was used as a control. Corneal NV was visualized by heart perfusion of Chinese ink and quantified as the percentage of corneal NV area to the whole corneal area. Human umbilical vein endothelial cells (HUVECs) were primarily cultured. The effects of AE on proliferation and tube formation of HUVECs were tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and in vitro angiogenesis assay. Pigment epithelium-derived factor (PEDF) in AE was detected by Western blot.
  
  
  RESULTS
  Relative corneal NV area in the control group was 56.6% +/- 9.9%, which was significantly reduced by 50 microg/mL AE (47.6% +/- 6.9%; P = 0.043) and 200 microg/mL AE (34.3% +/- 7.8%; P < 0.001) and by 0.1% dexamethasone (21.1% +/- 1.8%; P < 0.001). HUVEC cell proliferation was significantly decreased after treatment with AE at concentrations of 50 and 100 microg/mL compared with control (P = 0.036 and 0.001, respectively). The tube formation was significantly suppressed by 100 microg/mL AE (70.03% +/- 4.35%) compared with control (100% +/- 4.84%; P = 0.002). No expression of PEDF was detected in AE.
  
  
  CONCLUSIONS
  AE inhibits NV induced by alkali burn. This effect may be elicited at least in part through the inhibiting activity of blood vessel endothelial cells and is not associated with PEDF.
authors.affiliation: Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
authors.forename: Aihua
authors.initials: A
authors.lastname: Jiang
authors.forename: Chaoyang
authors.initials: C
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.forename: Jiaoyue
authors.initials: J
authors.lastname: Hu
authors.forename: Wenhui
authors.initials: W
authors.lastname: Kuang
authors.forename: Shangchen
authors.initials: S
authors.lastname: Hao
authors.forename: Wenzhao
authors.initials: W
authors.lastname: Yang
authors.forename: Chuanchao
authors.initials: C
authors.lastname: Xu
authors.forename: Guoquan
authors.initials: G
authors.lastname: Gao
authors.forename: Zhichong
authors.initials: Z
authors.lastname: Wang
authors.forename: Zuguo
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000650:Amnion / Q000737:chemistry*; D020533:Angiogenesis Inhibitors / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D001808:Blood Vessels / Q000187:drug effects; D015153:Blotting, Western; D002057:Burns, Chemical / Q000188:drug therapy; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D016510:Corneal Neovascularization / Q000188:drug therapy* / Q000209:etiology; D003907:Dexamethasone / Q000627:therapeutic use; D004730:Endothelium, Vascular / Q000187:drug effects*; D005126:Eye Burns / Q000139:chemically induced; D005136:Eye Proteins / Q000378:metabolism; D005260:Female; D006801:Humans; D009414:Nerve Growth Factors / Q000378:metabolism; D010865:Pilot Projects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015843:Serpins / Q000378:metabolism; D012972:Sodium Hydroxide; D013778:Tetrazolium Salts; D013844:Thiazoles; D014020:Tissue Extracts / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D014471:Umbilical Veins / Q000166:cytology; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D020533:Angiogenesis Inhibitors; D003907:Dexamethasone; D005136:Eye Proteins; D009414:Nerve Growth Factors; D015843:Serpins; D012972:Sodium Hydroxide; D013778:Tetrazolium Salts; D013844:Thiazoles; D014020:Tissue Extracts; D000099174:Pigment Epithelium-Derived Factor; C022616:thiazolyl blue