title: Effects of 17β-estradiol on human corneal wound healing in vitro.
abstract: OBJECTIVE
  To investigate the effect of 17β-estradiol on corneal wound healing, particularly on epithelial mitosis and migration.
  
  
  METHODS
  Immortalized human corneal epithelial cells (HCECs) were cultured in media with different concentrations of 17β-estradiol (10, 50, 100, and 200 pg/mL), Dulbecco modified Eagle medium: Nutrient Mixture F-12 (negative control), and serum-containing Dulbecco modified Eagle medium: Nutrient Mixture F-12 (positive control). After 6 or 24 hours of hormone treatment, to evaluate the migratory potential of 17β-estradiol, wound healing assays were conducted via the manual scraping of HCECs and western blot analysis of fibronectin and matrix metalloproteinase 9 (MMP9). The proliferative potential of 17β-estradiol was evaluated via a proliferation assay using western blot analysis for proliferating cell nuclear antigen. In addition, epidermal growth factor (EGF) was measured by reverse transcription-polymerase chain reaction, and for the inhibition of epidermal growth factor receptor (EGFR)-mediated signal transduction, a wound healing assay was conducted after HCECs cultured with EGFR small interfering RNA were stimulated with 100 pg/mL 17β-estradiol.
  
  
  RESULTS
  Wound healing assay rates were enhanced as 17β-estradiol increased, with statistically significant changes seen in 50, 100, and 200 pg/mL 17β-estradiol-treated and positive control cells, compared with negative control cells (P < 0.05, in each group). Western blot analysis revealed that the expression of the MMP9 gene was upregulated by 17β-estradiol, and the expression of the fibronectin gene was downregulated by 17β-estradiol. The mitosis assay via western blot analysis showed that the expression cell cycle-associated protein, proliferating cell nuclear antigen, increased gradually as a result of 17β-estradiol treatment. Reverse transcription-polymerase chain reaction showed that EGF was upregulated by 17β-estradiol, and the EGFR small interfering RNA did not totally block the wound healing of the 17β-estradiol-treated cells but statistically significantly reduced the wound healing rate (P = 0.031).
  
  
  CONCLUSIONS
  17β-Estradiol facilitated the maintenance of the beneficial effect on corneal epithelial migration and proliferation, and the promoting effect of 17β-estradiol is partially related to increased EGF in vitro.
authors: Department of Ophthalmology and Visual Science, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seocho-gu, Seoul, Korea.
authors: Tae-Hoon
authors: TH
authors: Oh
authors: Dong-Jin
authors: DJ
authors: Chang
authors: Jun-Sub
authors: JS
authors: Choi
authors: Choun-Ki
authors: CK
authors: Joo
mesh_terms: D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000235:genetics; D019573:Epithelium, Corneal / Q000166:cytology / Q000293:injuries* / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology*; D004967:Estrogens / Q000494:pharmacology*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D008954:Models, Biological; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D004967:Estrogens; D005353:Fibronectins; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D004958:Estradiol; D004815:Epidermal Growth Factor; D020780:Matrix Metalloproteinase 9