title: Low levels of hydrogen peroxide stimulate corneal epithelial cell adhesion, migration, and wound healing.
abstract: OBJECTIVE
  Intracellular reactive oxygen species have been reported to associate with growth factor and integrin signalings in promoting cell adhesion in many cell types. This study is to explore if exogenous H(2)O(2) at low levels can be beneficial to cell adhesion, migration, and wound healing.
  
  
  METHODS
  Primary rabbit corneal epithelial cells treated with 0-70 μM H(2)O(2) were tested for viability by MTT assay, adhesion by centrifugation assay, focal contacts of vinculin and F-actin by immunofluorescence, activated Src(pY416), EGF receptor (pY845), vinculin(pY1065), FAK(pY397), and FAK(pY576) by immunoblotting. Cell migration was examined with 0-50 μM H(2)O(2) using the scratch wound technique. Corneal wound healing of ex vivo pig model and in vivo mouse model was examined using H(2)O(2) with and without antioxidant N-acetylcysteine (NAC).
  
  
  RESULTS
  Compared with the untreated control, H(2)O(2) at 10-50 μM stimulated cell viability and facilitated adhesion and migration with clear induction of vinculin-rich focal adhesions and F-actin-containing stress fibers by increasing activated Src, FAK(pY576), and vinculin(pY1065). H(2)O(2) also increased phosphorylation of EGFR(Y845) parallel to that of activated Src, but both were eliminated by NAC and PP1 (Src inhibitor). Finally, H(2)O(2) induced faster wound healing in cornea both in vitro and in vivo, but the healing was diminished by NAC.
  
  
  CONCLUSIONS
  These findings suggest that H(2)O(2) at low levels promotes cell adhesion, migration, and wound healing in cornea cells or tissue, and the interaction of H(2)O(2) with Src plays a major role.
authors.affiliation: Department of Ophthalmology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Zhejiang, Hangzhou, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Pan
authors.forename: Wen-Ya
authors.initials: WY
authors.lastname: Qiu
authors.forename: Ya-Nan
authors.initials: YN
authors.lastname: Huo
authors.forename: Yu-Feng
authors.initials: YF
authors.lastname: Yao
authors.forename: Marjorie F
authors.initials: MF
authors.lastname: Lou
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D002470:Cell Survival; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000008:administration & dosage*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques; D010766:Phosphorylation; D011817:Rabbits; D013552:Swine; D016596:Vinculin / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology*
chemical_list: D000199:Actins; D016596:Vinculin; D006861:Hydrogen Peroxide; D066246:ErbB Receptors; D051416:Focal Adhesion Protein-Tyrosine Kinases