title: Bovine lactoferrin stimulates human corneal epithelial alkali wound healing in vitro.
abstract: OBJECTIVE
  The purpose of this study was to investigate the effect of bovine lactoferrin (BLF) on human corneal epithelial wound healing using an in vitro alkali-induced wound model and to understand its role in promoting wound healing.
  
  
  METHODS
  Confluent human corneal limbal epithelial (HCLE) cells wounded using 0.5 microL of 0.1 M sodium hydroxide were treated with BLF (0, 0.1, 1, 2.5, and 5 mg/mL) or anti-human interleukin-6 (IL-6) receptor neutralizing antibody (anti-IL-6 antibody; 1, 10, and 50 microg/mL) or tyrphostin AG1295 (an inhibitor of platelet-derived growth factor [PDGF] receptor kinase; 1 and 10 microM), IL-6, or PDGF-BB. The conditioned medium collected for BLF treatment (0 and 5 mg/mL) was analyzed using a protein array for a number of cytokines/growth factors involved in corneal wound healing. A preliminary animal study using mice was carried out to determine the effect of BLF on alkali wounds.
  
  
  RESULTS
  BLF at 2.5 and 5 mg/mL promoted wound healing (P<0.01). During wound closure, BLF upregulated PDGF-BB 180-fold and IL-6 10-fold compared with control. Treatment with tyrphostin AG1295 (10 microM; P<0.01) or anti-IL-6 antibody (50 microg/mL; P<0.01) in the presence of BLF inhibited wound closure, whereas the addition of exogenous IL-6 and PDGF-BB promoted wound closure. Preliminary animal studies have shown that BLF (5 mg/mL) promotes alkali wound healing in vivo.
  
  
  CONCLUSIONS
  These results suggest that BLF at >or=2.5 mg/mL stimulates HCLE wound healing, and this stimulation is mediated through the upregulation of PDGF or IL-6.
authors.affiliation: Institute for Eye Research, Sydney, New South Wales, Australia.
authors.forename: Ushasree
authors.initials: U
authors.lastname: Pattamatta
authors.forename: Mark
authors.initials: M
authors.lastname: Willcox
authors.forename: Fiona
authors.initials: F
authors.lastname: Stapleton
authors.forename: Nerida
authors.initials: N
authors.lastname: Cole
authors.forename: Qian
authors.initials: Q
authors.lastname: Garrett
mesh_terms: D000818:Animals; D002057:Burns, Chemical / Q000188:drug therapy* / Q000473:pathology; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000473:pathology; D005126:Eye Burns / Q000139:chemically induced* / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007781:Lactoferrin / Q000187:drug effects*; D016850:Limbus Corneae / Q000166:cytology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012972:Sodium Hydroxide / Q000633:toxicity; D020032:Tyrphostins / Q000494:pharmacology; D014945:Wound Healing / Q000187:drug effects*
chemical_list: C407689:6,7-dimethoxy-2-phenylquinoxaline; D016207:Cytokines; D005353:Fibronectins; D020032:Tyrphostins; D012972:Sodium Hydroxide; D007781:Lactoferrin