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Originally published In Press as doi:10.1167/iovs.08-1882 on December 5, 2008
 (Investigative Ophthalmology and Visual Science. 2009;50:1636-1643.)
© 2009 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.08-1882
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Bovine Lactoferrin Stimulates Human Corneal Epithelial Alkali Wound Healing In Vitro

Ushasree Pattamatta,1,2,3 Mark Willcox,1,2,3 Fiona Stapleton,1,2,3 Nerida Cole,1,2,3 and Qian Garrett1,2,3

1From the Institute for Eye Research, Sydney, New South Wales, Australia; 2Vision CRC, Sydney, New South Wales, Australia; and 3The School of Optometry and Vision Science, The University of New South Wales, Sydney, New South Wales, Australia.

PURPOSE. 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 µL 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 µg/mL) or tyrphostin AG1295 (an inhibitor of platelet-derived growth factor [PDGF] receptor kinase; 1 and 10 µM), 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 µM; P < 0.01) or anti-IL-6 antibody (50 µg/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 ≥2.5 mg/mL stimulates HCLE wound healing, and this stimulation is mediated through the upregulation of PDGF or IL-6.






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