The effect of therapeutic human serum drops on corneal stromal wound-healing activity.
OBJECTIVE
To investigate the effect of human serum, an ocular therapy, on human corneal fibroblast (HCF) wound-healing activities.


METHODS
The water soluble tetrazolium reagent-1, chemotactic chambers, fibroblast-populated type-I collagen gels, zymography, and Western blotting were used to assess HCF proliferation, migration, contraction, and matrix metalloproteinase (MMP) activity and levels, respectively. Fibroblasts were obtained from human donor corneas. Human serum, fibroblast culture medium (FCM; Dulbeccos Minimal Essential Medium/10% newborn calf serum) with and without calf serum supplementation, and 0.3% hypromellose were compared.


RESULTS
Proliferation and migration were maximal in 1% human serum. Relaxed gel contraction was maximal for fibroblasts cultured in 10%, 50%, and 100% serum and FCM. Whereas in stressed gels, maximal contraction was induced when fibroblasts were cultured in 50% and 100% serum. In low serum concentrations, greater MMP-2 activity was detected than MMP-1 and MMP-9.


CONCLUSIONS
Low concentrations of human serum stimulated HCF migration, proliferation, and MMP activity. High concentrations produced greater matrix contraction. This may have implications for the therapeutic use of autologous serum.
Watson
Stephanie L
SL
Cornea and External Diseases, Moorfields Eye Hospital, NHS Foundation Trust, London, United Kingdom. s.watson@unsw.edu.au
Secker
Genevieve A
GA
Daniels
Julie T
JT
D015153:Blotting, Western; D018929:Cell Culture Techniques; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D003319:Corneal Stroma / Q000166:cytology*; D004151:Dipeptides / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D053551:Keratin-12 / Q000378:metabolism; D053558:Keratin-3 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000378:metabolism; D018613:Microscopy, Confocal; D009883:Ophthalmic Solutions; D011480:Protease Inhibitors / Q000494:pharmacology; D044967:Serum / Q000502:physiology*; D014746:Vimentin / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology*
D004151:Dipeptides; D053551:Keratin-12; D053558:Keratin-3; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D009883:Ophthalmic Solutions; D011480:Protease Inhibitors; D014746:Vimentin; D020782:Matrix Metalloproteinases