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1 From the Ocular Surface and Tear Center, Department of Ophthalmology, Bascom Palmer Eye Institute; 2 Singapore National Eye Center, Singapore; and 3 Department of Cell Biology and Anatomy, University of Miami School of Medicine, Miami, Florida.
PURPOSE. To examine the expression patterns of extracellular matrix degrading enzymes in cultured primary pterygium body fibroblasts activated by cytokines and growth factors potentially derived from ocular surface epithelial cells and tears.
METHODS. EGF, TGF-
, PDGF-BB, IL-1ß, bFGF, TGF-ß1, TNF-, or IL-6 were
added at 10 ng/ml to early passaged primary pterygium body fibroblasts
(PBF) or normal human conjunctival fibroblasts (HJF) in a serum-free
medium. Expression of transcripts and proteins of MMP-1, MMP-2, MMP-3,
MMP-9, TIMP-1, TIMP-2, and uPA was determined by Northern
hybridization, ELISA, and Western blotting, respectively. Gelatin and
casein zymographies were performed in their serum-free conditioned
media with or without enzyme inhibitors to determine the activity of
MMP-2 and -3, respectively.
RESULTS. IL-1ß and TNF- dramatically increased the mRNA and protein
expression of MMP-1 and MMP-3 in cultured PBF when compared to normal
HJF and to their nonstimulated counterparts cultured in a serum-free
medium. EGF and TGF- also upregulated MMP-3 in PBF when compared to
HJF. The transcript levels of MMP-2 were high but stable for the two
cell types regardless of the cytokine treatment. Both TIMP-1 and TIMP-2
expressions were not influenced by the cell type or the cytokine
treatment. MMP-9 was not expressed in either of these two types of
fibroblasts. Both IL-1ß and TNF- induced a significant decrease in
uPA expression in PBF, whereas bFGF induced a slight increase in both
HJF and PBF.
CONCLUSIONS. Chronic inflammatory stimulation by IL-1ß and TNF-, which
potentially can be derived from the ocular surface and tears, may be
responsible for increased expression of MMPs in cultured PBF. These
data have clinical implications on progression of pterygium and
recurrence associated with incomplete excision of primary PBF under the
influence of ocular surface inflammation. Suppression of intraoperative
and postoperative inflammation may be a new strategy to prevent
pterygium recurrence.
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