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A. Solomon, F. Alviano, I. Anteby, J. Frucht-Pery, J. Pe'er, F. Levi-Schaffer, S.C. Tseng; Suppression of Inflammatory and Fibrotic Cytokines in Human Corneal Epithelial Cells and Conjunctival Fibroblasts Cultured on the Amniotic Membrane Stroma . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3764.
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Purpose: to evaluate the anti-inflammatory and anti-fibrotic effects of the amniotic membrane (AM) stoma on cultured human corneal epithelial cells and conjunctival fibroblasts. Methods: Human corneal epithelial cells or conjunctival fibroblasts were cultured from limbocorneal explants of donor eyes or from conjunctival biopsies from healthy volunteers, respectively, on plastic or on the AM stroma. Expression of IL-1 α, IL-1 ß, IL-1 receptor antagonist (RA) in corneal epithelial cells was compared with or without activation with lipopolysaccharide for 24 hours using RNAse protection assay. Protein production in the supernatant was analysed by ELISA. Activated human conjunctival fibroblasts were cultured on plastic or on the AM stroma. Expression of Transforming growth factor (TGF)-ß1 and of granulocyte-macrophafe colony stimulating factor (GM-CSF) in conjunctival fibroblasts was measured in conditioned media from these cultures by ELISA, and in RNA extracts by RT-PCR. Results: Expression of IL-1 α and IL-1 ß transcripts and proteins was significantly reduced by cells cultured on the AM stroma compared to plastic cultures whether lipopolysaccharide was added or not. Expression of IL-1 RA by cells cultured in the lipopolysaccharide-free medium was upregulated by AM stromal matrix. The ratio between IL-1 RA and IL-1 α protein levels in AM cultures was higher than in plastic cultures. The expression of TGF-ß1and of GM-CSF was significantly suppressed in activated conjunctival fibroblasts cultured on the AM compared to those cultured on plastic. Conclusions: The AM stromal matrix markedly suppresses the expression of inflammatory and fibrotic cyokines derived from cells of the ocular surface. These data may explain in part the effect of AM transplantation in reducing ocular surface inflammation.
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