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AbstractDO.02.13 MUC1, MUC4 and MUC16 expression and protein biosynthesis of cultivated limbal stem cells on amniotic Membrane in comparison to normal and pathological ocular surface tissue Kakkassery V., Pauklin M., Steuhl K.-P., Meller D.
Erkrankungen des vorderen Augenabschnittes, Zentrum für Augenheilkunde, Universitätsklinikum Essen, Essen Objective: Limbal stem cell deficiency is characterized by in-growing conjunctiva into the cornea (pannus), chronic inflammation, impaired vision and in most of the cases dry eye. Transplantation of limbal epithelium cultivated (cLE) on amniotic membrane has recently been developed to treat this condition. Membrane-associated mucins are an important component of the tear film. The objective of this study was to characterize the expression patterns of the membrane-associated mucins MUC1, MUC4 and MUC16 in cultivated limbal epithelium, cicatricial pannus tissue gained from in-growth after LSD and also healthy cornea and conjunctiva.
Methods: Total RNA and Protein were isolated from cLE and pannus samples. Healthy corneal and conjunctival tissue was used as control tissue. The expression of MUC1, MUC4 and MUC16 was analyzed by Real-Time PCR and Western blotting.
Results: Conjunctiva showed a significantly higher expression of MUC1 and MUC4 (p<0.001) than cornea, the expression of MUC16 was similar in both tissues. The expression of MUC1 was increased in cLE (p<0.01), MUC16 decreased (p<0.0001) and MUC4 showed no significant difference when compared to cornea. The expression of all studied genes was in conjunctiva significantly higher than in cLE (p<0.05), but was similar to pannus showing no significant differences. Pannus, like conjunctiva, revealed a significantly higher expression of studied genes when compared to cornea or cLE (p<0.001). Western blotting confirmed these results.
Conclusions: Conjunctiva and pannus revealed both a higher expression of studied membrane-associated mucins than cornea or cLE. The expression of studied mucins showed in cultured limbal epithelium more similarities to cornea and not too pannus or conjunctiva. The restoration of an expression pattern of membrane associated mucins similar to that of a healthy corneal epithelium may be an additional beneficial effect of cLE transplantation.
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