Effect of Extracellular Matrix on Proliferation and Differentiation of Porcine Lens Epithelial Cells
Hesse Y., Kampmeier J., Lang G. K., Lang G. E.
Department of Ophthalmology, University of Ulm
Purpose: Proliferation and differentiation of lens epithelial cells are important mechanisms of secondary cataract formation. After extracapsular cataract extraction the extracellular matrix (ECM) around remaining LECs is altered compared to the intact lens. This study investigated the effects of ECM on proliferation and alpha-smooth-muscle actin (a-SMA) expression, a marker for myofibroblasts, in cultured porcine LECs.
Method: Porcine LECs were cultured for 3 resp. 4 days on wells resp. glass cover slips coated with laminin, fibronectin, type I collagen or type IV collagen. LECs cultured on uncoated wells or cover slips served as control. Poliferative response were measured through [3H]-thymidine incorporation into DNA. Alpha-SMA was detected immunocytochemically with a mouse monoclonal antibody, and the relative numbers of a-SMA-positive cells were calculated. Statistical analysis was performed using Student´s unpaired t-test.
Results: Cell proliferation was significantly increased by coating with fibronectin (10320.5 CPM; p<0.0001) (mean), type I (12448 CPM; p<0.0001) and type IV collagen (9963.9 CPM; p<0.0001) compared to control (1753.1 CPM) whereas coating with laminin had no effect (1663.8 CPM; p=0.7679). The ratio of a-SMA-positive cultured on uncoated cover slips was 13.2%. This ratio was significantly increased by coating with fibronectin (24.4%; p=0.0028
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