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Z. Nathu, H. Sheardown, J. A. West-Mays; Effects of an MMP Inhibitor in an in-vitro Model of Posterior Capsule Opacification. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4223.
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We have previously shown that Matrix Metalloproteinases (MMPs) play a role in transforming growth factor beta (TGFß)-induced anterior subcapsular cataract (ASC) formation. ASC has many similar features to posterior capsule opacification (PCO) or ‘secondary’ cataracts, including the epithelial to mesenchymal transition (EMT) of lens epithelial cells. This study was designed to develop an in-vitro model of PCO to test the ability of MMP inhibitors to prevent TGFß-induced cellular changes characteristic of PCO.
To investigate the interaction of lens epithelial cells (LECs) and intraocular lenses (IOLs), human B3 LECs were seeded on silicone discs, or plastic tissue culture dishes. Following 4 days, cells were fixed and subjected to immunohistochemistry using an alpha smooth muscle actin (αSMA) antibody. To study the affect of TGFß and the MMP inhibitor, Ilomostat on cellular behaviour, human lens epithelial cells (FHL124) were cultured in M199 media (10% FBS) that was either supplemented with TGFß (200pg/ml, 500pg/ml and 1ng/ml), or TGFß (1ng/ml) + Ilomostat (25µM), Ilomostat alone, or left untreated (controls) for 6 days.
A proportion of the B3 LECs grown on plastic dishes exhibited immunoreactivity to αSMA, however a greater number of LECs adhering to the silicone IOLs were αSMA positive. FHL124 cells pre-treated with TGFß at 1ng/ml, 500pg/ml and 200pg/ml showed a reduction in the number of cells present on the tissue culture dish following 6 days of incubation, compared to controls, with fewer cells present at the highest concentration of TGFß (1ng/ml). In comparison, cells cultured in media pre-treated with TGFß (1ng/ml) + Ilomostat (25uM) showed a greater number of cells when compared to TGFß (1ng/ml) treated LECs. Cells cultured in media pre-treated with Ilomostat (25uM) alone resembled the control LECs.
These results indicate that pre-treatment of cells with increasing concentrations of TGFß resulted in fewer LECs being present following the 6 day culture period. However, when cells were pre-treated TGFß at the highest concentration along with Ilomostat the cell number was more comparable to controls. Further studies will be aimed at determining whether the reduction in cell number caused by TGFß is due to alterations in cell adhesion, proliferation or apoptosis and how Ilomostat may inhibit this.
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