Standardization of a new method for assessing the development of cataract in cultured bovine lenses

Abstract Purpose To standardize a new method for assessing cataractogenesis in isolated cultured bovine lenses using L-cysteine as the standard anti-cataract agent. Methods Intact bovine lenses were cultured in DMEM with L-cysteine in presence or absence of hydrogen peroxide (H 2 O 2 ). Lens opacity (transmittance) was determined using a plate reader. Lens homogenate glutathione (GSH) and superoxide dismutase (SOD) contents were measured using enzyme immunoassays kits. Results DMEM-cultured lenses exhibited a time-dependent loss in transmittance (230–710 nm) up to 120 h, achieving the highest reduction of 38.6 ± 0.09% at 420 nm ( p n  = 12). Compared to untreated lenses (time in hours [t] = 0), L-cysteine (10 −6  M and 10 −5  M) significantly ( p n  = 6) increased time-dependent transmittance (420 nm) by 31.6 ± 0.17% and 28.0 ± 0.07%( t  = 120), respectively. When compared to DMEM-cultured lenses ( t  = 0), H 2 O 2 (10 mM, 50 mM and 100 mM) significantly ( p n  = 12) reduced transmittance by 57.8 ± 0.1, 57.4 ± 0.04 and 87.7 ± 0.6%( t  = 120), respectively. Moreover, L-cysteine significantly ( p n  = 6) attenuated H 2 O 2 (50 mM)-induced decrease in transmittance by 12.5 ± 0.05%(10 −6  M), 13.0 ± 0.09%(10 −5  M), 14.5 ± 0.08%(10 −4  M) and 8.6 ± 0.11%(10 −3  M)( t  = 120), respectively. When compared to untreated lenses ( t  = 0), the time-dependent decrease ( p n  = 5) in lenticular total GSH content and total SOD activity of 46.1 ± 0.06% and 42.0 ± 1.65% ( t  = 120) was attenuated ( p n  = 5) by L-cysteine (10 −6  M) by 76.6 ± 0.06% and 7.4 ± 1.98%, respectively. Similarly, the H 2 O 2 (50 mM)-induced decline ( p n  = 5) in total GSH content and SOD activity of 82.6 ± 0.08% and 86.6 ± 0.66% ( t  = 120) was attenuated by L-cysteine (10 −4  M) by 74.7 ± 1.05% and 161.1 ± 4.9%, respectively. Conclusion Measurement of spectral transmission coupled with assessment of the activity of antioxidant enzymes in bovine cultured lens can provide a useful tool in studies of cataracts in an animal model of this disease.