Study of Genotoxic Effects of Artificially Induced UV-Radiation on Human Lens Epithelial Cell (HLEC) Culture by Using Dye Exclusion Assay
Author(s): Huddar MD and Paikrao VM*
Abstract
Cataract is one of major cause of blindness, 44% cases of blindness is due to cataract only. In developing countries such as India cataract is more common, and it manifest earlier in life than the most developed countries of the world. Cataract is more common in dry and hot area where sun (UV) exposure is more than prolong cloudy cover. In this study we are aimed to compare effects of UV-ray irradiation on cell viability of HLEC (Human lens epithelial cell) culture by using trypan blue dye exclusion assay against the unexposed control HLEC culture. Total 20 cataract patients from Dept. of Ophthalmology of Lata Mangeshkar Hospital, Nagpur were included in this study. The capsulorhexis sample from cataract surgery taken for study from eye OT. The capsulorhexis sample without any UV-ray exposure taken as control. For culture of HLEC’s RPMI 1640 medium containing 10% fetal calf serum used. The HLEC cultures were UV-ray irradiated with increasing time exposure PHILIPS® TUV 15W/G15 T8 ultraviolet UV rays tube. The Trypan blue dye exclusion assay was used to analyze cell viability. The slides were analyzed using Olympus® BX51 Research microscope with oil immersion and 100 were observed per sample. The cell viability was calculated using Epi info® (version 6.0), as subject means with standard deviation. Statistical significance will be defined at P≤0.001. Out of 100 cases the 12 were female and 8 were male. Mean age of both male and female patients were 62.45±3.18(SD). A positive linear correlation was found (r:0.819, p<0.001) between the time of UV-Ray exposure and death cells per 100 observed LECs.