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Sebastian Di Cesare, Bruno F. Fernandes, Matthew Balazsi, Tamara J. Granner, Patrick T. Logan, Miguel N. Burnier, Jr.; Transcriptional Changes in Human Lens Epithelial Cells Secondary to Intraocular Lens Exposure. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3065.
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© ARVO (1962-2015); The Authors (2016-present)
The epithelial to mesenchymal transition (EMT) has been previously reported to be an important step in transforming lens epithelial cells into myofibroblasts, which can cause significant visual loss in up to 20-30% of patients 2-3 years post-cataract surgery. The clinical process has been characterized as posterior capsule opacification (PCO) and few treatment options are available. The aim of this study is to characterize the transcriptional changes that occur in a human lens epithelial cell line (HLE-B3) when coated on three different types of intraocular lenses (IOLs).
The HLE-B3 cell line was used in this study. Three types of IOLs (AcrySof, Silicone, and PMMA) were evaluated. Five lenses of each type were seeded in a 24 well plate (one lens/well) and coated with 3.0x104 HLE-B3 cells. The cells were allowed to coat and proliferate on the lens for one week. Media was changed every second day carefully as to not disturb the coating. The IOLs were then placed in a fresh 24 well plate, trypsinized and pooled for RNA extraction. On column DNAase digestion was performed in order to ensure no gDNA contamination. RNA concentrations were normalized to 250ng/µl and processed for cDNA synthesis. The RT2 ProfilerTM PCR Array focused for the EMT pathway (86 genes) was used to evaluate the changes between the IOLs coated cells and normal cell control.
Significant results were considered when a greater than ±3-fold transcriptional change was observed. The AcrySof lens had no significant change for any of the genes located on the EMT gene panel. The PMMA lens had 15 genes with significant changes when compared to the control. The Silicone lens had the greatest number of significant changes (25 genes). Notable gene changes that matched both the PMMA and Silicone lenses include SPP1, SNAI2, SERPINE1, PDGFRB, MMP-2, MMP-3, MMP-9, KRT7, FN1, and CDH2.
AcrySof Natural lenses were less likely to cause an EMT transition in the HLE-B3 than the other two evaluated lenses. In addition, the Silicone and PMMA lenses induced many significant changes that may contribute to the development of PCO in vivo. The targets identified may become therapeutic candidates to reduce the fibrotic scarring that ultimately lead to visual loss in these patients. To the best of our knowledge, this is the first study using a focused gene array to conduct a side-by-side comparison of three widely used IOLs.
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