Abstract
Purpose: :
Research on cell damage and death as participating pathogenetic events in the development of cataract is to some extent conducted on human lens epithelial cells obtained during cataract surgery and with some discrepancy in results. We wanted to explore what effect variation in cataract incision size has on epithelium in the capsulotomy specimens obtained.
Methods: :
Consecutive capsulotomy specimens were retrieved through a 2,2 mm incision (n=8) or a 2,75 mm incision (n=8). Phase-contrast microscopy was performed on the specimens immediately after retrieval and after one week of incubation in DMEM-F12 with 15% FBS. Annexin V/Propidium Iodide was used as viability assay and ultrastructure examined by transmission electron microscopy (TEM).
Results: :
The capsulotomy specimens retrieved through a 2,2 mm incision showed a prominent cell loss, none of the cells produced outgrowth in vitro, and isolated cells or sheets of cells were positive for Annexin V/Propidium Iodide. Although necrotic cells were observed in the epithelium retrieved through a 2,75 mm section, all samples produced outgrowth. TEM showed a conspicuous autophagic activity in the cultured cells.
Conclusions: :
Significant variations in experimental results may be observed by a modest variation in a surgical procedural step such as the cataract incision size, and Micro Incision Cataract Surgery (MICS) with incision sizes down to 1,5 mm seem to reduce the ease of harvesting lens epithelium for research purposes.
Keywords: cataract • lesion study • small incision cataract surgery