Synchrotron‐based Fourier transform infrared microspectroscopy examined differences between the cultured and post‐operative human lens epithelial cells important for understanding posterior capsular opacification and lens regeneration

Abstract

Aims/Purpose: To compare the bio‐macromolecular composition and the differences between the human cultured and post‐operative lens epithelial cells (LECs) on lens capsules (LCs) for a better understanding of posterior capsular opacification (PCO)—the most common complication of cataract surgery, and lens regeneration.Methods: The explants of the anterior portion of the LC containing the LECs, obtained from cataract surgery and cultivated under adherent conditions, as well as the post‐operative LC with LECs, were analysed by using synchrotron radiation‐based Fourier transform infrared (SR‐FTIR) microspectroscopy, a vibrational spectroscopic technique that allows monitoring of the entire biochemical status of the biological processes. The SR‐FTIR microspectroscopy setup installed on the beamline MIRAS at the Spanish synchrotron light source ALBA was used, where measurements were set to achieve single‐cell resolution, with high spectral stability and high photon flux.Results: We found that the differences exist between the composites of cultured and post‐operative LECs on LCs on the level of all chemical constituents: proteins, lipids, nucleic acids and carbohydrates as well as oxidative stress. The strongest differences are found in protein secondary structure contribution where the composite of primary LECs cultures on LC have more α‐helix (1652 cm−1) and post‐operative LC have more β‐sheet (1624 cm−1 and 1697 cm−1) secondary structures. The oxidative stress is more expressed in the composite of primary LECs cultures on LC and relatively less in the composite of post‐operative lens epithelium LECs on LC.Conclusions: Our results obtained by SR‐FTIR increase the knowledge about the total proteins, lipids, and nucleic acids in human primary cultures LECs while offering also evidence that SR‐FTIR is sensitive to the pathologic processes of LECs transdifferentiation. We showed that the composite of primary cultures LECs on LC have a distinct bio‐macromolecular composition compared to the composite of post‐operative LECs on LC, which gives additional information for a better understanding of PCO and lens regeneration.