Advanced glycation end products in Descemet's membrane and their effect on corneal endothelial cell.

Purpose. The purpose of this study was to evaluate the effect of advanced glycation end products (AGEs) in Descemet’s membrane on the attachment and spreading of the corneal endothelial cells. Methods. An anti-AGEs monoclonal antibody (6D12), which recognizes a N e -carboxymethyl lysine (CML)-protein adduct as an epitope, was used for immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). Fresh bovine Descemet’s membrane was incubated for 4 weeks in the buffered solution with 500 mM of glucose-6-phosphate (G-6-P). In the incubated Descemet’s membrane, the immunohistochemical localization of CML was examined. Type I collagen-, type IV collagen-, fibronectin-, or laminin-coated 96-well plates were glycated by G-6-P. The amount of CML was determined by ELISA using 6D12. Cultured bovine corneal endothelial cells were seeded onto glycated or non-glycated extracellular matrix (ECM) in 96-well plates and allowed to attach for 3 hours. The number and the surface area of the attached cells were examin...