Effects of Fibrin Glue as a Three-dimensional Scaffold on Cultivated Adult Human Retinal Pigment Epithelial Cells

Purpose: To examine morphologic, genotypic and phenotypic alterations of cultured adult human retinal pigment epithelial (ahRPE) cells when encapsulated with different concentrations of fibrin glue.Methods: Cultivated ahRPE cells were encapsulated with different concentrations of fibrin Glue: FG1 (42mg / dl), FG2 (84 mg /dl ), FG3 (124 mg /dl), and FG4 (210 mg /dl), and then evaluated for cytomorphological changes, , and gene and protein expression profiles.Results: Cultured ahRPE cells demonstrated a dendritiform morphology in early days of encapsulation with FG. Moreover, an increasing inhibitory effect on cell growth was observed with increased concentration of FG. At the transcriptional level, gene expressions of MMP-2, PAX6, and integrin B1 in FG1-encapsulated cells were significantly higher than other treated groups and the expression of SMA gene was decreased in all FG-encapsulated groups as compared with the controls. FG2-encapsulated cells expressed cytokeratin 8.18, RPE65, and ZO-1 proteins on immunocytochemistry but no PAX6 expression.Conclusion: Autologous fibrin glue with the concentration of 84 mg /dl provides proper encapsulation of RPE cells, while preserving morphometric, genotypic, and phenotypic features of the cells. This 3D biopolymer can be considered as a reliable vehicle for RPE cells transplantation and further cell adhesion to Bruch s membrane