Construction and Optimization of the Fibrin Scaffold Fortified by Platelet lysate and investigating its effect on Cultivated 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 Scaffold Fortified by Platelet lysate.Methods: ahRPE cells were encapsulated with fibrin Glue fortified by different concentrations of platelets that were obtained from platelet-rich plasma. The mixtures includedMIX1= 150000plt/ul, MIX2= 240000plt/ul, MIX3= 300000 plt/ul, MIX4=390000 plt/ul, MIX5= 480000 plt/ul, and MIX6= 600000 plt/ul. The encapsulated cells were then evaluated for cytomorphological changes, gene and protein expression profiles.Results: Cultured ahRPE cells demonstrated dendritiform and epithelioid morphology in all mixtures. The results of Viability assay on the third day indicate a growth rate less than control in all mixtures, and this decrease in MIX1 than the control is significant. (P = 0.0001). However, with time and on the seventh day, there is no significant difference between the mixtures and the control. At the transcriptional level, gene expressions of PAX6, and RPE65 in MIX1 and MIX2-encapsulated cells were significantly higher than other treated groups and the expression of αSMA gene was decreased in all mixtures as compared with the controls. Moreover, all mixtures encapsulated cells expressed cytokeratin 8.18, RPE65protein while just MIX1 and MIX2 expressed PAX6 proteins on immunocytochemistry. Conclusion: Based on this study, fibrin glue fortified by platelet rich plasma especially MIX1 and MIX2 provide an appropriate scaffold for the preservation of identity of the cultivated human RPE cells.