Study of the effect of exendin-4 on mesenchymal stem cells osteogenic differentiation / Sarah Adel Habib Sayed ; Supervised Shohda Assem Elmaraghy , Mohamed Mostafa Kamal , Mahmoud Ahmed Senousy

Thesis (M.Sc.) - Cairo University - Faculty of Pharmacy - Department of Biochemistry

Mesenchymal stem cells (MSCs) have been tested for their ability to repair damaged bone tissues and repairing bone defects. Exendin-4 utilization increased the osteoblastic activity upon increasing its concentration and time of exposure and this improved bone mass and quality.The balance between bone formation and bone resorption is maintained by a balance between Receptor activator nuclear factor kappa beta(RANK), Receptor activator nuclear factor kappa beta ligand (RANKL) and Osteoprotegerin (OPG). However, little is known about this important axis in the osteogenic differentiation of MSCs.The purpose of this study was to investigate the effect of exendin-4 on the osteogenic differentiation of rat adipose tissue (Ad)-MSCs and examine if exendin-4 utilization can enhance the osteoblastic activity through augmenting the secretion of the OPG by Ad-MSCs.Isolation, characterization and differentiation of Ad-MSCs into osteocytes were examined with and without exendin-4. The effect of exendin-4 addition was detected by determination of the expression of osteogenic markers using RT-qPCR and westernblot.In addition, we examined its effect on the RANK/RANKL/OPG axis by determination of RANK, RANKL by western blot. While, the secreted OPG was determined in the conditional media of exendin-4 differentiated cells by commercially available ELISA kit.This study demonstrated successful isolation, characterization and differentiation of rat epididymal fat derived Ad-MSCs, into osteocytes with and without exendin-4 supplementation. The osteogenic differentiated cells stained with Alizarin-Red, with increased staining intensity upon exendin-4 addition

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