Long Term Sustained Growth of Insulin Producing Cells Using Agarose-Chitosan Coated Silver Nanocomposites

Abstract

Diabetes is a group of diseases characterized by high levels of blood sugar for an extended period. Despite newer and effective therapy, current treatment is riddled with fundamental challenges. To overcome the adverse effects of existing drugs, regenerative medicine has emerged as an essential treatment, for which tissue engineering may serve as a foundation for the repair of pancreatic cells secreting insulin. Different polymeric scaffolds have been explored for pancreatic tissue engineering. In the current study, a continuation of our preceding work we attempt to test the role of previously synthesized agarose-chitosan coated silver nanocomposite scaffold (AG-CHNp) for the long-term growth of pancreatic cells. Pancreatic cells were isolated from BALB/c mice and were characterized by dithizone (DTZ) staining, real time polymerase chain reaction (RT-PCR), western blotting, and flow cytometry for characteristic pancreatic markers. The isolated population of cells was grown on scaffolds and its effectiveness towards insulin secretion was studied. The isolated population was found to be positive for glucagon, PDX-1 and Pax-4, while a 200-fold change transcript level of insulin was observed. The cells upon seeding on the scaffolds exhibited sustained growth and insulin secretion as confirmed by western blotting. Overall, the study demonstrates the suitability and application of AG-CHNp for pancreatic tissue engineering.