“The role of growth medium in the potency of EPC-type 2 diabetes patients” (A study of ALDHbright as a marker of the potency of EPC)

Abstract

Background & Aim Background: Type 2 diabetes mellitus (T2DM) is a metabolic disease caused by insulin disruption, insulin secretion or both. Hyperglycemia occurs in diabetes, where long-term hyperglycemia causes an increase in oxidative stress, a decrease in antioxidants and an increase in pro-inflammatory factors that contribute to endothelial dysfunction. In these conditions where microenvironment is not right, the amount of available ATP is not adequate, causing a decrease in the number and potential of Endothelial Progenitor Cells (EPC) in playing its function and role in the decreased colonization process. In this preliminary study, the aim was to prove changes in microenvironment by adding growth media to improve the condition of the microenvironment (niche) in an in-vitro able to increase the number and potential of EPC Methods, Results & Conclusion Method: Circulating EPC counts were calculated using a combination of expressions from CD34+ and CD133+ antigen surfaces using BD FACS Canto II flow cytometry, and the potential of EPC was done by adding ALDH bright staining. Then the EPC was isolated and cultured on the growth medium, then cultured for seven days. After that, the number of EPC circulating and the EPC potential is recalculated Result The results of this study indicate that after the addition of growth medium and incubated for seven days, the number of EPCs increased by 413.20%, and 1,401.1% in the controlled and uncontrolled group. Also, the potential of EPC showed by express of ALDHbright was obtained at the end of incubation of 99.9%, in each group in diabetes. Discussion This study illustrates that by adding a growth medium, it can restore microenvironment conditions to be better characterised by the ability of EPC to express bright ALDH which increases very high. Bright ALDH activity shows the ability to re-populating and differentiation and illustrates the operation of retinoids that have a pro-angiogenic effect with modulation of endothelial cell cytokines, the impact of migration and vasculogenesis. Conclusion The addition of growth medium in EPC culture from peripheral blood improves microenviral conditions and can increase the amount of EPC and the potential of EPC associated with its role in the colonisation function.