Abstract Body: Introduction:
Diabetes is associated with impaired wound healing. Studies have demonstrated that endothelial progenitor cells (EPCs) and mature endothelial cells (EC) are susceptible to apoptosis in hyperglycemic environment and p53 gene silencing prevents cellular senescence and helps vascular regeneration. Previously we have shown that p53 silenced progenitor cell therapy helped tissue regeneration. Our goal was to test whether exosomes from conditioned media obtained from p53 silenced and SOD2 overexpressed human bone marrow derived stem cells (BM-MSC), done separately, can rescue endothelial damage in presence of hyperglycemia. Methods: Ad-human-P53 (TP53)-shRNA is used to silence P53, Ad-humanSOD2 is used to overexpress SOD2 and Ad-scrambled-null-shRNA was used as control in BM-MSCs. Exosomes were isolated from the condition medium(CM) collected from BM-MSCs transduced with either Ad-P53HS or Ad-SOD2 or scrambled (three variables) as described in our previous publications.
Endothelial regeneration was tested using endothelial wound healing assay kit ( from Cell Biolabs # CBA-120-T). We compared exosomes isolated from null, p53sh BM-MSCs and SOD2 BM-MSCs on human endothelial cells in high glucose conditions. Results: Number of cells in the scratch area increased significantly in both the exosomal treated wells as compared to null and non-transduced wells in p53-sh exosomes and SOD2-BMMSC derived exosomes. Moreover post SOD2 exosomal therapy, hyperglycemia exposed endothelial cells showed increase in mitochondrial function as assessed by Seahorse. Conclusion: Both p53sh and SOD2 exosomal therapy further improved the proliferation of human endothelial cells in high glucose conditions and may have therapeutic role in treating endothelial dysfunction related complications in diabetes. We are further analysing the differences between the two possible therapies.