Angiogenic effect of SHED-derived exosomes on hyperglycemia-induced endothelial cells

Aim or Purpose: This study aimed to investigate the angiogenesis of hyperglycemia-induced human umbilical vein endothelial cells (HUVEC) treated with SHED exosomes. Materials and Methods: SHED exosomes were extracted by differential centrifugation and then were characterized by nanoparticle tracking analysis, transmission electron microscope, and flow cytometry, for exosome size, ultrastructure, and for presence of exosome specific surface markers: CD9, CD63 and CD81, respectively. HUVEC was subjected to treatments with normal glucose level (5.5 mM), positive control group, and hyperglycemic level (25 mM). Afterwards, hyperglycemia-induced HUVEC was cultured with SHED exosomes. Through tube formation assay, reverse transcriptase polymerase chain reaction (RT-PCR) with GATA2, an angiogenic gene, and immunostaining of CD31, a vascular marker, the angiogenic potential of HUVEC was identified. Results: The characterization confirmed SHED exosomes: size ranged from 44-291 nm with mode of 133 nm, cup-shaped morphology and expressed CD9, CD63, and CD81. In comparison with the hyperglycemia-induced group, exosome treatment resulted in improved tube formation qualitatively, and exhibited statistically significantly (p<0.05) increased tube formation in the covered area, total branching points, total tube length and total loops. At 12 hours, there were improved expression of GATA2 and higher expression of CD31 in the exosome treated group in comparison to the hyperglycemia-induced group. Conclusions: SHED exosomes have the capability to enhance angiogenesis in hyperglycemia-induced endothelial cells. As a result, usage of SHED exosomes could offer a promising alternative for periodontal tissue regeneration for diabetic patients with periodontitis