Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCA026

Poster Communications

Sevoflurane increases plasma levels and angiogenic properties of human endothelial progenitor cells

A. Vlad2,1, L. Niculescu3, C. Stancu3, M. Popescu2,1, I. Stanca1, D. Corneci2,1, L. Ceafalan2, M. Gilca2, A. Popescu2,1, D. Dimulescu2,1

1. Elias University Emergency Hospital, Bucharest, Romania. 2. Carol Davila University of Medicine and Pharmacy, Bucharest, Romania. 3. Nicolae Simionescu Insititute of Cellular Biology and Pathology, Bucharest, Romania.

Plasma levels and functional properties of endothelial progenitor cells (EPCs) correlate to the efficacy of vessel expansion and repair after ischemia, whereas a consistent body of basic research suggest stimulating effects of anesthetic preconditioning (APC) on stem cells behavior. Therefore, we tested the ability of sevoflurane to increase the mobilization and to potentiate the angiogenic properties of these cells. EPCs levels were evaluated by flow-cytometry and in vitro culture assay before exposure to sevoflurane (baseline) and 24 hrs afterwards, in blood samples collected from CAD patients. Sevoflurane (1 MAC in 50% O2) was administered on a facial mask in a discontinuous manner (2 cycles of 15 min interspersed by 10 min washout; n = 16), while control subjects received only oxygen (n = 10). The levels of stromal derived factor 1 alpha (SDF1alpha) and vascular endothelial growth factor (VEGF), two chemokines involved in EPCs mobilization and homing, were assessed by ELISA in plasma samples at baseline, 4 and 24 hrs after sevoflurane administration. 7-days old cultured EPCs derived from human cord blood mononuclear cells were exposed in vitro to sevoflurane 2% or 4% in air/5% CO2, or only to air/5% CO2 (control) for 2 hrs in a modular chamber. The incorporation of preconditioned and control DilAcLDL-EPCs into tube-like structures of human umbilical vein endothelial cells (HUVEC) grown on Matrigel was evaluated 24 hrs later. Values are expressed as means ± SEM, compared by ANOVA. Plasma CD45dim/CD34+/KDR+, CD45dim/CD34+/KDR+/CD133+ and CD45dim/CD34+/CD309+/CXCR4+ mononuclear cell populations were increased in sevoflurane treated versus sham group at 24 hrs post-exposure (172.95 ± 73.74 vs. 153.84 ± 30.28, p = 0.006; 248.06 ± 37.4 vs. 198.62 ± 29.15, p < 0.001 and 153.87 ± 49.31 vs. 134.66 ± 37.95, p = 0.005 respectively, expressed as % at 24 hrs vs. baseline). Although not statistically significant, the number of adherent DilAcLDL+/FITC-UEAI+ cells in 7 days old EPCs cultures from preconditioned patients was slightly higher when compared to those of sham control subjects. SDF1alpha and VEGF levels were not significantly modified by sevoflurane. In the in vitro preconditioning setting, sevoflurane raised the number of incorporated DiIAcLDL-EPCs into tube-like HUVEC structures (n=3, p<0.01 for 4% sevoflurane) when compared to control. Augmented EPCs mobilization and ability of integrating themselves into vascular structures are candidate mechanisms underlying the regenerative and anti-ischemic processes of APC with sevoflurane. These results indicate a promising potential of preconditioning protocols for improving the endothelial and tissue repair following percutaneous coronary intervention or cardiovascular and non-cardiovascular surgery.

Where applicable, experiments conform with Society ethical requirements