Abstract

BackgroundAlthough hemodialysis is a highly effective treatment for diffusive clearance of low molecular weight uremic toxins, its effect on circulating extracellular vesicles and submicron particles is less clear. The purpose of this study was to examine the impact of hemodialysis on circulating levels of submicron particles.MethodsPlasma samples from patients were collected immediately before and after the mid-week hemodialysis session. Total submicron particles were assessed by nanoparticle tracking analysis and levels of endothelial (CD144+), platelet (CD41+), leukocyte (CD45+), and total (Annexin V+) membrane microparticles (MPs) were assessed by flow cytometry.ResultsTotal submicron particle number was significantly lower post-dialysis with reductions in particles < 40 nm, 40–100 nm, and 100–1000 nm in size. Circulating annexin V+ MPs, platelet MPs, leukocyte MPs, and endothelial MPs were all reduced following dialysis. Assessment of protein markers suggested that extracellular vesicles were not present in the dialysate, but rather adsorbed to the dialysis membrane.ConclusionsIn summary, hemodialysis is associated with reductions in circulating submicron particles including membrane MPs. Accordingly, there may be significant interdialytic variation in circulating submicron particles. Investigators interested in measuring extracellular vesicles in patients undergoing hemodialysis should therefore carefully consider the timing of biosampling.

Highlights

  • Hemodialysis is a highly effective treatment for diffusive clearance of low molecular weight uremic toxins, its effect on circulating extracellular vesicles and submicron particles is less clear

  • Assessment of submicron particles by nanoparticle tracking analysis In order to assess the effect of hemodialysis on extracellular vesicles and submicron particles, we examined pre and post-dialysis plasma samples by nanoparticle tracking analysis

  • Effects of hemodialysis on circulating large Extracellular vesicle (EV)/ microparticles To further assess the effect of hemodialysis on circulating EV levels, we performed flow cytometry analysis of MPs in plasma samples collected pre and post-dialysis

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Summary

Introduction

Hemodialysis is a highly effective treatment for diffusive clearance of low molecular weight uremic toxins, its effect on circulating extracellular vesicles and submicron particles is less clear. EVs are membrane-enclosed vesicles that are formed by all cells under physiological and pathophysiological conditions. Most studies do not directly assess the biogenesis of EVs, exosomes are conventionally described as arising from endosomal sorting processes and extracellular release following fusion of multivesicular bodies with the plasma membrane. In the context of chronic kidney disease circulating MPs arising from platelets, endothelial cells, leukocytes, or erythrocytes are increased and levels have been shown to correlate with measures of vascular injury [1,2,3,4,5,6,7,8]. Urinary exosomes have been extensively studied in chronic kidney disease (reviewed in [10]), information regarding levels of circulating small EVs in this condition is lacking

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