Abstract

During storage, red blood cells (RBCs) for transfusion purposes suffer progressive deterioration. Sialylated glycoproteins of the RBC membrane are responsible for a negatively charged surface which creates a repulsive electrical zeta potential. These charges help prevent the interaction between RBCs and other cells, and especially among each RBCs. Reports in the literature have stated that RBCs sialylated glycoproteins can be sensitive to enzymes released by leukocyte degranulation. Thus, the aim of this study was, by using an optical tweezers as a biomedical tool, to measure the zeta potential in standard RBCs units and in leukocyte reduced RBC units (collected in CPD-SAGM) during storage. Optical tweezers is a sensitive tool that uses light for measuring cell biophysical properties which are important for clinical and research purposes. This is the first study to analyze RBCs membrane charges during storage. In addition, we herein also measured the elasticity of RBCs also collected in CPD-SAGM. In conclusion, the zeta potential decreased 42% and cells were 134% less deformable at the end of storage. The zeta potential from leukodepleted units had a similar profile when compared to units stored without leukoreduction, indicating that leukocyte lyses were not responsible for the zeta potential decay. Flow cytometry measurements of reactive oxygen species suggested that this decay is due to membrane oxidative damages. These results show that measurements of zeta potentials provide new insights about RBCs storage lesion for transfusion purposes.

Highlights

  • red blood cells (RBCs) have sialylated glycoproteins which are responsible for a negatively charged membrane cell surface [1]

  • These results show that the presence of leukocytes enzymes is not responsible for the f decrease (p = 0.7 when days 8 and 15 for RBCs leukodepleted samples were compared to days 8 and 15 for RBCs non leukodepleted sample)

  • The results show that the f decay is not caused by the presence of leukocytes

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Summary

Introduction

RBCs have sialylated glycoproteins which are responsible for a negatively charged membrane cell surface [1]. In an electrolyte medium, such as in blood plasma, this induces the formation of a layer of surrounding medium ions of opposite charges rigidly bound around the cells, that creates a repulsive electrical zeta potential (f) between the RBCs. The f potential is an important property responsible to stabilize the RBCs colloidal suspension preventing cells to come too close and avoiding interactions between RBCs and other cells, and especially among themselves. Questions not answered yet are: RBCs membrane charges, the f potential, can change with storage time as well as the elasticity changes? During RBCs storage period, the leukocyte degranulation promotes enzyme release, which can reduce glycoprotein membrane expressions and probably can change the membrane electrical charges [9]. This raises the question if the leukocyte enzymes are capable to change the f potential. We measured RBCs elasticity, with an optical tweezers, and analyzed, by flow cytometry, the production of reactive oxygen species in standard stored RBCs units collected in the same preservative solution to correlate deformability with the f potential results

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