The Effect of the Erythrocyte Suspension Temperature on the Morphology and Nanostructure of Cell Membranes

The problem of unintentional hypothermia in the postoperative period is still an urgent one.

The purpose of the work: to assess the effect of the in vitro temperature on the morphology and nanostructure of erythrocyte membranes.

Material and methods. 4 containers with erythrocyte suspensions with different blood groups were used; the suspension was packed in sealed containers (400 mL) with CPD blood preservative. The erythrocyte suspension was kept at 4°С. Samples (15 ml) were placed in a thermostat at 20°C and 37°C; then they were placed on the BioRS-24 Mini-Rotator Bioscan (EU) rotator (6—8 rpm). The analysis of the acid-base balance was carried out using the I-510 ionometric converter (Russian Federation). The morphology and nanostructure of erythrocyte membranes were analyzed using an atomic force microscope (AFM) «NTEGRA Prima» (NT-MDT, Russian Federation), in a contact mode, on monolayers prepared using a method of sedimentation in liquid and air.

Results. Discocytes were present in monolayers of blood after 1-hour and 12-hour rotation (12±3%) on Day 19 of ES storage at temperature 20°С. The number of echinocytes was 14±2% after a 1-hour rotation and 40±7% after a 12%hour rotation. The number of spheroechinocytes was 74±2% after a 1-hour rotation and 42± 8% after a 12-hour rotation. At a temperature of 37°С, after a 1-hour rotation, four forms of erythrocytes were present in a monolayer: discocytes, echinocytes, spheroechinocytes, and ovalocytes. The distribution of cell forms changed after a 12-hour rotation, and the cells recovered their shapes to discocytes (97±2%).

Conclusion. The obtained findings confirm the effect of cooling during erythrocyte preservation. The shape of erythrocytes was changed. The morphological structure of frozen erythrocytes recovers slowly, within 12 hours. It means that the transfusion of red blood cells whose temperature is below 37°C may not be effective for at least 12 hours and the transfusion medium should be warmed to reduce the risk of transfusion of blood products. 

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