Evaluation of mechanical stability of dog erythrocytes under the influence of cryoprotectants

The mechanical stability of erythrocytes is a critical factor in ensuring their effective functioning during storage, transportation, and cryopreservation. The objective of this study was to ascertain the impact of diverse cryoprotectants, including glycerol, sucrose, dimethyl sulfoxide (DMSO), polyethylene glycol-1500 (PEG-1500), and hydroxyethyl starch (HES), on hemolytic damage to dog erythrocytes subjected to mechanical stress. For this purpose, dog erythrocytes were incubated in varying concentrations of cryoprotectants and NaCl. The cells were subjected to mechanical stress by stirring the suspension in a container filled with plastic beads at room temperature. The resulting hemolysis was evaluated spectrophotometrically. The results demonstrated that the most pronounced stabilization of erythrocyte membranes was observed during incubation with PEG-1500 and HES, while high glycerol concentrations caused membrane destabilization. Sucrose demonstrated a dual effect: at low concentrations, it exhibited protective properties for cellular membranes, while at higher concentrations, enhanced membrane vulnerability to stress. The results demonstrated that DMSO at all studied concentrations did not significantly change the mechanical stability of erythrocytes compared to the control group. Our findings indicate that an increase in salt concentration in the extracellular medium is associated with a reduction in the mechanical stability of dog erythrocytes. The effect of cryoprotectants on the mechanical stability of erythrocytes was found to be closely related to their physicochemical properties. This highlights the importance of precise selection of cryoprotectant concentrations to improve the results of red blood cell storage and transportation. The conclusions of this study are important for further optimization of technologies for the long-term storage of canine erythrocytes, in particular in cryobanks