The research object was the influence of anionic composition of the medium on hypertonic lysis, hypertonic cryohemolysis and posthypertonic lysis of human erythrocytes.
The aim of this research was to study the effect of lyotropic anions (ClO4- < Br- < Сl- < Ac- < F- < SO42-) on the adaptation of human erythrocytes to the conditions simulating the cryoinjury factors during freeze-thawing processes (hypertonic and posthypertonic shocks, hypertonic cryohemolysis).
Research methods were as follows: cryobiological (modeling of cryoinjury factors), spectrophotometric, microhematocritic, ionometric, osmometric, biophysical, light microscopy using a thermal assembly and statistical analysis of experimental data.
It was found that the position of the anion in the lyotropic series affected the level of sensitivity and morphology of cells by changing the hydration of their structures. According to the effect on cell damage under hypertonic conditions, the anions can be categorized in five types: strong (ClO4-, SCN-) and weak (Br-) chaotropic, lyotropic neutral (Cl-), as well as weak (Ac-) and strong (F-, SO42-) kosmotropic. Strong lyotropic effects of ions sensitize cells in hypertonic conditions. Weak kosmotropic Ac- protects erythrocytes during the hypertonic shock, especially at 900 mOsmol/kg, 0 °C and pH 7.4. During hypertonic cryo-hemolysis, weak chaotropic Br- and Ac- reduced the damage at prolonged exposure (60–120 min). In the course of rehydration after hypertension, the minimum level of hemolysis was observed in the presence of SO42- anions.
Lyotropic anions can be used to regulate cell adaptation under hypothermic storage.
For the first time, the influence of anions of the lyotropic series on adaptive behavior of human erythrocytes when simulating the main factors of cryoinjury was comprehensively studied. For the first time, the regularities of adaptation of cells to the osmolality shift and the medium temperature were established in the presence of chaotic and cosmotropic anions.
For the first time, the relationship between hypertonic lysis and hypertonic cryohemolysis has been identified. It consists in sensitization of erythrocytes to both types of stress after preliminary incubation in media of the same osmolality, individual for each type of ion. This critical osmolality varies in a "bell-shaped" manner according to the location of the anions in the lyotropic series, as well as the degree of hydration of cells in the presence of lyotropic anions. The phases of development of hypertonic cryohemolysis in time are most accurately manifested in the media with critical osmolality, which is individual for each lyotropic anion.
The increase in adaptive plasticity of cells under conditions of hypertonic cryohemolysis in the presence of weakly chaotropic Br-, which occurs with no membrane damage during prolonged pre-incubation, has been also demonstrated for the first time. For the first time, the nature of the protective effect of weakly cosmotropic Ac- was explained by increasing the degree of hydration of cells. Rise of osmotic sensitivity of erythrocytes to low pH (5.4) in the presence of strong chaotropic anions has been shown for the first time. The temperature effect on the temperature-osmotic adaptation of cells in the presence of chaotic and cosmotropic anion has been clarified. The decrease in the level of posthypertonic lysis of erythrocytes with increasing cosmotropic properties of anions, which correlates with the peculiarities of changes in morphological and volumetric characteristics of erythrocytes was for the first time established. In hypertensive lysis, the antihemolytic activity of orthovanadate nanoparticles of various erythrocyte shapes was evaluated for the first time.
The performed scientific studies substantiate the expediency of using the lyotropic anions effect to improve the cells adaptation to the freeze-thawing conditions. The scientific results obtained in this research will allow the selection of new cryoprotective agents and the creation of effective cryoprotective media. The identified patterns of influence of anions can be used for a more detailed investigation of the cryopreservation mechanisms and the development of approaches to protect the cells.
The findings can be recommended to be used in the educational and learning process for the training of specialists in the field of cryobiology, pharmacology, physiology and biophysics.
