Morphology, membrane nanostructure and stiffness for quality assessment of packed red blood cells

Abstract

Transfusion of packed red blood cells (PRBC) to patients in critical states is often accompanied by post-transfusion complications. This may be related with disturbance of properties of PRBC and their membranes during long-term storage in the hemopreservative solution. The purpose of our work is the study of transformation of morphology, membranes stiffness and nanostructure for assessment of PRBC quality, in vitro. By atomic force microscopy we studied the transformation of cell morphology, the appearance of topological nanodefects of membranes and by atomic force spectroscopy studied the change of membrane stiffness during 40 days of storage of PRBC. It was shown that there is a transition period (20–26 days), in which we observed an increase in the Young’s modulus of the membranes 1.6–2 times and transition of cells into irreversible forms. This process was preceded by the appearance of topological nanodefects of membranes. These parameters can be used for quality assessment of PRBC and for improvement of transfusion rules.

Insights into the morphological pattern of erythrocytes' aging: Coupling quantitative AFM data to microcalorimetry and Raman spectroscopy

“…Indeed, the ATP content is expected to influence strength and dynamics of the interaction between cytoskeleton and plasma membranes, and the roughness measurements allow its quantification. It is also worth noting that similar effects, namely, the appearance of topological defects on the surface over the ageing pattern, have been evidenced in blood exposed to hemin or in long‐term storage conditions . In the present case, because of the different environmental stimulus, the defects shown in Figure can have a quite different molecular origin (eg lesser involvement of Hb and stronger correlation to cytoskeleton degradation).…”

Atomic force microscopy study of red blood cell membrane nanostructure during oxidation‐reduction processes

“…In our earlier works, we described that irreversible changes in cell shape and changes in the membrane nanosurface of RBCs are associated with distortion of the spectrin matrix. It is suggested that the influences of various pharmaceuticals, long‐term storage of blood, ionizing radiation, and various diseases lead to irreversible changes in the spectrin network …”

“…The sedimentation method was used to prepare the RBC monolayer . To do this, 400 μL of RBC suspension were dropped onto glass with polylysine and left for 20 minutes at a temperature of 20°C.…”

“…The sedimentation method was used to prepare the RBC monolayer. 4,14 To do this, 400 μL of RBC suspension were dropped onto glass with polylysine and left for 20 minutes at a temperature of 20°C. Then, the glass was lowered into a solution of 1% glutaraldehyde for 20 seconds, washed in buffer, and dried in air.…”

“…Two ways of ROS influence: spectrin oxidation and lipid peroxidation (2) haemoglobin oxidation (3). Change of the spectrin network and local topological defects arising in the RBC membrane (4). Scheme of iron oxidation in the haemoglobin molecule and methaemoglobin arising (5).…”