Red blood cells (RBCs) are highly deformable and possess a robust membrane that can withstand shear force. Previous research showed that in diabetic patients, there is a changed RBC ultrastructure, where these cells are elongated and twist around spontaneously formed fibrin fibers. These changes may impact erythrocyte function. Ultrastructural analysis of RBCs in inflammatory and degenerative diseases can no longer be ignored and should form a fundamental research tool in clinical studies. Consequently, we investigated the membrane roughness and ultrastructural changes in type 2 diabetes. Atomic force microscopy (AFM) was used to study membrane roughness and we correlate this with scanning electron microscopy (SEM) to compare results of both the techniques with the RBCs of healthy individuals. We show that the combined AFM and SEM analyses of RBCs give valuable information about the disease status of patients with diabetes. Effectiveness of treatment regimes on the integrity, cell shape and roughness of RBCs may be tracked, as this cell’s health status is crucial to the overall wellness of the diabetic patient.

/pdf/changes-in-red-blood-cell-membrane-structure-in-type-2-57h38p1oej.pdf

Changes in red blood cell membrane structure in type 2 diabetes: a scanning electron and atomic force microscopy study

Identifying the causative relationship between the fatty acid composition of cell membranes and type 2 diabetes mellitus fundamentally contributes to the understanding of the basic pathophysiological mechanisms of the disease. Important outcomes of the reviewed studies appear to support the hypotheses that the flexibility of a membrane determined by the ratio of (poly)unsaturated to saturated fatty acyl chains of its phospholipids influences the effectiveness of glucose transport by insulin-independent glucose transporters (GLUTs) and the insulin-dependent GLUT4, and from the prediabetic stage on a shift from unsaturated towards saturated fatty acyl chains of membrane phospholipids directly induces a decrease in glucose effectiveness and insulin sensitivity. In addition, it has become evident that a concomitant increase in stiffness of both plasma and erythrocyte membranes may decrease the microcirculatory flow, leading ultimately to tissue hypoxia, insufficient tissue nutrition, and diabetes-specific microvascular pathology. As to the etiology of type 2 diabetes mellitus, a revised hypothesis that attempts to accommodate the reviewed findings is presented.

Lipid Composition of Cell Membranes and Its Relevance in Type 2 Diabetes Mellitus

Most non-communicable diseases involve inflammatory changes in one or more vascular systems, and there is considerable evidence that unliganded iron plays major roles in this Most studies concentrate on biochemical changes, but there are important biophysical correlates Here we summarize recent microscopy-based observations to the effect that iron can have major effects on erythrocyte morphology, on erythrocyte deformability and on both fibrinogen polymerization and the consequent structure of the fibrin clots formed, each of which contributes significantly and negatively to such diseases We highlight in particular type 2 diabetes mellitus, ischemic thrombotic stroke, systemic lupus erythematosus, hereditary hemochromatosis and Alzheimer's disease, while recognizing that many other diseases have co-morbidities (and similar causes) Inflammatory biomarkers such as ferritin and fibrinogen are themselves inflammatory, creating a positive feedback that exacerbates disease progression The biophysical correlates we describe may provide novel, inexpensive and useful biomarkers of the therapeutic benefits of successful treatments

/pdf/diagnostic-morphology-biophysical-indicators-for-iron-driven-27bearvuhn.pdf

Diagnostic morphology: biophysical indicators for iron-driven inflammatory diseases

Blood rheology tests are traditionally used for detection of organic disease and for monitoring disease activity. More recently they have been used for prediction of blood flow in vivo, not only in overt hyperviscosity syndromes but also in the covert hyperviscosity of low-flow states. The traditional ESR test result increases with red cell aggregation induced by increases in large, asymmetrical plasma globulins. However, small increases in haematocrit and large increases in plasma viscosity each decrease the ESR, reducing both its diagnostic utility and its ability to predict blood flow in vivo. The ESR should be corrected to a standard haematocrit, or else replaced by the ZSR or plasma viscosity, which are more rapid, simple, sensitive and independent of haematocrit. For prediction of blood flow in vivo, these tests can be supplemented by measurement of whole-blood viscosity, which can be performed simply and cheaply in capillary viscometers at high shear rates. Whole-blood viscosity is determined by plasma viscosity, haematocrit and red cell deformability at high shear rates. Its measurement is useful in overt hyperviscosity syndromes, particularly in estimating the effect of red cell transfusion in anaemic patients with plasma hyperviscosity, hyperleukocytic leukaemias or sickling disorders. Blood viscosity should be related to the haematocrit or haemoglobin concentration in order to estimate oxygen delivery to tissues. Changes in blood viscosity can be compensated readily in the normal circulation but not in the compromised, low-flow circulation. In these circumstances, systemic increases in plasma viscosity, haematocrit, whole-blood viscosity, red cell aggregation and in the numbers of circulating rigid red or white blood cells can perpetuate low-flow states and ischaemia. Red cell deformability in narrow vessels is best measured by micropore filtration systems, in which the effect of white cells has been eliminated. Red cell deformability is reduced by change in shape, decrease in the ratio of surface area to volume, decreased membrane flexibility and increased internal viscosity (MCHC and inclusions). White cells have negligible effects on bulk-blood viscosity but have important effects on blood flow in narrow vessels, due to their high internal viscosity and their adhesiveness when activated. White cell filterability is lowest for monocytes and for activated granulocytes and these adhesive and rigid cells may have important effects on microcirculatory blood flow in low-flow states.

1 Blood rheology in vitro and in vivo

Erythrocyte deformability was assessed in 40 diabetic patients, 24 insulin-dependent (IDD) and 16 non-insulin-dependent (NIDD), by measuring the initial filtration flow rate of whole blood, isolated red blood cells (RBC), and isolated RBC membranes with the Hanss hemorheometer, and its relationship to the plasma and ghost membrane lipid composition was investigated. RBC deformability was significantly reduced, whereas the deformability of the isolated RBC membranes did not differ significantly from the controls. In the plasma, the triglycerides were high, the high-density lipoprotein (HDL) cholesterol was reduced, and the ratio of total cholesterol over HDL cholesterol was high as compared with the controls. The RBC lipid composition expressed in μmol lipids/10 10 RBC showed significantly lower levels of free cholesterol, sphingomyelines, and phosphatidylcholine, which are the lipids principally located on the outer layer of the RBC membranes. These data suggest that in both IDD and NIDD patients, there may be a relation between these modifications in the RBC lipid composition and rheological impairment of the RBC.

E. Delatour-Hanss

Papers

Erythrocyte deformability in diabetes and erythrocyte membrane lipid composition.

Physico-chemical factors of erythrocyte deformability.

Altered rheological properties of RBCs in hemoglobinopathies

Rheological properties of desialylated erythrocytes studied by ultrasound backscattering and filtration