**2.2 Scanning electron microscopy**

*Apolipoproteins, Triglycerides and Cholesterol*

development of plaque instability in acute MI [7].

advanced treatments [10].

disorder in hypertension patients.

**2.1 Preparation of blood Smear**

**2. Morphological characterization**

maintenance of the surface electrical charge and stability of biological cellular system [3]. Erythrocytes and Erythrocyte membrane are more vulnerable to peroxidation due to constant exposure to high oxygen tension and richness in polyunsaturated fatty acid, respectively [4]. The interaction of Reactive oxygen species (ROS) with particularly with fatty acids available in membrane can result in undesirable irreversible changes in cellular membrane. The membranes are therefore naturally protected by available anti-oxidative enzymes like superoxide dismutase, catalase, and glutathione peroxidase and vitamins E and A from oxidative damages [5]. By-products of peroxidation have been shown to cause profound alterations in the structural organization and functions of the cell membrane including decreased membrane fluidity, increased membrane permeability, inactivation of membrane-bound enzymes and loss of essential fatty acids [6]. Reactive oxygen species are the responsible moiety that are involved in the generation and progression of atherosclerosis and that contribute to the

Blood viscosity is strongly affected by the surface charge of RBCs and is responsible for the spacing between them. A higher surface charge causes repulsive forces to increases distant RBCs, preventing their close aggregation, lowers the viscosity, and results into very low peripheral resistance to flow. Therefore, it can be hypothesized that stress like condition develops in hypertension generate ROS which can induce membrane deformity and as a result affect the membrane surface charge. The occlusive arterial disease resulting from RBC aggregation may develop due to comparable membrane deformity [8]. Besides vascular and cardiac tissue integrity, blood and especially the RBC, is critical to performing this critical assignment of CV risk assessment

considering it makes up over 90% of formed elements within blood. In cardiovascular diseases, impaired blood rheology has been observed. A direct relationship has been found between increased RBC deformability and increased risk for arterial hypertension [9]. Accurate risk stratification of patients with chronic heart failure is critically important to efficiently target the use of evidence-based therapies and identify high-risk patients who may benefit from

We tested the hypothesis that variations in zeta potential and deformation of erythrocytes were associated with risk of adverse cardiovascular outcomes in a population with hypertension that were free of symptomatic heart failure [11]. In the present work we envisaged to study and evaluate morphological changes taking place in RBCs, erythrocyte fragility, lipid per oxidation and zeta potential which can act as invaluable aid in the diagnosis of a hypertension and risk of Cardiovascular Disorder in hypertension patients. Hence, the aim of this study was to test the hypothesis of association of variation in RBC morphology, erythrocyte fragility and zeta potential in hypertension and its relation with the risk of cardiovascular

Blood smear was prepared with the aid of wedge method [12]. In this method a drop of blood was placed on base closed to one cease of the slide at least 1 cm away from the edged of the slide. Another slide with the smooth end was used as spreader and smear was prepared by moving spreader inclined at 30–45° angle to the blood.

**96**

To observe the morphological variations in the erythrocyte membrane structure in Erythrocytes of patients suffering with hypertension and MI, erythrocytes were analyzed by scanning electron microscopy. With this motive blood sample was taken in Eppendorf tube containing 10 μl of Heparin (5000 UI/ml) in 900 μl of pH 7.4 phosphate buffer saline. The blood suspension was then centrifuged (1000 rpm for 10 min) and washed with buffer three times. The supernatant was removed and replaced by same volume of buffer. One drop of these separated erythrocytes were then exposed to 500 μl of 2.5% Glutaraldehyde in distilled water overnight at 4°C to fix. Again samples were washed thrice with distilled water and centrifuged. About 40 μl of each sample was placed on glass covered studs and air dried at room temperature. The Scanning electron microscopy (SEM) analysis of prepared samples was accomplished using Jeol, Japan (Model—JSM 5610LV).
