The Role of Ozonized Erythrocytic Mass Transfusion in the Restoration of Myocardial Morphological Changes during Blood Loss (Experimental Study)

Purpose.To evaluate the role of ozonized erythrocytic mass transfusion in the restoration of damages in the architectonics of myocardium microvasculature and cardiomyocytes in case of a severe blood loss.Materials and Methods. Two batches of experiments were conducted, 17 white outbred rats in each. The animals were anesthetized with thiopental sodium (25 mg/kg). Blood loss was caused by taking 3ml of blood from the tail artery of rats, which is 35% of the circulating blood volume. One hour after the blood loss, transfusion of autoerythrocytes with normal saline solution and Ringer’s solution infusion in 1:1 ratio was performed in the control batch. In the experimental batch, 3 ml of autoerythrocytes treated with ozonized saline solution with ozone concentration of 2 mg/l and 3 ml of Ringer’s solution (1:1 ratio) were administered to rats. Erythrocyte mass was prepared from 3 ml of the autoblood harvested from the animals 3 days before the experiment. On a post-transfusion day5, in both batches and in five intact animals, hearts were removed following the intraperitoneal thiopental sodium injection (100 mg/kg). The left ventricle wall specimen from each heart was removed for examination. Histologic sections were stained with hematoxylin and eosin. The preparations were visually examined and morphometric studies were performed using microvisor Vizo-103.Results.It has been established that administration of ozonized erythrocyte for severe blood loss correction limited the decrease in numbers of capillary profiles and their diameters, formation of mixed and hyaline thrombi fully or partially occluding microvascular lumens, major hemorrhages and reduction of variation of nuclear profiles, decreased perivascular, pericellular, perinuclear, and endonuclear edema of myocardial tissue, cardiomyocyte overcontruction zone, and their ruptures.Conclusion.The positive trends for microcirculation indices, vascularization density and myocardial edematization as prognostic markers in assessing potential posthypoxic rehabilitation of damaged tissues upon blood loss correction with ozonized erythrocyte mass might be explained by the antihypoxic, antioxidant, and detoxifying actions of ozone on erythrocytes and/or its metabolites (ozonides) in the body post-transfusion.

Application of blood salvaging technologies in cardiosurgical patients operated under cardiopulmonary bypass

Aim. To evaluate the effectiveness of blood salvaging technologies in cardiosurgical patients operated under cardiopulmonary circulation. Methods. Analysis of perfusion protocols and transfusion cards of cardiosurgical patients operated using cardiopulmonary bypass during the period 2010-2011. Two groups of patients were identified: the first group - with the application of blood salvaging technologies (906 patients), the second group - without the application of this technique (122 patients). Blood from the operative wound was collected during the operation, processed, filtered through the «Cell-saver» machine from «Fresenius» company and was then re-introduced into the patient’s bloodstream. After completion of cardiopulmonary bypass the blood remaining in the circuit of the cell-saver machine and blood that was discharged through the drainage tubes was returned to the patient after being processed. Results. In the first group at the end of the operation 314.6±28.6 ml of washed red blood cells were returned to the patient from the heart-lung machine, while during the first postoperative day, the amount of the returned autologous erythrocytes from the drainage tubes was 72.8±12.5 ml. In this group, only 45 (4.9%) patients required donor erythrocyte mass transfusion, its volume per patient amounted to 172.3±31.8 ml. In the first group 182 (20.1%) patients required transfusion of fresh frozen plasma with the aim of achieving hemostasis, its volume amounted to 425±51.3 ml. In the second group 51 (41.8%) patients received erythrocyte mass transfusion in the volume of 346.7±31.1 ml, and 86 (70.5%) - received fresh frozen plasma in the amount of 568.7±41.2 ml. Hemoglobin content and hematocrit at the end of the operation and on the first postoperative day were significantly higher in the first group compared to the second group (p 0.01). These changes were associated with both intraoperative bleeding and with the loss of blood through the drainage tubes on the first postoperative day in the second group of patients. Compensation for these losses required transfusion of blood components significantly more frequently (p 0.001) and in larger volumes (p 0.01) than in the first group. Conclusion. Blood salvaging technologies using the «Cell-saver» machine during cardiac surgery under cardiopulmonary bypass significantly reduce blood loss, reduce the number of postoperative hospital days and complications, and are less expensive.

Stored erythrocytes have less capacity than normal erythrocytes to support primary haemostasis

SummaryPrimary haemostasis is mediated by platelet aggregation. Red blood cells (RBCs) are involved in this process. We hypothesised that stored RBCs could have less capacity to support primary haemostasis. This was tested with RBC units from 17 healthy volunteers stored for 45 days. Fresh citrated blood was taken again from the same donors and platelet-rich plasma was prepared, in which RBCs were resuspended with a constant haematocrit (40%), but changing fractions of stored versus fresh auto-logous RBCs (0, 25, 50, 75, and 100%, respectively). A platelet function analyser PFA-100® was used. In this instrument blood is aspirated through a membrane pore coated with collagen and either epinephrine (EPI) or ADP, which causes platelets to adhere, aggregate, and form an occluding plug, which stops blood flow and is measured as closure time (CT). We found that the CT increased with increasing fractions of stored blood. CT-EPI was 121 ± 17 seconds [s], 129 ± 32 s, 164 ± 45 s (p< 0.000, ANOVA), 214 ± 54 s (p< 0.0001), and 273 ± 36 s (p< 0.0001) for 0, 25, 50, 75, and 100% stored RBCs. For CT-ADP the values were 91 ± 22 s, 95 ± 12 s, 101 ± 13 s, 124 ± 44 s (p= 0.004), and 191 ± 72 s (p< 0.0001), respectively. We conclude that stored RBCs have less capacity than normal RBCs to support primary haemostasis by platelet aggregation in vitro, suggesting a decreased capacity of stored RBCs to bring platelets into close contact with the wall, which may contribute to sustained bleeding seen after mass transfusion.