Saul Yedgar

Blood banking–induced alteration of red blood cell flow properties

BACKGROUND: Blood banking procedures are associated with damage to red blood cell (RBC) membranes, which can impair their flow properties, namely, their deformability, aggregability, and adherence to endothelial cells (ECs) and thus possibly introducing a circulatory risk to recipients. This study was undertaken to comprehensively explore the effect of cold storage and gamma irradiation on RBC flow properties.

Alteration of red cell aggregability and shape during blood storage.

BACKGROUND: Storage of blood units (for 35–42 days, depending on the preservative solution) has been reported to induce changes (e.g., reduction of sialic acid level) in red cells that are expected to alter their aggregability.

The red blood cell in vascular occlusion.

Red blood cells (RBC) have unique flow-affecting properties--namely, aggregability, deformability and adherence to endothelial cells (EC)--which play major roles in blood flow. Under normal flow-induced shear stress RBC are dispersed, their adherence to EC is insignificant, and they are sufficiently deformable to enable tissue perfusion. However, in pathological conditions that are associated with low-flow states (e.g., trauma, ischemia), elevated plasma components (mainly fibrinogen), or altered RBC properties (e.g., hemoglobinopathies, oxidative stress, inflammation, diabetes), RBC flow properties are altered and present a circulatory risk.

Monitoring of erythrocyte aggregate morphology under flow by computerized image analysis

The morphology of red blood cell (RBC) aggregates was studied by direct visualization of RBC aggregation at different flow conditions in a computerized image analyzer. The aggregate morphology is expressed by an Aggregate Shape Parameter (ASP), defined as the ratio of the aggregate projected area to its square perimeter. Aggregation was induced by either dextran-70 (m.w. 70,000) or dextran-500 (m.w. 500,000), and compared to that in plasma. It was found that the aggregate morphology is a characteristic of the aggregating agent--in dextran-500, the RBC form rouleau aggregates as in plasma, while in dextran-70, they form clusters. In each system, while maintaining the overall typical morphology, the ASP decreases (i.e., the aggregate becomes longer) as the aggregate size is increased. The distribution of the ASP as a function of the aggregate size remains unchanged when the aggregate size is changed by modulation of the dextran concentration or the shear stress. Stretching of a rouleau aggregate by application of shear stress is reflected by a corresponding change in the ASP. It is suggested that the ASP is a characteristic of intercellular interactions. A theoretical model is proposed for evaluation of the deviation of aggregate shape from that of rouleau structure.

H2O2 renders cells accessible to lysis by exogenous phospholipase A2: a novel mechanism for cell damage in inflammatory processes

Phospholipase A2 (PLA2) and H2O2, secreted from activated inflammatory cells, play a central role in the tissue damage occurring in inflammatory processes. However, while exogenous PLA2 alone does not cause cell lysis, it readily does so when acting with H2O2. We have found that H2O2 degrades cell surface proteoglycans, thus rendering the membrane PL accessible to hydrolysis by exogenous PLA2. This novel mechanism introduces a role for cell surface proteoglycans in protection of cells from damage by pro‐inflammatory agents, and may assign a central role for the combined action of H2O2 and PLA2 in inflammatory and bacteriocidal processes.

Extracellular phospholipase A2 inhibitors suppress central nervous system inflammation.

Phospholipase A2 (PLA2) plays a key role in the production of proinflammatory mediators, namely the arachidonic acid‐derived eicosanoids, lysophospholipids, and platelet‐activating factor, and indirectly influences the generation of cytokines, nitric oxide (NO), and free radicals. Accordingly, regulation of its activity is important in the treatment of inflammation. Since the main site of PLA2 action in inflammatory processes is the cell membrane, we synthesized extracellular PLA2 inhibitors (ExPLIs) composed of N‐derivatized phosphatidyl‐ethanolamine linked to polymeric carriers. These membrane‐anchored lipid conjugates do not penetrate the cell and interfere with vital phospholipid metabolism or cell viability. The ExPLIs markedly inhibited central nervous system inflammation. This was reflected by the suppressed production and secretion of lipopolysaccharide‐induced sPLA2, prostaglandin E2, and NO by glial cells and by the amelioration of experimental autoimmune encephalomyelitis in rats and mice.

Role of red blood cell flow behavior in hemodynamics and hemostasis

The primary role of red blood cells (RBCs) is to transport oxygen to the tissues, which is performed predominantly in the blood capillaries. However, RBCs have unique flow-affecting properties that play a key role in blood flow in all blood vessel types and sizes. While RBCs as oxygen carriers have been studied extensively, their hemodynamic function has been examined less comprehensively. This review aims to bridge this gap, focusing on the role of RBC flow properties in hemodynamics, hemostasis and thrombosis.

Rejuvenation treatment of stored red blood cells reverses storage-induced adhesion to vascular endothelial cells.

BACKGROUND: Blood banking procedures are associated with elevated adherence of red blood cells (RBCs) to blood vessel wall endothelial cells (ECs), which can introduce a circulatory risk to recipients. This study was undertaken to examine the possibility of repairing this damage by a poststorage “rejuvenation” procedure before transfusion.

Effect of viscous macromolecules on peritoneal plasminogen activator activity: A potential mechanism for their ability to reduce postoperative adhesion formation

Activity of peritoneal plasminogen activator and its regulation by dextran and other macromolecules that clinically suppress postoperative adhesions was studied. Plasminogen activator activity was assayed by a two-stage globinolytic assay that monitors formation of plasmin, as well as by cleavage of a chromogenic peptide substrate (S-2444) in the presence of aprotinin (Trasylol). Plasminogen activator activity was located on the outer surface of human peritoneum. Incubation of peritoneal tissue with buffer in vitro (conditioning) prompted release of plasminogen activator into the conditioning medium. The released plasminogen activator formed a single band on sodium dodecyl sulfate-gel electrophoresis at an apparent molecular weight of 174,000 and was markedly suppressed by antiserum raised against human melanoma tissue-type plasminogen activator. Nonspecific proteolytic activity did not accumulate in the medium during conditioning. The presence of dextran 80 during conditioning of peritoneum reversibly suppressed tissue-bound plasminogen activator activity and reduced plasminogen activator activity in the spent medium. A similar inhibition of peritoneal plasminogen activator was induced by dextran 500, methyl cellulose, and polyvinylpyrrolidone. Dextran, when added to the medium after conditioning, had no direct inhibitory effect on plasminogen activator activity. Dextran did not induce peritoneal production of inhibitor(s) of trypsin, chymotrypsin, or urokinase. On the basis of these findings, two possible mechanisms for the effect of viscous polymers in the reduction of adhesion formation are proposed. These mechanisms consider the importance of peritoneal tissue-type plasminogen activator for removal of fibrin clots and suggest that polymer coating either prevents the shedding of plasminogen activator into the abdominal cavity or reduces the access of fibrin clots to the serosal surfaces.

Photodynamic Treatment of Red Blood Cell Concentrates For Virus Inactivation Enhances Red Blood Cell Aggregation: Protection with Antioxidants

Abstract— Photodynamic treatment (PDT) using phthalocyanines and red light appears to be a promising procedure for decontamination of red blood cell (RBC) concentrates for transfusion. A possible complication of this treatment may be induced aggregation of RBC. The production of RBC aggregates was measured with a novel computerized cell flow properties analyzer (CFA). The PDT of RBC concentrates with sulfonated aluminum phthalocy‐anine (AIPcS4) and the silicon phthalocyanine Pc 4 under virucidal conditions markedly enhanced RBC aggregation and higher shear stress was required to disperse these aggregates. The clusters of cells were huge and abnormally shaped, unlike the rouleaux formed by untreated RBC. This aggregation was prevented when a mixture of antioxidants was included during PDT. Addition of the antioxidants after PDT reduced aggregation only partially. It is concluded that inclusion of antioxidants during PDT of RBC concentrates prior to transfusion may reduce or eliminate the hemodynamic risk that the virucidal treatment may present to the recipient.