Platelet adhesion to smooth and rough hydrophobic and hydrophilic surfaces under conditions of static exposure and laminar flow.
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Biomedical subjects
Publications and source records attributed to D R Absolom.
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By slowly lowering the surface tension of the aqueous medium through the admixture of 47.5% dimethyl sulfoxide (DMSO) and 0.1% bovine serum albumin (BSA) and by raising the pH to 9, complete elution of A, D, and K antibodies from sensitized erythrocytes (RBC) could be achieved. The eluates comprising the antibodies were subjected to dialysis to remove the DMSO and to neutralize the pH, and to ultrafiltration to remove the excess water. Recovery of a sizeable proportion of the eluted RBC (of blood groups A and K) proved possible by slow and careful removal of the DMSO with phosphate-buffered saline containing 2% BSA.
Surface tensions of normal and pathological granulocytes were determined by (1) adhesion to solid substrates of different surface tensions while suspended in liquid media of different surface tensions, and by (2) measurement of cell-liquid-vapor contact angles obtained with sessile drops of saline water on cell monolayers. The results obtained by the two different methods were in close conformation with one another. With the cell adhesion method some residual leukocyte adhesion still persists even under conditions where there no longer is a van der Waals attraction between cells and solid substrate. At low ionic strength and by the abolishment of all multivalent cations through the admixture of EDTA, that residual cell adhesion virtually disappears (with normal as well as with pathological granulocytes), indicating that the earlier residual cell adhesion did indeed arise from electrostatic interactions mediated by multivalent cations (probably Ca2+). Comparison of the capacities for engulfment and the surface thermodynamics data of normal and pathological granulocytes obtained in this study leads to the novel observation that the phagocytic episode from half to complete engulfment of bacterial particles by granulocytes appears to be the crucial step from the thermodynamic point of view.
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Immunoglobulin G was present only in those Escherichia coli antisera causing coagglutination, as determined by immunoelectrophoretic patterns in eluates of antigen-antibody complexes.
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Adhesion of leukocytes and platelets to solid substrates of different surface tensions and hence different wettability is studied from a thermodynamic point of view. A simple thermodynamic model predicts that a cellular adhesion should increase with increasing surface tension of the solid substrate if the surface tension of the medium in which the cells are suspended is lower than the surface tension of the cells. If the surface tension of the suspending medium is higher than that of the cells, the opposite behavior is predicted. These predictions are borne out completely by neutrophil adhesion tests, where the surface tension of the aqueous suspending medium is varied by addition of dimethyl sulfoxide (DMSO). Platelet adhesion experiments also confirm these predictions, the only difference being that surface tensions of the suspending medium above that of the platelets cannot be realized, owing to exudation of surface active solutes from the platelets. Utilization of the thermodynamic prediction that cellular adhesion should become independent of the surface tension of the substrate when the surface tensions of the cells and that of the suspending medium are equal leads to a value of the surface tension of neutrophils of 69.0 erg/cm(2), in excellent agreement with the value obtained from contact angles measured on layers of cells.
Cellular adhesion of granulocytes and of platelets to solid substrates of different surface tensions has been studied from a thermodynamic aspect. A simple thermodynamic model predicts that cellular adhesion should increase as the surface tension of the solid substrate increases provided that the surface tension of the liquid medium in which the cells are suspended is lower than the surface tension of the cells themselves. If, however, the surface tension of the liquid medium is higher than the surface tension of the cells, then a decrease in cell adhesion with increasing substrate surface tension can be predicted. These predictions are completely substantiated by granulocyte adhesion tests in which the surface tension of the suspending liquid medium is varied through the addition of different volumes of dimethyl sulfoxide (DMSO). Platelet adhesion experiments also confirmed these predictions, the only difference being that it is not possible to obtain a suspending liquid medium with a surface tension higher than that of platelets themselves, as a consequence of the exudation of surface active substances by the platelets.
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The freezing front technique for solid surface tension measurements was used to obtain the surface tensions of glutaraldehyde-fixed human erythrocytes, and fresh human lymphocytes and granulocytes in aqueous media. The results agree well with the values obtained by other methods and indicate that the freezing front technique is sufficiently sensitive to detect small differences (of the order of 0.1 ergs/cm2) in surface tension. This property, along with a number of applications for which it is uniquely suited, makes the freezing front technique an important new approach to the measurement of the surface tensions of biological cells and of small particles in general.
A novel method for the isolation of granulocytes from nylon fibers is described. It is modification of filtration leukapheresis based on cellular surface thermodynamics. The system takes into account both electrical and van der Waals forces. The elution buffer contains a chelating agent and a surface active agent which lowers the surface tension of the liquid to a value intermediate between that of the surface tensions of the nylon fibers and of the granulocytes. Thus, the attractive van der Waals interaction between the fibers and the granulocytes becomes a repulsion. This results in a two- to threefold increase in cell yield. Phagocytosis, candicidal activity, and oxygen consumption did not appear to be affected through the isolation procedure. By both light and transmission electron microscopy the isolated granulocytes appeared to be morphologically intact.
An attempt is made to develop an understanding of the basic mechanism governing the interaction of the major blood proteins and cells with polymer surfaces with a focus on thermodynamic approach. It is believed that such investigations may be helpful to provide a rational for the design of suitable cardiovascular prosthetic materials and to choose materials in such a way that adsorption of certain proteins eg. 'fibrinogen' is thermodynamically unfavourable compared to adsorption of other proteins eg. 'albumin'.
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