PubMed HealthSearch

Biomedical subjects

I A Feuerstein

Publications and source records attributed to I A Feuerstein.

At least 19 recordsLinked to original sources

Platelet adherence and detachment with adsorbed fibrinogen: a flow study with a series of hydroxyethyl methacrylate-ethyl methacrylate copolymers using video microscopy.

The adhesion and detachment of platelets were studied on glass coatings of a series of copolymers of hydroxyethyl methacrylate (HEMA) and ethyl methacrylate (EMA) with preadsorbed fibrinogen. Observations of the interactions of acridine-orange-labeled washed platelets with these surfaces from a flowing (500 s-1 wall shear rate) suspension in Tyrode's solution containing albumin and red cells were made with epifluorescent video microscopy (EVM). In some cases preadsorbed materials were incubated for 24 h, during which little or no loss of protein occurred. Protein surface concentration, by itself, was a poor indicator of expected cell adhesion and morphology. Surface chemistry was a second important factor which must be considered. A third observation is that for the 100% EMA copolymer, 24 h of incubation led to a large reduction in platelet adhesion when compared to the 100% EMA material without incubation. For the 0% and 100% EMA polymers, the percentage of contacting platelets which adhere and detach is greater for the 24-h incubation cases than for those not incubated. These results led to the conclusion that our most hydrophilic surface favors adhesion with detachment, transient cell contact, over long-term adhesion, as does incubation of adsorbed protein. A brief discussion is presented of a possible connection between this behavior and platelet consumption in vivo for hydrogels.

Adsorption

Epifluorescent video microscopy (EVM) for platelet-biomaterial interactions: elimination of photoactivation and dye effects.

The use of two intracellular dyes for epifluorescent video microscopy (EVM) in observations of cell-surface interactions is evaluated and discussed. This methodology permits determinations of cell adhesion, detachment and movement at the surfaces of biomaterials in the presence of flow and physiological haematocrit. Two tests, one which examines for the effect of incident light on platelet adhesion and one which checks for sufficient light for accurate observation of cells, have been designed. Evaluations were made of the adhesion of platelets labelled with the fluorescent dyes mepacrine and acridine orange, used singly and in combination. The use of a number of light level-dye level combinations with glass and several polymers and the addition of a plasma level of fibrinogen did not show any photoactivation effects. This methodology paves the way for longer than previous exposures to light with our system, from 1 min up to 30 min now. Washed platelet suspensions are preferred; these allow for the selective labelling of specific cells and the removal of dye from the surface of the cell.

Acridine Orange

Platelet adherence and detachment: a flow study with a series of hydroxyethyl methacrylate-ethyl methacrylate copolymers using video microscopy.

The adhesion and detachment of platelets were studied on glass coatings of a series of copolymers of hydroxyethyl methacrylate (HEMA) and ethyl methacrylate (EMA). Observations of the interactions of mepacrine labelled washed platelets with these surfaces from a flowing (500 s-1 wall shear rate) suspension in Tyrode's solution containing albumin and red cells were made with epifluorescent video microscopy (EVM). Total platelet adhesion, including platelets which adhere on first contact and platelets which attach temporarily before adhesion, and the number of detaching platelets were minimal for the 0 and 20% EMA copolymers, reached a maximum for the 50% EMA copolymer and showed reduced values for the 80% and 100% EMA copolymers. For the 50, 80, and 100% EMA copolymers, the adhesion values expressed, as a percentage of total contacting platelets, were not different. Albumin adsorption to these copolymers shows a continuous increase from the 0% to the 100% EMA copolymer. It is likely that the peak in platelet adhesion at the 50% EMA composition is related to: low protein adsorption on the 0 and 20% EMA copolymers, too little albumin adsorption to block adhesion on the 50% EMA copolymer, and full-scale blocking on the 80 and 100% EMA copolymers due to greater albumin adsorption.

Adult

Platelets process adsorbed protein: a morphological study.

The nature of the adsorbed protein on a non-biological biomaterial surface is known to influence cell adhesion. This work deals with the contribution of adherent platelets towards the redistribution of preadsorbed proteins. A redistribution was only observed with spread platelets on adsorbed fibrinogen and fibronectin, but not with adsorbed beta-lactoglobulin. The labelled protein images showed a dark outer ring with an inner region having fluorescent areas, with greater intensity than regions not covered by platelets. The fluorescent patterns observed may be a result of the binding of adsorbed fluorescent protein to the glycoprotein IIb/IIIa receptors of platelets, followed by their redistribution within the plasma membrane.

Adsorption

Adhesion and aggregation of thrombin prestimulated human platelets: evaluation of a series of biomaterials characterized by ESCA.

Five materials of interest in blood contact applications (PVC, Silastic, Biomer and siliconized glass) were internally coated on glass tubes and exposed to suspensions of platelets and red cells. Uncoated glass was also examined. Thrombin was added (prestimulation) to some suspensions before exposure to the biomaterial surfaces. The three polymeric surfaces were characterized by electron spectroscopy for chemical analysis (ESCA). Prestimulation with thrombin leads to increased adhesion of single platelets only with Silastic. With a wide range of surface types, thrombin prestimulation consistently leads to higher levels of platelet accumulation in the form of aggregates; the PVC-coated material showed the highest levels. ESCA analysis of PVC, however, suggested our coating was impure or an oxidized material.

Biocompatible Materials

Adhesion of polymorphonuclear leukocytes to protein-coated and platelet adherent surfaces.

Since protein adsorption and platelet adhesion are likely to precede significant contact of leukocytes with the surfaces of artificial organs, we have chosen to study polymorphonuclear leukocyte (PMN) adhesion in a sequential manner. The work presented here deals with the effects of flow and surface type on PMN adhesion to fibrinogen- and albumin-coated glass. We compared direct adhesion to adsorbed protein with adhesion to adsorbed protein having adherent platelets. These experiments were designed to see if PMN's might preferentially adhere to albumin or fibrinogen and whether a particular morphological form of adherent platelet could promote PMN adhesion. The adhesion of PMN's to spread platelets on albumin or fibrinogen occurs to a greater extent than in the absence of platelets. Adhesion of PMN's to spread platelets may be an important mechanism for their depletion from the circulation during artificial organ use.

Adult

Microcomputer-based interactive tracking of blood cells at biomaterial surfaces.

A microcomputer-based system for analyzing the motion of human platelets and leukocytes at synthetic surfaces from a sequence of video frames on tape is described. The software is designed to provide convenient interaction with an operator to reduce the burden of manual analysis. In addition, the system computes and stores the cell movement data on disk for subsequent statistical analysis. Measurement include the number and nature of cell-to-surface collisions, residence times, and distances traveled.

Biocompatible Materials

Adhesion and aggregation of thrombin prestimulated human platelets: evaluation of surface-bound fibrinogen and surface-bound albumin.

The effect of prestimulation of human washed platelets with thrombin on their ability to adhere to and form aggregates on fibrinogen- and albumin-coated glass has been studied. Prestimulation of platelets does not alter platelet adhesion to surface bound albumin and fibrinogen. Our data for both protein coatings support a mechanism of adherence of single prestimulated platelets before formation of surface-bound aggregates from single platelets arriving from the flow. When whole blood contacts an artificial surface, thrombin will likely influence the platelet component of thrombus formation.

Cell Membrane

Platelet adherence and detachment--in what ways is surface-bound albumin different from surface-bound fibrinogen?

The adhesion and detachment of platelets on surface-bound albumin and surface-bound fibrinogen were studied. Fluorescent video-microscopy of platelets labelled with mepacrine was used to provide continuous information. Glass tubes (I.D. 1.3 mm) were precoated with either human albumin or human fibrinogen before exposure to a suspension of washed platelets and red cells. Observations were made 0.5 cm from the tube's entrance over a 1370 micron 2 portion of lumen. The rate at which cells leave the measurement area and the percent of initially attaching cells which leave are independent of protein coating but increase with flow rate. The percent of initially attaching platelets which permanently adhere is equal for both protein coatings but the pathways leading to this result can be different. For the lower shear rate studied, 80 s-1, the percent of cells which permanently adhere on first contact is less for albumin than for fibrinogen; the percent of initially attaching cells which adhere and then move before permanent adhesion is greater for albumin. The mechanism of detachment and reattachment leads to the equality of the overall adhesion efficiencies for the two protein coatings at 80 s-1. For the higher shear rate studied, 456 s-1, the adhesion pathways for both coatings were the same.

Chromium Radioisotopes

Blood platelet surface interactions on fibrinogen under flow as viewed with fluorescent video-microscopy.

The interaction of fluorescently labeled blood platelets with fibrinogen-coated glass was studied in Poiseuille flow at 3 wall shear rates, 40, 80 and 944 s-1. Observations were made via video-microscopy at a distance of 0.5 cm from a tube's entrance over a 1370 microns 2 portion of luminal area. The rates of arrival and detachment, and the net rate of adhesion of cells increased nonlinearly with flow rate. The fraction of arriving cells, first contacts, which adhered without subsequent movement and the fraction of arriving cells which adhered, moved to new positions and then remained adherent, were maximal at 80 s-1. For platelets which adhere and then move to a number of new positions, the likelihood of permanent adhesion is greater than 85 percent. The adhesion process is one in which 40-60 percent of cells permanently adhere on first contact with an additional 30 percent adhering after several moves along the surface. Cells contacting where a platelet was previously adherent had a greater chance of adhering than they would on an unaltered fibrinogen surface. The efficiency of platelet adhesion is greater for second contacts than for first contacts on unaltered fibrinogen coated surface.

Analysis of Variance

Platelet adhesion to fibrinogen-coated glass at an abrupt tubular expansion viewed with fluorescent video-microscopy.

The adhesion and detachment of human washed platelets was studied on the surface of the larger tube of a tubular expansion. Measurements were made within the vortex, at the reattachment point and downstream of the vortex. Fluorescent video-microscopy of mepacrine labelled platelets was used to record data continuously. Flow was from the smaller to the larger tube at Reynolds numbers (based on upstream conditions) of 75.4 and 212.2. Measurements of the adhesion efficiency for initially contacting cells and an overall adhesion efficiency were made. These efficiencies decreased with increasing Reynolds number. There was a pattern of variability for both efficiencies with respect to position and Reynolds number which is consistent with the generation of the unstable flow at the reattachment point.

Analysis of Variance

Platelet accumulation on collagen: drugs which inhibit arachidonic acid metabolism and affect intracellular cyclic AMP levels.

We have studied the accumulation of washed platelets on collagen-coated glass from flowing platelet-red blood cell suspensions in the presence and absence of drugs. Glass tubes were 10 cm long and the flow rate was 1 ml/min, 80 s-1. For all experiments, platelet accumulation was greatest near the tube's inlet with a continuous decrease to the exit. A common feature, of those drug treatments which lead to reduced accumulation at the inlet, was an increase in outlet accumulation when compared with outlet control values. Platelet-collagen adhesion resulted in maximal release of 3H-serotonin in the presence of agents that prevent platelet aggregation on collagen. Only drug treatment known to raise cAMP levels (PGE1 and dipyridamole) or prevent the formation of prostaglandins and thromboxanes (sulfinpyrazone, indomethacin and ASA) were found to inhibit aggregate growth. Platelet aggregation on collagen in the absence of thrombin likely proceeds through the liberation of prostaglandins and thromboxanes from surface-bound platelets into the flow where they may stimulate flow-born cells. An alternate hypothesis is that such treatments affect the release of alpha-granule components necessary for aggregation.

Alprostadil

Adhesion and aggregation of thrombin prestimulated human platelets on surface-bound fibrinogen: evidence for involvement of ADP and arachidonic acid pathways.

The purpose of this study was to examine the influence of substances released from human platelets upon their accumulation on human fibrinogen-coated glass tubes. After prestimulation with thrombin for one minute or in the absence of prestimulation, washed human platelets suspended in Eagle's medium with RBC were drawn through the tubes at 1 ml/min, 80 s-1, for 1, 2 or 6 min. Thrombin prestimulation (0.02, 0.05 or 0.25 U/ml) was followed by inactivation with hirudin (0.1, 0.25 or 1.25 U/ml) before flow. Singly adherent platelets were observed in the absence of thrombin or with thrombin for exposure times of 1 and 2 min. At 6 min after at least 0.05 U/ml of thrombin, surface-bound aggregates were observed. The initial rate of adhesion increased with the amount of thrombin used for prestimulation. For adhesion to fibrinogen in the absence of prestimulation, platelet-derived ADP was a stimulator. Adhesion was shown to be independent of the ADP and arachidonic acid pathways in response to prestimulation with a low level of thrombin, 0.02 U/ml. For adhesion and cohesion, aggregation, in the presence of sufficient thrombin for prestimulation, 0.05 U/ml, ADP, serotonin and substances from arachidonic acid metabolism acted jointly to stimulate platelets.

Adenosine Diphosphate

Maximum fluid concentrations of materials released from platelets at a surface.

We examine the estimation of local concentrations of materials that are released from the dense and alpha-granules of platelets during accumulation of platelets upon collagen-coated glass. Platelet/red blood cell suspensions were perfused through a 1.3-mm-ID tube. Empirical data were used in a calculation procedure, based on diffusion and convection, designed to yield an upper bound on the interfacial fluid concentration (IFC) for each substance considered. The necessary empirical data are the rate of platelet accumulation and the maximum amount of material in the platelet capable of secretion. It was found that the IFC is dependent on the shear rate at the surface (G) and is proportional to G0.27. This means that an eightfold increase in flow rate would increase the IFCs approximately twofold. Serotonin, pyrophosphate, adenosine 5'-monophosphate (AMP), and adenosine 5'-triphosphate (ATP) were found not to be present in sufficient quantities to produce IFCs that could influence platelet aggregation if used alone at the IFC. A second set of materials, fibrinogen, fibronectin von Willebrand factor, and calcium, had IFCs less than their concentrations normally found in plasma. A third category, containing adenosine 5'-diphosphate (ADP) alone, had an IFC close to those known to affect platelet aggregation. The role of metabolites of arachidonic acid, which may promote or inhibit platelet aggregation, awaits further description.

Adenosine Diphosphate

Experimental evaluation of streamline patterns and separated flows in a series of branching vessels with implications for atherosclerosis and thrombosis.

Flow conditions in four models representing the aortic bifurcation, iliac bifuraction, and a renal artery branch were investigated at volumetric flow rates corresponding to Reynolds numbers from 1000 to 4000 over the complete range of flow division between daughter vessels. Qualitative flow streamline patterns and quantitative definition of those flow conditions leading to disturbed flow (flow separation ) were determined primarily at steady flow with a limited set of pulsatie experiments. Under conditions of no flow separation, common characteristic streamline patterns not parallel to the center lines of parent or daughter tubes were found for all models. These effects were accentuated with increasing Reynolds number. Flow separation was inducible through alteration of flow division between daughter vessels or by an increase in flow rate. Each of the four models had distinct combinations of flow division ratio and flow rate which gave: (1) no flow separation, (2) flow separation at the outside of the right daughter tube, and (3) flow separation at the outside of the left daughter tube. Models representing the renal artery also had regions of simultaneous left- and righthand separation on the outside of their daughter tubes. The separated flows observed here displayed streamlines forming an open vortex with flows entering and leaving. These regions, which occur only at distinct combinations of flow rate and flow division, may be key centers where platelet aggregates may form, release constituents, and cause vessel injury.

Aorta, Abdominal