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Biomedical subjects

G H Ryu

Publications and source records attributed to G H Ryu.

10 recordsLinked to original sources

Plasma protein adsorption to sulfonated poly(ethylene oxide)-grafted polyurethane surface.

Adsorption of proteins (fibrinogen, albumin, and gamma globulin) from plasma onto surface-modified PUs (PU-PEO, PU-SO3, and PU-PEO-SO3) was evaluated. Adsorbed fibrinogen at steady state decreased in the order PU-SO3 > PU > PU-PEO-SO3 > PU-PEO, suggesting that sulfonate groups have specific high affinity to fibrinogen. The intermediate fibrinogen adsorption on PU-PEO-SO3 can be explained by the compensatory effect between the low protein binding affinity of the PEO chain and the high fibrinogen binding affinity of the sulfonate group. In addition, PU-PEO-SO3 showed a very fast fibrinogen adsorption due to the high accessibility of the sulfonate group to fibrinogen by the poly(ethylene oxide) (PEO) spacer. The kinetic profiles of their surfaces showed that as the adsorption time increases, fibrinogen initially adsorbed was decreased and a plateau reached, demonstrating that all the surfaces exhibited the Vroman effect (the fibrinogen displacement phenomenon). PU-PEO showed the least fibrinogen and albumin adsorption among PUs, confirming the known nonadhesive property of PEO chains. It is very interesting that PU-PEO-SO3 exhibited the highest adsorption of albumin and the lowest adsorption of IgG. Therefore, it may be concluded that such adsorption behaviors of proteins to PU-PEO-SO3 contribute to improved blood compatibility.

Adsorption

Surface characteristics and properties of lumbrokinase-immobilized polyurethane.

Potent and novel fibrinolytic enzymes (lumbrokinase [LK]) were extracted from the earthworm, Lumbricus rubellus. These enzymes were very stable and showed greater antithrombotic activity than other currently used fibrinolytic proteins. An LK fraction showing the most potent fibrinolytic activity was immobilized onto a polyurethane (PU) surface to investigate its enzymatic activity and antithrombotic activity. A methanol-extracted PU surface was coated with 3% (wt/vol) maleic anhydride methylvinyl ether copolymer (MAMEC)/tetrahydrofuran (THF) solution, and the surface was incubated in an LK solution/phosphate-buffered saline (PBS, pH 7.4). The surface properties were characterized by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), electron spectroscopy for chemical analysis (ESCA), and dynamic contact angle. The stability of immobilized LK was determined by caseinolytic activity assay and the specificity of immobilized LK on fibrinogen/fibrin was observed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The antithrombotic activity of immobilized LK was evaluated using an ex vivo rabbit A-A shunt experiment. LK immobilization was confirmed by ATR-FTIR and ESCA. Immobilized LK demonstrated stable proteolytic activity during various incubation periods. Immobilized LK proteolyzed fibrinogen and fibrin almost specifically, while it hardly hydrolyzed other plasma proteins including plasminogen and albumin. In the ex vivo A-A shunt experiment, the LK-immobilized surface significantly prolonged occlusion time over control surfaces. This is primarily due to the high thrombolytic activity of immobilized LK. In this work, a highly efficient surface modification method on the PU surface was developed, and this LK immobilization technique will be very useful in improving the blood compatibility of blood-contacting devices.

Animals

Cardioprotective drugs decrease the Na+ background current.

Cardiac dysfunctions such as myocardial functional failure and ventricular arrhythmia have been largely attributed to intracellular Ca2+ overload. One of the mechanisms of intracellular Ca2+ overload involves a rapid influx of Ca2+ via Na(+)-Ca2+ exchange during the reperfusion which utilizes the accumulation of Na+ in myocytes during ischemic cardiac arrest. Possible sources of the intracellular Na+ accumulation include Na+ channel, Na(+)-H+ exchange, Na(+)-Ca2+ exchange, and Na+ background current. In this study, we studied the role of the Na+ background current in intracellular Na+ accumulation during the cardiac arrest by measuring the Na+ background current in guinea pig ventricular myocytes with whole cell clamp method and evaluating the effects of cardioprotective drugs on the Na+ background current. The results were as follows: (1) The Na+ background inward current at -40 mV membrane potential was larger at Ca2+ free solution than 1.8 mM Ca2+ solution. (2) The Na+ background current was not affected by verapamil. (3) 2 microM O-(N, N-hexamethylene)-amiloride (HMA) decreased the Na+ background current at negative membrane potential. (4) The new cardioprotective drug, R 56865, decreased the Na+ background current. These results suggest that the Na+ background current plays a role in increasing the intracellular Na+ activity during high K+ cardioplegia and the blocking effect of myoprotective drugs, such as R 56865, on the Na+ background current may contribute to myocardial protection after cardioplegia.

Amiloride

Adhered platelet morphology in diabetes mellitus.

We compared the morphology of platelets obtained from diabetic patients in various stages of retinopathy and nephropathy with those of control patients. The platelets were collected on to polyethylene films, processed and observed under scanning electron microscopy. Different platelet morphologies were observed within the diabetic group, correlating with the severity of complications, whereas platelets appeared normal in the control group. After more extensive follow-up and comparative studies, these preliminary observations could provide another diagnostic tool for detecting and evaluating severe complications associated with diabetes.

Case-Control Studies

Antithrombogenicity of lumbrokinase-immobilized polyurethane.

Lumbrokinase is a potent fibrinolytic enzyme purified from the earthworm, Lumbricus rubellus. We immobilized 18 IU/cm2 of lumbrokinase to polyurethane using maleic anhydride methylvinyl ether copolymer (MAMEC) as an enzyme carrier, and the proteolytic and fibrinolytic activities of immobilized lumbrokinase were assayed. Immobilized lumbrokinase retained about 34% of its activity, compared with soluble lumbrokinase activity. Immobilized lumbrokinase showed stability against thermal inactivation and degradation and within a various pH range. The optimal pH of immobilized lumbrokinase shifted 1.0 pH unit upward compared with soluble enzyme. Upon exposure to the human whole blood, less amount of 125I-fibrinogen was adsorbed to lumbrokinase-immobilized surface than to the polyurethane control surface. The lumbrokinase-immobilized surface showed less platelet adhesion than did the MAMEC-grafted surface. At the early stage of platelet adhesion, the number of adhered platelets increased on the lumbrokinase-immobilized surface with increasing time; yet, the platelet number drastically decreased on the lumbrokinase-immobilized surface after 80 min incubation. This suggests that lumbrokinase-immobilized polyurethane digested the adsorbed fibrinogen and inhibited platelet adhesion on the surface, probably by inhibiting fibrinogen adsorption to be highly antithrombogenic. Clinical applications of this material to artificial organs should be developed in the near future.

Adsorption

Adsorption behavior of fibrinogen to sulfonated polyethyleneoxide-grafted polyurethane surfaces.

Fibrinogen adsorptions to surface modified polyurethanes (PU, PU-PEO, and PU-PEO-SO3) were studied from plasma in vitro. PU and PU-PEO surfaces demonstrated that initial adsorption increases with increasing plasma concentration in kinetic profiles and adsorption time in adsorption profiles as a function of plasma concentration, but after the plateau is reached, its adsorption amount decreases as plasma concentration (0.2-2.0%) and adsorption time (1-120 min) increase, respectively. In contrast, PU-PEO-SO3 showed that initial adsorption is almost same regardless of plasma concentration and adsorption time, which is due to the high affinity of surface sulfonate group to fibrinogen. All the surfaces indicated the Vroman effect at about 0.6% plasma concentration; however, the displacement was relatively low. Adsorbed amount of fibrinogen at steady state decreased in the order: PU > PU-PEO-SO3 > PU-PEO, regardless of adsorption time and plasma concentration. The adsorption behavior of PU-PEO-SO3 is attributed to both effect of low binding affinity of PEO chain and high affinity of pendant sulfonate group toward fibrinogen.

Adsorption

A moving-actuator type electromechanical total artificial heart--Part II: Circular type and animal experiment.

A new type of electromechanical total artificial heart (TAH) based on circular rolling-cylinder mechanism was developed to overcome critical problems in motor-driven artificial hearts such as large size and difficulties in fitting the heart to atrial remnants and arterial vessels. Its performance and reliability were evaluated in mock circulation and in an animal implant experiment. The total weight and volume of the pump is 650 g and 600 mL, respectively. This new pump was implanted in a calf for total heart replacement and 96 h of survival was achieved. The whole system, including pump, controller, and control algorithm performed well enough to improve the prospect of eventual clinical application of our TAH system.

Animals

Effect of shear stress on fibrinogen adsorption and its conformational change.

The composition and molecular organization of adsorbed protein films are strongly correlated with thrombogenesis on artificial surfaces. In particular, the antibody-detectable (that is, conformationally intact) bound fibrinogen, but not that the total amount of adsorbed fibrinogen, is correlated with platelet reactivity. In this work, the authors quantified the adsorbed plasma protein distribution inside the left ventricular assist device. They also evaluated the effect of wall shear stress on protein adsorption and conformational change of adsorbed fibrinogen. Conformational change of adsorbed fibrinogen was measured by exposing the fibrinogen preadsorbed polyurethane to three anti-fibrinogen monoclonal antibodies; the 134B-29 detectable alpha 566-580 domain of fibrinogen was increased with increasing concentration of adsorbed fibrinogen, whereas the other two fibrinogen domains were almost saturated when increasing the concentration of adsorbed fibrinogen. The adsorbed amounts of total fibrinogen and monoclonal antibody detectable fibrinogen was decreased with increasing shear rate. Results of in vivo plasma protein adsorption on polyurethane surfaces disclosed that the adsorbed amount of fibrinogen, as well as albumin and globulin, was also decreased with increasing shear rate. In conclusion, less protein was adsorbed in the higher shear region and the effect of shear level on fibrinogen adsorption and its conformational change was strongly dependent upon the surface characteristics of the biomaterials. The monoclonal antibody 134B-29 against the 566-580 domain of fibrinogen was the most reactive with the fibrinogen adsorbed on polyurethane surfaces in this experiment.

Adsorption

Antithrombotic activity of a lumbrokinase immobilized polyurethane surface.

Six fractions of strong and novel fibrinolytic enzymes (lumbrokinase, LK) were extracted from the earthworm Lumbricus rubellus. The enzymes in these fractions appeared to be very stable and showed greater antithrombotic activity than other currently used antithrombotics. The authors immobilized an LK fraction that shows the most potent fibrinolytic activity on a polyurethane (PU) surface to investigate its enzymatic and antithrombotic activity. The methanol extracted PU surface was treated with a 3% (wt/vol) maleic anhydride methylvinyl ether copolymer (MAMEC) solution and finally incubated in an LK solution in PBS (pH 7.4). The immobilized LK activity was estimated by the fibrin plate method and caseinolytic activity assay. The antithrombotic activity was evaluated by in vitro 125I-fibrinogen adsorption in fresh whole blood and 99mTc platelet adhesion tests. In addition, the occlusion time was determined through ex vivo rabbit A-A shunt experiments. The content and unit activity of immobilized LK were found to be 24 micrograms/cm2 and 18 IU/cm2, respectively. The relative activity ratio of immobilized LK to soluble LK was found to be approximately 34%. Immobilized LK was stable within a various pH range and resistant to inhibitors and thermal inactivation. Less fibrinogen was adsorbed and fewer platelets adhered on an LK-immobilized surface than on PU and PU-MAMEC controls. The ex vivo occlusion time of untreated PU and PU-MAMEC surfaces were only 32 and 42 minutes, respectively. But that of LK-immobilized PU was extended to 140 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

The fluid dynamic effect on protein adsorption in left ventricular assist devices.

Plasma protein adsorption onto an artificial surface is strongly influenced by not only the surface characteristics of materials, but also by the fluid dynamics inside the blood pump, and it would influence subsequent platelet adhesion or activation, which plays a major role in the initiation of thrombus formation at the blood-material interface in vivo. In vitro flow visualization of an electrohydraulic LVAD was performed by a video camera (CCD, Hitachi) and an image processor (PC VISION PLUS) with an IBM PC. The electrohydraulic LVADs were implanted in mongrel dogs of approximately 20 kg. The authors sectioned the blood contacted ventricle after animal death according to the level of shear rate. Because analysis of adsorbed protein might be influenced by the size of the ventricle segment, the number of segments was limited to eight per ventricle. Platelet adhesion and its morphology were observed by scanning electron microscopy (SEM). Adsorbed plasma proteins (fibrinogen, albumin, and IgG) on each segment were quantified by enzyme linked immunosorbent assay (ELISA). The specimens were soaked in 2% (wt/vol) SDS/PBS for 2 days and the released protein concentration assessed. A well developed large vortex was observed at the center of the artificial ventricle. Polyurethane blood pumps displayed different degrees of protein adsorption and subsequent platelet adhesion on each segment.

Adsorption