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Biomedical subjects

M K Dewanjee

Publications and source records attributed to M K Dewanjee.

At least 127 records · Page 7Linked to original sources
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Emboli from an extraluminal blood flow hollow fiber oxygenator with and without an arterial filter during cardiopulmonary bypass in a pig model.

The effect of an arterial filter on visceral emboli was quantified with autologous indium-111 labeled platelets (INPLT) during cardiopulmonary bypass (CPB) in Yorkshire pigs. Biodistribution of INPLT was determined in 12 control pigs (30-35 kg, unoperated control [n = 6] and sham operated control [n = 6]). CPB was carried out with (n = 6) and without (n = 6) an arterial filter in 12 pigs at a flow rate of 2.5-3.5 L/min. Platelets labeled with In-111 tropolone (650-780 microCi) were injected intravenously 24 hr before CPB. All pigs were systemically heparinized (activated coagulation time > 400 sec); CPB was instituted with a roller pump, an extraluminal blood flow oxygenator (Bentley Univox, 1.8 m2), and an arterial filter (0.25 m2) and continued for 3 hr. Platelet kinetics, pooling, and counts were monitored by a Geiger probe and a Coulter counter. The thrombi in the oxygenator and arterial filter and emboli in viscera and brain were imaged with a gamma camera and measured with an ion chamber and gamma counter. Percentage of INPLT (mean +/- SD) in organs, tissues, and components of the circuit in four groups of pigs was calculated. Flow cytometry with antibodies to CD61 (GPIIIa) and CD62P (GMP-140: control) of porcine platelets was carried out with blood samples taken before, during, and after CPB for estimation of circulating platelet aggregates and platelet microparticles. Pulmonary, renal, cardiac, and cerebral emboli in pigs undergoing CPB with and without a filter were similar (p < 0.1). The amount of filter adherent thrombi was small (0.04 +/- 0.01%); oxygenator adherent thrombus in both groups was similar (p < 0.1). Emboli were found in the cerebral medulla, hippocampus, and posterior cerebral cortex in both groups. During CPB, the arterial filter functioned minimally as a trap for platelet thrombi detached from the oxygenator and circulating emboli. Flow cytometry of blood demonstrated the shift of equilibria from single platelets to platelet aggregates and microparticles during CPB and their gradual reversal to single platelets after CPB; the loosely adherent emboli disaggregated and further shifted these equilibria to single platelets and smaller aggregates, probably through the action of endogenous nitric oxide and prostacyclin. The emboli were trapped in organs and tissues and microparticles were sequestered by the reticuloendothelial system.

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Tumor necrosis factor-alpha in plasma during cardiopulmonary bypass in a pig model: correlation with marginated neutrophils and cerebral edema by magnetic resonance imaging.

Tumor necrosis factor (TNFalpha) initiates the cytokine cascade via the nuclear factor (NFkappaB), increasing vascular permeability, inflammation, and edema during and after cardiopulmonary bypass (CPB). Cerebral edema before and after thoracotomy and CPB was evaluated by magnetic resonance imaging. TNF-alpha in plasma was measured in 12 Yorkshire pigs with an enzyme linked immunosorbent assay technique using a monoclonal antibody made against porcine TNFalpha. Blood samples were taken 30 min before and 1, 30, 60, 90, 120, 150, and 180 min after initiation of CPB. Plasma was separated from packed cells after centrifugation, and frozen at -80 degrees C. The level of TNFalpha, measured by color intensity, was read with a 96 well microtiter plate reader. Normal values in plasma were low (range, 0-17 pg/ml; mean, 4.7 pg/ml). In sham control pigs undergoing sternotomy, and pigs that had undergone CBP, TNFalpha reached a peak value at 120-150 min, and subsequently declined. TNFalpha in plasma increased in pigs that underwent thoracotomy and CPB. The increase was significantly higher in pigs that underwent CPB. Neutrophil mapping in the pig brain with 111In-labeled autologous neutrophils 180 min after CPB, suggested almost uniform distribution and accounted for the 0.02% of total circulating neutrophils in the brain, corresponding to a total of 8 x 10(5) neutrophils. Magnetic resonance imaging of the pig brain indicated the presence of cerebral edema after CPB, as depicted by the loss of structural details of the sulci, gyri, and ventricles. Activated neutrophils, monocytes, and other inflammatory cells may induce multi-organ edema and injury via TNFalpha and other regional cytokines.

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Quantification of device adherent, circulating, and organ pool of thrombin and fibrinogen after cardiopulmonary bypass in a pig model.

The pool of thrombin and fibrinogen in circulation, in organs, and on cardiopulmonary bypass devices was quantified during and after cardiopulmonary bypass in four groups of 24 Yorkshire pigs (weight, 30-35 kg); two groups of 10 unoperated pigs were used as controls. Thrombin-alpha and fibrinogen were iodinated with 125iodide using an iodogen transfer technique; 250-300 microCi of these tracers were injected intravenously 1 hr before cardiopulmonary bypass. All pigs were systematically heparinized (activated clotting time > 400 sec); cardiopulmonary bypass was performed at 2.5-3.5 L/min at 28 degrees C using a centrifugal pump, oxygenator (Bentley Univox 1.8 m2; Bentley Inc., Irvine, CA), arterial filter (0.25 m2), and cardiotomy reservoir (BMR 3500) for 90 min, followed by a 90 min reperfusion and 180 min of cardiopulmonary bypass. Iodinated thrombin-alpha and fibrinogen in intact organs and samples of blood, organs, tissues, and oxygenator-arterial filter-cardiotomy reservoir were quantified with an ion chamber and a gamma counter, respectively. The percent of injected iodinated thrombin-alpha and fibrinogen dose (mean +/- SD) in organs and cardiopulmonary bypass devices of all groups of cardiopulmonary bypass pigs was calculated. Thrombin generated at the small area of surgical wounds (0.016-0.038 m2), and fibrin deposited on surfaces of cardiopulmonary bypass devices (2.59 m2), initiate and propagate thrombus formation and embolization. The protein level reached saturation values on all cardiopulmonary bypass devices at 180 min. High levels of thrombin and fibrinogen-fibrin circulate in blood and organs, and are adsorbed on cardiopulmonary bypass devices; this large blood pool of pro-coagulants in the cardiac cradle, tissues, and perfused organs may account for thrombi and emboli during and after cardiopulmonary bypass.

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Quantitation of adsorption and conjugation of plasma proteins by residual glutaraldehyde in fixed collagenous tissue with radioiodinated plasma proteins.

Residual glutaraldehyde (GA) in collagenous cardiovascular tissue prostheses after multiple saline rinses remains in the prostheses and accounts for adsorption and conjugation of a variety of plasma proteins. This may account for later beneficial or adverse effects. Human serum albumin (SA), gamma globulin (GG), and fibrinogen (FB) were iodinated with 125I using the iodogen-transfer technique. Bovine pericardium (PC) was fixed with 0.5% GA for 24 hr and rinsed to remove excess GA. Fresh and GA-fixed PC (FRPC, GAPC: 1 x 1 cm2), in triplicate, were incubated with 0.5-1.0 microCi of tracers in human, porcine, or bovine blood (2 ml) for a period of 0.5, 1, 2, and 3 hr and washed (5x) with saline. Maximum adsorbed proteins per unit weight of collagen (pmol/mg of PC, mean +/- SD) at 3 hr on FRPC and GAPC were quantified with a gamma counter. Fixed PC absorbed significantly more plasma proteins from blood than fresh PC. These conjugated plasma proteins are tightly bound to fixed PC. The adsorbed and conjugated plasma proteins for GAPC and FRPC have the same sequence: SA > GG > FB vs SA > GG > FB. Protein conjugation may affect the remodeling of collagenous cardiovascular tissue prostheses post implantation.

Adsorption↗
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Effect of heparin reversal and fresh platelet transfusion on platelet emboli post-cardiopulmonary bypass in a pig model.

Heparin reversal by protamine and fresh platelet transfusion may decrease bleeding complications post-cardiopulmonary bypass (CPB) and may increase the level of organ trapped platelet emboli. Platelet emboli were quantified in two groups of 12 Yorkshire pigs (30-35 kg), where 111indium labeled autologous platelets (INPLT: 850-1,200 microCi) were injected intravenously before and after CPB (BCPB, ACPB), and the platelet emboli level in intact organs and their samples (brain, heart, kidneys, lung, liver, and spleen) was quantified with an ion chamber and a gamma counter, respectively. All pigs were systemically heparinized (ACT > 400 sec). CPB was carried out at 2.5-3.5 L/min at 28 degrees C using a centrifugal pump, an oxygenator (OX:Bentley Univox 1.8 m2), an arterial filter (AF:0.25 m2), and a cardiotomy reservoir (CR: BMR 250) for 90 min. Heparin was reversed with an equivalent dose of protamine. The percent of INPLT dose (ID%, mean +/- SD) in organs of BCPB and ACPB pigs was calculated. The sequence of platelet emboli on a unit weight basis (ID%/g) had the following order: Spleen > Liver > Lung > Kidneys > Heart > Brain. The presence of significantly higher levels of emboli in brain, heart, and kidneys in the ACPB than the BCPB group suggest that platelet transfusion after heparin reversal with protamine may increase the risk of platelet emboli. However, it is an acceptable risk for patients having bleeding complications post-CPB.

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Radioisotopic techniques for noninvasive detection of platelet deposition in bovine-tissue mitral-valve prostheses and in vitro quantification of visceral microembolism in dogs.

Platelet deposition on bovine pericardial-tissue mitral-valve prostheses in 11 dogs was observed noninvasively by use of 111In-labeled platelets and quantified after sacrifice at one (n = 3), 14 (n = 3), and 30 (n = 5) days postimplantation (300-400 microCi of labeled platelets having been injected 24 hours previously). Thrombosis on the sewing ring and pericardial leaflets at one and 14 days and on the leaflets at 30 days was delineated in scintiphotos. In vitro quantification (% injected dose) indicated that the leaflets, sewing ring, and perivalvular tissue retained 0.904% of labeled platelets at one day postimplantation, 0.198% at 14 days, and 0.040% at 30 days. Platelet half-life was reduced to 38 hours at 21 days postimplantation but returned toward the normal (50 hours) with fibrous ingrowth in the sewing ring. Microembolism in lung and kidney, as measured by tissue/blood radioactivity ratio, also was decreased significantly at 30 days. 111In-labeled platelets thus provide a sensitive marker for noninvasive imaging and in vitro quantification of platelet deposition on valvular prostheses and microemboli trapped in viscera, although histochemical confirmation will be necessary to correlate the increase in tissue/blood ratio with the presence of microembolism.

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Quantitation of comparative thrombogenicity of dog, pig, and human platelets in a hemodialyzer.

Comparative platelet thrombogenicity was quantified with In111 labeled platelets. The platelets collected from the blood of Beagle dogs, Yorkshire pigs, and human volunteers were labeled with detergent free In111 tropolone, and sham hemodialysis (SHD) was performed with a hollow fiber dialyzer in a flow loop at 37 degrees C, with flow rates of 7, 150, and 270 ml/min. After SHD, hemodialyzer radioactivity was measured with an ionization chamber, gamma counter, and it was imaged with a gamma camera. The mean values of hemodialyzer-adherent platelet radioactivity were calculated. Canine platelets are more thrombogenic than porcine and human platelets. Quantitation of comparative thrombogenicity with In111 platelets may provide an estimation of prosthesis induced thrombogenicity of human platelets from animal studies.

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Platelet preservation during cardiopulmonary bypass with iloprost and Duraflo-II heparin-coated surfaces.

To test the hypothesis that temporary platelet inhibition during cardiopulmonary bypass (CPB) with surface heparinized systems may result in platelet preservation, nine Yorkshire pigs were placed on CPB for 3 hours. Platelet labeling was done in all pigs with Indium-111 tropolone. CPB was instituted with a roller pump, a hollow fiber membrane oxygenator (Bentley CM-50 [Baxter-Bentley Laboratories, Irvine, CA]), and an arterial filter. The extracorporeal perfusion systems were surface-coated with the Duraflo-II heparin complex. Group A pigs (n = 5) were systemically heparinized (activated coagulation time longer than 400 sec). Group B pigs (n = 4) were placed on CPB without systematic heparinization, but have received the stable prostacyclin-analog Iloprost (ZK36374) at 1 ng/kg/min i.v. from 30 min before and during CPB. Platelet counts declined in group A pigs at 5 min, 1 hr, 2 hr, and 3 hr of CPB to 79.8% (mean), 66.5%, 71.3%, and 69.0% of pre-CPB values, respectively (p less than 0.05). In group B pigs, mean platelet count during CPB was higher than 90% of control value. Percentage of injected radioactivity detected in the oxygenator was 2.82% in group A pigs versus 0.73% in group B pigs (p = 0.0541). Surface heparinization with the Duraflo II heparin coating complex in combination with Iloprost-induced temporary platelet inhibition resulted in platelet count preservation during CPB in the pig model.

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Cardiopulmonary bypass with a surface-heparinized extracorporeal perfusion system.

To evaluate the effect of surface heparinization on platelet consumption during cardiopulmonary bypass (CPB) ten pigs were placed on CPB for 3 hours. All pigs were injected with autologous Indium-111 labeled platelets (300-420 uCi) 24 hours prior to CPB and were systemically heparinized prior to cannulation for CPB. CPB was established with a roller pump, a hollow fiber membrane oxygenator (HFMO, Bentley CM-50) and an arterial filter (AF, Bentley 1025). In six pigs the extracorporeal system was untreated whereas in four pigs it was surface heparinized with the Duraflo-II method. Cardiotomy suction was not used. Percent of injected radiation dose in HFMO and AF at 3 hours of CPB in the nontreated systems was 1.53 +/- 1.12 and 0.88 +/- 0.63%, whereas in the surface heparinized systems was 2.45 +/- 1.71 and 0.49 +/- 0.39% respectively (NS). (Values are mean +/- SD). Blood loss during (CPB) was 225 +/- 179 ml in the nontreated systems, and 263 +/- 103 ml in the surface heparinized systems (NS). Platelet counts were reduced by 12% or 21.8% at 3 hours of CPB in the two groups of pigs respectively (NS). No difference was observed in platelet consumption (in HFMO and in AF) or in platelet count reduction between the two groups of pigs. Surface heparinization did not improve platelet preservation in systemically heparinized pigs at 3 hours of CPB.

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Increase of intraplatelet free calcium ion during extracorporeal circulation with a hollow fiber oxygenator: arterial filter and dynamics of platelet thrombosis on oxygenator and filter in a pig model.

Intraplatelet free calcium (IPFC) ions provide a common pathway for platelet activation leading to thrombosis and embolization. IPFC levels were determined by chlorotetracycline fluorometry during extracorporeal circulation (ECC) with systemic heparin in eight Yorkshire pigs (weighing 30-40 kg; 3 control and 5 ECC); the ratio of slow phase organelle calcium sequestration to fast phase platelet-membrane binding is an index of free calcium. During 3 hr of ECC with a hollow fiber oxygenator (HFO) (Bentley CM-50) and AF (Bentley 1025), seven blood samples were collected 5 min before and during ECC. The platelet deposition (CPM/microCi) on HFO (PDHFO) was simultaneously measured with In-111-labeled autologous platelets (300-400 microCi) and a Geiger probe detector at -5, 0, 5, 30, 45, 60, 120, and 180 min. During ECC, IPFC and HFO thrombus increase significantly (p less than 0.05) at 45 min with respect to control IPFC values of 0.4 +/- 0.1, suggesting direct participation of calcium activated platelets in thrombosis on HFO. The decline of IPFC is due to extrusion and sequestration by dense granules, and decline in HFO thrombus is due to embolization. On the other hand, the embolus in the arterial filter was trapped in a linear fashion, with a consistent increase with time of ECC.

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The dynamics of platelet thrombus formation rate, thrombus retention time, and rate of embolization on a control and heparin bonded polyurethane angio-catheter.

The dynamics of platelet deposition and embolization from control and heparin bonded polyurethane catheters (CPC and HBPC) was evaluated with In-111 labeled autologous platelets (IN-PLT) and a computerized gamma camera (CGC). Ten non-heparinized dogs (18-25 kg) were catheterized in both femoral arteries with 10 cm of CPC and HBPC (5 Fr., Cordis) 24 hr after injection of 300-420 microCi of In-PLT, and imaged for 3 hr with the computerized gamma camera. The regional platelet deposition curves (RPDC) indicated multiple peaks and valleys; the curves were analyzed for early rate of thrombus formation (upswing), thrombus retention time (full width at half maxima of the RPDC-peak), and rate of embolization (downswing) on both catheters. The four parameters (mean +/- SD) of thrombosis on catheters and integral of the radioactivity time curve for the 3 hr duration of imaging were calculated from normalized counts/sec. The rate of thrombus formation and rate of embolization are higher for the control than HBPC, suggesting that heparin-bonding decreases the early rate of thrombosis and embolization. The thrombus adhesivity and retention time appear shorter for the control catheter, indicating that the control thrombogenic catheter forms multiple thrombi and emboli than HBPC. The integral appears larger for the control catheter than HBPC. In vivo (dynamic) studies, in vitro studies, and critical analyses of the radioactivity time curve were essential for complete evaluation of thrombogenicity of catheters and other cardiovascular prostheses.

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Effect of diphosphonate binding to collagen upon inhibition of calcification and promotion of spontaneous endothelial cell coverage on tissue valve prostheses.

A noncalcifying drug, aminodiphosphonate, was immobilized on pericardial collagen to inhibit calcification. The parameters of ADP binding were optimized by using labeled tracer. The effects of the detergents, Triton X-100 and sodium dodecyl sulfate, pH, and temperature on ADP binding to collagen in fresh pericardium, and stabilization of ADP-collagen bond by borohydride (BH) reduction, were optimized. These studies indicate that pretreatment of pericardium with SDS and TX-100 increases ADP binding three- to fivefold. Three-step cross-linking via Schiff-base reactions between collagen, ADP, and cross-linking via Schiff-base reactions between collage, ADP, and glutaraldehyde gives a high value of 30 to 35 molecules of ADP per collagen in pericardium. Twenty-five millimeter tissue valves were made with detergent (1% SDS, 1% TX-100) treated ADP-bound pericardium and implanted in the mitral annuli in calves, which were killed at 60 days post implantation. Regional calcium deposition was measured with atomic absorption spectrometry. The calcium level (micrograms/mg of tissue) in components of control and four processed valves were determined. Regional platelet deposition on zones of leaflets and components of tissue valves were quantified with 111In-labeled autologous platelets. The calcium levels in thrombus and flexion zones of treated valves are lower than that in the control valve. SEM studies of leaflet surfaces at 60 days indicate that these treatment processes reduce calcification and promote spontaneous cell coverage on leaflets around the smooth surface of the outflow tract.

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Early and late comparison of umbilical vein and PTFE grafts in control and piroxicam-treated dogs.

For comparative analysis of small vessel prostheses, 78 dogs underwent femoral artery replacement with autogenous vein, polytetrafluoroethylene (PTFE) or human umbilical vein (HUV) grafts. Thirty-six animals received antiplatelet therapy with piroxicam orally once a day. Three-hour deposition of 111In-labeled platelets was less on HUV than on PTFE grafts but still greater than on autogenous vein grafts. Accordingly, 3-hour patency rates were lowest with PTFE and highest with autogenous vein, although the difference between HUV and PTFE grafts was not statistically significant. One month following implantation, PTFE grafts had a higher patency rate than HUV grafts (p less than 0.05). Treatment with piroxicam decreased 3-hour platelet deposition, improved 1-month patency of both PTFE (p less than 0.05) and HUV (p less than 0.01) grafts, and decreased pseudointimal thickening on PTFE grafts (p less than 0.01). This experimental study did not demonstrate any clearcut difference in the performance of PTFE and HUV grafts. Piroxicam treatment improved the results with both grafts, which would justify future clinical trials with this simple antiplatelet regimen.

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Polyvinyl alcohol sponge for embolotherapy: particle size and morphology.

The size and morphology of particulate polyvinyl alcohol (PVA) sponge prepared by a blender technique for embolotherapy were analyzed by light and scanning electron microscopy. Techniques for preparation and sizing of PVA sponge particles are described. Two types of particulate material, differing in both size and morphology, were found. Large particles (greater than 50 micron) had irregular shapes and sharp, jagged edges. Smaller particles (2-50 micron) were more numerous. Attempts to characterize the smaller particles were unsuccessful. Preparation of PVA sponge particles using the blender technique is a viable choice in external carotid neuroembolization procedures. However, the potential effects of small particles possibly causing cranial nerve palsies does need consideration.

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Migration and granulomatous reaction after periurethral injection of polytef (Teflon).

Although patients with urinary incontinence have been treated successfully by periurethral injection of polytef paste, this study in continent animals demonstrates migration of polytef particles from the injection site. We injected polytef paste periurethrally into female dogs and male monkeys. Particles were found at 50 to 70 days in pelvic nodes in six of seven animals and lungs in four of seven (the kidneys and brain were not studied); and at 10 1/2 months in pelvic nodes, lungs, and brain in seven of seven; kidneys in four of seven; and spleen in two of seven. X-ray microanalysis confirmed that the particles were polytef. At 10 1/2 months, polytef granulomas were found at all injection sites and some sites of distant migration. Since these granulomas signify chronic foreign-body reaction, we believe that until the long-term effects in humans are known, polytef paste should not be used in children or young adults with normal life expectancy.

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Comparative study of reticuloendothelial system pools of the normal and immunologically activated rabbit synovium with radiocolloids.

A quantitative study of the uptake of radiocolloids by the joint synovium relative to other reticuloendothelial system organs was performed in the rabbit model. The radiocolloids of 198Au and 99mTc-sulfide colloid were administered intravenously and distribution in synovial pad, liver, spleen, bone marrow, blood, lung, and other tissues was determined in a gamma counter. The uptake of radiocolloids by both normal and immunologically activated synovial pad was very small in comparison with the reticuloendothelial cells in liver, spleen, and bone marrow.

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