Coagulation monitoring: current techniques and clinical use of viscoelastic point-of-care coagulation devices.

Perioperative monitoring of blood coagulation is critical to better understand causes of hemorrhage, to guide hemostatic therapies, and to predict the risk of bleeding during the consecutive anesthetic or surgical procedures. Point-of-care (POC) coagulation monitoring devices assessing the viscoelastic properties of whole blood, i.e., thrombelastography, rotation thrombelastometry, and Sonoclot analysis, may overcome several limitations of routine coagulation tests in the perioperative setting. The advantage of these techniques is that they have the potential to measure the clotting process, starting with fibrin formation and continue through to clot retraction and fibrinolysis at the bedside, with minimal delays. Furthermore, the coagulation status of patients is assessed in whole blood, allowing the plasmatic coagulation system to interact with platelets and red cells, and thereby providing useful additional information on platelet function. Viscoelastic POC coagulation devices are increasingly being used in clinical practice, especially in the management of patients undergoing cardiac and liver surgery. Furthermore, they provide useful information in a large variety of clinical scenarios, e.g., massive hemorrhage, assessment of hypo- and hypercoagulable states, guiding pro- and anticoagulant therapies, and in diagnosing of a surgical bleeding. A surgical etiology of bleeding has to be considered when viscoelastic test results are normal. In summary, viscoelastic POC coagulation devices may help identify the cause of bleeding and guide pro- and anticoagulant therapies. To ensure optimal accuracy and performance, standardized procedures for blood sampling and handling, strict quality controls and trained personnel are required.

Timing and reproducibility of access flow measurements using extracorporeal temperature gradients.

The duration, accuracy, and reproducibility of a new access flow measuring technique was analyzed in a series of in vitro experiments using an extracorporeal line switch that allowed for almost instantaneous reversal of extracorporeal blood flow without disconnecting the blood lines. Access flow was modeled from the magnitude and time course of extracorporeal temperature changes caused by switching the blood lines. Ten tests were done with access flows covering a range from 410 to 1500 ml min. The coefficient of variation of triplicate access flow identifications was 3.8 +/- 1.5%. The mean bias between measured and modeled access flows was 54 +/- 54 ml min and independent of the range of measured access flows. The average time constant for temperatures to stabilize after switching the blood lines was 0.68 +/- 0.11 min. These results show that the instantaneous change in the direction of blood flow in proximal parts of the extracorporeal circulation produces a smooth change in extracorporeal temperatures that can be explained by a mathematical model incorporating access flow and that a reproducible measure for access blood flow can be obtained as one of the model parameters from that fit within a few minutes of switching the blood lines without the injection of indicator.

Comparative evaluation of two newly developed devices for capillary viscometry.

Viscometry is an often applied method in clinical chemistry. A variety of studies demonstrate an association of parameters related to blood viscosity with human pathology of varying origin. Whole blood and plasma viscosity are considered to be clinically useful indicators in the diagnostic workup and therapy monitoring of certain diseases. In this study, we compare the "Waegeviskosimeter" (WV) described in previous publications with a newly developed device, the "Reverse Flow Viscometer" (RFV). Both viscometers are capillary flow viscometers. Both overcome the disadvantage of common viscometers of the Ubbelohde and Cannon-Fenske type which require large amounts of plasma and which can be only applied to Newtonian fluids. The accuracy of the measurements of both viscometers, requiring less than 1.0 ml sample volume, is superior to most conventional methods. The major distinction in the functionality of the WV and the RFV is that the WV measures the kinematic viscosity whereas the RFV directly estimates dynamic viscosity without the requirement of additional density measurement. We found good reproducibility of viscosity with coefficient of variation CV < or =1.1% for both viscometers. Quality assurance measures have been carried out. Because no quality assurance scheme according to the guidelines proposed by the German Medical Association exists for plasma or whole blood viscosity, we tested reference material Lyphochek Unassayed Chemistry Control Level 1 and Level 2 (Bio-Rad Laboratories). We determined the viscosities 1.40 mPa s and 1.08 mPa s (37 degrees C) and the between-run precision from daily quality control runs with CV of 1.4% and 1.2% for the WV, and 1.7% and 1.4% for the RFV. For direct comparison reasons, we determined the viscosity in seventy human plasma and serum samples by both methods. Using the regression analysis described by Passing-Bablok, the RFV and the WV methods are highly correlated and show only little variations (r = 0.990, tau = 0.896). The regression equation is y(WV) = 1.035x(RFV) - 0.056 with a mean deviation of 0.4+/-3.6%. We conclude that both new devices for viscosity assessment fulfill all quality requirements as prescribed for clinical chemical laboratories. One advantage RFV is to measure the dynamic viscosity directly.

Evidence that rehydrated, lyophilized red blood cells are sufficiently deformable for normal microcirculation transit.

A method was developed for the preparation of rehydratable lyophilized red blood cells (RL RBCs) that hold promise as cell-based oxygen carriers for transfusion medicine. The maintenance of normal cellular deformability is essential for the successful development of cell-based oxygen delivery systems. Improper deformability of RBCs can lead to hemolysis if too fragile or microvascular occlusion if too rigid. We developed an aldehyde stabilization method that is based on the use of paraformaldehyde polymers that complement the function of spectrin as a structural unit with conformational flexibility. Three types of in vitro deformability studies (filter transit, pipette aspiration, and atomic force microscopy) and in vivo intravital microscopy were performed to characterize the deformability of RL RBCs. When considered with safety data from previously reported studies in dogs, the results of these studies indicate that paraformaldehyde-modified RL RBCs have visco-elastic deformability properties that are in the nonpathological range.

What is the maximum duration to perform the hemorheological measurement for the human and mammals.

The hemorheological performance of mammals and human will change with the time after the blood is withdraw from the subject. The whole blood apparent viscosity, erythrocyte deformability and aggregation of some mammals and human were measured. Our results showed that the hemorheological characteristics of mice, rats, guinea pigs, dogs and human began to change respectively after the threshold time of 0.5, 1, 3, 7, 12 h when the blood were stored in vitro. Moreover there exist a relationship: t=3.18W(0.32) between the threshold time (t) and the animal weight (W) and the related coefficient is 0.992.

Determination of regional bone blood flow by means of fluorescent microspheres using an automated sample-processing procedure.

The determination of regional blood flow utilizing fluorescent microspheres (FMs) is an established method for numerous organs. Recent progress, in particular the automation of sample processing, has further improved this method. However, the FM method (reference sample technique), which allows repetitive measurement of regional organ blood flow, has so far not been used for the determination of blood flow in bone. The aim of the present study was to establish FM for the quantification of regional bone blood flow (RBBF). Female, anesthetized New Zealand rabbits (n = 6) received left ventricular injections of different amounts of FM at six subsequent time points. In order to examine the precision of RBBF determination, two different FM species were injected simultaneously at the sixth injection. At the end of the experiments the femoral and tibial condyles of each hind limb were removed and the fluorescence intensity in the tissue samples was measured by an automated procedure. In an in vitro study we have shown that acid digestion of the crystalline matrix has no effect on the fluorescence characteristics of FM. The determination of the number of spheres per tissue sample revealed that depending on the tissue sample size up to 3 x 10(6) spheres/injection were necessary to obtain about 400 microspheres in the individual bone samples. RBBF values of the tibial and femoral condyles did not differ at various injection intervals. The tibial blood flow values varied between 6.6 +/- 1.1 and 8.5 +/- 1.4 ml/min/100 g and were significantly higher than those of the femur (4.3 +/- 1.1 to 6.0 +/- 1.8 ml/min/100 g). The bone blood flow values obtained by simultaneous injection of two FM species correlated significantly (r = 0.96, slope = 1.06, intercept = 0.05), the mean difference was 0.39 +/- 1.11 ml/min/100 g. Our data demonstrate that the measurement of RBBF by means of FM allows a valid determination of RBBF.

Whole blood viscosity, plasma viscosity and erythrocyte aggregation in nine mammalian species: reference values and comparison of data.

In this study species-specific values for whole blood viscosity (WBV), plasma viscosity (PV) and erythrocyte aggregation (EA) were determined in a total of 360 animals. We used 40 individual adult animals of nine mammalian species: horse, pig, dog, cat, rat, cattle, sheep, rabbit and mouse. WBV measurements were carried out using a LS30 viscometer, PV was measured using OCR-D and EA was measured using a Myrenne aggregometer and the LS30 (aggregation index at low shear rate). At low shear rates (0.7 s(-1) and 2.4 s(-1)) haematocrit (Hct)-standardized (40 % Hct) samples showed a higher value of WBV and EA in horse, pig, dog and cat. In cattle, sheep, rabbit and mouse, EA and WBV were markedly decreased and EA was almost undetectable, although the plasma fibrinogen concentration was higher in these animals. Rats showed the highest WBV at low shear rate in native blood and WBV was not different from horse in Hct-standardized blood; however, EA was very low in the rat, a result that might be explained by mechanical or geometrical properties of the red blood cell. EA correlated with the plasma protein concentration in each species except dog and mouse. In horse, cattle and pig, EA correlated with the plasma fibrinogen concentration. At high shear rate (94 s(-1)), WBV was higher in cattle than cat and rat, and dog had higher values than horse, suggesting specific interspecies differences depending on low shear and high shear values of WBV, as a result of mechanisms that influence RBC flexibility. PV was highest in cattle and lowest in rabbit and mouse and did not correlate with WBV. Haemorheological parameters differed between the species. Each species has its own rheological fingerprint. The physiological significance of these variations among mammalian species has not yet been established. Viscosity contributes to endothelial cell shear stress. While haemorheological parameters differ across the species it may be postulated that factors influencing flow-mediated endothelial cell signal transduction are different among the species.

Fluid-solid interaction: benchmarking of an external coupling of ANSYS with CFX for cardiovascular applications.

Fluid-solid interaction is a primary feature of cardiovascular flows. There is increasing interest in the numerical solution of these systems as the extensive computational resource required for such studies becomes available. One form of coupling is an external weak coupling of separate solid and fluid mechanics codes. Information about the stress tensor and displacement vector at the wetted boundary is passed between the codes, and an iterative scheme is employed to move towards convergence of these parameters at each time step. This approach has the attraction that separate codes with the most extensive functionality for each of the separate phases can be selected, which might be important in the context of the complex rheology and contact mechanics that often feature in cardiovascular systems. Penrose and Staples describe a weak coupling of CFX for computational fluid mechanics to ANSYS for solid mechanics, based on a simple Jacobi iteration scheme. It is important to validate the coupled numerical solutions. An extensive analytical study of flow in elastic-walled tubes was carried out by Womersley in the late 1950s. This paper describes the performance of the coupling software for the straight elastic-walled tube, and compares the results with Womersley's analytical solutions. It also presents preliminary results demonstrating the application of the coupled software in the context of a stented vessel.

In vitro hemocompatibility testing of biomaterials according to the ISO 10993-4.

The development of synthetic materials, textured polymers and metals and their increasing use in medicine make research of biomaterials' hemocompatibility very relevant. Problems arise from the polymorphism and diversity of the different materials, the static and dynamic test models and the patients' individual biologic factors. First, methods, models, tests as well as preanalytical factors have to be standardized according to the current knowledge in medicine laid down in the ISO 10993 part 4. The routine controls used in clinical chemistry and hematology have to be performed. Information about normal ranges (mean value, standard deviation, 95% confidence interval) should be provided. Tests have to be performed within a minimal delay of usually 2 h since some properties of blood change rapidly following collection. Various conditions (depending on the wall shear rate) were simulated within the centrifugation system and a Chandler system. Qualities and aspects of hemocompatibility such as platelet activation, oxidative burst, hemolysis, fibrinolysis, fibrin formation, generation of thrombin, contact activation, and complement activation were analysed and the results were entered non-dimensionally into a non-dimensional score system, where 0 points stand for the best and 65 points for the worst evaluation. We found a good correlation between the total score and contact activation, thrombin generation and leukocyte activation in a low shear stress system and a good correlation between the total score and thrombin generation, hemolysis and platelet activation in the high shear stress system. Further on the effect of additives and sterilization procedures can be measured. The concepts presented underline the relevance/importance of an efficient diagnostic approach to hemocompatibility that takes account of clinical and socio-economic concerns.

Transcutaneous flow related variables measured in vivo: the effects of gender.

BACKGROUND: The identification of potential sources of error is a crucial step for any new assessment technique. This is the case for transcutaneous variables, such as flow and arterial gases, which have been applied as functional indicators of various aspects of human health. Regarding gender, a particular subject-related determinant, it is often claimed that women present higher transcutaneous oxygen pressure (tcpO2) values than men. However, the statistical significance of this finding is still uncertain. METHODS: The haemodynamical-vascular response to a local reactive hyperaemia procedure (the tourniquet cuff manoeuvre) was studied in two previously selected group of volunteers (n = 16; 8 women and 8 men). The effect of gender was assessed under standardised experimental conditions, using the transcutaneous flow-related variables tcpO2-tcpCO2 and Laser-doppler Flowmetry (LDF). RESULTS: Regarding tcpO2, statistically significant differences between genders were not found, although higher values were consistently found for the gases in the female group. Regarding LDF, high statistically significant differences (p < 0.005) were found, with the men's group presenting the highest values and variability. Other derived parameters used to characterise the vascular response following the cuff-deflation (t-peak) were similar in both groups. CONCLUSIONS: The relative influence of gender was not clearly demonstrated using these experimental conditions. However the gender-related LDF differences suggest that further investigation should be done on this issue. Perhaps in the presence of certain pathological disparities involving peripheral vascular regulation, other relationships may be found between these variables.

A reevaluation and discussion on the threshold limit for hemolysis in a turbulent shear flow.

One major factor of red blood cell damage induced by artificial heart valves is the magnitude of turbulent shear stresses in the flow field. An often-cited threshold for hemolysis is 400 N/m(2) (Sallam and Hwang, Biorheology 21 (1984) 783). This value, however, was measured with a one-component laser Doppler anemometer and was not calculated from the major principal Reynolds shear stress, therefore underestimating the threshold. This study performed flow field measurements under similar conditions, using a two-component laser Doppler anemometer and determining major principal Reynolds shear stress, to reevaluate the hemolytic threshold as 800 N/m(2) with an exposure time of 1 ms. The Kolmogorov length scales, approximately 9 microm, are capable of inflicting direct damage to the red blood cells. These results will serve as a more accurate reference in future heart valve design and testing.

In vitro model of intravenous fluid administration: analysis of vein resistance to rapid fluid delivery.

Rapid fluid administration is the cornerstone of successful trauma resuscitation of patients in a state of shock. Intravenous (IV) fluid delivery is a physical intrusion into a vein which results in a complex interaction between the rigid catheter and the compliant vein. We present an experimental model of IV infusion into a vein-like compliant tube that (a) demonstrated the interdependence between fluid administration and blood flow in a compliant tube and (b) allowed investigation of the contribution of the central venous system (between the infusion site and the heart) to the total resistance to infusion flow rate. The results show that in cases with very high resistance in the central venous system a significant increase of infusion flow rate cannot be achieved just by increasing the infusion pressure. Similarly, in cases of small veins when only small catheters can be used, infusate flow rate may be increased only by using two independent infusion ports. In cases with increased tissue pressure due to edema, gravity-driven infusion may not produce sufficient perfusion of the vascular compartments. It was also shown that the vein valves do not always close, and that peripheral blood flow may continue together with the infusate fluid (e.g., when there is a small downstream resistance and infusion with a small catheter).

Preferred technique for blood flow volume measurement in cerebrovascular disease.

BACKGROUND AND PURPOSE: A noninvasive reliable technique that can reveal cerebral blood flow volume could be a valuable tool in screening programs for stroke prevention. In diagnostic ultrasonography, spectral Doppler imaging (SDI) is popular among sonologists and vascular technologists to estimate blood flow volume despite its documented inaccuracy and the availability of the more accurate technique of color velocity imaging (CVI). The aim of the present study was to demonstrate the discrepancy of blood flow volume estimation with CVI and SDI with use of an "internal" standard. METHODS: The common, internal, and external carotid arteries of 50 healthy subjects (22 men, 28 women, age range 19 to 54 years) were examined with CVI and SDI. The total blood flow volume of the internal and external carotid arteries was then compared with the ipsilateral common carotid artery flow. An accurate technique would demonstrate no difference. The difference (expressed as a percent inconsistency) was therefore a measure of the accuracy of the method. RESULTS: The mean+/-SD inconsistency was found to be 10.6+/-8.3% for CVI and 27.9+/-14.3% for SDI. The difference in inconsistency between CVI and SDI in measurement of carotid blood flow volume was statistically significant (P<0.01). CONCLUSIONS: CVI is more accurate than SDI in the determination of blood flow volume in the carotid arteries. For noninvasive clinical estimation of cerebrovascular blood flow volume, CVI quantification should be the preferred technique.

Measurement of leukocyte rheology in vascular disease: clinical rationale and methodology. International Society of Clinical Hemorheology.

The measurement of leukocyte rheology in vascular disease is a recent development with a wide range of new opportunities. The International Society of Clinical Hemorheology has asked an expert panel to propose guidelines for the investigation of leukocyte rheology in clinical situations. This article first discusses the mechanical, adhesive and related functional properties of leukocytes (especially neutrophils) which influence their circulation, and establishes the rationale for clinically-related measurements of parameters which describe them. It is concluded that quantitation of leukocyte adhesion molecules, and of their endothelial receptors may assist understanding of leukocyte behaviour in vascular disease, along with measurements of flow resistance of leukocytes, free radical production, degranulation and gene expression. For instance, vascular cell adhesion molecule (VCAM-1) is abnormally present on endothelial cells in atherosclerosis, diabetes mellitus and inflammatory conditions. Soluble forms of intercellular adhesion molecule (ICAM-1) or VCAM can be found elevated in the blood of patients with rheumatoid arthritis or infections disease. In the second part of the article, possible technical approaches are presented and possible avenues for leukocyte rheological investigations are discussed.

Chronic animal health assessment during axial ventricular assistance: importance of hemorheologic parameters.

Chronic testing of the Nimbus/UOP Axial Flow Pump was performed on 22 calves for periods of implantation ranging from 27 to 226 days (average, 74 days). The following parameters were measured: plasma free hemoglobin, blood and plasma viscosity, erythrocyte deformability and mechanical fragility, oxygen delivery index (ODI), blood cell counts, hematocrit, hemoglobin, blood urea nitrogen, creatinine, bilirubin, total protein, fibrinogen, and plasma osmolality. Most of the above parameters were stable during the full course of support. Compared with baseline, statistically significant differences during the entire period of implantation were only found in: hematocrit (p<0.001), hemoglobin (p<0.005), red blood cell (RBC) count (p<0.001), and whole blood viscosity (p<0.01). Plasma viscosity and ODI were mostly stable during the period of implantation. In some animals, an acute increase in fibrinogen concentration, plasma and blood viscosity, and a decrease in ODI were found to be early signs of the onset of infection. A small (10%) decrease in deformability of RBCs was found during the first 2 weeks after implantation. This alteration in RBC deformability was highly correlated (r = 0.793) with changes in total plasma protein concentration that fell more than 15% (p<0.001) during the same period. Mechanical fragility of RBCs was found to be slightly increased after implantation. Plasma free hemoglobin remained close to baseline level (p>0.2). After the first 2 weeks of the postoperative period, pump performing parameters for all animals were consistent and stable. In general, the Nimbus/UOP Axial Flow Pump demonstrated basic reliability and biocompatibility and did not produce significant alterations in the mechanical properties of blood or animal health status. The pump provided adequate hemodynamics and was well tolerated by the experimental animal for periods as long as 7.5 months. Monitoring rheologic parameters of blood is very helpful for evaluation of health during heart-assist device application.

Flow measurement in coronary surgery.

BACKGROUND: Many of the modern less invasive approaches to coronary artery bypass grafting (CABG) are performed without the use of the heart lung machine and cardiac asystole. Even after the introduction of mechanical stabilizers, the ability to achieve a technically perfect anastomosis is less certain in beating heart bypass surgery. Our group has begun to assess the surgical results of beating heart CABG using Transit Time Flow Measurement (TTFM). Our experience indicates that a meticulous and controlled method of assessing the results of intraoperative flow measurements can improve the quality of information and increases the accuracy of diagnosing technical problems with newly constructed bypass grafts. For this reason, we developed a standard algorithm for using and interpreting intraoperative TTFM. METHODS: From January to August of 1998, 161 patients underwent off-pump CABG with a total of 323 distal anastomoses (2.0 grafts per patient). All completed grafts were tested intraoperatively with TTFM and the decision to accept or revise any individual graft was based on a decision nomogram using key values readily available from the TTFM output. RESULTS: Thirty-two grafts (9.9%) were surgically revised based on unsatisfactory flow curves, the Pulsatile Index, or both. All revised grafts were found to have a significant technical error, such as an intimal flap, thrombus, conduit kinking, or dissection. There were no major complications, myocardial infarctions, or deaths in the entire series of patients. CONCLUSIONS: Based on our favorable use of TTFM, we strongly recommend that patency of every graft be assessed whether the operation is performed off pump or on cardiopulmonary bypass. Guidelines for performing and interpreting TTFM ensure a high degree of confidence in the completed graft. The decision to revise a graft can be made based on simple parameters easily acquired from the TTFM device. Any concern about quality or quantity of flow should prompt immediate revision.

A method of endoscopic investigation of vascular structures directly through flowing blood.

BACKGROUND: Noninvasive cardiovascular diagnosis has improved immensely due to key technological refinements such as digital subtraction angiography, ultrasonography, Doppler flow analysis, and magnetic resonance imaging. Each of these methodologies provides a unique image of the cardiovascular system but will not permit surgical maneuvers or repairs during real time imaging. Our group has developed a new method of endoscopic visualization of the luminal surface of blood vessels directly through flowing blood without interference of the blood or vessel wall. This opens new possibilities in both diagnosis and surgical interventions. METHODS: Transluminal imaging through flowing blood was performed in normal animals using laser frequency light delivered and retrieved via conventional fiberoptic angioscopic instruments. The reflected light energy was reconstructed into a viewable image using a specialized method of optical data processing and filtering systems. Unlike conventional angioscopy, displacement of flowing blood was not needed as the images were obtained with higher frequency laser light. RESULTS: A total of 20 canine experiments were performed between 1996 and 1997 using our endoluminal imaging system. The images obtained revealed details of luminal surfaces, although primitive and low resolution with this first generation of technology. Images of the topography of the femoral, axillary, and subclavian arteries and veins, as well as several intracardiac structures (aorta and aortic valve) were successfully obtained without trauma or physiologic consequence to the animal. CONCLUSIONS: Using conventional fiberoptic angioscopes coupled with laser light of differing wavelengths, it was possible to image the interior of vascular structures through flowing blood. This method visualizes the intraluminal surface in real time and is dependent only on the delivery and capacity of the endoscope. The implications for future cardiovascular diagnosis and corrective surgical procedures are widespread.

[Study on a new rheological equation of state for blood].

Blood behaves like a Non-Newtonian fluid under low shear rate. A number of constitutive equations for blood have been proposed in the literature, but yet a general equation which demonstrates all aspects of blood's Non-Newtonian characteristics has not been established. With this problem we analysed the Oldroyd Equation as a possible model for blood. Theory analysis indicated that the phenomena of blood flow under low shear rate, such as shear thinning, stress overshoot and phase delay in periodic flow, can be explained by introducing the equation with four parameters, suggesting it to be an appropriate model for blood. We tested the parameters. Stress versus time curves under rectangular step of shear rate were also tested and the results were compared with those of theory prediction.