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Geometric distribution of chordae tendineae: an important anatomic feature in mitral valve function.

BACKGROUND AND AIM OF THE STUDY: This study examined the geometric distribution of chordae tendineae and their importance in compensating for papillary muscle (PM) displacement. METHODS: Anatomic, chordal mechanics and hemodynamic measurements were performed with porcine mitral valves. For hemodynamic measurements, physiological pulsatile flow conditions were maintained, and PM positions varied. Leaflet coaptation was documented by 2-D echocardiography, and regurgitation measured directly. RESULTS: Anatomic measurements showed the sum of marginal leaflet and marginal chordal lengths to exceed basal chordal length (1.8+/-0.4 versus 2.8+/-0.7 cm for anterior leaflets; 1.6+/-0.3 versus 2.5+/-0.6 cm for posterior leaflets). Triangular structures existed between basal chordae and marginal chordae with the marginal leaflet as the third side. Basal chordae resisted apical PM displacement in static experiments, while marginal chordae governed leaflet closure in hemodynamic experiments. Under pulsatile flow conditions, apical PM displacement decreased leaflet coaptation length and increased regurgitation (9.4+/-2.1 versus 4.0+/-1.6 ml). When marginal chordae were fused to the basal chordae, eliminating the role of the marginal chordae, severe regurgitation resulted (28.5+/-5.0 ml with apical PM displacement). CONCLUSION: Based on triangular structures involving the basal and marginal chordae, a compensatory mechanism was described which explains how the severity of mitral regurgitation can vary following PM displacement. Basal chordae provide a constant connection between the annulus and papillary muscles, while marginal chordae maintain marginal leaflet flexibility, governing proper valve closure. This study relates chordal distribution to normal valve function, and provides a better understanding of breakdown in valve function under pathophysiological conditions.

Animals↗

Pulsatile versus oscillatory shear stress regulates NADPH oxidase subunit expression: implication for native LDL oxidation.

Shear stress regulates endothelial nitric oxide and superoxide (O2-*) production, implicating the role of NADPH oxidase activity. It is unknown whether shear stress regulates the sources of reactive species production, consequent low-density lipoprotein (LDL) modification, and initiation of inflammatory events. Bovine aortic endothelial cells (BAECs) in the presence of 50 microg/mL of native LDL were exposed to (1) pulsatile flow with a mean shear stress (tau(ave)) of 25 dyne/cm2 and (2) oscillating flow at tau(ave) of 0. After 4 hours, aliquots of culture medium were collected for high-performance liquid chromatography analyses of electronegative LDL species, described as LDL- and LDL2-. In response to oscillatory shear stress, gp91phox mRNA expression was upregulated by 2.9+/-0.3-fold, and its homologue, Nox4, by 3.9+/-0.9-fold (P<0.05, n=4), with a corresponding increase in O2-* production rate. The proportion of LDL- and LDL2- relative to static conditions increased by 67+/-17% and 30+/-7%, respectively, with the concomitant upregulation of monocyte chemoattractant protein-1 expression and increase in monocyte/BAEC binding (P<0.05, n=5). In contrast, pulsatile flow downregulated both gp91phox and Nox4 mRNA expression (by 1.8+/-0.2-fold and 3.0+/-0.12-fold, respectively), with an accompanying reduction in O2-* production, reduction in the extent of LDL modification (51+/-12% for LDL- and 30+/-7% for LDL2-), and monocyte/BAEC binding. The flow-dependent LDL oxidation is determined in part by the NADPH oxidase activity. The formation of modified LDL via O2-* production may also affect the regulation of monocyte chemoattractant protein-1 expression and monocyte/BAEC binding.

Animals↗

The measurement of cardia output by the thoracic impedance method.

The aim of our experiments was to study the thoracic electrical impedance method as a method for measuring cardiac output in anesthetized dogs. Four electrodes were placed around the neck and thorax. A 50 kHz, 1 mA electric current was applied to the outer two electrodes and the two inner electrodes were used to measure impedance changes related to the stroke volume during the cardiac cycle at end-expiratory apnea. The cardiac output obtained by the impedance method was compared to the cardiac output measured by isotope dilution and by the electromagnetic flowmeter. Either mean cardiac output or cardiac output determined beat-to-beat from the pulsatile flow was measured with the latter method. Significant correlations were obtained between the impedance and the isotope dilution method (r = 0.8799), and between the impedance and the electromagnetic (mean) flow measurements (r = 0.7330). The comparison of impedance cardiac output to that determined from the pulsatile flow (beat-to-beat) also showed a good correlation (r = 0.7618). The effect of changing the fluid and air contents in the chest on the basal thoracic impedance was also studied.

Animals↗

Relative performance of prosthetic heart valves based on power measurements.

The in vitro steady and pulsatile flow results of tests of a series of valves are reported. Fifteen mechanical and two tissue valves from 23 mm to 27 mm nominal size from six manufacturers, plus two prototype experimental valves, were tested in the aortic position in a circulation system simulator. Steady-flow pressure drop/flow rate measurements were direct-recorded on an X-Y plotter. Pulsatile flow rate, LVP, AoP, pressure drop, and pressure-drop power were recorded. Input (left ventricle pumping) power was also recorded directly as the multiplier output from the product of flow rate and LVP. Results are presented in tables and graphs. Power loss at 70/min varied from 3% to 6%; backflow from 2.5% to 11%. Valve evaluation is based on the relative valve power loss that the heart must overcome. Backflow is shown to be as important as pressure drop in determining valve performance. A performance index (PI) equal to the percentage pressure-drop power loss plus the percentage backflow is presented as a means for performance rating. PI values from 6 to 17 were obtained on the 19 valves tested.

Aortic Valve↗

Evaluation of vascular compromise in the injured extremity: a photoplethysmographic technique.

A monitoring technique in which an infrared photoplethysmograph (PPG) is used to assess the vascular status of injured extremities is described. PPG has been correlated with blood flow in the forearm muscle and direct measurements of compartment pressure. These studies show a significant decrease in amplitude of the PPG signal with diminished blood flow in muscle and with compartment pressures greater than 40 mm Hg. We used PPG clinically on 55 extremities injured by direct vascular trauma, crushing forces, or severe burns. Although clinical signs suggested use of escharotomy in six of 29 burned extremities, normal PPG signals were present and conservative management resulted in no long-term morbidity. Three limbs had moderate pulsatile flow shown by PPG that significantly diminished over a period of 12 to 24 hours even though Doppler flow signals remained strong. Good pulsatile flow was restored in all cases after early escharotomy . PPG monitoring of vascular status is simple and reproducible and accurately reflects levels of ischemia in injured extremities.

Adult↗

Wall shear stress and early atherosclerotic lesions in the abdominal aorta in young adults.

OBJECTIVES: To study the correlation between wall shear stress and early atherosclerotic lesions in the abdominal aorta. DESIGN: Blinded histomorphometric studies. Comparison with in vitro data. MATERIALS: Abdominal aortic haemodynamics were simulated in a realistic pulsatile flow model. Abdominal aortas from 10 young adults with no signs of atherosclerotic disease were obtained during autopsy. METHODS: Quantitative wall shear stresses were measured at rest and exercise in one suprarenal and two infrarenal positions using laser Doppler anemometry. Intimal thickening indices were measured blindly at the corresponding locations using histomorphometric methods, and compared to wall shear stress variables using linear regression analysis. RESULTS: Intimal thickness index increased significantly with age. Intimal thickness index was significantly lower in the suprarenal than the infrarenal aorta, and higher at the distal posterior vessel wall compared to the anterior wall. Intimal thickness index correlated significantly with mean, minimum and oscillating wall shear stresses measured at rest. CONCLUSION: Intimal thickness in the undiseased abdominal aorta correlated significantly with mean, minimum and oscillating wall shear stresses at rest measured in a pulsatile flow model. No correlations were found with maximum shear stress parameters. Exercise changed the local wall shear stresses away from the characteristics associated with intimal thickness index.

Adolescent↗

Isolation of endothelial cells and their progenitor cells from human peripheral blood.

PURPOSE: We have developed techniques to isolate endothelial cell (EC) progenitors from human peripheral and umbilical cord blood. METHODS: Human adult peripheral and umbilical cord blood monocytes were isolated by centrifugation, and progenitor cells were separated with the use of magnetic polystyrene beads that were coated with a monoclonal antibody specific for the CD34 cell-membrane antigen. Cells were propagated in selective media, and developing cultures were immunostained for CD31, CD34, factor VIII, and vascular endothelial growth factor cell receptors. ECs that developed were transfected with a gene for prourokinase and used to line ePTFE grafts, which were evaluated in vitro in a pulsatile flow system. RESULTS: Umbilical cord monocyte cultures demonstrated colonies that resembled ECs at approximately 2 weeks, with growth being best supported by EC growth media plus 20% calf serum with iron. Immunostaining of colonies was positive for CD31 and factor VIII. After 18 days in culture, CD34(+) cells from adult peripheral blood were noted, which had the typical cobblestone appearance of ECs and immunostained positively for CD31 and factor VIII-related antigens. Cultures of umbilical cord-derived cells and adult peripheral blood-derived cells developed complex line formations within 1 week in culture that stained positively for vascular endothelial growth factor receptor-2. Urokinase-transfected ECs were shown to overexpress urokinase. Prosthetic grafts lined with transfected cells showed 87.33% +/- 4.97% cell adherence after 2 hours in a pulsatile flow system at clinically relevant shear stress. CONCLUSION: We conclude that endothelial progenitor cells can be isolated from human adult peripheral and umbilical cord blood and developed into EC cultures as a source of cells for vascular graft seeding and gene therapy.

Adult↗

Impacts of pulsatile systemic circulation on endothelium-derived nitric oxide release in anesthetized dogs.

BACKGROUND: The effects of pulsatile flow on endothelium-derived nitric oxide-mediated vasodilation are not fully elucidated in an in vivo model. METHODS: A left ventricular assist device was established in 10 anesthetized dogs with a centrifugal pump and an air-driven pneumatic pump. The systemic circulation was subjected to step changes in the frequency of pulse (0, 30, 60, and 120 bpm with a fixed pulse pressure of 50 mm Hg), and in the amplitude of pulse (0, 20, and 50 mm Hg with a fixed pulse rate of 120 bpm). Hemodynamic variables and calculated total systemic vascular resistance were compared before and after the administration of N(G)-Nitro-L-arginine Methyl Ester (L-NAME) (20 mg/kg). Plasma NO2-/NO3- concentration levels were also measured. RESULTS: Total systemic vascular resistance significantly decreased while plasma NO2-/No3- concentration increased in response to the rise in both pulse rate and pulse pressure. However, L-NAME significantly diminished these effects of pulsatile flow. CONCLUSIONS: Both the frequency and the amplitude of pulse wave in the systemic circulation are significant independent stimuli for endothelium-derived nitric oxide-mediated vasodilation in vivo.

Animals↗

In vitro fabrication of a tissue engineered human cardiovascular patch for future use in cardiovascular surgery.

BACKGROUND: One approach to tissue engineering has been the development of in vitro conditions for the fabrication of functional cardiovascular structures intended for implantation. In this experiment, we developed a pulsatile flow system that provides biochemical and biomechanical signals in order to regulate autologous, human patch-tissue development in vitro. METHODS: We constructed a biodegradable patch scaffold from porous poly-4-hydroxy-butyrate (P4HB; pore size 80 to 150 microm). The scaffold was seeded with pediatric aortic cells. The cell-seeded patch constructs were placed in a self-developed bioreactor for 7 days to observe potential tissue formation under dynamic cell culture conditions. As a control, cell-seeded scaffolds were not conditioned in the bioreactor system. After maturation in vitro, the analysis of the tissue engineered constructs included biochemical, biomechanical, morphologic, and immunohistochemical examination. RESULTS: Macroscopically, all tissue engineered constructs were covered by cells. After conditioning in the bioreactor, the cells were mostly viable, had grown into the pores, and had formed tissue on the patch construct. Electron microscopy showed confluent smooth surfaces. Additionally, we demonstrated the capacity to generate collagen and elastin under in vitro pulsatile flow conditions in biochemical examination. Biomechanical testing showed mechanical properties of the tissue engineered human patch tissue without any statistical differences in strength or resistance to stretch between the static controls and the conditioned patches. Immunohistochemical examination stained positive for alpha smooth muscle actin, collagen type I, and fibronectin. There was minor tissue formation in the nonconditioned control samples. CONCLUSIONS: Porous P4HB may be used to fabricate a biodegradable patch scaffold. Human vascular cells attached themselves to the polymeric scaffold, and extracellular matrix formation was induced under controlled biomechanical and biodynamic stimuli in a self-developed pulsatile bioreactor system.

Absorbable Implants↗

Shear-stress preconditioning and tissue-engineering-based paradigms for generating arterial substitutes.

In situ tissue engineering using shear-stress preconditioning and adhesive biomolecules is a new approach to autologous tissue engineering. In the present study, novel tissue-engineering grafts (TEGs) were preconditioned within an in vitro pulsatile flow circuit, with and without the addition of fibronectin (FN), to establish whether low-shear-stress conditions promoted endothelial cell (EC) retention and differentiation. TEGs ( n =24) were generated by the contraction and compaction of collagen(I) by porcine aortic smooth-muscle cells (SMCs) on to a compliant polyester graft scaffold. ECs were radiolabelled with [(111)In]indium tropolonate and seeded on to the luminal surface of the TEGs. Following organ culture in a bioreactor (7 days), TEGs were split into four groups ( n =six TEGs per group): Group A acted as controls with TEGs unmodified and seeded with radiolabelled ECs; Group B underwent luminal pre-coating with FN (75 microg/ml) prior to EC seeding; Group C underwent preconditioning within a pulsatile flow circuit at 10-20 microN (1-2 dyn)/cm(2) for 7 days prior to EC seeding, and Group D TEGs were preconditioned for 7 days at 1-2 dyn/cm(2), followed by luminal pre-coating with FN prior to EC seeding. The resistance to physiological shear stress of the seeded ECs was assessed using a gamma-radiation counter within a physiological flow circuit producing an arterial waveform with a mean shear stress of 93.2 microN (9.32 dyn)/cm(2). Environmental scanning electron microscopy (ESEM) was used to determine the distribution and degree of differentiation of the attached Ecs, and tissue-type-plasminogen-activator (tPA) assays provided a measure of function and viability. EC resistance to shear stress at 93.2 microN/cm(2) was significantly enhanced by a period of preconditioning (Group C) at 10-20 microN/cm(2), surface modification with FN (Group B), or both (Group D) when compared with control grafts (Group A). However, TEGs coated with FN whether preconditioned (Group D) or not (Group B) demonstrated the best results for EC retention. ESEM demonstrated near-confluent differentiated flattened ECs in both these cases. EC function was demonstrated by a steady increase in tPA production. Low-shear-stress preconditioning of TEGs enhances EC retention in vitro with an additional advantage demonstrated by pre-treatment with FN prior to endothelialization. These findings may be exploited in the development of tissue-engineered constructs to maintain a confluent endothelial lining.

Adaptation, Physiological↗

Cine MR in the evaluation of normal and abnormal CSF flow: intracranial and intraspinal studies.

Evaluation of intracranial and intraspinal CSF flow was accomplished by the use of cardiac gated gradient echo magnetic resonance (MR) technique. Normal patterns of pulsatile flow within the ventricles, cisterns and cervical subarachnoid space were established by this technique and these observations were compared to prior description of CSF flow. With systole there is downward (caudal) flow of CSF in the aqueduct of Sylvius, the foramen of Magendie, the basal cisterns and the dorsal and ventral subarachnoid spaces while during diastole, upward (cranial) flow of CSF in these same structures is seen. The relationships between the cardiac cycle and the CSF pulsations are demonstrated on both magnitude reconstruction and phase reconstruction MR images. Calculations of actual fluid velocity within CSF containing spaces can be obtained from the phase reconstruction images and holds promise for a more accurate analysis of CSF flow. In conditions which result in alterations of flow, cine MR dramatically shows either obstruction or excessively turbulent flow within the CSF pathways. The site of obstructed flow whether in the third ventricle, aqueduct, fourth ventricle, or subarachnoid space can be appreciated by changes in or absence of the normal hypointense signal. Cystic cord lesions such as congenital syringohydromyelia and posttraumatic spinal cord cysts may show pulsatile flow of CSF, a fact which can relate to progressive enlargement of these cysts. The distinction between myelomalacia and cyst formation in the cord is facilitated by the technique. Although the use of cine MR for the analysis of CSF flow is in its infancy, our experience indicates that this technique is useful in a wide range of pathological conditions including, but not limited to, conditions resulting in hydrocephalus or cystic cord lesions.

Brain↗

Numerical analysis of flow in an elastic artery model.

Oscillatory and pulsatile flows of Newtonian fluids in straight elastic tubes are simulated numerically with the aid of Ling and Atabek's "local flow" assumption for the nonlinear convective acceleration terms. For the first time, a theoretical assessment of the local flow assumption is presented, and the range of validity of the assumption is estimated by comparison with perturbation solutions of the complete flow problem. Subsequent simulations with the local flow model indicate that the flow field and associated wall shear stress are extremely sensitive to the phase angle between oscillatory pressure and flow waves (impedance phase angle). This phase angle, which is a measure of the wave reflection present in the system, is known to be altered by arterial disease (e.g., hypertension) and vasoactive drugs. Thus, the paper elucidates a mechanism by which subtle changes in systemic hemodynamics (i.e., phase angles) can markedly influence local wall shear stress values.

Aorta↗

Local hemodynamics affect monocytic cell adhesion to a three-dimensional flow model coated with E-selectin.

Monocyte adhesion to the endothelium depends on concentrations of receptors/ligands, local concentrations of chemoattractants, monocyte transport to the endothelial surface and hemodynamic forces. Monocyte adhesion to the inert surface of a three-dimensional perfusion model was shown to correlate inversely with wall shear stress, but was also affected by flow patterns which influenced the near-wall cell availability. We hypothesized that (a) under the same flow conditions, insolubilized E-selectin on the model's surface may mediate adhesive interactions at higher wall shear stresses, compared to an uncoated model, and (b) pulsatile flow may modify the adhesion profile obtained under steady flow. An axisymmetric flow model with a stenosis and a sudden expansion produced a range of wall shear stresses and a separated flow with recirculation and reattachment. Pre-activated U937 cells were perfused through the model under either steady (Re = 100, 140) or pulsatile (Remean = 107) flow. The velocity field was characterized through computational fluid dynamics and validated by inert particle tracking. Surface E-selectin greatly increased cell adhesion in all regions at Re = 100 and 140, compared to an uncoated model under the same flow conditions. In regions where the cells near the wall were abundant (taper and stenosis), adhesion to E-selectin correlated with the reciprocal of local wall shear stress when flow was steady. Pulsatile flow distributed the adherent cells more evenly throughout the coated model. Hence, characterizing both the local hemodynamics and the biological activity on the vessel wall is important in leukocyte adhesion.

Cell Adhesion↗

[A clinical study of cerebral perfusion during pulsatile and nonpulsatile cardiopulmonary bypass].

The purpose of this study was to determine the effect of pulsatile flow on cerebral perfusion under cardiopulmonary bypass (CPB). Twenty-three patients who underwent cardiac operations were divided into two comparable groups: Group A (N = 11) had standard nonpulsatile flow, while in Group B (N = 12), a pulsatile pump was used. The blood flow of left common carotid artery and radial arterial pressure were continuously monitored during cardiac operation in both groups and cerebral vascular resistance was calculated. In Group B, the perfusion pressure of left common carotid artery was monitored and compared with that of radial artery. Arterial and internal jugular venous blood were sampled and the difference of cerebral A.V O2 contents and cerebral oxygen consumption was calculated. Cerebral vascular resistance in Group B (54.0 +/- 11.2% of the value of before-CPB) significantly decreased compared to that in Group A (72.2 +/- 11%) at the end of CPB (p less than 0.05). Pulse pressure following pulsatile CPB flow was 15.1 +/- 5.8 mmHg monitored in radial artery and it reduced to 8.5 +/- 5 mmHg in left common carotid artery. Although there was no significant difference in cerebral oxygen consumption of both groups during and just after CPB, the difference of cerebral A-V O2 contents of Group B was greater than Group A just after CPB. These data suggest that pulsatile flow may minimize the cerebral microcirculatory shunt during CPB, resulting from the reduction of cerebral vascular resistance.

Aged↗

Flow investigations in a model of a three-dimensional human artery with Newtonian and non-Newtonian fluids. Part I.

Together with biochemical factors, fluid mechanical factors play a role in atherogenesis and the deposition of blood platelets at bends and bifurcations in human arteries. Hence, flow patterns were investigated in a simplified 3-dimensional model of a human renal artery bifurcation using Newtonian (aqueous glycerol) and non-Newtonian (aqueous solution of polyacrylamide) fluids. Studies were carried out in steady as well as pulsatile flow at inflow Reynolds numbers of 498 and 951 with flow rate ratios main tube V1: right branch V4: left branch V3 of 1: 0.25: 0.25 and 1: 0.18: 0.18 respectively. The velocity distribution proximal and distal to the bifurcations was measured using a laser-Doppler anemometer. In steady flow, zones of flow separation and reverse flow were observed distal to the bifurcations. In pulsatile flow using non-Newtonian fluids, there was a significant enlargement of these zones. Differences between the Newtonian and non-Newtonian fluids occurred especially distal to the bifurcations. Shear stresses along all measuring positions were computed from the velocity gradients.

Acrylic Resins↗

Hydro- and hemodynamic effects of catheterization of vessels. II. Model experiments comparing circular and annular lumen area reduction.

Experiments and a simple theoretical analysis have been performed concerning the flow through circular (stenosis) and annular (catheter piece) lumen area reduction of rigid- and elastic-walled vessels. Both steady and pulsatile flow at various volumetric flow rates were investigated. A good agreement between the experimental and the analytical results has been found. A critical annular lumen area reduction exists on a par with the well-known critical stenosis. The strongest reduction in pressure and in volumetric flow rate was caused by the catheter pieces, compared with the stenoses. The per cent reduction in pulse mean pressure and in volumetric mean velocity at pulsatile flow was equal to the per cent reduction in pressure and volumetric mean velocity at steady flow. The influence of the geometrical shape of the stenoses and the catheter pieces is further discussed.

Arteries↗

Evaluation of pulsatile and nonpulsatile flow in microvessels of the bulbar conjunctiva in the goat with an undulation pump artificial heart.

This study has three purposes, as follows. The first is to develop a microscopic system to observe the microcirculation of animals implanted with an artificial heart. The second is to investigate the influence of flow pattern change from pulsatile to nonpulsatile on the microcirculation. The third is to study the effects of pulsatility in blood flow on endothelium-derived nitric oxide release in the microvasculature. When the flow pattern was changed from pulsatile to nonpulsatile, the velocity of erythrocytes in many capillaries dropped and remained at a low level, and the number of perfused capillaries decreased. After the flow pattern was returned to pulsatile, the velocity of erythrocytes recovered to the initial level. In many cases, the flow of nonperfused capillaries recovered to the initial level as well. Also, the pulsatile flow enhances the basal and flow-stimulated endothelium-derived nitric oxide release in microvessels.

Animals↗

In vitro flow dynamics of four prosthetic aortic valves: a comparative analysis.

The velocity fields downstream of four prosthetic heart valves were mapped in vitro over the entire cross-section of a model aortic root using laser Doppler anemometry. THe Björk-Shiley 60 degrees convexo-concave tilting disc valve, the Smeloff-Cutter caged ball valve, the St. Jude Medical bileaflet valve, and the Ionescu-Shiley standard bioprosthesis were examined under both steady and pulsatile flows. Velocity profiles under steady flow conditions were a good approximation for pulsatile profiles only during midsystole. The pulsatile flow characteristics of the four valves showed variation in large scale flow structures. Comparison of the valves according to pressure drop, shear stress and maximum velocities are also provided.

Aortic Valve↗