The glycocalyx and its significance in human medicine.

Cells are covered by a surface layer of glycans that is referred to as the 'glycocalyx'. In this review, we focus on the role of the glycocalyx in vascular diseases (atherosclerosis, stroke, hypertension, kidney disease and sepsis) and cancer. The glycocalyx and its principal glycosaminoglycans [heparan sulphate (HS) and hyaluronic acid (HA)] and core proteins (syndecans and glypicans) are degraded in vascular diseases, leading to a breakdown of the vascular permeability barrier, enhanced access of leucocytes to the arterial intima that propagate inflammation and alteration of endothelial mechanotransduction mechanisms that protect against disease. By contrast, the glycocalyx on cancer cells is generally robust, promoting integrin clustering and growth factor signalling, and mechanotransduction of interstitial flow shear stress that is elevated in tumours to upregulate matrix metalloproteinase release which enhances cell motility and metastasis. HS and HA are consistently elevated on cancer cells and are associated with tumour growth and metastasis. Later, we will review the agents that might be used to enhance or protect the glycocalyx to combat vascular disease, as well as a different set of compounds that can degrade the cancer cell glycocalyx to suppress cell growth and metastasis. It is clear that what is beneficial for either vascular disease or cancer will not be so for the other. The overarching conclusions are that (i) the importance of the glycocalyx in human medicine is only beginning to be recognized, and (ii) more detailed studies of glycocalyx involvement in vascular diseases and cancer will lead to novel treatment modalities.

Vascular permeability in experimental diabetes is associated with reduced endothelial occludin content: vascular endothelial growth factor decreases occludin in retinal endothelial cells. Penn State Retina Research Group.

Blood-retinal barrier (BRB) breakdown is a hallmark of diabetic retinopathy, but the molecular changes that cause this pathology are unclear. Occludin is a transmembrane component of interendothelial tight junctions that may regulate permeability at the BRB. In this study, we examined the effects of vascular endothelial growth factor (VEGF) and diabetes on vascular occludin content and barrier function. Sprague-Dawley rats were made diabetic by intravenous streptozotocin injection, and age-matched animals served as controls. After 3 months, BRB permeability was quantified by intravenous injection of fluorescein isothiocyanate-bovine serum albumin (FITC-BSA), Mr 66 kDa, and 10-kDa rhodamine-dextran (R-D), followed by digital image analysis of retinal sections. Retinal fluorescence intensity for FITC-BSA increased 62% (P < or = 0.05), but R-D fluorescence did not change significantly. Occludin localization at interendothelial junctions was confirmed by immunofluorescence, and relative protein content was determined by immunoblotting of retinal homogenates. Retinal occludin content decreased approximately 35% (P < or = 0.03) in the diabetic versus the control animals, whereas the glucose transporter GLUT1 content was unchanged in rat retinas. Additionally, treatment of bovine retinal endothelial cells in culture with 0.12 nmol/l or 12 nmol/l VEGF for 6 h reduced occludin content 46 and 54%, respectively. These data show that diabetes selectively reduces retinal occludin protein expression and increases BRB permeability. Our findings suggest that the elevated VEGF in the vitreous of patients with diabetic retinopathy increases vascular permeability by downregulating occludin content. Decreased tight junction protein expression may be an important means by which diabetes causes increased vascular permeability and contributes to macular edema.

Effect of fluid flow on smooth muscle cells in a 3-dimensional collagen gel model.

A 3D collagen gel model was developed to simulate interstitial fluid flow and to assess the importance of this flow on the biochemical production rates of vascular smooth muscle cells (SMCs). Rat aortic SMCs were suspended in type I collagen, and the gel was supported by nylon fibers that allowed a 9-cm length of the SMC-gel model to withstand 90 cm H(2)O differential pressure over a 6-hour period without significant compaction. Up to 1 dyne/cm(2) shear stress on the suspended SMCs could be induced by the pressure-driven interstitial flow. The suspended SMCs were globular, had a diameter of approximately 10 microm, and were distributed uniformly throughout the gel. The collagen fibers formed a network that was connected randomly with the surface of SMCs and nylon fibers. The diameter of the collagen fibers was approximately 100 nm, and the concentration of collagen was 2.5 mg/mL. Using these parameters, fiber matrix theory predicted a Darcy permeability coefficient (K:(p)) of 1.22x10(-)(8) cm(2), which was close to the measured value of K:(p). The production rates of prostaglandin (PG) I(2) and PGE(2) were used as markers of biochemical responsiveness of SMCs to fluid shear stress. Both PGI(2) and PGE(2) production rates under 1 dyne/cm(2) shear stress were significantly elevated relative to static (no-flow) controls. The production rates, however, were approximately 10 times lower than observed when the same cells were plated on collagen-treated glass slides (2D model) and exposed to the same level of shear stress by use of a rotating disk apparatus. The results indicate that interstitial flow can affect SMC biology and that SMCs are more quiescent in 3D cultures than in 2D cultures. The 3D collagen gel model should be useful for future studies of interstitial flow effects on SMC function.

Computational model of collagen turnover in carotid arteries during hypertension.

It is well known that biological tissues adapt their properties because of different mechanical and chemical stimuli. The goal of this work is to study the collagen turnover in the arterial tissue of hypertensive patients through a coupled computational mechano-chemical model. Although it has been widely studied experimentally, computational models dealing with the mechano-chemical approach are not. The present approach can be extended easily to study other aspects of bone remodeling or collagen degradation in heart diseases. The model can be divided into three different stages. First, we study the smooth muscle cell synthesis of different biological substances due to over-stretching during hypertension. Next, we study the mass-transport of these substances along the arterial wall. The last step is to compute the turnover of collagen based on the amount of these substances in the arterial wall which interact with each other to modify the turnover rate of collagen. We simulate this process in a finite element model of a real human carotid artery. The final results show the well-known stiffening of the arterial wall due to the increase in the collagen content.

Effect of VEGF on retinal microvascular endothelial hydraulic conductivity: the role of NO.

PURPOSE: Vascular endothelial growth factor (VEGF) increases microvascular permeability in vivo and has been hypothesized to play a role in plasma leakage in diabetic retinopathy. Few controlled studies have been conducted to determine the mechanism underlying the effect of VEGF on transport properties (e.g., hydraulic conductivity [Lp]). This study was conducted to determine the effect of VEGF on bovine retinal microvascular endothelial LP and the role of nitric oxide (NO) and the guanylate cyclase/guanosine 3', 5'-cyclic monophosphate/protein kinase G (GC/cGMP/PKG) pathway downstream of NO in mediating the VEGF response. METHODS: Bovine retinal microvascular endothelial cells (BRECs) were grown on porous polycarbonate filters, and water flux across BREC monolayers in response to a pressure differential was measured to determine endothelial LP RESULTS: VEGF (100 ng/ml) increased endothelial LP: within 30 minutes of addition and by 13.8-fold at the end of 3 hours of exposure. VEGF stimulated endothelial monolayers to release NO and incubation of the BRECs with the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA; 100 microM) significantly attenuated the VEGF-induced LP increase. It was observed that incubation of the monolayers with the GC inhibitor LY-83583 (10 microM) did not alter the VEGF-mediated LP: response. Addition of the cGMP analogue 8-br-cGMP (1 mM) did not change the baseline LP over 4 hours. Also, the PKG inhibitor KT5823 (1 microM) did not inhibit the response of BREC LP to VEGF. CONCLUSIONS: These experiments indicate that VEGF elevates hydraulic conductivity in BRECs through a signaling mechanism that involves NO but not the GC/cGMP/PKG pathway.

Effect of pressure on hydraulic conductivity of endothelial monolayers: role of endothelial cleft shear stress.

Significant changes in transvascular pressure occur in pulmonary hypertension, microgravity, and many other physiological and pathophysiological circumstances. Using bovine aortic endothelial cells grown on porous, rigid supports, we demonstrate that step changes in transmural pressure of 10, 20, and 30 cmH(2)O induce significant elevations in endothelial hydraulic conductivity (L(p)) that require 5 h to reach new steady-state levels. The increases in L(p) can be reversed by addition of a stable cAMP analog (dibutyryl cAMP), and the increases in L(p) in response to pressure can be inhibited significantly with nitric oxide synthase inhibitors (N(G)-monomethyl-L-arginine and nitro-L-arginine methyl ester). The increase in L(p) was not due to pressure-induced stretch because the endothelial cell (EC) support was rigid. It is unlikely that the increase in L(p) was due to a direct effect of pressure because exposure of the cells to elevated pressure (25 cmH(2)O) for 4 h had no effect on the volume flux driven by a transmural pressure of 10 cmH(2)O. We hypothesize that elevated endothelial cleft shear stress induced by elevated transmural flow in response to elevated pressure stimulates the increase in L(p) through a nitric oxide-cAMP-dependent mechanism. This is consistent with recent studies of the effects of shear stress on the luminal surface of ECs. We provide simple estimates of endothelial cleft shear stress, which suggest magnitudes comparable to those imposed by blood flow on the luminal surface of ECs.

A numerical study of flow in curved tubes simulating coronary arteries.

Numerical simulations of pulsatile flow in coronary arteries which take into account the curvature associated with the bending of arteries over the surface of the heart are presented for resting, excited and drug induced states. The study was motivated by reported observations of atherosclerotic plaque localization on the inner curvature of coronary arteries. The simulated flow field appears quasi-steady under resting conditions with wall shear stress always highest on the outside wall and only a single secondary flow vortex in the half tube. However, reversal of wall shear stress direction at the inside wall does occur under resting flow conditions and this is not a quasi-steady characteristic. The flow field is markedly unsteady under excited conditions with wall shear stress sometimes peaking on the inside wall and an increase in the magnitude of wall shear stress reversal on the inside wall. However, only a single secondary flow vortex in the half tube is observed. Implications of the simulations for the role of fluid mechanics in coronary artery atherosclerosis are also discussed.

Pressure drop and flow rate measurements in a human aortic bifurcation cast for steady and pulsatile flow.

Pressure drop and flow rate measurements in a rigid cast of a human aortic bifurcation under both steady and physiological pulsatile flow conditions are reported. Integral momentum and mechanical energy balances are used to calculate impedance, spatially averaged wall shear stress and viscous dissipation rate from the data. In the daughter branches, steady flow impedance is within 30% of the Poiseuille flow prediction, while pulsatile flow impedance is within a factor of 2 of fully developed, oscillatory, straight tube flow theory (Womersley theory). Estimates of wall shear stress are in accord with measurements obtained from velocity profiles. Mean pressure drop and viscous dissipation rate are elevated in pulsatile flow relative to steady flow at the mean flow rate, and the exponents of their Reynolds number dependence are in accord with available theory.

A study of the wall shear rate distribution near the end-to-end anastomosis of a rigid graft and a compliant artery.

The development of intimal hyperplasia in the anastomotic region of a vascular graft which does not match the compliance of the parent artery may be related to altered wall shear rates near the anastomosis. The purpose of this study is to determine the effect of radial wall motion and the phase angle between pressure and flow waves (impedance phase angle) on the wall shear rate distribution near an end-to-end vascular graft anastomosis model incorporating a rigid graft and a compliant artery. The wall shear rate is determined from near-wall velocity profiles obtained by a flow visualization method using a photo-chromic dye for different locations near the anastomosis. The results show that the mean wall shear rate under pulsatile flow conditions is 15-30% lower than under steady flow conditions at the same mean flow rate. The effect of the impedance phase angle on the mean wall shear rate is shown to be small, but its effect on the amplitude of the wall shear rate is not negligible. For our anastomosis model which has well-matched diameters at the mean pressure, the mean shear rates at the distal sites are lower than at the proximal sites by 15-23%. We suppose that the differences in the mean wall shear rate between the proximal and distal sites are related to the convergent/divergent geometry caused by the mismatch of the compliance. Since the distal side of the anastomosis is more prone to intimal hyperplasia, lower shear rates near the distal anastomosis favor the hypothesis that low/oscillatory wall shear rates lead to intimal hyperplasia.

Compliance and diameter mismatch affect the wall shear rate distribution near an end-to-end anastomosis.

The development of intimal hyperplasia near the anastomosis of a vascular graft to an artery may be related to changes in the wall shear rate distribution. Mismatches in compliance and diameter at the end-to-end anastomosis of a compliant artery and a rigid graft cause shear rate disturbances that may induce intimal hyperplasia and ultimately graft failure. The goal of this study is to determine how compliance mismatch, diameter mismatch, and impedance phase angle affect the wall shear rate distribution in end-to-end anastomosis models under sinusoidal flow conditions. Wall shear rates are obtained through flow visualization using a photochromic dye. In a model with a well-matched graft diameter (6% undersized), the compliance mismatch causes low mean wall shear rates near the distal anastomosis. Considering diameter mismatch, the wall shear rate distributions in 6% undersized, 16% undersized, and 13% oversized graft models are markedly different at similar phase angles. In the two undersized graft models, the minimum mean shear rate occurs near the distal anastomosis, and this minimum is lower in the model with greater diameter mismatch. The oversized graft model has a minimum mean shear rate near the proximal anastomosis. Thus in all three models, the minimum mean wall shear rate is observed at the site of the divergent geometry. The impedance phase angle, which can be altered by disease states and vasoactive drugs, has a minor effect on the wall shear rate amplitude far from the anastomosis but a more pronounced effect closer to the anastomosis. Mean wall shear rates under sinusoidal flow conditions are significantly lower than under steady flow conditions at the same mean flow rate, but they are fairly insensitive to phase angle changes. In order to avoid the divergent geometry that may cause lower wall shear rates, we recommend that compliance mismatch be minimized whenever possible and that graft diameter be chosen to match the arterial diameter at the relevant physiologic pressure, not at the reduced pressure present when the graft is implanted.

Numerical simulation of unsteady laminar flow through a tilting disk heart valve: prediction of vortex shedding.

Heart valves induce flow disturbances which play a role in blood cell activation and damage, but questions of the magnitude and spatial distribution of fluid stresses (wall shear stress and turbulent stress) cannot be readily addressed with current experimental techniques. Therefore, a numerical simulation procedure for flow through artificial heart valves is presented. The algorithm employed is based on the Navier-Stokes equations in generalized curvilinear coordinates with artificial compressibility for coupling of velocity and pressure. The algorithm applies a finite-difference technique on a body-conforming composite grid around the heart valve disk on which the numerical simulations are performed. Steady laminar flow over a backward-facing step and unsteady laminar flow inside a square driven cavity are computed to validate the algorithm. Two-dimensional, time-accurate simulation of flow through a tilting disk valve with a steady upstream Reynolds number as high as 1000 reveals the complex behavior of 'vortex shedding'. By scaling the results at the Reynolds number of 1000 to peak systolic flow conditions, the maximum value of shear stress on the valve disk is estimated to be 770 dyn cm-2. The 'apparent' Reynolds stress associated with vortex shedding is estimated to be as high as 3900 dyn cm-2 with a vortex shedding frequency of about 26 Hz. The 'apparent' Reynolds stress value is of similar magnitude as reported in experiments but would not be expected to damage blood cells because the spatial scales associated with vortex shedding are much larger than blood cell dimensions.

In vitro studies of gas bubble formation by mechanical heart valves.

BACKGROUND AND AIM OF THE STUDY: Recent clinical research using transcranial Doppler ultrasonography has shown the presence of emboli in the cranial circulation of some mechanical heart valve patients. Due to the high-intensity signals produced by these emboli, it has been suggested that they are small gas bubbles. Meanwhile, transesophageal echocardiography of mechanical heart valve patients has shown images of bright, mobile particles (also considered to be gas bubbles) near the valve. Motivated by these reports, a series of in vitro studies was performed to investigate the relationship between dissolved gas concentration and the incidence of bubble formation after valve closure. METHODS: A mock circulatory loop was used to study a Medtronic Hall tilting disc valve in the mitral position of the Penn State Electrical Ventricular Assist Device (EVAD). The valve was videotaped as it operated in saline with various levels of dissolved CO2. A Doppler ultrasound probe served as a bubble detector on the outflow side of the EVAD. Measurements of vaporous cavitation intensity with a high-fidelity pressure transducer were also made. Similar experiments were then performed in porcine blood, using an imaging ultrasound transducer to detect bubbles. RESULTS: Bubbles were seen moving off the valve in the retrograde direction just after closure. The ultrasound probe detected these bubbles downstream, indicating a bubble lifetime on the order of seconds. It was observed with high-speed video that bubble formation and cavitation are separate events and occur at different times during valve closure. Bubbles were more likely to be observed when CO2 levels were higher. Experiments in blood provided images of bubbles near the valve, predominantly at higher CO2 levels and high valve loading conditions. CONCLUSIONS: These results show that stable gas bubbles can form during mechanical heart valve operation. The bubbles likely form from the combined effects of gaseous nuclei formed by cavitation, low-pressure regions associated with regurgitant flow, and the presence of CO2, a highly soluble gas.

A comparison of the cavitation potential of prosthetic heart valves based on valve closing dynamics.

BACKGROUND AND AIMS OF THE STUDY: This study compares the cavitation potential of prosthetic heart valves based on valve closing dynamics. METHODS: A laser sweeping technique measured valve closing dynamics (average closing velocity and deceleration) immediately before valve closure. A high-fidelity, piezoelectric pressure transducer was mounted proximal to the mitral valve and measured the high-frequency pressure fluctuations caused by cavitation bubble formation and collapse after valve closure. The band-pass filtered root mean squared (RMS) value of the mitral pressure signal was used as a measure of cavitation intensity. The combination of these two techniques allowed the direct correlation of valve dynamics and cavitation intensity for each valve closure. The effects of three parameters on prosthetic heart valve dynamics and cavitation were examined: valve geometry (Medtronic Hall and Björk-Shiley Monostrut), occluder material (pyrolytic carbon and Delrin), and gap width between the occluder and housing. A dimensional analysis was also performed to investigate the general form of the relationship between valve dynamics and cavitation intensity. RESULTS: For all of the valves investigated in this study, the RMS pressure increased (signifying an increase in cavitation) as the average closing velocity and deceleration increased. In order to compare the cavitation potential of the valves, the RMS pressure was estimated at specific closing velocities using the linear regression of RMS pressure versus average closing velocity for each valve. The effects of valve geometry, occluder material and gap width were then examined at high valve loading conditions (closing velocity of 4.0 m/s). For both pyrolytic carbon and Delrin, the Medtronic Hall valves had significantly higher RMS pressures than did the Björk-Shiley Monostrut valves. For a given valve geometry, the pyrolytic carbon occluder had a significantly higher RMS pressure than the Delrin occluder. The valve gap width did not have a significant effect on RMS pressure. The dimensional analysis revealed the general relationship among average closing velocity, occluder material properties and cavitation intensity. CONCLUSIONS: The results presented here contribute to our fundamental understanding of cavitation on mechanical heart valves.

Mitral heart valve cavitation in an artificial heart environment.

BACKGROUND AND AIMS OF THE STUDY: The formation and subsequent collapse of vaporous cavities in the fluid around mechanical heart valves at valve closure can create stresses large enough to damage both the valve itself and blood cells. Improved understanding of cavitation mechanisms should lead to a reduction in the cavitation potential of future valve designs. MATERIALS AND METHODS: This study compares eight mechanical mitral valves of two different geometries (Monostrut and Medtronic Hall), occluder housing gaps (tight, medium, and leaky), and occluder materials (Delrin and pyrolytic carbon). The valves were evaluated in a model ventricle of the Penn State Electric Ventricular Assist Device (EVAD) operating within a mock circulatory loop. The EVAD represents one half of a total artificial heart. The mock loop consisted of silicone tubing connected to elements designed to mimic the compliant and resistant properties of the natural circulation. Cavitation was controlled by varying the degree of filling of the ventricle: low filling caused higher valve closing velocities resulting in greater cavitation intensities than complete filling of the ventricle. The intensity of cavitation was quantified using a parameter derived from the high frequency fluctuations in the mitral pressure that occur around the valve during cavitation events. The shape of the cavitation pressure signature and that of the power spectrum of the cavitation pressure signature were used in addition to the cavitation intensity parameter to make comparisons between valves. RESULTS: Of the three valve characteristics studied, occluder material showed the most significant influence on cavitation intensity: valves with pyrolytic carbon occluders demonstrated greater cavitation than did those with Delrin discs. CONCLUSION: It is hypothesized that the dominant form of cavitation on the valves studied is related to vortex formation and that occluder material influences the intensity of cavitation through the strength of the tension wave generated at valve closure, while geometry and gap have only secondary effects. Future studies are planned to incorporate this technique in an in vivo environment.

An in-vitro investigation of prosthetic heart valve cavitation in blood.

Vapor cavities produced by low pressure fluid flow conditions have been observed in the vicinity of mechanical heart valves for many years. As cavities collapse during pressure recovery, they can produce stresses large enough to cause pitting of the valve occluders and lysing or activation of blood cells. To date, no method has been presented for the quantification of mechanical heart valve cavitation in blood because it has only been detected optically in transparent blood analog fluids. This paper describes a novel method for quantifying cavitation intensities in opaque fluids such as blood. It is based on the detection of high frequency pressure oscillations (35-350 kHz) at a location 4.5 cm proximal to a Björk-Shiley monostrut mitral valve in a mock circulatory loop driven by a Penn State Electric Ventricular Assist Device. The pressure oscillations which result from cavity collapse are used to quantify cavitation intensities in blood. One time domain and three frequency domain parameters have been developed to quantify cavitation intensity during a single valve closure event and over an ensemble of closure events. The time domain parameter is the Root Mean Squared (RMS) value of the pressure signal after it has been high-pass filtered at 35 kHz. The other three parameters are derived from the power spectrum of the pressure signal. One is the maximum value of the power spectrum between 100 and 200 kHz, another is the area under the power spectrum between 35 and 400 kHz, and the last is the volume under a 3-dimensional time vs. frequency vs. power spectrum plot. The parameters are averaged over a random sample of pressure traces to determine an average cavitation intensity for each operating condition studied. In addition, cavitation pressure fluctuations and hemolysis rates were determined simultaneously at several different mock flow loop operating conditions using porcine blood, and the relationships between various measures of cavitation intensity and the associated index of hemolysis have been established. Hemolysis was shown to increase with cavitation intensity.

Effect of shear stress on the hydraulic conductivity of cultured bovine retinal microvascular endothelial cell monolayers.

The shear stress of flowing blood on endothelial cells increases water transport (hydraulic conductivity, Lp) in several vascular beds in vivo and has been hypothesized to play a role in elevating vascular transport in ocular diseases such as diabetic retinopathy. The purpose of this study is to determine the response of Lp to varying levels of shear stress using an in vitro model of the blood-retinal barrier: bovine retinal endothelial cells (BRECs) grown on polycarbonate filters. The study also addresses the role of nitric oxide (NO) and other downstream effectors in mediating shear-induced changes in water transport. A step change in shear stress of 10 dyn/cm(2) did not produce a significant change in Lp over 3 hours, whereas a 20 dyn/cm(2) step change elevated Lp by 14.6-fold relative to stationary controls at the end of 3h of shear exposure. 20 dyn/cm( 2) of shear stress stimulated the endothelial monolayers to release nitric oxide in a biphasic manner and incubation of the BRECs with a nitric oxide synthase (NOS) inhibitor, L-NMMA, significantly attenuated the shear-induced Lp response. These experiments demonstrate that NO is a key signaling molecule in the pathway linking shear stress and Lp in BRECs. A widely studied pathway downstream of NO involves the activation of guanylate cyclase (GC), guanosine 3', 5' -- cyclic monophosphate (cGMP) and protein kinase G (PKG). It was observed that incubation of BRECs with the GC inhibitor, LY83583 (10 microM) or the PKG inhibitor, KT5823 (1 microM) did not significantly alter the shear-induced Lp response. Also the cGMP analogue, 8-br-cGMP (1mM), did not affect the baseline Lp over 4h. These results demonstrate that shear stress elevates hydraulic conductivity in BRECs through a signaling mechanism that involves NO but not the GC/cGMP/PKG pathway.

Physiological transport properties of cultured retinal microvascular endothelial cell monolayers.

PURPOSE: To characterize baseline transport properties: hydraulic conductivity (Lp), albumin permeability (Pe), and transendothelial electrical resistance (TER) of bovine retinal microvascular endothelial cells (RMEC) in the development of an in vitro model of the blood-retinal barrier (BRB). METHODS: RMEC were grown on porous, polycarbonate filters for determination of the number of days required to achieve minimal transport rates. Lp, Pe, and TER were measured by utilizing a bubble tracking spectrophotometer, by quantifying the diffusional movement of fluorescein isothiocyanate-labeled albumin, and by utilizing a Millipore electrical resistance meter, respectively. RESULTS: Lp decreased significantly from 7.82 +/- 0.85 x 10(-7) (mean +/- SEM) cm/sec/cm H2O at post-plating Day 5 to 1.44 +/- 0.26 x 10(-7) cm/sec/cm H2O at Day 9. Pe of the monolayer also decreased progressively with days post-plating from 3.44 +/- 0.53 x 10(-6) cm/sec at Day 7 to a minimum of 1.95 +/- 0.29 x 10(-6) cm/sec at Day II. Peak TER fluctuated until Day 7, when it began to steadily increase from 17.14 ohm-cm2 to a peak value of 25.42 ohm-cm2 at Day 10, decreasing from then on to 22.24 ohm.cm2 on Day 12. Known disrupters of the BRB, NECA and VEGF, elicited significant increase in RMEC Lp showing the sensitivity of this model to pharmacological alterations. CONCLUSIONS: Our data indicate that RMEC grown on polycarbonate filters form a restrictive monolayer of cells, which exhibit dynamic alterations in response to pharmacological agents, thus demonstrating an in vitro model of the BRB. Future studies with the model may offer insights into the pathogenesis of retinal vascular diseases and allow convenient testing of pharmacological interventions.

Computational simulation of flow in the end-to-end anastomosis of a rigid graft and a compliant artery.

Implanted vascular grafts often fail because of the development of intimal hyperplasia in the anastomotic region, and compliance mismatch between the host artery and graft exacerbates the problem. This study focused on the effects of radial artery wall motion and phase angle between pressure and flow waves (impedance phase angle [IPA]) on the wall shear rate (WSR) behavior near end-to-end vascular graft anastomoses models connecting rigid grafts and compliant arteries. A finite element model with transient flow and moving boundaries was set up to simulate oscillatory flow through a 16% undersized (mean) diameter graft model. During the simulations, different artery diameter variations (DVs) over a cycle (DV) and IPAs were simulated in the physiologic range for an oscillatory flow (mean Re = 150, peak Re = 300, unsteadiness parameter alpha = 3.9). The results show that for normal physiologic conditions (DV = 6%, IPA = -45 degrees) in a 16% undersized graft, the minimum distal mean WSR is reduced by 60% compared to steady flow at the mean Re; the minimum distal WSR amplitude increases 50% when IPA changes from -5 degrees to -85 degrees, and increases 60% when DV changes from 2% to 10%. This indicates that compliance mismatch induces lower mean WSR and more oscillatory WSR in the distal anastomotic region, which may contribute to intimal hyperplasia. In addition, the convergent-divergent geometry of the 16% undersized graft model can significantly affect the force pattern applied to the local endothelial cell layer near the anastomosis by altering the local phase angle between the flow induced tangential force (synchronous with WSR) and the radial artery expansion induced cyclic hoop strain (synchronous with DV). This local phase angle is decreased by 65 degrees in the distal divergent geometry, while increased by 15 degrees in the proximal convergent geometry.

Relative blood damage in the three phases of a prosthetic heart valve flow cycle.

Blood flow through a prosthetic heart valve operating in a ventricular assist device can be subdivided into three phases: a) forward flow through an open valve, b) rapid valve closure, and c) regurgitant back flow through a closed valve. Recent studies of fluid stresses in the Penn State Electric Left Ventricular Assist Device (PS LVAD) operating under physiologic conditions indicate that Reynolds stresses of possibly hemolytic magnitude may exist in the valve area. Although several studies have been made of the fluid stresses seen in forward flow through an open valve, few have looked at valve closure or backflow, and none have related these stresses directly to blood damage. In this study, novel in vitro blood flow loops were developed to allow for the separate analysis of the three flow phases of a Bjork-Shiley monostrut Delrin disk valve operating in a PS LVAD. Forward flow through fully open aortic and mitral valves and backflow through closed valves are studied separately in flow loops driven by a roller pump with the LVAD acting as a valve housing and compliance vessel. Valve closure is investigated with a PS LVAD operating in a low volume mock circulatory loop characterized by cavitation potential through stroboscopic videography of this mock loop, using saline as the working fluid. Rate of hemolysis, characterized by the index of hemolysis, IH, is determined for each of the three flow loops charged with fresh porcine blood.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo observation of cavitation on prosthetic heart valves.

In this study, a method to determine the existence of prosthetic heart valve cavitation in vivo is presented. Pennsylvania State University Left Ventricular Assist Devices (LVADs) were implanted in two separate calves for this study. Björk-Shiley Monostrut (Irvine, CA) 27 mm and 25 mm valves with Delrin occluders were used in the mitral and aortic positions, respectively. A high fidelity, piezoelectric pressure transducer was mounted approximately 1.25 cm proximal to the mitral valve and measured the high frequency pressure fluctuations caused by cavitation bubble formation and collapse after valve closure. The root mean square (RMS) value of the mitral pressure signal during a 5 ms interval after valve closure was used as a measure of cavitation intensity. The pressure signals observed in vivo were similar to ones observed in vitro with the same type of pressure transducer and were associated with the visually observed cavitation. The percentage of beats with cavitation increased from 20.3% to 67.7% when pump filling was decreased by increasing beat rate. A blood test conducted during post-operative days 1-3 showed a significant increase in plasma hemoglobin during the low filling condition. However, blood tests conducted later (post-operative days 7-44) did not show a significant change in plasma hemoglobin during low filling conditions.