The effects of extremely low shear stress on cellular proliferation and neointimal thickening in the failing bypass graft.

OBJECTIVE: Previous studies demonstrating a correlation between low shear stress (tau = 5-15 dyne/cm(2)) and experimental vein graft neointimal thickening (NIT) support the role of low tau in vein graft failure. However, a simple linear relationship between low tau and NIT would underestimate the degree of NIT evident in high-grade occlusive lesions of failing human vein grafts. In this study we used a new experimental model that maintains patency at low tau (< 2 dyne/cm(2)), to delineate possible deviations from linearity in the low tau --> NIT hypothesis. METHODS: Thirty-two New Zealand White rabbits underwent creation of a common carotid vein patch with a segment of ipsilateral external jugular vein. Very low tau was created in 13 patches by ligation of the distal common carotid artery, leaving the only outflow through a small muscular branch. Normal tau was created in 11 patches by leaving the common carotid artery outflow intact. High tau was created in eight patches by ligation of the contralateral common carotid artery. Six patches were harvested after 2 weeks for measurement of cell cycle entry by proliferating cell nuclear antigen (PCNA) immunohistochemistry. The remaining 26 patches were harvested after 4 weeks, perfusion fixed, and excised for morphometric analysis. RESULTS: Mean blood flow and tau at implantation ranged from 0.5 to 41 mL/min and 0.07 to 15 dyne/cm(2), respectively. At the time of harvest, 30 of 32 patches remained patent, and the artificially created aberrations in blood flow were maintained (range, 0.7-41 mL/min). After 2 weeks PCNA immunohistochemistry showed a significantly higher level of cell cycling in patches exposed to low tau (40 +/- 5 vs 1.6 +/- 0.3 PCNA-positive cells per high-power field; P <.001), which is equivalent to approximately 20% of the total cells present. In patches harvested after 4 weeks, NIT ranged from 42 to 328 microm and significantly correlated with mean tau at implantation. Patches with very low tau exhibited histologic characteristics similar to those of failing human bypass grafts, including laminar thrombus and flow-limiting luminal stenosis. The relationship between tau and NIT was nonlinear in that extremely low tau (< 2 dyne/cm(2)) resulted in NIT beyond that predicted by a simple linear correlation (P =.003). CONCLUSION: Extremely low tau (< 2 dyne/cm(2)) stimulates high rates of smooth muscle cellular proliferation in arterialized vein patches. NIT is accelerated in these regions of low tau far beyond that predicted by a simple linear model. The nonlinear nature of the cellular proliferative response and NIT at tau less than 2 dyne/cm(2) may explain the rapid progression of neointimal lesions in failing bypass grafts.     

Modulation of vascular remodeling induced by a brief intraluminal exposure to the recombinant R7020 strain of Herpes simplex-1

OBJECTIVE: Vascular remodeling in response to injury or low shear stress (or both) is characterized by neointimal hyperplasia and luminal contraction. When profound, the response leads to restenosis after percutaneous endovascular intervention as well as to de novo stenosis in vein grafts. It has recently been reported that exposure of vein patches to neurovirulence-attenuated Herpes simplex virus-1 (HSV-1) decreases neointimal hyperplasia and increases luminal area. This experiment tested the hypothesis that R7020, a more highly attenuated mutant of HSV-1, would modulate the vascular remodeling response of experimental vein grafts chronically exposed to low shear stress. METHODS: The external jugular veins of 31 New Zealand white rabbits were clamped and intraluminally exposed to vehicle (phospate-buffered saline solution, n = 11), R7020 2.5 x 10(8) plaque forming units [PFU]/mL (n = 8), or R7020 2.5 x 10(9) PFU/mL (n = 12) for 10 or 30 minutes at an average pressure of 80 mm Hg. After exposure, an end-to-side distal external jugular-to-common carotid artery anastomosis was created, resulting in a widely patent arteriovenous fistula. The external jugular was suture-ligated just proximal to the thoracic inlet, distal to a small 10- to 50-microm venous tributary, creating a reversed vein "graft" segment immediately and abruptly exposed to arterial pressure (48 +/- 3 mm Hg) and low shear stress (0.12 +/- .02 dyne/cm(2)). In the 29 animals (N = 31) that survived to harvest, 26 grafts were found to be patent and were analyzed further. Nine grafts were harvested within the first week after operation, snap frozen in liquid nitrogen, and assayed for the presence of the Herpes viral immediate-response protein ICP0 by Western blot analysis. The 17 remaining grafts were perfusion-fixed, excised, stained, and analyzed morphometrically by digital planimetry. RESULTS: In patent grafts, the hemodynamic environment of low shear stress was maintained (shear stress at harvest, 0.26 +/- .06 dyne/cm(2)). Western blot analysis revealed the presence of ICP0 in R7020-exposed vein grafts after 2, 3, 7, and 14 days; ICP0 was not detected in unexposed vein grafts or adjacent carotid arteries. After 4 weeks, vein grafts exposed to R7020 exhibited a statistically significantly increased ratio of luminal radius to wall thickness, indicating altered remodeling (vehicle, 6.7 +/- 1.3; R7020 2.5 x 10(8), 9.1 +/- 1.3; R7020 2.5 x 10(9) ratio, 11.3 +/- 1.4; P < .05 for high dose compared with vehicle). CONCLUSION: A brief exposure of the neurovirulence-attenuated HSV-1 strain R7020 results in an increased ratio of luminal radius to wall thickness in experimental vein grafts chronically exposed to low shear stress.     

Optimal graft diameter: effect of wall shear stress on vascular healing

Arterial walls tend to adapt to maintain a specific wall shear stress. The formation of neointimal hyperplasia and endothelial cell healing of polytetrafluoroethylene grafts may also be governed by wall shear stress, which suggests that an optimal graft diameter may exist. To test this, 40 polytetrafluoroethylene grafts with internal diameters of 3, 6, and 8 mm were inserted end to end in the femoral and carotid arteries of 10 mongrel dogs. Total flow and diameter were measured, and grafts were stained with Evans blue dye, fixed by pressure perfusion, and analyzed by computer for anastomotic neointimal thickening, graft pseudointimal thickening, and degree of endothelial coverage. Mean calculated shear stress was 41 dyne/cm2 for the 3 mm grafts, 7 dyne/cm2 for the 6 mm grafts, and 3 dyne/cm2 for the 8 mm grafts. Fifteen weeks later the patency rate was 0 of 10 for the 3 mm grafts, 16 of 20 for the 6 mm grafts, and 7 of 10 for the 8 mm grafts. The mean graft shear stress was calculated to be 10 dyne/cm2 for the 6 mm grafts and 4 dyne/cm2 for the 8 mm grafts. Pseudointima lining the graft was composed of disorganized protein and cell remnants. The rough surface contained no overlying endothelium. Anastomotic neointima contained a layer of well-organized smooth muscle cells covered by a single layer of polygonal-shaped endothelial cells. A transition zone of thrombus, which is sandwiched by a wedge of smooth muscle cells near the graft surface and covered by endothelial cells, is described. Mean thickness of pseudointima of the patent 8 mm grafts was 150 microns thicker than that of the 6 mm grafts. Anastomotic neointimal thickness was 110 microns thicker in the 8 mm grafts compared with the 6 mm grafts. Among the 6 mm grafts, the carotid grafts had an average initial shear stress of 10 dyne/cm2, whereas the femoral grafts averaged a lower 5 dyne/cm2 and yielded pseudointima and neointima that were 40 microns thicker. The percent graft surface area covered with neointima did not differ among the grafts of differing diameter either proximally or distally. Lower shear stresses produced greater amounts of pseudointimal thickening within polytetrafluoroethylene grafts and neointimal thickening at their anastomoses. Conversely, the high shear stress from small-diameter grafts was associated with poor graft patency. These results suggest that an optimal graft diameter may help to prevent neointimal hyperplasia and graft thrombosis.     

Remodeling and suppression of intimal hyperplasia of vascular grafts with a distal arteriovenous fistula in a rat model.

PURPOSE: The purpose of this study was to evaluate the effects of a distal arteriovenous fistula (dAVF) on the morphologic changes occurring in arterial bypass grafts by the use of a novel experimental model. METHODS: Aortofemoral bypass grafts with or without dAVFs were constructed in 36 Sprague-Dawley rats with a microsurgical technique. The bypass graft material consisted of deendothelialized autogenous tail artery (length, 25 mm; inside diameter, 0.5 mm). In 18 rats, dAVFs were constructed at the distal anastomosis. After 6 weeks, flow rates and shear stress were determined, and grafts were then harvested. Luminal, intimal, and medial cross-sectional areas were measured with computer imaging. Desmin, alpha-smooth muscle actin, and von Willebrand factor (vWF) were identified with immunohistochemistry. Endothelialization was evaluated with SEM. RESULTS: All bypass grafts remained patent at the time of graft harvest. Grafts with dAVFs showed increased flow rates (11.5 +/- 0.6 mL/min) compared with grafts without dAVFs (2.1 +/- 0.3 mL/min; P < .01). Shear stress was also increased in the dAVF group (340.9 +/- 23.4 dyne/cm(2) vs 113.7 +/- 12.5 dyne/cm(2); P < .01), with a corresponding suppression of intimal hyperplasia (0.059 +/- 0.011 mm(2) for dAVF grafts vs 0.225 +/- 0.009 mm(2) for non-dAVF grafts; P < .01). Staining for vWF was found in both the reendothelialized flow surface and the neointimal extracellular matrix. Remodeling of the grafts was characterized by a 50% increased luminal area, 70% decreased intimal area, and a 25% decreased medial area when a dAVF was constructed. CONCLUSION: A small animal experimental model of an arterial bypass graft can enable the evaluation of a variety of factors that influence graft patency. Increased blood flow velocity and shear stress induced by a dAVF are associated with a decrease in intimal and medial areas, which may reflect changes in cell proliferation, apoptosis, migration, or matrix deposition. Deposition of vWF was also found both in the endothelium and throughout the hyperplastic intima. These findings suggest that the hemodynamic and morphologic changes associated with dAVF may potentiate graft patency and function.     

Slower Onset of Low Shear Stress Leads to Less Neointimal Thickening in Experimental Vein Grafts

Vein grafts respond to low flow and shear stress (τ_w) by generating thicker walls and smaller lumens through the processes of neointimal hyperplasia and remodeling. Clinically, however, vein grafts with obviously low τ_w, such as those distal to high-grade proximal obstructions, are not infrequently found to be widely patent and pliable. One possible explanation for this phenomenon may be that vein grafts remodel more favorably in response to changes in shear that occur gradually over time compared to abruptly. This hypothesis was tested in an experimental animal model in this report. Two separate models of experimental vein graft failure were created, causing either immediate exposure to ultralow τ_w (<1 dyne/cm²) or delayed exposure to ultralow τ_w. Under general anesthesia and using a sterile technique, the right external jugular (EJ) veins of 28 New Zealand white rabbits were surgically exposed and isolated. An end-to-side distal EJ/common carotid artery anastomosis was created, resulting in a widely patent arteriovenous fistula. For the immediate exposure group (n = 5), the EJ was suture-ligated just proximal to the thoracic inlet, distal to a small 10-50 μm venous tributary. This created a reversed vein segment immediately and abruptly exposed to high wall tension (2.0 ± 0.3 × 10⁴ dyne/cm) and ultralow τ_w (0.15 ± 0.08 dyne/cm²). For the delayed exposure group (n = 22), the EJ was ligated over a 0.035 guidewire, leaving a small aperture to sustain some measure of blood flow and τ_w. This predictably resulted in slightly less wall tension (1.4 ± 0.2 × 10⁴ dyne/cm) and higher τ_w (0.68 ± 0.21 dyne/cm²) than the immediate exposure group. During the first week, the small outflow aperture in the delayed exposure grafts thrombosed, eventually exposing them to the same low level of τ_w as the immediate exposure grafts. Thus, the only difference in the two models was that delayed exposure grafts enjoyed a slower decline in τ_w than immediate exposure grafts. Fourteen rabbits in the delayed exposure group were harvested over the first 7 days to define the patency curve of the restricted outflow channel. As expected, the small aperture had thrombosed in all animals by 7 days. The remaining 14 grafts were harvested after 4 weeks, and 13/14 remained patent. Examination of the hemodynamic parameters at the time of death confirmed that wall tension and τ_w had equalized (wall tension 0.9 ± 0.1 vs. 1.1 ± 0.1 × 10⁴ dyne/cm, τ_w 0.45 ± 0.12 vs. 0.30 ± 0.08 dyne/cm²). Histological examination revealed less neointimal hyperplasia in the delayed exposure group compared to the immediate exposure group (wall thickness 266 ± 16 vs. 180 ± 24 μm, p = 0.025) as well as a slightly greater luminal diameter (0.30 ± 0.02 vs. 0.40 ± 0.02 cm, p = 0.038). The results of this experiment suggest that slow exposure to reduced τ_w results in more favorable remodeling (less thickening) than abrupt exposure. This finding may explain the occasional clinical observation of a widely patent vein graft even in the face of proximal arterial obstruction and very low flow; the change in τ_w presumably occurred slowly mitigating the remodeling response. Presented at the Vascular Society 2004 Annual Meeting, Anaheim, CA, June 5, 2004.     

Dialysis access failure: A sheep model of rapid stenosis.

PURPOSE: Intimal hyperplasia at the venous anastomosis of dialysis access grafts causes early failure, although increased flow inhibits intimal hyperplasia in arterial grafts and after vessel injury. We designed a sheep model to study this process. METHODS: Polytetrafluoroethylene (PTFE) grafts were placed in the necks of sheep from the carotid artery to the external jugular vein. Grafts were harvested after perfusion fixation at 4, 8, and 12 weeks and submitted for histologic and immunohistochemical examination, including morphometry of neointimal lesions. RESULTS: The venous anastomoses developed thick neointima within the PTFE graft by 4 weeks. Lesions at the venous end were significantly thicker than those at the arterial end by 8 weeks (1.2 +/- 0.1 vs 0.38 +/- 0.05 mm, P <.02) and had greater cross-sectional area at both 4 (0.32 +/- 0.21 vs 3.6 +/- 0.8 mm(2), n = 7, P <.02) and 8 weeks (9.8 +/- 1.9 vs 1.1 +/- 0.7 mm(2), n = 7, P <.02). Only one of the four grafts (25%) in the 12-week group remained patent. Lesions were composed of smooth muscle cells, matrix, and thrombus of various ages. Cellular proliferation was prominent in neointima adjacent to thrombus and in granulation tissue surrounding the graft. Organizing thrombus contributed significantly to luminal narrowing. CONCLUSION: The sheep model of dialysis access reliably produces venous stenosis within 4 weeks. Lesions develop in the absence of uremia, graft puncture, or dialysis, suggesting that these factors are not necessary for graft failure. The continued presence of thrombus and high rates of cellular proliferation suggest ongoing injury is an important cause of lesion formation. This model allows study of the cellular mechanisms of dialysis failure.     

The potassium channel opener levcromakalim causes expansive remodelling of experimental vein grafts

BACKGROUND: Maintenance of luminal area is essential for the optimal performance of venous bypass grafts. However, injury and response to the arterial circulation evoke vascular remodelling that favors intimal hyperplasia, with luminal encroachment and inward remodelling. Potassium channel-opening drugs reduce tissue workload and peripheral vascular resistance and through these mechanisms could favor outward or expansive remodelling of vein grafts. We tested the hypothesis that levcromakalim, a potassium channel opener, would enhance expansive remodelling in vein grafts. METHODS: A randomized, double-blind, placebo-controlled trial was conducted in 33 rats with vena cava-to-aorta bypass grafts. Drugs were administered via osmotic pump for 7 days after surgery. Half the cohort had bromodeoxyuridine (BrdU) infused at day 6. Morphometric analysis was conducted of pressure perfusion-fixed grafts harvested at 1 week and 4 weeks. RESULTS: At 1 week, lumen area was similar in both groups (1.82 +/- 0.39 mm(2) placebo vs 1.85 +/- 0.36 mm(2) levcromakalim), although medial cell density and BrdU staining were significantly increased in the placebo group. At 4 weeks, lumen area was unchanged in the placebo group (1.88 +/- 0.51 mm(2)) but had increased to 2.32 +/- 0.46 mm(2) in the levcromakalim group (P = .039 vs 1 week), with a very significant reduction in the intimal area (levcromakalim, 0.06 +/- 0.02 mm(2) vs placebo, 0.33 +/- 0.17 mm(2); P = .001). CONCLUSIONS: Early, short-term treatment with levcromakalim favors expansive remodelling of experimental vein grafts to mimic the effect of external stenting. This expansive remodelling was associated with a reduction in medial cell proliferation at 1 week. CLINICAL RELEVANCE: Critical limb ischemia can be treated by bypass surgery or angioplasty, but inward remodelling with restenosis is a common problem. There has been little previous experimental work to identify treatments associated with expansive remodelling, which would increase the chances of vessel patency. Here, in a randomized trial, we show that short-term treatment with a potassium channel opener (a class of drug that can be used to treat hypertension) results in strong, expansive remodelling, with increases the lumen area and graft size of experimental vein grafts by >25%.     

Effect of adenoviral titer and instillation pressure on gene transfer efficiency to arterial and venous grafts ex-vivo

OBJECTIVE: Adenoviral-mediated gene transfer to arterial and venous grafts has potential in the treatment of a number of vascular diseases. Despite widespread use of these vectors to mediate gene transfer to blood vessel walls, the optimal transduction conditions for each type of vessel has yet to be determined. Our objective was to study the effect of adenoviral titer and instillation pressure on efficiency of gene transfer to arterial and venous grafts ex-vivo. METHODS: Jugular vein and carotid artery segments of 8 cm were harvested from Yorkshire Cross pigs. Tissue culture media or different titers of an adenoviral vector encoding human placental alkaline phosphatase (hpAP) were instilled into venous and arterial grafts at 0 mm Hg or 80 to 100 mm Hg of pressure and bathed externally in the same solution at 37 degrees C for 30 minutes. The grafts were rinsed, opened longitudinally, and incubated in culture media at 37 degrees C for 48 hours. Grafts were fixed and stained for hpAP transgene expression to quantitate percent luminal transduction or homogenized for alkaline phosphatase (AP) activity to determine total transmural transduction. RESULTS: For venous grafts, the percent luminal area stained for hpAP was greatest with 10(8) plaque-forming units/mL at 0 mm Hg (81% +/- 7%) and decreased with increasing titers (53% +/- 9% at 10(9) pfu/mL and 44% +/- 11% at 5 x 10(9) pfu/mL; n = 7; P <.05). No increase in percent luminal area stain was achieved with an instillation pressure of 80 to 100 mm Hg at any viral titer. The inverse finding was observed in arterial grafts. For arterial grafts, the greatest percent luminal area stained was achieved with 5 x 10(9) pfu/mL at 80 to 100 mm Hg (76% +/- 7%). An instillation pressure of 80 to 100 mm Hg increased the percent luminal area stained at 10(8) pfu/mL from 31% +/- 9% to 66% +/- 8% (n = 8; P =.01). For venous grafts, total AP activity peaked with 10(9) pfu/mL at 0 mm Hg and decreased with an instillation pressure of 80 to 100 mm Hg (30.6 +/- 9.7 U/mg versus 10.9 +/- 2.5 U/mg; n = 7; P <.01). However, for arterial grafts, total AP activity peaked with 5 x 10(9) pfu/mL (0 mm Hg) and increased with an instillation pressure of 80 to 100 mm Hg (32.8 +/- 9.9 U/mg versus 63.4 +/- 20.5 U/mg; n = 8; P <.05). CONCLUSION: High transduction efficiency can be achieved with adenoviral-mediated gene transfer of arterial and venous grafts. Gene transfer with the vascular graft's physiologic pressure conditions improved transduction efficiency for the artery (80 to 100 mm Hg) and vein (0 mm Hg). Comprehensive analysis of adenoviral transduction conditions is important to realize the full promise of adenoviral-mediated gene transfer.     

Quantitation of vascular outflow by measurement of impedance

One of the most important determinants of graft patency is the degree and character of vascular outflow. This study was designed to evaluate input impedance as a functional assessment of the outflow bed of vascular grafts. Four distinct outflow environments were created for external jugular vein conduits in 42 New Zealand white rabbits. Vein grafts (n = 14) were fashioned as end-to-side common carotid interposition bypass grafts. Arteriovenous fistulas (n = 15) were created by side-to-side anastomosis of the distal common carotid artery and linguofacial vein. Arteriovenous fistulas with outflow obstruction (n = 7) were fistulas with a metal clip partially obstructing the distal outflow channel (1 mm lumen). Vein graft/arteriovenous fistula combinations (n = 6) consisted of a vein graft and arteriovenous fistula in series. Pressure and flow in the external jugular vein were measured, and input impedance spectra were calculated by Fourier methods. By use of a PC-based acquisition and processing system, impedance results for 20 cardiac cycles could be obtained in approximately 10 minutes. The results revealed that vein grafts typically demonstrated high resistance to steady state flow (Rin = 235 +/- 50 x 10(3) dyne . sec/cm-5) and steadily decreasing impedance to pulsatile flow resulting in a characteristic impedance (Z0; average of fourth to tenth harmonics) of 35.5 +/- 8.0 x 10(3) dyne . sec/cm-5. Phase angle values were usually negative, especially at low harmonics (first harmonic phase angle = -1.11 +/- 0.10 radians) indicating that flow led pressure. In contrast, arteriovenous fistula Rin was minimal (6.3 +/- 1.4 x 10(3) dyne . sec/cm-5; p less than 0.05 compared to vein graft, and the impedance was flat across the frequency spectrum (Z0 = 8.5 +/- 1.5 x 10(3) dyne . sec/cm-5; p less than 0.05) with pressure and flow nearly in phase (first harmonic phase angle = -0.05 +/- 0.10 radians). Creation of outflow obstruction in arteriovenous fistulas resulted in significantly elevated Rin (136 +/- 41 x 10(3) dyne/sec . cm-5; p less than 0.05 compared to arteriovenous fistula and Z0 (23 +/- 9 x 10(3) dyne . sec/cm-5, p less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)     

Endothelial cell seeding of expanded polytetrafluoroethylene vena cava conduits: effects on luminal production of prostacyclin, platelet adherence, and fibrinogen accumulation

The blood-surface interface of 12 mm ID x 5 cm ePTFE vena cava conduits, unseeded (n = 8) and seeded (n = 8) with enzymatically derived autologous endothelial cells, was studied in a canine model at 4 and 12 weeks after graft implantation. Acetylsalicylic acid (325 mg each day) and Coumadin (prothrombin time 1.4 to 1.7 times the control value) were administered preoperatively and continued 4 weeks postoperatively. Platelets labeled with 111In and 125I-labeled fibrinogen were administered 24 hours before graft removal. Luminal platelet adherence, expressed as 10(6) platelets/cm2 of graft surface, was 8.9 +/- 5.6 vs. 56.4 +/- 8.0 (p less than 0.008) and 4.0 +/- 0.9 vs. 12.4 +/- 2.3 (p less than 0.005) in seeded vs. unseeded grafts at 4 and 12 weeks, respectively. Luminal fibrinogen deposition, expressed in micrograms per square centimeter of graft surface, was 11.8 +/- 2.2 vs. 32.0 +/- 2.0 (p less than 0.06) and 6.1 +/- 2.4 vs. 12.4 +/- 6.3 (p less than 0.005) in seeded vs. unseeded grafts at 4 and 12 weeks, respectively. Cumulative 4- and 12-week luminal production of 6-keto-PGF1 alpha from seeded and unseeded grafts represented 11% and 5%, respectively, of that produced from the native iliac vein. Luminal endothelial cell coverage was 71% +/- 22% vs. 33% +/- 9% and 79% +/- 8% vs. 55% +/- 8% (p less than 0.05) in seeded and unseeded grafts at 4 and 12 weeks, respectively. Although endothelialization was not complete in seeded vena cava grafts, it is clear that seeded prostheses exhibited improved thromboresistance compared with unseeded conduits.     

Effect of wall shear stress on intimal thickening of arterially transplanted autogenous veins in dogs

To determine whether or not changes in wall shear stress play a determinant role in the induction of hyperplasia of intimal tissue of arterially transplanted vein grafts, we developed two models of canine femoral arteries. Wall shear stress was defined by variation of wall shear stress (tau-variation) in the cardiac cycle, with the use of a newly designed computational flow waveform analyzer. In the group I model autogenous vein grafts were implanted under flow conditions of 79.7 +/- 3.2 ml/min of the normally high flow rate with 33.1 +/- 1.9 dynes/cm2 of low tau-variation. In the group II model grafts were implanted under conditions of 2.9 +/- 1.8 ml/min of low flow rate with 178.8 +/- 11.0 dynes/cm2 of normally high value of tau-variation. The intimal thickness of 259 +/- 36 microns 4 weeks after implantation in group I was statistically significant compared with that of 31 +/- 14 microns in group II (p less than 0.005). Our study revealed that change in wall shear stress and not the rate of blood flow is the essential hemodynamic factor related to intimal hyperplasia.     

Theoretical hydraulic consequences of vein graft taper

OBJECTIVE: Internal diameter is a strong predictor of patency of infrainguinal vein grafts. However, most vein grafts are tapered, with variable diameter along their length. It is unknown which diameter is most important in determining graft resistive properties, that is, its mean diameter, minimum diameter, or some geometric combination thereof. The purpose of this analysis was to examine the hydraulic consequences of vein graft tapering, with longitudinal impedance (Z(L)), a conduit-specific measure of pulsatile resistance along straight rigid tubes. METHODS: Proximal and distal graft pressure, pressure gradient (DeltaP), and blood flow (Q) were measured intraoperatively in a 100 cm bypass graft and digitally recorded for 10 seconds at 200 Hz. With the Womersley solution for fully developed fluid flow in a rigid tube, a series of DeltaP waveforms were generated for graft diameters ranging from 1.2 to 8.2 mm. With an axisymmetric form of the Navier-Stokes equations, a second series of DeltaP waveforms were computed for grafts with long smooth symmetric tapers ranging from 0% to 90%, with geometric mean diameter of 3.2, 4.2, and 5.2 mm (%Taper = 100 x [proximal diameter - distal diameter]/proximal diameter). For each set of DeltaP and Q, Z(L) was calculated as DeltaP/Q, plotted over a range of 8 Hz, and integrated over 4 Hz to yield integral Z(L). RESULTS: The architecture of the calculated DeltaP and Z(L) waveforms closely approximated their measured counterparts, validating the method. As expected, Z(L) was highly diameter-dependent in a nonlinear fashion. With a clinically relevant boundary of less than 50 x 10(3) dyne/cm(5) as "acceptable," the minimum acceptable diameter of nontapered 100 cm bypass conduits was 4.3 mm. Analysis of graft taper revealed that small amounts of taper in large conduits were well-tolerated. For example, introduction of 32% taper in a 5.2 mm graft (6.2 mm --> 4.2 mm) caused only an 8% increase in integral Z(L) (from 32 to 35 x 10(3) dyne/cm(5)). More pronounced taper in smaller conduits rendered them unacceptable. For example, 53% taper of a 4.2 mm graft (5.7 mm --> 2.7 mm) created a conduit with integral Z(L) of 70 x 10(3) dyne/cm(5), well above the acceptable limit. The relationship between Z(L) and percent taper was nonlinear and strongly dependent on mean diameter. CONCLUSIONS: The relationship between Z(L) and diameter in vein grafts is nonlinear; thus Z(L) increases rapidly in conduits smaller than 4 mm. Tapered vein grafts behave hydraulically like nontapered grafts, provided their geometric mean is greater than 4 mm and their degree of taper is less than 40%. Tapered veins are satisfactory conduits for long-segment bypass grafts, provided their mean diameter is acceptable.     

Immunocytochemical study on endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery with the use of vascular mayo stripers. A randomized controlled trial.

OBJECTIVES: The aim of this study is to compare the endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery and veins harvested conventionally for coronary artery bypass surgery in 200 participants who were assigned to interventions by using random allocation. DESING: Randomized controlled trial. Methods. Immunocytochemistry with anti-CD 31 antibodies and anti-nitric oxide synthase (NOS) antibodies were employed to identify the endothelial integrity. RESULTS: The CD 31 immunostaining showed that the endothelial cell integrity of the minimally invasive harvested veins was preserved in 82+/-13% of the circumference of luminal endothelium, while in conventionally harvested grafts it was reduced to 64+/-15% (p=0.05).> This was associated with the lack of CD 31 expression in vasa vasorum (10 and 18%) in both groups, respectively, (p=0.02). The NOS immunostaining revealed that the endothelial integrity of the minimally invasive harvested grafts was preserved in 96+/-4% of the luminal endothelium circumference as compared to 74+/-10% in conventionally harvested grafts (p=0.05). The percentage of cases with the lack of NOS expression in all vasa vasorum was 12 and 21%, in G1 and G2, respectively, (p=0.02). CONCLUSION: The endothelial integrity of saphenous vein grafts harvested by minimally invasive surgery is better preserved than with the grafts obtained by the conventional manner. This could play an important role in improving vein graft patency rates.     

Effects of distention and short-term grafting on vasoreactivity in rabbit external jugular veins.

The relative effects of distention, intraluminal pressure, and wall tension on venous smooth muscle and endothelial cell function were examined in 40 external jugular veins from New Zealand white rabbits. Vein grafts (n = 5) were interposed in the common carotid artery and explanted after 10 minutes. Distended veins were inflated in vitro with modified Krebs' solution at 37 degrees C for 10 minutes at pressures of either 20 mm Hg (D-20; n = 5) or 80 mm Hg (D-80; n = 5). Externally supported veins (ES-80; n = 5) were inflated at 80 mm Hg pressure, but distention was prevented by covering with a 3 mm internal diameter polytetrafluoroethylene sleeve. Bradykinin-induced in vitro maximal tension was attenuated significantly in vein grafts (0.13 +/- 0.04 g) and D-80 rings (0.27 +/- 0.07 g) compared with D-20 rings (1.20 +/- 0.14 g), ES-80 rings (0.99 +/- 0.13 g), or nondistended control rings (n = 40; 1.19 +/- 0.10 g; p less than 0.001). The attenuation in contraction in the vein graft and D-80 groups was nonspecific (i.e., similar results were obtained with respect to other smooth muscle agonists). Contractile function was inversely associated with wall tension, the product of pressure and radius (r2 = 0.7438; p = 0.06). In contrast, there were no differences in endothelium-dependent or endothelium-independent relaxation among the five groups. It is concluded that, in this experimental system, (1) venous smooth muscle function is significantly attenuated after short-term in vitro distention or grafting although endothelial function is largely preserved, and (2) the decrement in contraction is due to elevated wall tension.     

Pressure traps in femoro-popliteal reversed vein grafts. Are valves culprits?

BACKGROUND: Stenosis is a major cause of vein graft failure in peripheral arterial surgery. Our goal is to determine whether vein valves play a role in this process by creating a "pressure trap". METHODS: Seventeen patients with femoro-popliteal reversed saphenous vein grafts were studied intraoperatively. Flow and pressure in the grafts were measured, while the graft outflow was gradually occluded and released for 2-4 seconds. In 3 patients the graft flow was reduced by compressing calf muscles. RESULTS: Patients heart rates were 54-84 BPM, blood pressures 170/80-110/55 mm Hg, and normal graft flow was 40-180 ml/min. In 12 patients with competent vein valves, at reduced flow (<30 m/min) the valves opened and closed in each cardiac cycle. At each closure the pressure was "trapped" distal to the valve producing diastolic hypertension. Also the flow was stagnant for a considerable portion of the cardiac cycle. Maximum diastolic pressure gradient across the valve ranged from 35 to 60 mm Hg and the level of pressure trapped was inversely proportional to the graft flow. CONCLUSIONS: In patients in whom reversed vein grafts with competent valves are placed in the femoro-popliteal positions a "pressure-trap" develops in the distal segment. This segmental hypertension combined with the flow stagnation could play an important role in the graft thickening and stenosis.     

Low-pressure environment and remodelling of the forearm vein in Brescia-Cimino haemodialysis access.

BACKGROUND: The aim of the study was to determine which, and to what extent, haemodynamic parameters contribute to the remodelling of the venous limb of the Brescia-Cimino haemodialysis access. METHODS: The dimensions of the radial artery and the venous limb of the haemodialysis access were measured by an echo-tracking technique. In six ESRD patients undergoing primary arteriovenous fistula (AVF) formation, vessel diameter, wall thickness, blood pressure and blood flow were measured after the operation, and at 1 and 3 months follow-up. The contralateral forearm vessels in their native position served as baseline values for comparison. RESULTS: The diameter of the proximal antecubital vein progressively increased over the study period without reaching significant differences (4430, 5041 and 6620 microm at weeks 1, 4 and 12 respectively), whereas the intima-media thickness remained unchanged. The venous dilatation was associated with a reduction of the mean shear stress that culminated after the operation and progressively returned to normal venous values at 3 months (24.5 vs 10.4 dyne/cm(2), P<0.043). Thus the venous limb of the AVF undergoes eccentric hypertrophy as demonstrated by the increase in wall cross-sectional area (4.42 vs 6.32 mm(2) at week 1 vs week 12, P<0.028). At the time of the operation, the blood pressure in the AVF was 151+/-14/92.4+/-11 mmHg vs 49+/-19/24.5+/-6 mmHg (means+/-SEM) for the radial artery and the venous limb of the vascular access, respectively. One year after the operation the blood pressure in the venous limb had not changed: 42+/-14/25.3+/-7 mmHg (means+/-SEM). Under these conditions, the systolo-diastolic diameter changes observed in the radial artery and the antecubital vein were within a similar range at all time points: 56+/-17 vs 90+/-26 microm (means+/-SEM) at week 12. CONCLUSIONS: The increased circumferential stress resulting from the flow-mediated dilatation rather than the elevation of blood pressure appears to represent the main contributing factor to the eccentric hypertrophy of the venous limb of Brescia-Cimino haemodialysis access.     

Glomerular endothelial dysfunction in chronic kidney disease

A dysfunctioning glomerular endothelium was demonstrated in chronic kidney disease (CKD) patients by means of in vitro endothelial cell cytotoxicity test and of in vivo intrarenal hemodynamic study. An enhanced endothelial cell cytotoxicity in CKD patients was 26.5 +/- 12% as compared to 0.4 +/- 1% of control. An altered intrarenal hemodynamics revealed 1) a reduction in renal plasma flow, 190 +/- 67 mL/min/1.73 m2 versus control 595 +/- 45 mL/min/1.73 m2, and in peritubular capillary flow, 149 +/- 55 mL/min/1.73 m2 versus control 479 +/- 46 mL/min/1.73 m2, 2) an elevated intraglomerular hydrostatic pressure, 55 +/- 2 mmHg versus control 51 mmHg, elevated afferent arteriolar resistance, 13184 dyne x s x cm(-5) versus control 2443 +/- 154 dyne x s x cm(5), and elevated efferent arteriolar resistance, 13591 +/- 7591 dyne x s x cm(-5) versus control 3062 +/- 177 dyne x s x cm(-5). Both enhanced endothelial cell cytotoxicity and altered intrarenal hemodynamics reflect glomerular endothelial dysfunction which is likely responsible for the renal disease progression in CKD.     

Experimental microvascular autogenous vein grafts for arterial defects: a study of anastomotic sites

Autogenous femoral vein grafts with an average external diameter of 1.5 mm and an average length of 2.87 cm have been used to bridge defects in the contralateral femoral artery of 15 adult New Zealand white rabbits. An experimental microvascular technique to minimise trauma to the graft was performed, by which clamps were never applied to the graft itself. Patency was assessed over a 12-week period, and the overall graft success rate, excluding one technical failure, was 86%. Each of the three failures resulted from thrombosis. Histologic examination of the patent anastomoses showed marked medial damage at 1 week, complicated by fibrosis and calcification by 4 weeks; thereafter the intima developed prominent fibroelastic thickening. This led to a degree of luminal narrowing by 12 weeks. However, the underlying medial damage, attributable to operative trauma, did not seem to diminish luminal patency.