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.     

Histogenesis of arterial intimal hyperplasia and atherosclerosis

This article briefly reviews the histological evidence for the genesis of intimal hyperplasia and atherosclerosis in arteries. It concentrates upon the origin, structure and behaviour of vascular smooth muscle cells in the intimal (subendothelial) layer. The interactions between these intimal muscle cells, the endothelium and the blood are discussed with particular reference to their role in the histogenesis of intimal hyperplasia and atherosclerosis.     

Inhaled carbon monoxide prevents graft-induced intimal hyperplasia in swine

BACKGROUND: Arteriovenous grafts often fail due to stenosis caused by venous anastomotic intimal hyperplasia (IH) and vascular smooth muscle cell (VSMC) proliferation. We examined the effects of inhaled carbon monoxide (CO), a product of heme-oxygenase-1 degradation of heme, on IH in a porcine arteriovenous graft model. MATERIALS AND METHODS: Eighteen Yorkshire pigs were divided into three groups (N = 6/group): (1) CO 100 ppm preoperatively for 1 h; (2) CO 250 ppm preoperatively for 1 h and intraoperatively; and (3) air-treated controls. Animals underwent end-to-side placement of polytetrafluoroethylene grafts connecting the common femoral artery and vein in both groins. Intimal thickness of the venous anastomosis at 30 days was measured blinded. The effect of CO on pig VSMC proliferation was studied in cell culture using [(3)H]thymidine incorporation. RESULTS: Pigs in the group receiving CO 250 ppm showed significantly less IH compared to animals in the group receiving 100 ppm and the air-treated group (267.5 +/- 21.4, 824 +/- 145.8, and 914.8 +/- 133.7 pixels, respectively, P < 0.0001). This effect was not observed when comparing the 100 ppm group to the air-treated group. COHb levels were significantly elevated in the 100 ppm and 250 ppm compared to air-treated pigs (5.8 +/- 0.47, 13.2 +/- 1.0 versus 2.3 +/- 0.11%, respectively, P < 0.001). Oxygen saturation, respiratory rate, and hemodynamics were not significantly different between the groups. CO induced VSMC growth arrest compared to air in vitro (11.9 +/- 4 versus 20.3 +/- 5 10(3) counts/min/well, P < 0.01). CONCLUSION: A single exposure to a low concentration of inhaled CO (250 ppm) confers protection against intimal proliferation of VSMCs when given perioperatively in a clinically relevant model of arteriovenous grafts. These data are the first to suggest, in a clinically relevant model, the potential role for CO in clinical applications.     

Vascular closure staples reduce intimal hyperplasia in prosthesis implantation

Background: Vascular surgery, like the various other surgical specialities, has seen an increasing demand toward faster and more minimally invasive procedures. One such need is to create a reliable vascular anastomosis that is faster, easier and less damaging to the tissue. The vascular closure staples (VCS*) device provides such characteristics but, to date, no studies have investigated its effectiveness in reducing intimal hyperplasia when used for vascular prosthesis implantation. The present study evaluated its effectiveness compared with suturing of a graft in vascular prosthesis implantation. Methods: Twelve female Merino sheep underwent gelatin sealed Dacron patch graft implantation into the left and right common carotid artery. Grafts were randomly allocated so that one carotid artery and graft was anastomosed using sutures and the other with VCS*. The two techniques were compared for operation time, clip/suture numbers and blood loss during the implantation procedure. After a 4‐week period, the sheep were killed and the grafts were harvested for intimal hyperplasia (IH) assessment. Results: There was a significant reduction in the amount of IH seen in the VCS* group (mean ± SD: 0.278 ± 0.079 mm²/mm) when compared with the sutured group (0.575 ± 0.331 mm²/mm) (P < 0.05). There was also significant reduction in anastomosis time (mean ± SD: 14 ± 4.4 min) and fewer points of contact (23 ± 1.4) using the VCS* compared with suturing (22 ± 3.2 min, P < 0.01; 27 ± 3.3, P < 0.05, respectively). Conclusions: In this model, the VCS* shows several distinct advantages over suturing with significant time saving at operation and, most importantly, the reduction of IH seen at 1 month.     

Altered ubiquitin/proteasome expression in anastomotic intimal hyperplasia

OBJECTIVE: Anastomotic intimal hyperplasia remains a leading mechanism of prosthetic arterial graft failure. Recent studies using messenger RNA differential display have demonstrated altered proteasome gene expression at the anastomoses in an expanded polytetrafluoroethylene canine carotid model. However, this technique is technically limited because of a paucity of available hyperplastic tissue at early time periods after arterial injury. Microarray gene chip technology offers a new and sensitive technique to assay early gene expression, requiring far less tissue for analysis. The purpose of this study was to screen for altered proteasome gene expression at 48 hours and 14 days after prosthetic arterial grafting. METHODS: Expanded polytetrafluoroethylene grafts (6-mm diameter, n = 9) were implanted into 25-kg mongrel dogs. The normal intervening carotid artery was used as control. At 48 hours and 14 days, RNA was extracted from the perianastomotic tissue and compared with RNA from the control carotid. Messenger RNA was then hybridized to microarray genomes screening for differential gene expression. RESULTS: Two 26S proteasome genes and five ubiquitin pathway genes were significantly underexpressed at 48 hours, among several hundred significantly expressed clones. The two 26S proteasome genes were 26S proteasomal subunit p55 (0.26), and 26S proteasomal subunit p40.5 (0.13). The underexpressed ubiquitin genes included ubiquitin (0.31), Nedd-4-like ubiquitin-protein ligase (0.30), ubiquitin conjugating enzyme UbcH2 (0.25), putative ubiquitin C-terminal hydrolase UHX1 (0.11), and ubiquitin-conjugating enzyme UbcH7 (0.12). At 14 days, six ubiquitin genes were underexpressed, and 17 26S proteasome genes were significantly downregulated. CONCLUSIONS: This study shows decreased expression of the ubiquitin/proteasome pathway 48 hours after graft implantation and similar diminished expression patterns after 14 days. This early and sustained underexpression after arterial bypass may lead to altered cell cycle control and matrix protein signaling, contributing to the unregulated proliferation of smooth muscle cells and extracellular matrix in anastomotic intimal hyperplasia after prosthetic arterial grafting.     

Inhibitory effect of brachytherapy on intimal hyperplasia in arteriovenous fistula

PURPOSE: To determine whether endovascular radiation can inhibit intimal hyperplasia in a swine model of hemodialysis access. MATERIALS AND METHODS: Polytetrafluoroethylene arteriovenous grafts (6 mm in diameter) were placed between the common carotid artery and the external jugular vein bilaterally in 8 pigs. Two days after the surgery, fistulography was performed. Gamma radiation (12 Gy) was delivered endovascularly to one of the grafts at the venous anastomosis by using an iridium(192) source. Thus, the other graft in each pig served as an untreated control. Fistulas were evaluated with fistulography or venography 6 week after radiation. All grafts were then harvested for histological and immunohistochemical examination. RESULTS: Seven grafts on the treated side and 5 grafts on the control side remained patent for at least 6 weeks. Angiography demonstrated that the percentage stenosis at the venous anastomosis was significantly lower for the treated group (15.9 +/- 14.1 versus 32.6 +/- 16.7%, P = 0.045). Histopathologic analyses revealed that the mean intimal area and maximal intimal thickness were significantly lower with reduced smooth muscle cell proliferation at the venous anastomosis on the treated side compared to the control side (0.68 +/- 0.30 versus 1.06 +/- 0.29 mm(2), P = 0.017, and 0.18 +/- 0.08 versus 0.26 +/- 0.07 mm, P = 0.004, respectively). The residual lumen was significantly greater for the treated group (1.59 +/- 0.42 versus 1.06 +/- 0.37 mm(2), P = 0.031). No significant differences were found in the area, nor maximal thickness in the vein either proximal or distal to the anastomosis between the two groups. CONCLUSIONS: In an animal model of hemodialysis access, brachytherapy with iridium(192) delivered 2 days after graft implantation reduces intimal hyperplasia and stenosis at the venous anastomosis. The reduced smooth-muscle cells found in the radiated veins suggests that brachytherapy may exert its effect on neointimal formation by inhibition of smooth muscle cell proliferation.     

The role of hyaluronic acid in atherosclerosis and intimal hyperplasia

Atherosclerosis is a chronic inflammatory condition of the blood vessel wall that can lead to arterial narrowing and subsequent vascular compromise. Although there are a variety of open and endovascular procedures used to alleviate the obstructions caused by atherosclerotic plaque, blood vessel instrumentation itself can lead to renarrowing of the vessel lumen through intimal hyperplasia, wound contracture, or a combination of the two. While the cell types involved in both atherosclerosis and vessel renarrowing after surgical intervention are largely characterized, current research has shown that components of the extracellular matrix are also important in the pathogenesis of the aforementioned processes. One such component is hyaluronic acid (HA). The objective of this review, therefore, is to examine the involvement of HA in these pathologic processes. Literature on the structure and function of HA was reviewed, with particular attention given to the role of HA in the processes of atherogenesis, intimal hyperplasia, and wound contracture after blood vessel instrumentation. HA interacts with vascular smooth muscle cells (VSMCs), endothelial cells (ECs), and platelets to promote atherogenesis. In particular, VSMCs manufacture large amounts of HA that form "cable-like" structures important for leukocyte adhesion and rolling. Additionally, transmigration of leukocytes across the EC layer is mediated by HA. Platelets cleave large molecules of HA into fragments that up-regulate leukocyte production of chemokines and cytokines. HA also has a role in both intimal hyperplasia and wound contracture, the two processes most responsible for vessel renarrowing after vascular instrumentation. HA has a complex, and sometimes conflicting, role in the pathologic processes of atherogenesis and vessel wall renarrowing after surgical intervention.     

Development of intimal hyperplasia in six different vascular prostheses.

OBJECTIVES: to compare six vascular prostheses for the development of intimal hyperplasia (IH) in a sheep model. MATERIAL AND METHODS: prostheses tested were gelatin sealed Dacron (GSD), fluoropassivated Dacron (FPD), FluroIpassiv TM (FD), expanded polytetrafluoroethylene (ePTFE), carbon-lined expanded polytetrafluoroethylene (CL-ePTFE) and vascular access graft (VAG). Sixty-two adult female Merino sheep (35-45 kg) were used. Elliptical graft patches were implanted into the left common carotid artery using one of the six graft types: GSD (n=10), FPD (n=10), FD (n=12) VAG (n=10), ePTFE (n=10), or CL-ePTFE (n=10). Four weeks later grafts were removed for histopathological assessment and measurement of the degree of IH obtained on a computerised image analysis system. RESULTS: IH indices were significantly less for FPD (0.191+/-0.095, p<0.05), FD (0.199+/-0. 081, p<0.05), ePTFE (0.213+/-0.078, p<0.05) and CL-ePTFE (0.161+/-0. 066,p<0.01), compared to the GSD group (0.287+/-0.077). The VAG group (0.257+/-0.091) showed no difference compared to GSD. There was no significant difference between the FPD, FD, ePTFE and CL-ePTFE grafts. CONCLUSION: this study indicates that less IH occurred in the two-ePTFE grafts and two fluoropolymer coated Dacron grafts than in gelatin sealed Dacron or polyurethane grafts.     

Distal anastomotic intimal hyperplasia: biogenesis and etiology

Anastomotic intimal hyperplasia occurred exclusively at the heel and the toe plus the floor of the distal end-to-side anastomosis of canine autologous femoro-femoral bypass (n = 14) and not in the end-to-end carotid or femoral interposition graft (n = 14). The occurrence of anastomotic intimal hyperplasia in the absence of compliance mismatch in an autologous bypass suggests that the geometry of the end-to-side anastomosis is primarily responsible for intimal hyperplasia formation. It is believed that because an end-to-side distal anastomosis is not a natural occurrence it is conductive to turbulent flow. The latter causes endothelial injury which in turn allows platelet growth factor to incite subendothelial myoblasts in extracellular matrix synthesis and intimal hyperplasia formation. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) identify myofibroblasts and fibrocollagenous matrix as the dominant cellular and extracellular substances in anastomotic intimal hyperplasia.     

Macrophage depletion alters vein graft intimal hyperplasia.

BACKGROUND: The principal cause of vein graft failure is intimal hyperplasia (IH); however, its etiology remains unclear. In a rat model of vein graft IH we have observed prolonged transmural macrophage infiltration, leading us to hypothesize that these cells regulate IH. To test this, we used liposome-encapsulated dichloromethylene bisphosphonate (L-Cl2MBP) to deplete rat macrophages and observed the effects on IH. METHODS: Epigastric vein-to-femoral artery grafts were microsurgically placed in male Lewis rats that had been intravenously injected with L-Cl2MBP, phosphate-buffered saline solution liposomes, or phosphate-buffered saline solution alone 2 days before surgery. Several animals in each group received a second equivalent dose at 2 weeks. Grafts, contralateral epigastric veins, spleens, and livers were harvested at 1, 2, and 4 weeks for histologic examination, immunohistochemistry, and transmission electron microscopy. RESULTS: In the L-Cl2MBP-treated animals splenic and hepatic macrophages were greatly reduced, confirming the efficacy of the agent. At 1 to 2 weeks graft macrophages were significantly decreased, and there was a trend toward decreased IH. At 4 weeks macrophage numbers were normal and IH development had resumed. In contrast, the 4-week grafts treated with 2 doses of L-Cl2MBP had fewer macrophages and displayed severely attenuated IH. CONCLUSIONS: The results indicate a suppression of IH as macrophages are depleted, with a resumption of the process as macrophages repopulate the graft.     

Aspirin and dipyridamole decrease intimal hyperplasia in experimental vein grafts

Release from platelets of a factor mitogenic for smooth muscle cells is a postulated mechanism for the pathogenesis of vascular intimal hyperplasia. In this study the effect of antiplatelet therapy was evaluated. Aspirin (165 mg twice daily) and dipyridamole (25 mg twice daily) were administered to six rhesus monkeys and six were given placebo only. Bilateral vein bypass grafts were placed in the iliac arteries. In addition, to evaluate the relative contribution of adventitial dissection and intimal injury, on one side the carotid artery and femoral vein were stripped of adventitia and on the other side the intima of these vessels were injured by the single passage of an inflated balloon tipped catheter. Animals were killed after 16 weeks. In grafts relative luminal area was determined by a photographic gravimetric method at three standard locations. Femoral veins and carotid arteries were classified as histologically normal or as exhibiting hyperplasia. All vessels with adventitial stripping were normal. All vessels with intimal injury in the placebo group except one exhibited intimal hyperplasia compared to the drug treated group in which over half were normal. Relative intimal area was significantly less in grafts from drug treated animals at all three locations and luminal area greater in two. These data suggest that vascular intimal hyperplasia can be reduced by treatment with antiplatelet agents.     

Colchicine inhibits intimal hyperplasia and leukocyte VEGF expression in dogs

BACKGROUND: Restenosis due to intimal hyperplasia following percutaneous transluminal angioplasty limits its long-term efficacy. We evaluated the effect of colchicine on the development of intimal hyperplasia following balloon angioplasty and on the vascular endothelial growth factor (VEGF) expression in leukocytes. MATERIAL AND METHODS: Adult dogs underwent balloon angioplasty of the right iliofemoral artery. Group 1 served as control, while groups 2 and 3 (six animals per group) received 0.1 and 0.5 mg/kg/d of colchicine p.o., respectively, starting 2 d before angioplasty and continued for 14 d. Before angioplasty and at day 14, blood samples were collected for drug toxicity analysis and the determination of leukocyte expression of VEGF. Animals were euthanized and iliofemoral arteries were perfusion fixed in situ and processed for histological and morphometric analysis. RESULTS: Balloon angioplasty without colchicine resulted in 446% (P < 0.001), 111% (P = 0.7), and 267% (P < 0.001) increase in intimal and medial thickness and intima/media ratio compared with contralateral uninjured iliofemoral arteries. Low-dose and high-dose colchicine resulted in 32% and 58% reduction in intima/media ratio, respectively (both P < 0.001). VEGF expression in leukocytes of control group was up-regulated (40%), but was down-regulated by 12% and 55%, respectively, in low-dose and high-dose colchicine groups at 2 wk after angioplasty compared with preangioplasty expression. The results of complete blood count and serum transaminases and creatinine were within normal range. CONCLUSION: This study demonstrates that oral colchicine for 2 wk significantly reduces intimal hyperplasia following balloon angioplasty in dogs through down-regulation of leukocyte VEGF expression and without apparent toxicity.