Aneurysm formation in arteriovenous grafts: associations and clinical significance

Aneurysms are a common complication of arteriovenous grafts in hemodialysis patients, resulting from repetitive needle sticks in the graft material. Although aneurysms are thought to contribute to graft failure, there are no prospective studies evaluating their risk factors or impact on graft survival. The present study evaluated aneurysms in 117 hemodialysis outpatients with upper extremity grafts at a university-affiliated dialysis center. An arterial aneurysm was defined as a cannulation site defect diameter (difference between arterial cannulation site diameter and normal graft diameter) above the median value for the study population (0.63 cm). Subsequent graft outcomes were determined by retrospective analysis of a prospective vascular access database. Thrombosis-free graft survival was compared among patient subgroups using Cox proportional hazards models. Patients with an arterial aneurysm had significantly longer median graft age, when compared with those not having a aneurysm (888 vs. 588 days, p = 0.01). However, the two groups did not differ in patient age, sex, diabetes, body mass index, or graft location. The hazard ratio for graft thrombosis was 0.45 (95% confidence interval, 0.25-0.82, p = 0.009) for grafts with an arterial aneurysm, when compared with those without a defect (1-year graft survival of 71 vs. 50%). Graft age was not associated with the likelihood of graft thrombosis (p = 0.12). In contrast to the prevailing wisdom, arterial aneurysms are associated with improved graft survival.     

Colonisation of prosthetic grafts by immunocompetent cells in a sheep model

The present study examined the distribution of immunocompetent cells in synthetic vascular grafts in an experimental sheep model. Sixty-two adult Merino sheep underwent synthetic patch closure of a longitudinal arteriotomy in the left common carotid artery. The synthetic patch materials used were gelatin sealed Dacron (n=10), fluoropassivated Dacron (n=10), Fluoropassiv (n=12), polyurethane (n=10), expanded polytetrafluoroethylene (n=10) and carbon-lined expanded polytetrafluoroethylene (n=10). The sheep were sacrificed after four weeks when the prosthetic patches were harvested and fixed in 10% neutral buffered formalin. Transverse sections were taken along the graft and paraffin embedded. Serial sections were stained with cell type specific antibodies to identify T-lymphocytes (CD3(+)), dendritic cells (S-100(+)), endothelial cells (von Willebrand factor(+)) and smooth muscle cells (smooth muscle alpha-actin(+)). All six graft types contained CD3(+) and S-100(+) cells in the neointima, within the synthetic matrix and in the perigraft layer. Three different tissue responses to synthetic materials were observed and the grafts were classified accordingly into three groups: (1) gelatin sealed Dacron, fluoropassivated Dacron and Fluoropassiv; (2) expanded polytetrafluoroethylene and carbon-lined expanded polytetrafluoroethylene; (3) polyurethane. The three synthetic materials in Group 1 showed almost identical reactions with least accumulation of immunocompetent cells within the synthetic material but greater accumulation of immuno-inflammatory infiltrates in the perigraft vascular tissue. In this group, new vessels penetrated into the synthetic material and there was prominent formation of foreign body (giant) cells. Group 2 showed greater accumulation of immunocompetent cells within the synthetic material itself but only sparse immuno-inflammatory infiltrates in the perigraft tissue. Group 3 showed a high degree of inflammatory response within both the synthetic material and the perigraft vascular tissue. These observations demonstrate that immunocompetent cells colonise the synthetic matrix of grafts and accumulate in the perigraft tissue, but inflammatory responses vary in different graft types.     

Nitric oxide-containing neurons in long-term grafts in a rat model of Parkinson's disease

The role that nitric oxide may play in modulating graft function in long-term fetal ventral mesencephalic grafts in an animal model of Parkinson's disease was investigated. Mature grafts harvested from the entire fetal ventral mesencephalon possessed a large number of neuronal nitric oxide synthase (nNOS)/NADPH-diaphorase-containing neurons throughout the graft intermingled with dopaminergic neurons. The morphological and neurochemical characteristics of these NADPH-diaphorase neurons resembled those in centers adjacent to the substantia nigra of adult brain but not that of the striatum. Pretreatment with the nNOS blocker, 7-nitroindazole, resulted in contralateral rotations following methamphetamine challenge in long-term grafted animals that previously showed normalized rotational behavior. In contrast, mature grafts derived from fetal ventral mesencephalon without the midline areas possessed only a few nNOS-containing neurons within the grafts, and a similar methamphetamine challenge following 7-nitroindazole pretreatment in long-term grafted rats that previously showed normalized rotational behavior resulted in random movements. Our results indicate that nitric oxide-containing neurons inadvertently included during grafting may affect graft function, and excluding the midline areas of the ventral mesencephalon during tissue harvesting may minimize this effect.     

Nitric oxide synthase isoform expression in a porcine model of granulation tissue formation

BACKGROUND: This study was designed to determine whether the nitric oxide (NO) pathway is involved in wound granulation tissue formation. METHODS: A section of the pig abdominal wall (excluding the skin) was excised, creating an incisional hernia. The resulting defect was repaired with silicone sheeting in a manner that mimics a temporary abdominal wall closure. During the 14-day experimental period, porcine omentum adhered to the peritoneal edges of the defect and a highly vascularized granulation tissue formed on both sides of the sheeting. Granulation tissue thickness and wound fluid volume were monitored by ultrasonography and epigastric artery flow velocity was monitored by color Doppler flow analysis at days 2, 4, 7, 9, 11, and 14. Fluid was serially harvested from the wound compartment at days 2, 4, 7, 9, 11, and 14 for nitrite/ nitrate (NOx) analysis. Finally, granulation tissue was harvested at day 14 for immunohistochemical and molecular analyses. RESULTS: There was a significant increase in granulation tissue thickness and wound fluid volume during the 14-day study period. Blood flow to the wound increased significantly by day 4 and returned toward baseline by day 14. Wound fluid NOx levels significantly increased from days 7 to 11 and then decreased to near baseline values by day 14. Wound fluid arginine levels significantly decreased when compared with peritoneal fluid and plasma levels at day 14, while wound fluid ornithine levels significantly increased. Immunohistochemical analysis of granulation tissue at day 14 revealed nitric oxide synthase (NOS) 2 was present in the majority of the cells in the granulation tissue. NOS 3 was expressed in endothelial cells only, and NOS 1 expression was not observed in the granulation tissue. CONCLUSIONS: This study suggests that NO, NOS 2, and arginine may play critical roles in granulation tissue formation and wound healing. Arginase and NOS 2 may compete for available arginine as a substrate, thereby limiting later NO production in favor of sustained ornithine synthesis.     

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.     

他克莫司抑制兔耳瘢痕增生的作用及其机制研究

目的：他克莫司是临床广泛应用的一种药物,其对增生性瘢痕是否产生作用并无相关报道,为此提出并证实了他克莫司可以抑制兔耳瘢痕增生。方法：建立兔耳增生性瘢痕模型,选10只新西兰大耳白兔双耳腹侧用打孔器制作直径1cm圆形创面,伤后14天创面上皮化后给药,每只兔左耳为空白对照组涂等剂量凡士林软膏,右耳为他克莫司治疗组。分别在伤后14天、21天2、8天3、5天和49天采集标本,行HE染色,观察形态学差异;Real-t i me PCR检测纤维化相关因子TGF-β1、TGF-β2、Col l agen-α1等的表达。结果：HE染色可见他克莫司组成纤维细胞数量及胶原纤维较对照组明显减少,PCR结果可见TGF-β1、TGF-β2及Col l agen-α1表达较对照组在各时间点均减少。结论：实验组较对照组瘢痕明显减轻,证明他克莫司显著抑制兔耳瘢痕增生,可作为临床上治疗及预防瘢痕增生的全新疗法。     

Preclinical study of patient-specific cell-free nanofiber tissue-engineered vascular grafts using 3-dimensional printing in a sheep model

Background

Tissue-engineered vascular grafts (TEVGs) offer potential to overcome limitations of current approaches for reconstruction in congenital heart disease by providing biodegradable scaffolds on which autologous cells proliferate and provide physiologic functionality. However, current TEVGs do not address the diverse anatomic requirements of individual patients. This study explores the feasibility of creating patient-specific TEVGs by combining 3-dimensional (3D) printing and electrospinning technology.

Paclitaxel-coated expanded polytetrafluoroethylene haemodialysis grafts inhibit neointimal hyperplasia in porcine model of graft stenosis.

BACKGROUND: The main pathology of haemodialysis graft stenosis is venous neointimal hyperplasia at graft-venous anastomoses. Neointimal hyperplasia is also observed in cases of coronary artery in-stent restenosis. Paclitaxel is a chemotherapeutic agent used to treat cancer, and has been proven to inhibit neointimal hyperplasia of coronary artery in-stent restenosis. In this study, we examined whether a paclitaxel-coated haemodialysis graft could inhibit neointimal hyperplasia and prevent stenosis. METHODS: We dip-coated paclitaxel on expanded polytetrafluoroethylene (ePTFE) grafts at a dose density of 0.59 microg/mm(2). In vitro release tests showed an initial paclitaxel burst followed by a long-term slow release. Using ePTFE grafts with (coated group, n = 8) or without a paclitaxel coating (control group, n = 11), we constructed arteriovenous (AV) grafts connecting the common carotid artery and the external jugular vein in Landrace pigs. RESULTS: After excluding seven pigs for technical failure, cross-sections of graft-venous anastomoses obtained 6 weeks after placing the AV grafts were analysed. Percentage luminal stenosis, ratios of intima to media in whole cross-sections, areas of intima in the peri-junctional areas (within 2 mm above and 2 mm below the graft-venous junction), and the mean thickness of intima within venous sides of cross-sections, were 60.5% (range, 41.5-60.7), 13.0 (range, 8.6-20.4), 23.7 mm(2) (range, 10.8-32.1) and 2.1 mm (range, 1.1-3.0), respectively, in the control group, whereas corresponding median values in the coated group were 10.4% (range, 1.0-17.8), 1.0 (range, 0.7-5.1), 1.6 mm(2) (range, 0.2-8.0) and 0.3 mm (range, 0.1-2.2). All parameters were significantly different between the two groups (P<0.05 by Mann-Whitney test). CONCLUSION: Paclitaxel-coated ePTFE grafts could prevent neointimal hyperplasia and the stenosis of AV haemodialysis grafts.     

Intraperitoneal onlay mesh: an experimental study of adhesion formation in a sheep model

Purpose  

Current hernia literature shows that the use of mesh in ventral hernia repair reduces the risk of recurrence significantly. In laparoscopic repair, the mesh is placed intraperitoneally. Accordingly, the close contact between mesh and viscera involves a risk of adhesion formation. In this experimental study, we examined the degree of de novo adhesion formation over time to currently available meshes.     

Forskolin impregnation of small calibre PTFE grafts lowers early platelet graft sequestration and improves patency in a sheep model

Synthetic vascular grafts have a thrombogenic surface which plays a role in graft failure. Systemic pharmacologic interventions have been used to lower platelet sequestration onto the graft surface but are associated with side effects. In this communication we describe the results of a new therapeutic principle of applying forskolin, a powerful cyclic adenosine monophosphate stimulator (cAMP) to the inner surface of PTFE vascular grafts. The grafts were evaluated with Indium-III-oxine labelled platelets and by graft patency on 3 consecutive days after implantation at 1 month and 3 months. Forskolin significantly lowered early platelet sequestration onto the treated graft surface when compared with controls. Graft potency at 1 and 3 months was also significantly higher in the forskolin treated grafts.     

Pathogenesis of thrombus formation in varicose veins

Under observation there were 60 patients operated upon for varicose dilatation of the lower extremity veins. It was established that the reconstruction of the walls of the altered vessels had a substantial influence on processes of the parietal microthromboformation and facilitated local alteration of hemodynamics and elevation of the aggregation properties of thrombocytes.     

The hemodynamics of thrombus formation in arteries

Alterations in arterial blood flow are thought to predispose to thrombus formation, but the exact relationships have not been fully elucidated. The effect of varying blood flows on the accumulation of thrombotic material within arteries was investigated, with use of shear rate as an index of flow across the luminal surface. Partially denuded rabbit aortas were perfused with fresh nonanticoagulated human blood for 3 minutes, with an in vitro recirculating apparatus, Indium 111-labeled platelets, and fibrinogen I 125. Shear rates ranged from zero to 1500 sec-1, correlating with the hemodynamics of various segments of the human arterial tree. A significant correlation was observed between shear rate and platelet deposition, ranging from 5.2 +/- 2.8 x 10(6) platelets/cm2 of vessel surface area at zero shear to a maximum of 64.7 +/- 8.3 x 10(6) platelets/cm2 at a shear rate of 1500 sec-1 (F = 5.01, p less than 0.05). Fibrin deposition paralleled that of platelets, ranging from 28.2 +/- 7.6 micrograms/cm2 at zero shear to 354.1 +/- 62.7 micrograms/cm2 at a shear rate of 1500 sec-1 (F = 5.91, p less than 0.05). These results suggest that shear rate is a most important determinant of platelet and fibrin deposition on altered arterial surfaces.     

Autoaugmentation peritoneocystoplasty in a sheep model

OBJECTIVE: To report our experience with autoaugmentation peritoneocystoplasty (AAPC) in a sheep model, and to compare the results with autoaugmentation gastrocystoplasty (AAGC) in a sheep model and in paediatric patients. MATERIALS AND METHODS: Ten 6-month-old male lambs underwent bladder augmentation by detrusorotomy. A flap of parietal peritoneum, dissected from the anterior abdominal wall, was used to cover the bladder mucosa. The sheep were evaluated by urodynamics 6 months after surgery. Bladder compliance (bladder volume/intravesical pressure) was calculated for the bladder capacity at leakage. The urodynamic results were compared with age-matched control sheep and with 12 sheep that had undergone AAGC; the results were assessed using the Mann-Whitney U-test. RESULTS: In two of the 10 sheep, bladder volumes after AAPC increased by > 100%, although for the group, the mean (range) bladder volume after augmentation, at 159 (42-261) mL, was not significantly different from that before surgery (mean 143 mL). Bladder volumes after AAPC were not significantly different from those in the control sheep (mean 205 mL) but were significantly less than in the AAGC group (mean 317 mL; P < 0.05). Bladder compliance at leak capacity in the AAPC group (mean 5.4 mL/cmH2O) was also not significantly different from the controls (mean 9.1 mL/cmH2O), but was lower than the in the AAGC animals (median 14.6 mL/cmH2O; P < 0.05). CONCLUSIONS: AAPC in a sheep model does not result in a reliable increase in bladder volume or compliance. The volume and compliance are inferior to those found in bladders augmented by AAGC.     

Autoaugmentation omentocystoplasty in a sheep model

Objective. To develop a sheep model of autoaugmentation omentocystoplasty and study the histologic appearance, and to compare the urodynamic results with a control group.Methods. Ten male lambs underwent a bladder autoaugmentation reinforced with an omental patch. Three were culled early, to study urothelial survival and inflammatory changes. One was sacrificed at six months to assess late histologic changes; five sheep had a urodynamic study at that stage and two died of unknown causes without further investigation. A group of seven six-month-old male sheep, of similar weights, formed the control urodynamic group.Results. The urothelial lining remained viable under the omentum, but marked inflammation and heterotopic calcification were seen within the subepithelial tissues in most of the animals. The histologic changes were reflected in the bladder dynamic data, which were no better than the control group; the average compliance value was 9.2 ± 6.4 mL/cm H₂O, compared to the control group figure of 11.8 ± 5.2 mL/cm H₂O.Conclusions. It would appear that autoaugmentation alone does not usually produce bladder augmentation in the sheep.     

Inflow stenosis in arteriovenous fistulas and grafts: a multicenter, prospective study

BACKGROUND: Traditionally, arteriovenous hemodialysis access inflow stenosis has been reported to occur infrequently (0% to 4%). In contrast, recent reports have suggested a significantly higher incidence (14% to 42%). Interpretation of these studies has been complicated by the presence of one or more confounding factors such as retrospective study design, small sample size, arteriovenous fistulas grouped with grafts to determine the incidence of inflow stenosis, inclusion of fistulas that had failed primarily, failure to provide adequate definition of inflow stenosis, and the technique of retrograde angiography. This is a report of a prospective, multicenter study to examine the incidence of inflow stenosis separately in arteriovenous fistulas and grafts. METHODS: Patients were referred to interventional nephrology either for percutaneous balloon angioplasty or thrombectomy procedures. Angiography to evaluate access inflow (arterial anastomosis and adjacent vascular structures) was performed in all cases. This was accomplished by retrograde angiography using either manual occlusion of the venous side and/or advancing a diagnostic catheter across the arterial anastomosis. Multiple images using digital subtraction angiography were recorded in multiple planes. An inflow stenosis was defined as stenosis within the arterial system, artery-graft anastomosis (graft cases), artery-vein anastomosis (fistula cases) and juxta-anastomotic region (the first 2 cm downstream from the arterial anastomosis). Vascular stenosis was defined as >/=50% reduction in luminal diameter judged by comparison with either the adjacent vessel or graft. A standardized definition for anastomotic stenosis was applied. RESULTS: Two hundred and twenty three consecutive procedures (grafts, 122; fistulas, 101) were performed in 158 patients. Inflow stenosis occurred in 36/122 (29%) in graft cases. All had a coexisting stenosis on the venous side. In fistula cases, 41/101 (40%) had inflow stenosis. Of these, 22 (54%) had a coexisting lesion on the venous side. Overall, inflow stenosis occurred in 77/223 procedures (35%). CONCLUSION: This prospective, multicenter study demonstrates that access inflow stenosis occurs in one third of the cases referred to interventional facilities with clinical evidence of venous stenosis or thrombosis. This is much higher than has been traditionally reported.