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The pathophysiologic effects of infusing unstable lipid emulsions are unclear, but these were shown to cause reticuloendothelial system (RES) dysfunction in animals and humans. We investigated the effects of unstable lipid emulsions in RES organs defined by two levels of the percent fat5 microm (percentage of fat, PFAT5 microm) in a guinea pig model. Two identical injectable lipid emulsions with differing (stable versus unstable) PFAT5 microm levels, were infused for over 24h into two groups of animals (n=5/group). The PFAT5 microm concentration was measured before and at the end of the infusion to ascertain the dose range of enlarged fat globules in each group. Animals were euthanized and specimens from the upper, middle and lower lung, and a single liver sample were examined histologically and for micromolar concentrations of malondialdehyde (MDA) per gram (micromol(-1)g) of wet tissue. The PFAT5 microm concentrations pre-infusion were 0.004+/-0.001 and 2.418+/-0.273 for the stable and unstable injectable lipid emulsions respectively. At 24 h, the PFAT5 microm level increased in both the groups (stable: 0.161+/-0.008; unstable: 7.861+/-0.291). MDA concentrations were significantly higher in the lungs of animals receiving the unstable (47.2+/-26.2 micromol(-1)g) versus stable (32.4+/-11.2 micromol(-1)g) injectable lipid emulsions (P=0.033), but was not different for the liver specimens (stable: 16.9+/-7.6 micromol(-1)g versus unstable: 17.7+/-2.2 micromol(-1)g, P=0.944). These preliminary data suggest that infusion of unstable injectable lipid emulsions has pathological consequences showing greater evidence of oxidative stress in the lungs.

Background Following prosthetic arterial grafting, cytokines and growth factors released within the perianastomotic tissues stimulate smooth muscle cell proliferation and matrix production. While much in vitro work has characterized this response, little understanding exists regarding the sequential up- and down-regulation of genes following prosthetic arterial grafting. This study evaluates temporal gene expression at the distal anastomosis of prosthetic arterial grafts using microarray analysis. Methods Expanded polytetrafluoroethylene (ePTFE) carotid interposition grafts ( n = 12) were surgically implanted into mongrel dogs. Distal anastomotic segments were harvested at 7, 14, 30, or 60 days. Contralateral carotid artery served as control. Total RNA was isolated from the anastomotic tissue and paired controls. Samples were probed with oligonucleotide microarrays consisting of approximately 10,000 human genes to analyze differential gene expression at each time point. Results Forty-nine genes were found to be up-regulated and 37 genes were found to be down-regulated at various time points. Six genes were found to be consistently up-regulated at all time intervals, including collagen type 1 α-1 and α-2, 80K-L protein (MARCKS), and osteopontin. Six genes were found to be consistently down-regulated, including smoothelin and tropomyosin 2. RT-PCR and immunohistochemistry confirmed the microarray data. Conclusions This study uses microarray analysis to identify genes that were temporally up- and down-regulated after prosthetic arterial grafting. Genes with similar patterns of expression have been identified, providing insights into related cellular pathways that may result in the formation of anastomotic intimal hyperplasia.

Background— We have previously shown that endothelial function is impaired not only in diabetes but also in subjects at risk of developing type 2 diabetes. We hypothesized that changes in the expression or activity of the endothelial isoform of nitric oxide synthase (eNOS), the receptor for advanced glycation end products (RAGE), and poly(ADP-ribose) polymerase (PARP) are related to this impairment. Methods and Results— We included a control group of 21 healthy subjects, a group of 22 healthy individuals with parental history of type 2 diabetes, a group of 23 subjects with impaired glucose tolerance, and a group of 21 type 2 diabetic patients. Two 2-mm forearm skin biopsies were taken from each participant and used for measurements. The percentage of PARP-positive endothelial nuclei was higher in the group with parental history of type 2 diabetes and diabetic patients compared with the controls ( P P Conclusions— PARP activation is present in healthy subjects at risk of developing diabetes as well as in established type 2 diabetic patients, and it is associated with impairments in the vascular reactivity in the skin microcirculation.

Objective: The retinoblastoma gene product is a cell cycle control protein that when inhibited allows cell proliferation to progress by releasing E2F. Retinoblastoma manipulation has been attempted to prevent intimal hyperplasia (IH) in injured native vessels by arresting vascular smooth muscle cell proliferation. However, no studies have identified the role, if any, of retinoblastoma in anastomotic IH formation after prosthetic arterial grafting. The goal of this study was to describe the relation of retinoblastoma and E2F to anastomotic IH with analyzing retinoblastoma/E2F levels, retinoblastoma phosphorylation, and transcription of retinoblastoma and E2F in prosthetic arterial grafting. Methods: Six-mm-diameter expanded polytetrafluoroethylene carotid interposition grafts (n = 12) were implanted in 25-kg mongrel dogs. The intervening arterial segments were harvested as controls. The distal anastomoses were harvested at 14 and 30 days after implantation for immunoblot, messenger RNA (mRNA), and immunohistochemistry analyses. Tissue homogenate was separated with sodium dodecylsulfate-polyacrylamide gel electrophoresis and probed with antibody to total retinoblastoma, phosphorylated retinoblastoma at serine 795, serine 780, and serine 807/811, and E2F-1. Bands at each time point were quantitated and compared with control artery (n = 12). Each lane was standardized with reprobing with antibody to β-tubulin. Immunohistochemistry was performed with antibody to retinoblastoma. Retinoblastoma and E2F mRNA expression levels in anastomotic IH and control artery were analyzed with an oligonucleotide microarray. Results: Total retinoblastoma, from immunoblot analysis, was decreased at the 14-day and 30-day distal anastomoses by 35.7% and 33.6%, respectively, compared with control (P

Background. Apolipoprotein J (ApoJ) is expressed after vascular injury and remodeling and may inhibit endothelial cell activation in the vascular wall. Recently, ApoJ was identified as upregulated in hyperplastic lesions after prosthetic arterial grafting. This study analyzed the effect of ApoJ on human umbilical vein endothelial cell (HUVEC) migration, adhesion, and proliferation. Methods. Cell migration towards ApoJ + fetal bovine serum (FBS) or vascular endothelial growth factor (VEGF) was evaluated with the use of a microchemotaxis chamber with or without a fibronectin-coated membrane. For migration that involved fibronectin, cells were exposed to ApoJ before or after placement on the membrane. Cell adhesion to fibronectin was studied similarly but without stimulant. The vital dye alamar blue assessed proliferation of ApoJ + FBS− or VEGF-stimulated HUVECs. Results. ApoJ alone did not cause migration or proliferation of HUVECs. Without fibronectin, ApoJ decreased the migration of HUVECs towards FBS or VEGF. When fibronectin was introduced, ApoJ decreased cell migration toward FBS or VEGF and decreased adhesion only when HUVECs in solution were exposed to ApoJ before the placement on fibronectin. ApoJ had no effect on FBS- or VEGF-induced proliferation. Conclusions. ApoJ inhibits HUVEC migration and adhesion. By altering endothelial function during vascular injury, ApoJ appears to regulate, in part, the early development of intimal hyperplasia after prosthetic arterial grafting. (Surgery 2001;130:204-9.)

The safety of a percutaneous method and streamlined catheter system to access the normal pericardial space via the right atrial appendage for drug delivery and diagnostic sampling was demonstrated in 20 anesthetized pigs. Access was successfully accomplished in all animals within 3 min of guide catheter positioning and was documented by fluoroscopic imaging and pericardial fluid sampling. The animals were sacrificed at 24 hr (n = 10) and 2 weeks (n = 10) for histopathologic analysis. Mean pericardial hematocrit was 1.1% +/- 0.3% at initial sampling, 4.3% +/- 1.4% at 24 hr (P = 0.005 vs. baseline), and 0.4% +/- 0.2% at 2 weeks (P = 0.13 vs. baseline). At 24 hr, there was local inflammatory reaction in the atrial wall and a small thrombus at the site of puncture. At 2 weeks, no significant inflammatory changes or pericarditis were evident. The technique is well tolerated with no apparent adverse complications. Advances in intrapericardial therapeutics and diagnostics will direct the clinical application of this novel approach in human subjects.

Glucose enters the heart via GLUT1 and GLUT4 glucose transporters. GLUT4-deficient mice develop striking cardiac hypertrophy and die prematurely. Whether their cardiac changes are caused primarily by GLUT4 deficiency in cardiomyocytes or by metabolic changes resulting from the absence of GLUT4 in skeletal muscle and adipose tissue is unclear. To determine the role of GLUT4 in the heart we used cre-loxP recombination to generate G4H–/– mice in which GLUT4 expression is abolished in the heart but is present in skeletal muscle and adipose tissue. Life span and serum concentrations of insulin, glucose, FFAs, lactate, and β-hydroxybutyrate were normal. Basal cardiac glucose transport and GLUT1 expression were both increased approximately 3-fold in G4H–/– mice, but insulin-stimulated glucose uptake was abolished. G4H–/– mice develop modest cardiac hypertrophy associated with increased myocyte size and induction of atrial natriuretic and brain natriuretic peptide gene expression in the ventricles. Myocardial fibrosis did not occur. Basal and isoproterenol-stimulated isovolumic contractile performance was preserved. Thus, selective ablation of GLUT4 in the heart initiates a series of events that results in compensated cardiac hypertrophy. J. Clin. Invest. 104:1703–1714 (1999).

Purpose: Anastomotic intimal hyperplasia remains a significant cause of delayed prosthetic arterial graft failure. Prior studies have identified several genes with altered expression within the hyperplastic region at the downstream polytetrafluoroethylene arterial anastomosis as compared with normal arteries. The purpose of the current study was to determine the sequence of early gene-related events at the distal anastomosis of an in vivo prosthetic arterial graft model. Messenger RNA (mRNA) differential display was used to screen for alterations in gene expression between anastomotic sites and control arterial segments. Methods: Six carotid interposition 6-mm expanded polytetrafluoroethylene grafts were placed in mongrel dogs, with the intervening carotid artery segment serving as the baseline control. Five days after graft implantation, the distal anastomotic artery segments were harvested and total RNA was isolated from both the intervening normal arteries and anastomotic segments. Differential mRNA display was used to identify candidate complementary DNA (cDNA) clones with expression that differed in anastomotic segments as compared with normal intervening arteries. Northern blot analysis confirmed alteration of gene expression. The cDNA clones were sequenced, and gene databases were searched. Novel sequences were used as probes for screening human cDNA libraries. Results: Approximately 7000 mRNA species were screened, and 26 candidate clones were obtained. Northern blot analysis showed altered gene expression in 10 (38%) of the clones, undetectable signals in 13 (50%), and nonregulation in 3 (12%). Seven clones with 92% homology at the nucleotide level to human α 1 (III) procollagen gene and a novel sequence were expressed only at the distal anastomosis. A clone representing apolipoprotein J and a novel sequence had increased expression at the distal anastomosis of 364% ± 236% and 156% ± 47%, respectively (mean percentage, control ± standard deviation). Conclusions: These studies identified genes with expressions that increased or were exclusive to the distal anastomosis of healing prosthetic arterial grafts in an in vivo prosthetic arterial graft model. Type III collagen may contribute significantly to the composition of the extracellular matrix associated with intimal hyperplasia by increasing lesion volume. Apolipoprotein J, through its association with proteases, may modulate some of the matrix changes seen early after grafting. (J Vasc Surg 1998;28:157-66.)

Purpose: Autologous veins used as arterial bypass grafts undergo initial loss of the endothelial cell (EC) lining, which is followed by reendothelialization. We characterized the expression of the EC-specific angiogenic mitogen, vascular endothelial growth factor (VEGF), in vascular grafts to help elucidate the molecular and cellular events after bypass procedures.Methods: Cephalic vein–femoral artery interposition grafts were placed in mongrel dogs. Vein grafts and arteries were harvested at either 48 hours or 4 weeks after bypass, the total RNA was isolated, and the VEGF mRNA expression was evaluated by Northern blot analysis. Tissue segments from each time period were evaluated by immunohistochemical analysis using anti-VEGF antibodies.Results: VEGF mRNA expression in vein grafts as compared with control veins was increased 2.5-fold 48 hours after bypass grafting (p = 0.02) but returned to initial control levels in grafts removed at 4 weeks. Distal arterial segments, which included the anastomotic site without attached vein graft, had a 21.4-fold increase in VEGF expression at 48 hours (p = 0.02) and a 6.6-fold increase at 4 weeks (p < 0.01) as compared with control arterial segments. Vessels subjected to arteriotomy or ischemia alone also demonstrated increased VEGF expression. Immunohistochemical analysis revealed VEGF protein within ECs and smooth muscle cells of the venous bypass graft, with maximal levels observed within intimal hyperplasia at the arterial anastomosis.Conclusions: After arterial reconstruction procedures using venous conduits, VEGF is significantly increased at 48 hours in the vein graft and arterial anastomosis. VEGF expression in the vein graft normalizes within 4 weeks but remains significantly elevated in the adjacent arterial segment. Increased VEGF production after arterial grafting may facilitate reendothelialization, thus partially accounting for optimal patency rates achieved with autologous vein grafts. (J Vasc Surg 1997;26:79-86.)

Background: The current use of collagen vascular hemostasis devices to percutaneously seal femoral arteriotomy sites is limited by a significant incidence of vascular complications. The purpose of the present study was to assess the efficacy of new collagen plug specifically designed to avoid these complications by accurately gauging the depth of the femoral artery and by minimizing the risk of intra-arterial collagen deployment. Methods: The hemostasis device used in this study consists of a multicomponent collagen plug with an external rigid collagen tube lined by an inner layer of absorptive sponge collagen. Placement of this plug is facilitated with a specialized two-lumen dilator, which localizes the arterial surface using a “bleedback” mechanism from one of the lumens, and prevents the placement of collagen through the arteriotomy site. The acute efficacy of this device was assessed in 26 adult dogs in which 36 collagen plugs were used to seal 8Fr femoral arteriotomies. An additional 16 arteriotomies treated with standard manual compression served as study controls. Following plug placement or manual compression, all puncture sites were observed for bleeding and hematoma formation over a 45-minute period. Patency of each instrumented artery was assessed with serial femoral angiography and localization of each plug was confirmed with surgical cutdown at the puncture site. The chronic efficacy of this device was assessed in three pigs in which three collagen plugs were used to seal 8Fr femoral arteriotomies. The three animals were subsequently sacrificed at 7,14, and 30 days, respectively, for histologic analysis. Results: In acute animals, all 36 plugs were successfully placed without evidence of vascular compromise or intra-arterial collagen deployment. In nonanticoagulated animals, hemostasis was achieved within 5 minutes in 10 of 10 plugs placed with a skin-to-arlery distance > 1.8 cm, in 4 of 7 plugs with a skin-to-artery distance of 1.2–1.7 cm, and in 0 of 9 plugs with a skin-to-artery distance 1.8 cm; small hematomas occurred in the remaining two animals in the setting of a PTT > 100 seconds. Time to hemostasis was significantly less for collagen plugs than manual compression in both nonanticoagulated animals (plug 17 ± 16 minutes; manual compression 28 ± 5 minutes; P 1.8 cm and the anticoagulation profile was not excessive. At 1 month postplacement, there is no histologic difference between plug use and standard manual compression. (J Interven Cardiol 1996;9:25–33)

Purpose: To produce biodurable small diameter microporous vascular grafts with self-sealing properties for vascular access, for peripheral vascular and potentially for coronary artery bypass. The prosthesis should retain compliance and pulsatile flow in situ using a unique modus operandi permitting wall compression which accommodates changes in volume. Method: We have utilised efficient low temperature coagulation technology to develop a unique range of small diameter microporous vascular grafts using ChronoFlex, a biodurable polycarbonate urethane. Results: Grafts have been subjected to a range of in vitro and in vivo testing, demonstrating excellent physical and mechanical characteristics, self-sealing, maintenance of compliance and pulsatile flow in situ and patency up to twenty-two weeks.

The quality of the anastomosed vessels berore vascular grafting has been shown to influence long-term patency rates in arterial reconstructive procedures. A study was designed to assess vascular graft quality during microvascular artery bypass procedures for impotence, correlate identified vasculopathy with the clinical history and gain insight into vascular pathophysiological mechanisms. A total of 194 donor or recipient vessel segments was biopsied m 111 patients with impotence who underwent microvascular artery bypass surgery during a 10-year period. A prospective histological grading system was used m a blinded randomized fashion. A preexisting vascular pathological condition was identified m 48% of the patients. Proliferative lesions above the internal elastic lamina with luminal stenosis were identified in 38 of 69 dorsal penile artery segments (55%) and 8 of 69 inferior epigastric artery segments (12%). Venous hypertrophy or sclerosis was observed in 13 of 56 deep dorsal vein segments (23%)...

Purpose: The purpose of this study was to explore new strategies for enhancing specific cell type attachment to biomaterials using immobilized lectins for cell surface glycoconjugates. The lectin Ulex europaeus I (UEA I) has a high affinity for human vascular endothelial cell surface glycoconjugates. Methods: UEA I was covalently bound to polyethylene terephthalate (Dacron) with the cross-linking agent 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride to achieve oligosaccharide-mediated endothelial cell attachment to this otherwise nonadherent surface. Results: Experiments with radiolabeled UEA I demonstrated covalent linkage of as much as 1.35 μg/cm 2 . The lectin binding site is available after the reaction, as demonstrated in experiments a neoglycoprotein. Adhesion studies reveal a 100-fold increase in endothelial cell attachment for the UEA I/polyethylene terephthalate surface (99.7 ± 29.6 cells/high-power field) when compared with untreated (0.7 ± 0.5), crosslinking agent (0.4 ± 0.3), and denatured UEA I (1.2 ± 1.1) control groups. Five vascular endothelial cell lines adhered to the UEA I/polyethylene terephthalate surface, whereas monocytes, smooth muscle cells, and fibroblasts did not. Conclusion: These results imply new strategies for endothelialization of prosthetic grafts and promotion of selective cell adherence to biomaterials, with emphasis on carbohydrate interactions. Moreover, this experimental system offers a model for exploring the biologic significance of the endothelial cell-UEA I ligand. (J VASC SURG 1993;18:486-94.)

This is a prospective study of the relationship between graft preparation technique and the subsequent morphologic fate of vein grafts. Paired vein grafts (optimal vs injury prepared) were placed in a canine model and removed over time. Vein grafts intentionally injured by warm saline storage demonstrated endothelial and smooth muscle cell damage. In the acute postimplantation period, platelet adhesion and white cell infiltration of the graft were present. By 7 days, the endothelium had “healed,” but the underlying smooth muscle cells had modulated and were of the transitional or synthetic phenotype. This persisted at 30 days, but by 60 days the graft wall had remodeled to a contractile smooth muscle cell phenotype. Changes in the extracellular matrix were greatest at 30 days corresponding to changes in the smooth muscle cell phenotype. None of these injurious responses were noted in optimally prepared, papaverine treated vein grafts. The combined intima and media (lumen to adventitial edge) was measured at baseline and at graft excision with use of digitized graphic techniques. The intimal/medial thickness of injured vein graft walls was always greater than that of pair-matched optimally treated vein grafts (p J Vasc Surg 1992;16:225–31.)

This is an investigation of the relationship between graft preparation techniques and the subsequent fate of vein grafts. Vein grafts intentionally injured by warm saline storage demonstrated endothelial and smooth muscle cell damage. In the acute postimplantation period, platelet adhesion/activation and white cell infiltration were present. By 7 days the endothelium had “healed,” but the underlying smooth muscle cells had modulated and were of the synthetic phenotype. This persisted at 30 days, but by 60 days the graft wall remodeled with smooth muscle cells that were of the contractile phenotype, with an organized extracellular matrix. None of these injurious responses were noted in optimally prepared papaverine-treated vein grafts. Optimal preparation of vein grafts is effective in minimizing endothelial and smooth muscle cell injury before and after arterial reconstruction. Prevention of vein graft injury during harvesting prevents the morphologic changes characteristic of the “arterialization response.” (J T horac C ardiovasc S urg 1992;103:671-7)

23-year-old man originally from Angola presented to anoutlying hospital with a 1-day history of palpitations,dyspnea, and near-syncope. In the ambulance, he developedrecurrent ventricular tachycardia. In the emergency room, hewas started on intravenous metoprolol and amiodarone. Hisphysical examination was within normal limits. His ECGshowed underlying normal sinus rhythm with runs of ventric-ular tachycardia (Figure 1). His laboratory values weresignificant for a white blood cell count of 17.4310

Various models have been proposed to examine blood-prosthetic materials interactions in terms of the effect of the prosthetic material on platelet structure and function, blood coagulation and fibrinolysis, and tissue infiltrates (cellular or acellular). In addition, these modeles have been used to examine the change in the graft surface over time. Particular difficulties in examining graft-materials interactions include species differences, short residence time for blood-materials interactions with commonly employed short grafts, and length of study limitations with ex vivo shunts. In this paper we report a canine, carotid-aorta subcutaneous prosthetic graft model. The specific advantages of this model are the length of the graft, which allows prolonged contact of blood with the prosthetic surface; the subcutaneous location of the graft, which allows repeated sampling of blood along the graft; and the healing characteristics of canine grafts. We selected the canine model because the healing characteristics are morphologically similar to those in humans in that endothelialization of the prosthetic surface is limited. Other models, such as the pig, are favored for use when examining blood coagulation, platelet, or fibrinolytic studies; however, these models can fully endothelialize prosthetic surfaces.

The reliability, rapidity, and safety of nonsurgical, transatrial pericardial access for local cardiac therapy have been demonstrated in healthy animals. Since many patients take aspirin or have increased right-sided pressures, we evaluated the procedure's safety under these conditions. Transatrial pericardial access was performed in anesthetized pigs following aspirin administration (162 mg p.o., n = 6) or during experimental pulmonary artery hypertension (n = 4 different animals) and required only 3 minutes following guide catheter positioning. Platelet aggregability testing with arachidonic acid confirmed aspirin effectiveness. Mean pericardial fluid hematocrit was 0.1 +/- 0.1% after 2 days of aspirin therapy and 1.9 +/- 1.1% at sacrifice 24 hours later (NS). Mean pericardial fluid hematocrit was 1.0 +/- 0.5% after 45 minutes of pulmonary artery hypertension and 4.3 +/- 0.8% at sacrifice 30 minutes later (NS). Histologic analysis in both groups revealed a small thrombus and localized inflammation at the site of puncture. Neither aspirin use nor pulmonary artery hypertension causes significant bleeding into the pericardial space following transatrial access and thus does not preclude this route for local cardiac drug delivery.

Phenotypic variation of vascular smooth muscle cells (VSMC) may result in altered biological behavior and responses. Within the vessel wall, arterial VSMC have a greater propensity to form atherosclerotic lesions as compared to venous VSMC. In this study the rates of proliferation, adhesion, and migration were compared between VSMC of atherosclerotic arterial and venous origin.Human VSMC cultures were isolated from 18 infragenicular arteries at the time of below knee amputation and from 20 saphenous veins during lower extremity revascularization surgery. Cell cultures were isolated from the media of each specimen and maintained in distinct cell lines for all assays. Cells from passages 2 and 3 were assayed for their proliferative capacity using total DNA fluorescence photometry and for adhesion and migration using a modified Boyden chamber.Patient age and the incidence of atherosclerotic risk factors did not vary significantly between the arterial and the venous patient groups. VSMC of atherosclerotic arterial origin demonstrated greater proliferation (arterial, 162 +/- 59 absorption units, vs. venous, 106 +/- 56 absorption units, P0.001), adhesion (arterial, 74.1 +/- 22.6 cells/microscopic field, vs. venous, 41.3 +/- 12.8 cells/microscopic field, P0.001) and migration (arterial, 427 +/- 185 cells/microscopic field, vs venous, 119 +/- 101 cells/microscopic field, P0.001) than VSMC of venous origin.Human atherosclerotic arterial VSMC exhibit significantly increased rates of proliferation, adhesion, and migration as compared to human venous VSMC. These observations of VSMC in culture are consistent with the clinical predilection for the hyperplasic responses that result in the development of atherosclerosis in the arterial wall. Possible intrinsic differences in VSMC phenotype should be considered in designing methods to limit atherosclerosis.

Purpose: Patients with diabetes mellitus (DM) experience progressive macrovascular atherosclerosis and intimal hyperplastic restenosis with increased frequency as compared with nondiabetic patients. These observations suggest that vascular smooth muscle cells (VSMCs) behave in a phenotypically different and more aggressive manner in diabetic patients. In this study, we compared the in vitro rates of proliferation, adhesion, and migration of human VSMCs obtained from diabetic and nondiabetic patients. Methods: Human VSMC cultures were isolated from 23 diabetic patients (9 artery, 14 vein) and 15 nondiabetic patients (9 artery, 6 vein) with extensive lower extremity atherosclerosis. All patients were between 61 and 78 years of age (average: 68.4 years [diabetic]; 67.3 years [nondiabetic]). All diabetic patients had type 2 DM. Vascular specimens were obtained at the time of amputation from infragenicular arteries and during arterial revascularization from saphenous veins. Cells from passages 2 and 3 were assayed for their proliferative capacity with total DNA fluorescence photometry and for adhesion and migration with a modified Boyden chamber. Results: The average duration of diabetes was 11.6 ± 4.1 years. The average number of diabetic complications (retinopathy, neuropathy, nephropathy, coronary artery disease) was 2.8 ± 0.7 per patient. Diabetic VSMCs exhibited abnormal morphology in cell culture with loss of the normal hill and valley configuration. Proliferation was significantly increased in VSMCs of diabetic origin (156 ± 57 absorption units) as compared with those of nondiabetic origin (116 ± 42 absorption units) (P

Present prosthetic arterial conduits continue to suffer the clinically and economically catastrophic complication of infection. We recently described a novel technique for binding quinolone antibiotics to Dacron based on principles of textile chemistry. This thermofixation procedure ("pad/heat") utilizes the limited fibrophilic characteristics of the quinolone antibiotic ciprofloxacin (Cipro) to permit pad/heat application and allowed controlled, sustained release from Dacron in several in vitro assays. The objective of this study was to test this infection-resistant prosthetic vascular graft material in an in vivo model. Dacron segments (1 cm2, either plain, dipped into antibiotic immediately prior to implantation, or Cipro pad/heat treated) were implanted in the dorsal subcutaneous tissue of the rabbit and directly contaminated with 10(6) Staphylococcus aureus. After 1 week, the samples were sterily harvested. Wounds were blindly graded on a scale from 1 (no evidence of infection, good tissue incorporation) to 4 (suppurative infection extending outside of the graft pocket, no gross tissue incorporation). Plain Dacron was easily infected in this model (mean grade 3.1 +/- 0.6, 92% culture positive). Notably, however, a significant (P0.05) wound grade difference between the dipped (2.3 +/- 1.0) and pad/heat (1.4 +/- 0.6) samples was demonstrated. Determination of adherent bacteria present on the implanted Dacron pieces by sonication and culture studies again revealed a significant difference between the dipped (56% culture positive) and pad/heat (12% culture) groups (P0.025). Histologic studies confirmed good tissue incorporation of the pad/heat samples. This project opens new avenues in the development of infection-resistant biomaterials.

The precise location and progression of anastomotic hyperplasia and its possible relationship to flow disturbances was investigated in femoro-femoral Dacron grafts in 28 dogs. In 13 grafts, the outflow from the end-to-side downstream anastomosis was bidirectional (BDO), and in 15 it was unidirectional (UDO) (distally). Grafts were electively removed at intervals of two to 196 days or at the time of thrombosis. Each anastomosis and adjacent artery was perfusion-fixed and sectioned sagittally. The mean sagittal section was projected onto a digitized pad, and the total area of hyperplasia internal to the arterial internal elastic lamina and within the adjacent graft was integrated by computer. The location of the hyperplasia was compared with previously established sites of flow separation and stagnation. The observation was made that hyperplasia is significantly greater at the downstream, as compared with the upstream, anastomosis in both groups (BDO = p less than 0.001 and UDO = p less than 0.001) (analysis of variance for independent groups). Furthermore, this downstream hyperplasia was progressive with time (BDO p less than 0.01) (UDO p less than 0.01); Spearman Rank Correlation. There was no significant increase in the extent of downstream hyperplasia where flow separation was known to be greater (BDO). Five grafts failed (three BDO, two UDO), as a result of complete occlusion of the downstream anastomosis by fibrous hyperplasia. Transmission electron microscopy showed the hyperplasia to consist of collagen-producing smooth muscle cells. Anastomotic hyperplasia is significantly greater at the downstream anastomosis, is progressive with time, and is the primary cause of failure of Dacron arterial grafts in this model. Quantitative analysis of downstream anastomotic hyperplasia may be a valuable measure of the biocompatibility of Dacron grafts.

Boundary layer separation in a plexiglass model carotid bifurcation was investigated in relation to the origin of atherosclerotic plaque clinically found in this region. Our model was comparable to a human carotid in both dimensions and geometry. Water flowed through the model at Reynolds numbers from 200 to 1200 under steady and pulsatile flow conditions, with outflow through the external and internal branches varied. The near-wall flow was visualized by slow injection of dye through ports machined in the model. Under steady flow at a physiological Reynolds number of 500 and a flow split at the bifurcation similar to that of a human carotid at rest, boundary layer separation was found to occur in a carotid sinus across from the external carotid origin, forming a shell of slowly moving fluid around the bifurcation. The rapidly moving mainstream impinged directly on the flow divider. The location of atherosclerotic plaque correlates best with the low shear region of separation and not with the region of high shear at the flow divider. Preliminary studies with pulsatile flow demonstrated little change from the steady flow results.

Spasm and endothelial injury during harvesting of saphenous vein grafts can be prevented by a protocol that involves perivenous infiltration with 37 degrees C papaverine hydrochloride solution prior to skin incision, initial distention of the vein with the same warm papaverine solution, storage of the distended vein graft in a cold papaverine solution, and maintenance of the vein in the distended state throughout the operative procedure. This technique preserves endothelial integrity in human reversed saphenous vein grafts.