Effects of external crushing forces on a novel below-the-knee vascular implant.

BACKGROUND: The Tack Endovascular System® is a novel vascular implant designed to focally treat dissections with low radial force and minimal metal burden. As there are currently no approved below-the-knee (BTK) implants in the USA, a unique, 3-stage model was developed to characterize crush deformation and fracture potential of the Tack Endovascular System in BTK arteries. METHODS: First, 35 Tack® implants were deployed bilaterally in the posterior tibial, anterior tibial, and peroneal arteries of 3 cadavers, and clinically relevant external forces were applied to simulate BTK crushing deformation including focal load, leg crossing, and leg bending. Intravascular ultrasound images of the implanted vessels were used to assess the magnitude of artery deformation. Outputs of the cadaver testing were input into a finite element analysis (FEA) model to determine the appropriate conditions for subsequent bench testing. Tack implants were then subjected to increasing crush forces at 30Hz for up to 650,000 cycles at 25% flat plate deformation within the worst-case FEA test condition. RESULTS: Crush deformation across all arteries ranged from 0% to 23.1%. The posterior tibial artery and large male cadaver exhibited the most vulnerability to external crush forces, while the small female model exhibited the most resistance. No fractures were observed during cadaver or bench testing. CONCLUSIONS: This study characterized deformation forces in tibial arteries during various loading conditions. Tack implants withstood the loading conditions without fracture within the limits of this ex-vivo human vascular model and in-vitro bench testing.

An Animal Model of Human Peripheral Arterial Bending and Deformation.

BACKGROUND: Designing peripheral arterial stents has proved challenging, as implanted devices will repetitively and unpredictably deform and fatigue during movement. Preclinical testing is often inadequate, given the lack of relevant animal models. The purpose of this study was to test the hypothesis that deformation of the human peripheral vasculature could be qualitatively and quantitatively modeled using an experimental animal. METHODS: Anteroposterior contrast angiography was performed in domestic Landrace-Yorkshire farm pigs. Images were obtained with the hind limbs naturally extended then repeated, (1) flexed approximately 90° at the hip and knee, (2) overflexed in a nonphysiological fashion. Quantitative vascular angiographic analysis was utilized to measure arterial diameter, length, and deformation. Percent axial arterial compression and bending were assessed. RESULTS: Eight iliofemoral arteries in four animals were imaged. Mean luminal diameters of the iliac and femoral segments in the neutral position were 5.4 ± 0.5 mm and 4.6 ± 0.5 mm. Hind limb physiologic flexion induced profound arterial compression, 17 ± 8% and 29 ± 6% and bending, 36°±10° and 76° ± 13° within the iliac and femoral segments, respectively. With extreme flexion, the femoral artery could be reliably bent >90°. The observed findings exceeded the deformation observed historically within the human superficial femoral (∼5% compression and 10° bending) and popliteal artery (∼10% compression and 70° bending). CONCLUSIONS: Significant nonradial deformation of the porcine iliofemoral arteries was observed during manual hind limb flexion and exceeded that typically observed in humans. This model constitutes a "worst case" scenario for testing deformation and fatigue of intravascular devices indicated for the human peripheral vasculature.

Calcified plaque modification alters local drug delivery in the treatment of peripheral atherosclerosis.

BACKGROUND: Calcific atherosclerosis is a major challenge to intraluminal drug delivery in peripheral artery disease (PAD). OBJECTIVES: We evaluated the effects of orbital atherectomy on intraluminal paclitaxel delivery to human peripheral arteries with substantial calcified plaque. METHODS: Diagnostic angiography and 3-D rotational imaging of five fresh human lower limbs revealed calcification in all main arteries. The proximal or distal segment of each artery was treated using an orbital atherectomy system (OAS) under simulated blood flow and fluoroscopy. Explanted arterial segments underwent either histomorphometric assessment of effect or tracking of 14C-labeled or fluorescent-labeled paclitaxel. Radiolabeled drug quantified bulk delivery and fluorescent label established penetration of drug over finer spatial domain in serial microscopic sections. Results were interpreted using a mathematical model of binding-diffusion mediated arterial drug distribution. RESULTS: Lesion composition affected paclitaxel absorption and distribution in cadaveric human peripheral arteries. Pretreatment imaging calcium scores in control femoropopliteal arterial segments correlated with a log-linear decline in the bulk absorption rate-constant of 14C-labeled, declining 5.5-fold per calcified quadrant (p=0.05, n=7). Compared to controls, OAS-treated femoropopliteal segments exhibited 180µm thinner intima (p<0.001), 45% less plaque calcification, and 2 log orders higher paclitaxel bulk absorption rate-constants. Correspondingly, fluorescent paclitaxel penetrated deeper in OAS-treated femoropopliteal segments compared to controls, due to a 70% increase in diffusivity (p<0.001). CONCLUSIONS: These data illustrate that calcified plaque limited intravascular drug delivery, and controlled OAS treatment of calcific plaques resulted in greater drug permeability and improved adjunct drug delivery to diseased arteries.

Assessment of self-expanding nitinol stent deformation after chronic implantation into the femoropopliteal arteries.

AIMS: The purpose of this prospective clinical investigation was to quantify the degree and range of compressive and bending deformations sustained by self-expanding nitinol stents when implanted into the femoropopliteal arteries of patients with symptomatic peripheral vascular disease (PVD). METHODS AND RESULTS: Twenty-three nitinol self-expanding stents (Absolute; Abbott Vascular, Santa Clara, CA, USA) with diameters ranging from 5-10 mm and lengths ranging from 40-100 mm were implanted in 19 lesions in 18 extremities of 17 patients. Two days following implantation, in vivo stent compression and bending were assessed by measurement of stent length and deflection angle via lateral view radiographs. The results showed that leg flexion was associated with significant stent bending; popliteal stents bent almost 90° while SFA stents bent only minimally. Leg flexion was also associated with stent shortening (compression), the greatest amount being observed for stents implanted into the popliteal artery (popliteal 8.5% ± 3.2%; SFA/prox pop 5.3% ± 0.5%; SFA 3.1% ± 1.8%). After a mean follow-up of 7.1 ± 1.3 months, the degree of stent deformation during leg flexion was essentially unchanged as compared to immediate post-procedure levels. CONCLUSIONS: Indwelling nitinol stents are routinely bent and compressed during leg flexion, with most bending observed in stents implanted near the popliteal artery. The degree of deformation observed immediately after implantation appears to be predictive of the chronic state, as repeat measurements obtained after a mean of seven months were essentially unchanged from baseline.

Mechanisms of tissue uptake and retention in zotarolimus-coated balloon therapy.

BACKGROUND: Drug-coated balloons are increasingly used for peripheral vascular disease, and, yet, mechanisms of tissue uptake and retention remain poorly characterized. Most systems to date have used paclitaxel, touting its propensity to associate with various excipients that can optimize its transfer and retention. We examined zotarolimus pharmacokinetics. METHODS AND RESULTS: Animal studies, bench-top experiments, and computational modeling were integrated to quantify arterial distribution after zotarolimus-coated balloon use. Drug diffusivity and binding parameters for use in computational modeling were estimated from the kinetics of zotarolimus uptake into excised porcine femoral artery specimens immersed in radiolabeled drug solutions. Like paclitaxel, zotarolimus exhibited high partitioning into the arterial wall. Exposure of intimal tissue to drug revealed differential distribution patterns, with zotarolimus concentration decreasing with transmural depth as opposed to the multiple peaks displayed by paclitaxel. Drug release kinetics was measured by inflating zotarolimus-coated balloons in whole blood. In vivo drug uptake in swine arteries increased with inflation time but not with balloon size. Simulations coupling transmural diffusion and reversible binding to tissue proteins predicted arterial distribution that correlated with in vivo uptake. Diffusion governed drug distribution soon after balloon expansion, but binding determined drug retention. CONCLUSIONS: A large bolus of zotarolimus releases during balloon inflation, some of which pervades the tissue, and a fraction of the remaining drug adheres to the tissue-lumen interface. As a result, the duration of delivery modulates tissue uptake where diffusion and reversible binding to tissue proteins determine drug transport and retention, respectively.

Inhibition of experimental neointimal hyperplasia and neoatherosclerosis by local, stent-mediated delivery of everolimus.

INTRODUCTION: A novel self-expanding, drug-eluting stent (DES) was designed to slowly release everolimus in order to prevent restenosis after percutaneous peripheral intervention. The purpose of this experimental animal study was to test the hypothesis that long-term local, stent-mediated delivery of everolimus would reduce neointimal hyperplasia in porcine iliac arteries. METHODS: The iliac arteries of 24 Yucatan mini-swine were percutaneously treated with overlapping 8- × 28-mm self-expanding nitinol stents loaded with everolimus (225 µg/cm2 stent surface area) formulated in a poly(ethylene-co-vinyl alcohol) copolymer intended to deliver the drug in a sustained fashion over about 6 months (DES). Bare nitinol self-expanding stents (bare metal stent [BMS]) were implanted in an identical fashion on the contralateral side to serve as controls. After 3, 6, or 12 months, the animals were sacrificed and the stented arteries perfusion-fixed for histomorphometric analysis. RESULTS: The chronic presence of everolimus in arterial tissue reduced stent-induced inflammation after 3 months (inflammation score: BMS 2.29±0.44 vs DES 0.17±0.17; P=.001) and 6 months (BMS 2.06±0.43 vs DES 0.50±0.5; P=.007), although some late inflammation was observed after drug exhaustion (BMS 1.00±0.25 vs DES 2.56±0.62 after 12 months; P=not significant [NS]). Treatment with locally delivered everolimus significantly reduced neointimal hyperplasia after 3 months (neointimal thickness: BMS 0.79±0.20 vs DES 0.37±0.04 mm; P=.03) and 6 months (BMS 0.73±0.14 vs DES 0.41±0.08 mm; P=.05), although the effect had dissipated after 12 months (BMS 0.68±0.11 vs DES 0.67±0.11 mm; P=NS). Remarkably, stent-induced neoatherosclerosis, characterized by the histologic presence of foamy macrophages and cholesterol clefts, was significantly attenuated by treatment with everolimus (atherogenic change scores at 3 months: BMS 0.56±0.15 vs DES 0.04±0.04; P=.003; 6 months: BMS 0.84±0.23 vs DES 0.00±0.00; P=.004; and 12 months: BMS 0.09±0.10 vs DES 0.19±0.19; P=NS). CONCLUSIONS: In this experimental animal model, local arterial stent-mediated delivery of everolimus inhibited the formation of neointimal hyperplasia and neoatherosclerosis during the first 6 months. The effect was transient, however, as arterial morphology and histology appeared similar to control stented arteries after 12 months.

Vascular response to zotarolimus-coated balloons in injured superficial femoral arteries of the familial hypercholesterolemic Swine.

BACKGROUND: Drug-coated balloons are rapidly emerging as a therapeutic alternative for the interventional treatment of peripheral vascular disease. The purpose of this study was to test the hypothesis that an angioplasty balloon coated with the mTOR inhibitor zotarolimus (ZCB) would inhibit neointimal hyperplasia in a novel injury-based superficial femoral artery model in the familial hypercholesterolemic swine. METHODS AND RESULTS: A total of 44 familial hypercholesterolemic swine were included (12 designated to study tissue pharmacokinetics and 32 to study safety and efficacy). Fogarty balloon denudation was performed in all superficial femoral artery segments, followed by balloon angioplasty. In the pharmacokinetic study, a total of 24 ZCBs (300 µg/cm(2)) were used. Zotarolimus was detected in arterial tissue at 5 minutes (162 ng/mg of tissue), 24 hours (5.9 ng/mg of tissue), and 28 days (0.007 ng/mg of tissue) after ZCB inflation. In the safety and efficacy study, superficial femoral artery segments were randomized to either high-dose (600 µg/cm(2), n=16), low-dose (300 µg/cm(2), n=16), or paired uncoated balloons (high-dose ZCB control, n=16; low-dose ZCB control, n=16). At 28 days, the percentage of angiographic stenosis was similar among all tested groups. Histological analysis demonstrated a reduction in neointimal formation in both ZCB groups compared with controls (high-dose ZCB 44% reduction, P=0.007; low-dose ZCB 22% reduction, P=0.08). There was no evidence of delayed arterial healing or vascular toxicity in any of the ZCB groups. CONCLUSIONS: The single delivery of zotarolimus via coated balloon is feasible, and therapeutic levels are maintained up to 28 days. The ZCB technology appears to be effective in the reduction of neointimal proliferation in the superficial femoral artery of the familial hypercholesterolemic swine.

Respiration-induced kidney motion on cobalt-chromium stent fatigue resistance.

During normal breathing, the kidneys move up and down due to the diaphragm motion and the renal artery subsequently experiences bending at or close to its point of fixation to the aorta. The impact of this kidney motion on implanted stent fatigue performance was not well understood in the past. Previous study from the authors on an 18-mm long single cobalt-chromium stent showed that the change in bending angle was minor during simulated respiration-induced kidney motion on cadavers. Finite Element Analysis revealed excellent fatigue resistance of the studied cobalt-chromium stent under simulated respiratory motion for the single stent configuration. In this article, the study was extended further to the overlapped stent configuration where a physician deploys two stents overlapping at the stent ends to fully cover a long lesion. Fluoroscopic images showed that the change in bending angle during simulated respiration-induced kidney motion on cadavers was more significant when two cobalt-chromium stents were overlapped. Calculated data of the Goodman analysis for the overlapped stents migrated toward the Goodman diagram failure line, indicating lower fatigue resistance during respiration when compared to a single stent.

Late stent expansion and neointimal proliferation of oversized Nitinol stents in peripheral arteries.

For peripheral endovascular intervention, self-expanding (SE) stents are commonly oversized in relation to target arteries to assure optimal wall apposition and prevent migration. However, the consequences of oversizing have not been well studied. The purpose of this study was to examine the effects of SE stent oversizing (OS) with respect to the kinetics of late stent expansion and the long-term histological effects of OS. Pairs of overlapped 8 x 28-mm Nitinol SE stents were implanted into the iliofemoral arteries of 14 Yucatan swine. Due to variations in target artery size, the stent-to-artery ratio ranged from 1.2:1 to 1.9:1. Lumen and stent diameters were assessed by quantitative angiography at the time of implantation. Following angiographic assessment at 6 months, stented arteries were perfusion-fixed, sectioned, and stained for histological analysis. Immediately following implantation, the stents were found to be expanded to a range of 4.7-7.1 mm, largely conforming to the diameter of the recipient target artery. The stents continued to expand over time, however, and all stents had enlarged to nearly their 8-mm nominal diameter by 6 months. The histological effects of OS were profound, with marked increases in injury and luminal area stenosis, including a statistically significant linear correlation between stent-to-artery ratio and area stenosis. In this experimental model of peripheral endovascular intervention, oversized Nitinol SE stents are constrained by their target artery diameter upon implantation but expand to their nominal diameter within 6 months. Severe OS (stent-to-artery ratio >1.4:1) results in a profound long-term histological response including exuberant neointimal proliferation and luminal stenosis.

Fracture of self-expanding nitinol stents stressed in vitro under simulated intravascular conditions.

OBJECTIVE: The use of intravascular stents in the superficial femoral artery (SFA) continues to be controversial due to the potential for compression and fracture in the tortuous physical environment of the adductor canal. The purpose of this study was to (1) characterize the types and ranges of stent distortion theoretically produced by extremity movement and (2) use these ranges as parameters for in vitro long-term fatigue testing of commercially available self-expanding nitinol stents. METHODS: Nitinol self-expanding stents were placed in the SFAs of cadavers and lateral view radiographs were obtained with the limb in various degrees of hip and knee flexion. The measured degrees of axial shortening and bending of the stent were estimated by planimetry and used for in vitro fatigue testing, which was undertaken using specially designed equipment. Six different commercially available nitinol self-expanding stents-Protégé EverFlex (EV3, Minneapolis, Minn), S.M.A.R.T. Control (Cordis/Johnson & Johnson, Miami Lakes, Fla), Luminexx (C.R. Bard, Murray Hill, NJ), LifeStent FlexStar (Edwards Lifesciences, Irvine, Calif), and Xceed and Absolute (Abbott Vascular, Santa Clara, Calif)-were mounted in elastic silicone tubing, bathed in phosphate buffered saline at 37 degrees +/- 2 degrees C, and examined for fracture after 10 million cycles of chronic deformation. RESULTS: For unstented arteries, the distal SFA/proximal popliteal artery exhibited the greatest axial compression (23%) vs the middle SFA (9%) or popliteal artery (14%) at 90 degrees /90 degrees knee/hip flexion. For stented arteries, the popliteal artery exhibited the most axial compression (11%) vs the middle SFA (3%) or distal SFA/proximal popliteal artery (6%) at 90 degrees /90 degrees knee/hip flexion. Axial compression of the stented popliteal artery at 70 degrees /20 degrees knee/hip flexion was 6% with a deflection angle of 33 degrees . These parameters were used for chronic in vitro fatigue testing, which produced a range of responses in commercially available stents. Chronic 5% axial compression resulted in high rates of fracture of Luminexx (80%) and LifeStent FlexStar (50%), with lower fracture rates for Absolute (3%), Protégé EverFlex (0%), and S.M.A.R.T. Control stents (0%). Chronic 48 degrees bending deformation resulted in high rates of fracture in Protégé EverFlex (100%), S.M.A.R.T. Control (100%), and Luminexx stents (100%), with lower rates in Absolute (3%) and LifeStent FlexStar (0%). CONCLUSION: Nitinol self-expanding stents undergo both axial and bending deformation when implanted into the superficial femoral and popliteal arteries. Commercially available stents exhibit a variable ability to withstand chronic deformation in vitro, and their response is highly dependent on the type of deformation applied.

Sustained high-level expression of human factor IX (hFIX) after liver-targeted delivery of recombinant adeno-associated virus encoding the hFIX gene in rhesus macaques.

The feasibility, safety, and efficacy of liver-directed gene transfer was evaluated in 5 male macaques (aged 2.5 to 6.5 years) by using a recombinant adeno-associated viral (rAAV) vector (rAAV-2 CAGG-hFIX) that had previously mediated persistent therapeutic expression of human factor IX (hFIX; 6%-10% of physiologic levels) in murine models. A dose of 4 x 10(12) vector genomes (vgs)/kg of body weight was administered through the hepatic artery or portal vein. Persistence of the rAAV vgs as circular monomers and dimers and high-molecular-weight concatamers was documented in liver tissue by Southern blot analysis for periods of up to 1 year. Vector particles were present in plasma, urine, or saliva for several days after infusion (as shown by polymerase chain reaction analysis), and the vgs were detected in spleen tissue at low copy numbers. An enzyme-linked immunosorption assay capable of detecting between 1% and 25% of normal levels of hFIX in rhesus plasma was developed by using hyperimmune serum from a rhesus monkey that had received an adenoviral vector encoding hFIX. Two macaques having 3 and 40 rAAV genome equivalents/cell, respectively, in liver tissue had 4% and 8% of normal physiologic plasma levels of hFIX, respectively. A level of hFIX that was 3% of normal levels was transiently detected in one other macaque, which had a genome copy number of 25 before abrogation by a neutralizing antibody (inhibitor) to hFIX. This nonhuman-primate model will be useful in further evaluation and development of rAAV vectors for gene therapy of hemophilia B.