[show abstract][hide abstract] ABSTRACT: Percutaneous coronary interventions play a major role in the management of patients affected by coronary artery diseases. However, their efficiency is impaired by restenosis, defined as a reduction of the vessel lumen, occurring a few months after the procedure. A low-molecular-weight fraction of fucoidan, a vegetal heparin-like sulphated polysaccharide, was recently shown to greatly reduce in-stent restenosis after angioplasty in rabbits. To better understand the in vivo anti-restenotic effects of this polymer, we used fractions of fucoidan and compared to heparin and dextran of different sizes. We carried out in vitro growth inhibition experiments on vascular smooth muscle cells, performed an in vivo pharmacokinetic study, and locally delivered fluorescently-labeled polysaccharides in rabbit iliac arteries after angioplasty with a non-occlusive catheter. The results indicated that (i) preparation of well-characterized fractions from natural fucoidan is compulsory for in vitro and in vivo studies, (ii) antiproliferative activity of sulphated polysaccharides on cultured smooth muscle cells is not a major predictive factor for the reduction of restenosis in vivo and (iii) pharmacokinetic parameters and binding of low-molecular-weight fucoidan on angioplasty-induced injured vascular walls are important local and general factors controlling its mechanisms of action.
[show abstract][hide abstract] ABSTRACT: To prospectively evaluate in rats whether magnetic cell labeling can be used to noninvasively assess the technical success of endovascular cell therapy for abdominal aortic aneurysms (AAAs).
The study was approved by an institutional animal care and use committee. Vascular smooth muscle cells (VSMCs) labeled with iron oxide nanoparticles were seeded endovascularly in already formed AAAs. T2*-weighted gradient-echo and T2-weighted spin-echo magnetic resonance (MR) imaging was performed in vivo at 1.5 T before and 30 minutes after the injection of iron-loaded VSMCs (14 rats), nonlabeled VSMCs (three rats), or iron-free particles (three rats). Ten rats were euthanized shortly after the injection (day 0). Of the 10 remaining rats, which were seeded with iron-loaded cells, three were imaged on day 7 after cell delivery; three, on day 14; and four, on day 28; then they were euthanized. Ex vivo high-field-strength MR imaging of two AAAs was performed 28 days after cell delivery. Histologic examination of cross sections of all AAAs was performed. Statistical evaluations were performed with a nonparametric Wilcoxon correlation test.
Magnetic cell labeling did not alter the capability of VSMCs to stabilize the diameter of the aneurysms. T2*-weighted gradient-echo images showed areas of hypointense signal within the aortic wall immediately and up to 1 month after cell therapy. The mean signal intensity decreased significantly after cell delivery (from 2362 +/- 244 [standard deviation] before to 434 +/- 275 after delivery, P < .001). Areas of hypointense signal and iron-loaded VSMCs were colocalized in the area of aortic wall reconstruction at both high-field-strength MR imaging and histologic analysis.
MR imaging with magnetic cell labeling can be used to document endovascular cell delivery in already formed AAAs in rats.
[show abstract][hide abstract] ABSTRACT: To perform a quantitative analysis of anionic maghemite nanoparticle-labeled cells in vitro and determine the effect of labeling on signal intensity at magnetic resonance (MR) imaging.
The study was approved by the institutional animal care and use committee at Hôpital Bichat. In vitro cell proliferation, iron content per cell, and MR signal intensity of cells were measured in agarose phantoms for 0-14 days of culture after labeling of rat smooth muscle cells with anionic maghemite nanoparticles. Next, iron oxide-labeled smooth muscle cells were injected into healthy hearts and hearts with ischemic injury in seven live Fisher rats. Ex vivo MR imaging experiments in excised hearts 2 and 48 hours after injection were performed with a 1.5-T medical imaging system by using T2-weighted gradient-echo and spin-echo sequences. Histologic sections were obtained after MR imaging. Correlation analyses between division factor of iron load and cell amplification factor and between 1/T2 and number of labeled cells or number of days in culture were performed by using linear regression.
Viability of smooth muscle cells was not affected by magnetic labeling. Transmission electron micrographs of cells revealed the presence of iron oxide nanoparticles in vesicles up to day 14 of culture. Intracellular iron concentration decreased in parallel with cell division (r2 = 0.99) and was correlated with MR signal intensity (r2 = 0.95). T2*-weighted MR images of excised rat hearts showed hypointense signal in myocardium at 2 and 48 hours after local injection of labeled cells. Subsequent histologic staining evidenced iron oxide nanoparticles within cells and confirmed the presence of the original cells at 2 and 48 hours after implantation.
Magnetic labeling of smooth muscle cells with anionic maghemite nanoparticles allows detection of cells with MR imaging after local transplantation in the heart.
[show abstract][hide abstract] ABSTRACT: The aim of our study was to evaluate the feasibility, safety, and potential role of the contrast agent gadoterate meglumine for digital subtraction angiography as a single diagnostic procedure or before percutaneous transluminal angioplasty of malfunctioning native dialysis fistulas.
Over a 20-month period, 23 patients (15 women, eight men) with an age range of 42-87 years (mean, 63 years) having end-stage renal insufficiency and with recent hemodialysis fistula surgical placement underwent gadoterate-enhanced digital subtraction angiography with a digital 1024 x 1024 matrix. Opacification was performed on the forearm, arm, and chest with the patient in the supine position using an injection (retrograde, n = 14; anterograde, n = 8; arterial, n = 1) of gadoterate meglumine into the perianastomotic fistula segment at a rate of 3 mL/sec for a total volume ranging from 24 to 32 mL. Percutaneous transluminal angioplasty was performed in three patients and required an additional 8 mL per procedure. Examinations were compared using a 3-step confidence scale and a two-radiologist agreement (Cohen's kappa statistic) for diagnostic and opacification quality. Tolerability was evaluated on the basis of serum creatinine levels and the development of complications.
No impairment of renal function was found in the 15 patients who were not treated with hemodialysis. Serum creatinine level change varied from -11.9% to 11.6%. All studies were of diagnostic quality. The presence of stenosis (n = 14) or thrombosis (n = 3) in arteriovenous fistulas was shown with good interobserver agreement (kappa = 0.71-0.80) in relation to opacification quality (kappa = 0.59-0.84). No pain, neurologic complications, or allergiclike reactions occurred. Three percutaneous transluminal angioplasty procedures (brachiocephalic, n = 2; radiocephalic, n = 1) were successfully performed.
Gadoterate-enhanced digital subtraction angiography is an effective and safe method to assess causes of malfunction of hemodialysis fistulas. It can also be used to plan and perform percutaneous transluminal angioplasty.
American Journal of Roentgenology 11/2002; 179(4):1023-8. · 2.90 Impact Factor
[show abstract][hide abstract] ABSTRACT: Smooth muscle cell (SMC) proliferation within the intima is regulated by heparan sulfates. We studied a low molecular weight (LMW) fucoidan (sulfated polysaccharide from brown seaweed) on SMC proliferation in vitro and intimal hyperplasia in vivo.
In vitro study revealed that LMW fucoidan reduces rabbit SMC proliferation and is internalized in SMC perinuclear vesicles. On rabbit iliac arteries perfused in vivo with fluorolabeled LMW fucoidan after angioplasty, the labeling was mainly located on sites of injury. Pharmacokinetic studies showed that LMW fucoidan exhibited in rats an elimination half-life of 56+/-25 minutes (n=8) after intravenous administration and a constant plasma rate for > or =6 hours after intramuscular administration. After stent implantation in their iliac arteries, rabbits were also treated with LMW fucoidan (5 mg/kg IM twice a day). Histomorphometric analysis at day 14 indicated that LMW fucoidan reduced intimal hyperplasia by 59% (1.79+/-0.4 versus 0.73+/-0.2 mm2, P<0.0001) and luminal cross-sectional area narrowing by 58% (0.38+/-0.08 versus 0.16+/-0.04, P<0.0001). Blood samples showed no anticoagulant activity due to LMW fucoidan.
This natural polysaccharide with high affinity for SMCs and sustained plasma concentration markedly reduced intimal hyperplasia, suggesting its use for the prevention of human in-stent restenosis.
[show abstract][hide abstract] ABSTRACT: Intimal smooth muscle cell (SMC) hyperplasia is a main component of the arterial wall response to injury. We have investigated the capacity of a water-soluble nonanticoagulant functionalized dextran (E9) in inhibition of SMC growth in vitro and in vivo.
E9 was obtained with chemical substitutions with anionic and hydrophobic groups on the dextran backbone. SMC proliferation (cell counting, thymidine uptake, cell cycle analysis) was followed in culture in the presence of E9. Western blot analysis against phosphorylated mitogen-activated protein kinase (MAPK), extracellular signal-regulated protein kinase 1/2, and assessment of MAPK activity on serum-stimulated SMCs also were investigated. Binding/displacement experiments, electron microscopy, and cell fractionations were used to follow the binding and internalization of radiolabeled and fluorescentlabeled E9. New Zealand white rabbit iliac arteries were injured with balloon dilatation and stent deployment. Animals were treated for 14 days with saline solution or E9 (5 mg/kg injected subcutaneously, twice daily). Morphometric analyses were carried out in each group (n = 6 arteries, 18 sections).
Nonanticoagulant E9 inhibited SMC proliferation in vitro. Tyrosine phosphorylation of MAPK 1/2 and MAPK activity were inhibited with E9 within 5 minutes of incubation. The binding and rapid cytoplasmic internalization of the synthetic compound was evidenced, but, in contrast to heparin, we did not detect any nuclear localization of the antiproliferative E9. In the in vivo model, qualitative modifications of neointimal structure with a thinner fibrocellular neointima were noticed after E9 treatment. Morphometric analyses of stented arteries in E9-treated animals indicated an important reduction (P <.01) of intimal growth: 33% and 45% for intimal area and intima/media ratio, respectively.
Cytoplasmic internalization of the synthetic polysaccharide correlated to the SMC growth inhibition that involved the MAPK pathway. In vivo inhibition of intimal instent hyperplasia with this nonanticoagulant derived dextran is shown providing a new candidate for a potential selective treatment of SMC proliferation.
Journal of Vascular Surgery 05/2002; 35(5):973-81. · 2.88 Impact Factor