(Circulation. 1997;96:636-645.)
© 1997 American Heart Association, Inc.
Articles |
Paclitaxel Inhibits Arterial Smooth Muscle Cell Proliferation and Migration In Vitro and In Vivo Using Local Drug Delivery
From the Departments of Cardiology and Surgery (G.K.), University of Tübingen, Germany.
Correspondence to Dorothea I. Axel, PhD, Dept of Cardiology, University of Tübingen, Otfried-Müller St 10, D-72076 Tübingen, Germany. E-mail dorothea.axel{at}uni-tuebingen.de
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Background The antineoplastic compound paclitaxel (Taxol) causes an increased assembly of extraordinarily stable microtubules. The present study was designed to characterize the effects of paclitaxel on proliferation and migration of human arterial smooth muscle cells (haSMCs) in vitro and on neointima formation in an in vivo experimental rabbit model.
Methods and Results Both monocultures of haSMCs and cocultures with human arterial endothelial cells (haECs) were used. Cell growth after 4, 8, and 14 days was determined in the absence or presence of platelet-derived growth factor-AB (PDGF-AB), basic fibroblast growth factor (bFGF), or thrombin. Nonstop paclitaxel exposure, as well as single-dose applications of paclitaxel for 24 hours or even 20 minutes (0.1 to 10.0 µmol/L), caused a complete and prolonged inhibition of haSMC growth up to day 14, with an IC50 of 2.0 nmol/L. Mitogens or cocultures with stimulating haECs did not significantly attenuate paclitaxel-induced effects. Immunohistochemistry showed characteristic cytoskeletal changes predominantly in the microtubule network. Additionally, in 20 male New Zealand White rabbits, intimal plaques were produced by electrical stimulation. In 10 animals, paclitaxel was locally applied by use of microporous balloons. Histologically, the intima wall area, wall thickness, and degree of stenosis were reduced significantly in paclitaxel-treated animals compared with controls.
Conclusions Our data show that paclitaxel inhibits haSMC proliferation and migration in a dose-dependent manner in monocultures and cocultures even in the presence of mitogens. Furthermore, paclitaxel prevents neointima formation in rabbits after balloon angioplasty. The long-lasting effect after just several minutes' exposure time makes this lipophilic substance a promising candidate for local antiproliferative therapy of restenosis.
Key Words: restenosis • paclitaxel • pharmacology • muscle, smooth • endothelium
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Microtubules are important parts of the cytoskeleton, especially of the mitotic spindle. They are involved in diverse cellular functions, such as cell division, cell motility, and the maintenance of cell shape. In addition, they play a role in signal transduction (eg, from mitogen receptors), intracellular transport mechanisms, and gene activation.1 2 The novel diterpenoid paclitaxel (Taxol), a potent antineoplastic drug, has been shown to be a promising agent for the therapy of ovarian, breast, and other cancers.3 In contrast to the molecular mechanism of the Vinca alkaloids colchicine and Colcemid (demecolcine), which prevent the polymerization of tubulin into microtubules, paclitaxel stabilizes polymerized microtubules and enhances microtubule assembly.4 As a result, cell replication is inhibited predominantly in the G0/G1 and G2/M phases of the cell cycle.5 Morphologically, paclitaxel-treated cells form abnormal, disorganized microtubules, leading to an epithelioid cell shape.6 Previous experimental in vitro studies with paclitaxel have been performed in tumor cell lines,7 epithelial cells,8 and fibroblasts, predominantly 3T3 cells.9
Unlike other antiproliferative agents, paclitaxel also has several properties that make it a good candidate for local drug therapy of excessive arterial smooth muscle cell proliferation in restenosis after balloon angioplasty or stent implantation; these properties have been tested in vitro,10 in animal models,11 and in clinical studies12 thus far. First, the highly lipophilic character of paclitaxel promotes a rapid cellular uptake by enabling it to easily pass through the hydrophobic barrier of cell membranes.13 Second, the unique mode of action supports a long-lasting antiproliferative action even after a brief, single-dose application at very low concentrations, as previously shown in tumor cells.1 14 An antiproliferative effect of paclitaxel on vascular cells has been shown in vitro in rat VSMCs as well as in vivo in the rat carotid artery injury model.15 Paclitaxel was found to interfere with VSMC proliferation and migration at nanomolar levels in vitro and to prevent neointimal VSMC accumulation in the carotid artery in vivo. However, the failure of nearly all human studies in the prevention of restenosis after testing a broad spectrum of antiproliferative drugs showed that promising results of animal cell cultures and in vivo studies cannot be transferred directly to the human situation.12 Thus, the current study was designed to analyze the effects of paclitaxel on haSMC and haEC growth and structure of the cytoskeleton. Detailed experiments comparing very brief (20-minute) with nonstop incubation periods at several time points up to 14 days were designed to exclude rebound effects after drug removal between the first wave of cell proliferation and migration after injury,11 eg, as described for heparins.12 Additional studies with growth factor–stimulated haSMCs and with our previously described transfilter coculture system16 17 were performed to imitate the complexity of in vivo cell-cell interactions, which certainly influences the efficacy of antiproliferative drugs.
Finally, the in vitro results were substantiated by in vivo studies that used locally applied paclitaxel to prevent neointima formation after balloon dilatation of plaques that were induced by electrical stimulation of rabbit carotid arteries.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Cell Isolation
Human iliac and renal arteries were obtained from organ donors (approved by the local ethics committee). haSMCs and haECs were isolated and passaged according to techniques described previously.17 haSMCs were grown in a 1:1 mixture of Waymouth MB 752/1 medium and Nutrient Mixture F12 Ham (1:1 vol/vol) supplemented with 10% FCS, 100 U/mL penicillin, and 100 µg/mL streptomycin (Gibco BRL). Postconfluence haSMCs grew in the characteristic "hill-and-valley" growth pattern. Smooth muscle origin was confirmed by immunocytochemical stainings with monoclonal antibodies against smooth muscle
-actin (Progen) and heavy-chain
myosin (Sigma).
haECs were subcultured onto collagen-coated plastic culture dishes
(Iwaki Glass) and were grown in an endothelial medium
kit (EMK-1; Sigma) containing endothelial cell growth
factor and 2% FCS. Subcultured haECs were characterized by
immunocytochemical stainings with polyclonal antibodies against the von
Willebrand factor (Boehringer Mannheim), and
contamination by haSMCs was excluded by additional double stainings
using antibodies against smooth muscle -actin. Monolayers showed the
characteristic cobblestone morphology. Cells were fed every third day
and used for experiments in low passages (2 through 6).
To exclude contamination by mycoplasma, DAPI stainings (Boehringer Mannheim) were performed routinely in several passages.18
Drugs
Paclitaxel (Sigma) was dissolved in 100% ethanol and sterile
filtered. Taxol was obtained from Bristol-Myers Squibb. It consisted of
7.0 mmol/L paclitaxel dissolved in a lipoid vehicle, a 1:1 mixture
of polyethoxylated castor oil (Cremophor EL), and absolute ethanol. To
obtain different test concentrations between 0.1 nmol/L and 10.0
µmol/L, serial dilutions of a stock solution (2.5 mmol/L) were
prepared with 0.9% NaCl solution. Both vehicles were also
analyzed separately for antiproliferative or cytotoxic
effects.
Test Assays
Cell Proliferation Assays
Cells were rinsed with PBS, trypsinized (Gibco BRL), counted
with a Coulter counter (CASY I; Schärfe Systems), and seeded onto
six-well plates at a density of 5x103
cells/cm2. After cell attachment (24 hours), the number of
intact cells and the mitotic index were determined and used as a
baseline for day 1.
Two modes of drug application were performed: (1) nonstop incubation with the addition of paclitaxel every third day in combination with medium changes and (2) single-dose application, adding paclitaxel only once for a period of either 24 hours or 20 minutes, followed by a washout process with PBS and addition of fresh, drug-free culture medium.
After 2, 4, 6, 8, and 14 days, final cell numbers were measured by cell counting, mitotic indexes were determined by BrdU-ELISAs according to the manufacturer's instructions (colorimetric cell proliferation ELISA, Boehringer Mannheim),19 and MTT tests (Sigma) were performed according to previously described techniques.20 The MTT test serves as an indirect marker for proliferation and cell viability by measuring the mitochondrial activity of cells.
Growth Factors
haSMCs were incubated with either 20.0 ng/mL PDGF-AB (R&D
Systems), 15.0 ng/mL bFGF (R&D Systems), or 1.0 U/mL thrombin (Sigma)
in addition to FCS. Paclitaxel was added for 24 hours. Next,
drug-containing medium was rinsed with PBS and replaced by fresh
culture medium (+10% FCS). Growth factors were supplemented throughout
the cultivation period. The described concentrations showing maximally
stimulatory effects on haSMC growth were found in pretests in which
increasing concentrations (0.1, 10.0, 15.0, 20.0, and 40.0 ng/mL) of
PDGF-AB and bFGF as well as thrombin (0.5 and 1.0 U/mL) were
examined.
Each test consisted of six measurements for each concentration of paclitaxel (0.1, 1.0, and 10.0 nmol/L and 0.1, 1.0, and 10.0 µmol/L) and was performed three times under identical conditions.
Transfilter Coculture System
Transfilter cocultures were prepared by coating polycarbonate
filters with collagen according to techniques described
previously.16 With this system, coated filters are
inserted between an inner and an outer polycarbonate frame, and
different cell types can be cultured in separate culture media on a
growth area of 8 cm2, as previously
described.17 First, haECs were seeded onto the lower side
of the filter at a density of 2.5x104
cells/cm2. On the following day, haSMCs
(2.5x104 cells/cm2) were seeded onto the
opposite side of such filters, and both cell types were cocultured for
14 days. Different concentrations of paclitaxel (1.0 nmol/L to
10.0 µmol/L) were added either for 24 hours or for 20 minutes to
the lower compartment (containing haECs) to imitate local drug
application at the luminal side of the vessel. Culture medium in both
compartments was then replaced by normal, paclitaxel-free culture
medium, which was replaced by fresh medium every third day. After 14
days, cell numbers on both sides of the filters were determined
separately by cell counting after disaggregation with trypsin/EDTA and
compared with controls that were treated with equivalent concentrations
of the vehicle (ethanol without paclitaxel). Each concentration was
tested in a total of six cocultures.
Filter pieces were also fixed for 3 hours with 4% paraformaldehyde in PBS and embedded in araldite (Serva) as described previously.16 17 Semithin sections (4 µm) were taken on poly-L-lysine–coated coverslips. Cells were then stained with toluidine blue (Merck), and cell numbers on both filter sides were determined under a light microscope.
Immunocytochemical and DNA Stainings
Cytoskeletal proteins. Cells were seeded on
coverslips and treated with drug as described above. After 1, 12, and
24 hours and 4 days, cells were fixed with methanol at -20°C and
labeled with the following primary antibodies: monoclonal
anti–ß-tubulin (Sigma; 1.0 µg/mL), monoclonal anti-vimentin
(Sigma; 1.25 µg/mL), monoclonal anti–smooth muscle -actin
(Progen; 2.0 µg/mL), or anti–von Willebrand factor
(Boehringer Mannheim; 2.0 µg/mL). Secondary labeling was
achieved with FITC-conjugated goat anti-mouse IgG (40.0 µg/mL),
Cy3-conjugated sheep anti-rabbit IgG (10.0 µg/mL), or Cy3-conjugated
sheep anti-mouse IgG (10.0 µg/mL) (all from Sigma).
DNA staining. For additional nucleus staining, cells were rinsed and incubated for 30 minutes with DAPI dissolved in methanol (250 ng/mL) before incubation with the described primary antibodies.
Detection of apoptosis. Apoptosis was
detected by one of two methods: (1) DNA staining by
DAPI.18 Cells were rinsed with PBS, fixed with methanol,
incubated for 30 minutes with the DNA dye, and rinsed again several
times with PBS. (2) TUNEL method.21 Proliferating cells
were seeded on coverslips (5x104 cells/mL), incubated for
1 day routinely, and treated with paclitaxel for 6 hours. Then, the In
Situ Cell Death Detection Kit, AP (Boehringer Mannheim) was
used according to the manufacturer's instructions. The total cell
number, TUNEL-stained cells, and cells with 
5 DAPI-stained nucleus
fragments were determined microscopically: 1000 total cells/cover slide
were counted in six different sets of cultures of paclitaxel-treated
haSMCs and haECs. The apoptotic index was determined as the
number of positively stained cells divided by the total cell number
counted x100.
Rabbit Carotid Artery Injury Model
Animal model. Intimal plaques were produced in 20
male New Zealand White rabbits (Thomae; Biberach, Germany) by use of
the electrostimulation model as described previously.22 In
brief, this model is based on the local and transmural electrical
stimulation of the carotid artery under standardized conditions to
produce plaques of comparable size before intervention.
Drug preparation. For local delivery, the described paclitaxel stock solution was diluted to a final concentration of 10.0 µmol/L in a volume of 4 mL sterile 0.9% NaCl solution.
Local drug-delivery device. A 2.1-mm microporous infusion catheter (Cordis) consisting of a porous membrane with both 0.4- and 0.8-µm pore diameters was used. The device has a proximal port for balloon inflation and a shaft diameter of 3.5F.23 The balloon was inflated with a low-pressure pump (DVI). A volume of 4.0 mL was delivered locally with the use of an inflation pressure of 2 atm.
Study protocol. After the stimulation period, transluminal
balloon angioplasty of the preformed plaque was performed in a total of
20 male rabbits (2.6 to 3.2 kg body weight). For angioplasty, a 2.0-mm
balloon catheter (Micro-Hartzler, ACS) was introduced into the exposed
vessel by direct arteriotomy and then advanced into the region of
plaque formation. The balloon was inflated to 5 atm for 60 seconds. The
rabbits were then randomized in a pharmacologically nontreated control
group in which balloon angioplasty was performed alone (BA group, n=10)
and a local drug-delivery group (LDD group, n=10). In the LDD group,
the porous balloon was advanced into the region of the predilated
plaque, and 4 mL of the paclitaxel solution was delivered with a
pressure of 2 atm over a time period of 
30 seconds. The rabbits were
killed 28 days after balloon treatment. The carotid arteries were
perfused in situ with 500 mL of a 0.1 mol/L cacodylate-buffered 2%
paraformaldehyde solution under
physiological pressure. A 3- to 4-cm segment of
the treated carotid artery with a
polytetrafluoroethylene cuff on it was
excised, and the proximal and distal ends were marked with sutures.
Immunohistochemical stainings. The technique of the
immunohistological examination of vessel sections has
been described previously22 and is based on stainings with
monoclonal antibodies against BrdU and smooth muscle -actin using an
avidin-biotin-peroxidase complex. To confirm the
endothelial origin of the luminal cell lining, sections
were additionally stained with a polyclonal antibody against von
Willebrand factor (goat anti-human factor VIII–related
antigen, Atlantic Antibodies).24
Quantitative histopathology. All sections were quantitatively analyzed by computerized morphometry, as described previously,25 by an independent investigator who was blinded to the type of treatment protocol. In sections stained with the elastica van Gieson stain, luminal perimeter (equal to the length of the endothelial lining), intimal area, and intimal and medial thicknesses were traced manually under stereoscopic control by use of standard software (Bioquant, Bilany Consulting). To obtain the percentage of proliferative cells, the number of BrdU-positive cells was determined and related to the absolute number of intimal and medial smooth muscle cells. For the quantification of endothelial regeneration, the number of endothelial cells was counted and related to the length of the endothelial layer.
Statistical Analysis
Results of the experiments are expressed as mean±SD. The
significance of differences between BA and LDD vessels of the in vivo
study, as well as the in vitro results of single treatment groups that
were related to controls, were proved by the Student unpaired
two-tailed t test unless otherwise stated. In vitro results
in which two or more treatment groups were compared (ie, Fig 2) were
tested by means of two-way ANOVA. By both the Student t test
and the Tukey test/Scheffé's F test, probability values of <.05
were considered to be significant; values of P<.001 were
considered to be highly significant.26
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Effects of Paclitaxel on haSMCs
Nonstop Incubation
Paclitaxel caused a dose-dependent inhibition of haSMC proliferation after 1, 2, 4, 6 (data not shown), and 8 days of incubation time (Fig 1A
). All three assays yielded
comparable results with respect to the IC50 and the
ICmax. The cell-counting assay resulted in an
IC50 of 2.1 nmol/L and an ICmax of 1.0
µmol/L after 8 days' incubation, the BrdU-ELISA resulted in an
IC50 of 2.3 nmol/L and an ICmax of 0.1 to
1.0 µmol/L, and the MTT test resulted in an IC50 of
1.6 nmol/L and an ICmax of 0.1 to 1.0 µmol/L. Within
the ICmax range of 0.1 to 10.0 µmol/L, an almost
complete growth inhibition was observed compared with controls
(100.0±5.1%) (cell counting: 0.3±2.1%, P<.001;
BrdU-ELISA: 1.6±4.9%, P<.001; and MTT tests: 9.5±4.2%,
P<.001 versus controls).
|
Single-Dose Application
When exposure time was restricted from 8 days nonstop to 24 hours
or 20-minute single-dose treatment, the dose-dependent
growth-inhibitory effect of paclitaxel persisted after 4,
8, and 14 days' incubation (Fig 1B
). Almost complete growth inhibition
was again observed by 1.0 to 10.0 µmol/L paclitaxel with nearly
identical ICmax. Shortening of application times, however,
resulted in a slight shift of the inhibition curves toward higher
concentrations (Fig 1B), reflected by an increased IC50
(IC50 by BrdU-ELISAs after 8 days: nonstop, 2.3 nmol/L; 24
hours, 13.0 nmol/L; 20 minutes, 41.2 nmol/L). No significant rebound
phenomena were observed after drug removal.
To differentiate the specific mechanism of the effect of paclitaxel on
microtubules from other unspecific effects, cell toxicity and
apoptosis were studied. Cell counting at day 2 (after 24
hours' paclitaxel treatment) showed that even at 10.0 µmol/L,
cell numbers did not fall below those measured before the addition of
paclitaxel. Viability tests (MTT tests, trypan blue stainings)
confirmed that no cell loss occurred and that haSMCs remained viable
within the first 24 hours after drug addition as well as after 2, 4, 6,
and 8 days' incubation (data not shown). Microscopically, no cell
rounding or cell detachment was observed. Remarkable unspecific cell
toxicity was seen only at very high concentrations (100.0
µmol/L paclitaxel), when even the vehicle ethanol exerts toxic
effects. Fluorescence microscopic examinations of cells stained
with the DNA dye DAPI and stainings by the TUNEL technique showed that
paclitaxel in concentrations between 0.1 nmol/L and 10.0 µmol/L,
which may be a potentially favorable range for local delivery, did not
significantly induce apoptosis. A remarkable number of
disrupted, fragmented cell nuclei in the cytoplasm of
paclitaxel-treated haSMCs, indicating an apoptotic
process,27 was observed only at very high doses (50.0 to
100.0 µmol/L). At 50.0 µmol/L, an apoptotic index
of 80% haSMCs positively stained by the TUNEL method and 25%
DAPI-stained, totally fragmented cell nuclei were found, whereas lower
doses (0.1 nmol/L to 10.0 µmol/L) showed apoptotic
indexes <5% (cell numbers are not shown). Ethanol showed no
apoptotic effects. In haEC cultures, fragmented cell nuclei
were found only sporadically. Assays performed with Taxol (paclitaxel
dissolved in 50% ethanol/50% Cremophor EL) yielded results comparable
to those described for paclitaxel (data not shown).
The vehicle ethanol, used in concentrations equivalent to 0.1 nmol/L to 10.0 µmol/L paclitaxel, exerted no significant growth-inhibitory effects on haSMCs and haECs. In contrast, high doses of Cremophor EL, as used in Taxol preparations, exhibited independent inhibitory (1.0 to 10.0 µmol/L) and even toxic (10.0 to 100.0 µmol/L) effects on haSMCs and haECs (data not shown).
Effect of Paclitaxel on Growth Factor–Stimulated haSMCs
Continuous incubation of haSMCs with PDGF-AB (20.0 ng/mL) in the
presence of 10% FCS led to a significant growth stimulation after 4
days' incubation (162.3±7.9% versus 100.0±6.9% for controls;
P<.001). bFGF (15 ng/mL) showed similar growth-stimulatory
effects (141.2±9.7% versus 100.0±6.9% for controls;
P<.001). The results correspond to those obtained in
pretests. With the addition of paclitaxel for 24 hours, cell
proliferation was inhibited dose dependently (BrdU-ELISA after 4 days:
IC50 of 9.4 nmol/L with PDGF-AB and 6.9 nmol/L with bFGF).
In contrast to PDGF-AB and bFGF, thrombin (1.0 U/mL) showed only
moderate growth-stimulatory effects on haSMCs (105.3±4.0%). Cell
growth of thrombin-treated cells was also inhibited by paclitaxel in a
dose-dependent fashion (BrdU-ELISA after 4 days: IC50 of
21.9 nmol/L). Comparable data were measured by use of the BrdU-ELISA
(Fig 2) or the MTT test (data not shown). Statistical
analysis showed highly significant effects within and between
all three different growth factor treatment groups, all paclitaxel
concentrations, and each paclitaxel concentration (between 0.01 and
10.0 µmol/L) compared with the control group that was also
treated with the corresponding growth factor [C(+GF) in Fig 2].
|
Thus, growth stimulation by the addition of PDGF-AB, bFGF, or thrombin did not significantly attenuate growth inhibition of paclitaxel.
Effects of Paclitaxel on haECs
Similar studies were performed to test the influence of paclitaxel
on haEC growth. Paclitaxel was added to proliferative haEC cultures in
the log phase of cell growth. Nonstop and single-dose (24-hour)
applications were performed, and cell proliferation was determined
after 6 days by use of cell counting, BrdU-ELISA, and MTT tests. A
dose-dependent, significant growth inhibition occurred at high
concentrations (0.01 to 1.0 µmol/L; P<.001 versus
controls), whereas lower paclitaxel doses (0.1 to 1.0 nmol/L) did not
inhibit haEC growth significantly (Fig 3). Furthermore,
no unspecific cytotoxic effects were observed within this concentration
range. Immunocytochemistry showed normal expression of the von
Willebrand factor (data not shown). In control experiments,
ethanol concentrations used in the 10.0-µmol/L paclitaxel
preparations (concentration of ethanol corresponds to 0.4%
[vol/vol]) stimulated haEC proliferation significantly
(P<.05 versus controls) and partially counterbalanced the
effects of paclitaxel.
|
Effects of Paclitaxel on Transfilter Cocultures
The transfilter coculture system is a cell culture model that
imitates the architecture of a normal vessel wall. We have previously
shown that haECs can influence haSMC growth in transfilter
cocultures.16 Stimulated by proliferating but not
confluent haECs, haSMCs migrate into the opposite (haEC) compartment,
forming a cell-rich, neointima-like
structure.17
On the basis of these data, the effect of paclitaxel on smooth muscle
cell proliferation and migration in coculture with proliferative haECs
was evaluated (Fig 4). Neither cell type was confluent
immediately after seeding (day 1). haSMCs of controls reached
confluence after 5 to 7 days. Single-dose applications of paclitaxel
into the lower (haEC) compartment for 24 hours or even 20 minutes dose
dependently reduced total cell numbers per dish (sum of the lower and
upper compartments), reaching significance at 0.01 to 10.0
µmol/L (P<.001 versus controls). Cell numbers, however,
did not fall below those measured on day 1 (Fig 4), indicating that
paclitaxel exerted no cytotoxic effects on cocultures during the entire
cultivation period. By microscopic examination, no cell rounding and no
cell detachment characteristic of cell death was observed during the
entire cultivation period (not shown). Separate analysis of
cell numbers on the upper and lower sides of the filter showed that
single doses of 0.01 to 10.0 µmol/L paclitaxel in the lower
compartment (haEC side) significantly inhibited haSMC proliferation in
the upper compartment. Furthermore, paclitaxel induced a dose-dependent
inhibition of cell numbers in the lower compartment compared with
controls (Fig 4). The total cell number on the lower side of the filter
represents the number of haECs and the result of haSMC
migration from the upper to the lower filter side, followed by
proliferation of subendothelial haSMCs. Because the
number of haECs remains stable after reaching
confluence,16 our results show that haSMC migration
through the filter pores and after proliferation was inhibited
significantly by a brief, single dose of paclitaxel (P<.001
versus controls).
|
These findings were confirmed by histological
observations on semithin sections of several filter segments in which
haECs and haSMCs were identified with the use of specific antibodies
against von Willebrand factor as well as smooth muscle
-actin, respectively. Stainings with toluidine blue showed the
formation of three to five cell layers on both sides of the filter in
untreated controls, indicating migration and proliferation of haSMCs on
the lower filter side. These processes were nearly totally inhibited by
high paclitaxel concentrations (0.1 to 10.0 µmol/L), resulting
in a single subconfluent cell layer on both filter sides.
Cytoskeletal Changes
Microscopic observations were performed to analyze
morphological alterations of treated haSMCs that confirmed the specific
mode and extent of paclitaxel effects. Even short-term treatment with
0.1 to 10.0 µmol/L paclitaxel for 1, 12, or 24 hours, as well as
continuous treatment for 8 days, led to typical alterations of cell
morphology. The cell shape was round instead of elongated, and cell
size was smaller (Fig 5A and 5B).
Immunofluorescence stainings were used to
characterize paclitaxel-induced changes in the intracellular
distribution of three of the main cytoskeletal protein classes:
ß-tubulin, the intermediate filament vimentin, and the contractile
filament smooth muscle -actin.
|
Immunocytochemical stainings for ß-tubulin showed numerous densely
packed groups of short and decentralized microtubules that were not
connected to any perinuclear center (Fig 6B) as observed
in controls (Fig 6A) according to results described by
others.9 28 The distribution of -actin filaments was
heterogeneous: some treated cells showed a disarrangement
of -actin fibers with circumferential bundles, whereas the majority
showed the normal formation of straight filaments extending throughout
the entire cytoplasm (Fig 7A and 7B).6 8 29
Vimentin structures showed the normal molecular
structure8 30 (Fig 7C), but the distribution within the
cytoplasm was altered similar to the ß-tubulin organization (Fig 7D).6
|
|
Results of the In Vivo Study
Quantitative Histopathology
Mean intimal wall area 28 days after balloon angioplasty (Fig 8) was significantly smaller in the LDD group
(0.21±0.11 mm2 ) than in the BA group
(0.36±0.29 mm2; P=.01).
Histological examination (Fig 9A and 9B)
also revealed a significant difference in intimal wall thickness 28
days after angioplasty, with a thickness of 110±39 µm in the
LDD group compared with 138±85 µm in the BA group
(P=.04; Fig 8). The degree of stenosis was 26±8%
in the LDD group compared with 34±20% in the BA group
(P=.01; Fig 8).
|
|
On histological examination, one thrombus formation was found in each of the two groups and excluded from further quantitative histological evaluation.
Quantification of Smooth Muscle Cell Proliferation
The percentage of proliferative cells in the intimal layer was
0.67±0.6% 28 days after local drug delivery and 0.58±0.34% after
balloon angioplasty alone (P=NS). Quantification of the
proliferative activity in the media also did not show any significant
differences between the LDD and BA groups (0.81±0.5% versus
0.7±0.46%; P=NS).
Examination of Endothelial Regeneration
In all animals studied, regenerated endothelium
was present in the balloon-injured areas. In the animals that
underwent angioplasty alone (BA group),
immunohistological quantification showed 45±17
endothelial cells/mm2 compared with 47±12
endothelial cells/mm2 in the LDD group
after local paclitaxel delivery (P=NS).
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
In the present study, it could be shown that paclitaxel, the first representative of a new class of microtubule-affecting drugs, exerts very potent, dose-dependent, inhibitory effects on haSMC proliferation and migration. We found that paclitaxel dissolved in ethanol inhibits haSMC growth at least as potently as described for other cell types, such as tumor cells, fibroblasts, or rat smooth muscle cells.15 31 32 This favors paclitaxel as a promising drug for local application to prevent restenosis after percutaneous transluminal coronary angioplasty. The antiproliferative potential of this very lipophilic compound is characterized by a sustained effect over a period of 14 days after a brief (20-minute), single-dose treatment of haSMCs in a wide concentration range without showing rebound or cytotoxic effects. The IC50 in monocultures of haSMCs continuously incubated with paclitaxel for a period of 8 days was 1.6 to 0.3 nmol/L, as determined by three different test assays. In regard to the growth-inhibitory potency of paclitaxel, the IC50 in haSMCs was half as much as described for rat smooth muscle cells (5.8 nmol/L).15 haSMCs respond 2- to 50-fold more sensitively to paclitaxel than several tumor cell lines.13 31 33 IC50 in colon cancer, gliosarcoma, leukemia, or ovarian tumor cells, for example, has been shown to vary between 4.0 and 100.0 nmol/L.13
Cell counting 24 hours after paclitaxel exposure did not reveal a
significant decrease in cell numbers due to acute toxicity or induction
of apoptotic cell death. Only at very high concentrations
(50.0 µmol/L) did we find evidence of apoptosis.
Approximately 80% of haSMCs showed DNA breaks, and 25% of haSMCs
demonstrated significant cell nucleus fragmentation, both of which
might be indicators of apoptosis.27 33 34 Thus,
the specific effect of paclitaxel on microtubules is not overlapped by
apoptosis at therapeutically relevant doses and thus does not
have any important consequence for further in vivo studies.
The effect of paclitaxel on haEC growth was also studied because reendothelialization plays an important role after vascular injury in vivo. At low doses of paclitaxel (<0.1 µmol), growth of haEC monocultures was less inhibited than haSMC growth, whereas high doses of paclitaxel (0.1 to 10.0 µmol/L) exerted comparable effects in both haECs and haSMCs. These results are in concordance with results achieved in an endothelial wound repair model using porcine aortic endothelial cell cultures in which lower doses, such as 1.2 nmol/L paclitaxel, did not affect endothelial cell growth. Only higher doses resulted in a remarkable reduction of endothelial cell proliferation and migration.35 Comparable results were obtained with the use of the commercially available preparation Taxol instead of paclitaxel dissolved in ethanol. The use of Taxol, however, carries concerns because the lipoid vehicle and stabilizer Cremophor EL causes independent effects on vascular cell growth. Furthermore, it may cause hypersensitive reactions in humans and may be incompatible with catheter materials.1 13 36
In contrast to other compounds of the colchicine type, which inhibit
microtubule assembly,32 37 paclitaxel shifts the balance
of microtubule assembly and disassembly toward microtubule assembly,
forming numerous unorganized and decentralized microtubules inside the
cytoplasm.4 9 38 Although both paclitaxel and colchicine
inhibit cell division in the M phase, the opposite biological
mechanisms involved seem to be of functional relevance. It has been
shown previously that microtubules modulate the response of cells to
several mitogens and cytokines by affecting the transmembrane
signal pathways and other surface processes.39 40 41
Activation of protein kinases such as mitogen-activated protein
kinases by growth factors is associated with microtubule
depolymerization42 and is inhibited by
paclitaxel.43 In addition to inhibiting
mitogen-activated protein kinase activity, paclitaxel reduces
growth factor–stimulated release of transcription factors such as
nuclear factor-
B44 and thus influences the expression
of proto-oncogenes such as c-jun and c-myc at
different stages.45 Microtubule disassembly induced by
colchicine in combination with cytokines such as insulin,
epidermal growth factor, fibroblast growth factor, or serum can result
in synergistic growth-stimulatory effects and increased DNA
synthesis.32 46 This might be one reason why colchicine
failed in preventing in vivo smooth muscle cell proliferation in
restenosis47 despite the fact that initial results
in vitro showed growth-inhibitory effects of colchicine on
unstimulated smooth muscle cells.48 In contrast,
stimulation with bFGF, PDGF, or thrombin as performed in the
present study did not significantly attenuate the antiproliferative
effects of paclitaxel. The growth factor doses used in the present
study were found to exert optimal growth-stimulatory effects, and our
results are in accordance with those described for other
cells.49 50 Our results with growth-stimulated haSMCs also
confirm observations by others, eg, in mouse embryonic cells, that
paclitaxel inhibits initiation of DNA synthesis induced by several
growth factors.51
In addition to the experiments in monocultures, the effect of paclitaxel on our previously described transfilter coculture system consisting of haSMCs and haECs was studied.17 This model extends the findings in monocultures and the chemoinvasion studies of Sollott et al15 in several ways. First, proliferative haECs exert a stimulatory effect on haSMC proliferation, as previously shown.16 These interactions are transmitted directly by cell-cell contacts and by the secretion of growth factors.52 Second, the transfilter coculture model provides information about cell migration, because untreated haSMCs regularly migrate across the filter membrane into the haEC compartment when stimulated by proliferative haECs, forming a cell-rich in vitro neointima.17 53 Third, the transfilter coculture imitates normal arterial vascular architecture,16 54 enabling experiments that simulate local drug delivery, eg, by applying the substance during a short period onto the endothelial side.
In the transfilter coculture system, a brief (20-minute) addition of
paclitaxel applied solely to the endothelial cell
compartment resulted in an inhibition of haSMC proliferation and
migration for a period 
14 days. This may be due to the very
lipophilic structure of paclitaxel, which facilitates a rapid cellular
uptake and onset of action, whereas a subsequent strong binding to the
ß-subunit of tubulin leads to a long-lasting effect on the
arrangement of the cytoskeleton.55 56 Initial effects of
paclitaxel on spindle organization have been observed in several cell
types within 3 to 5 minutes at concentrations of 5.0 to 50.0
µmol/L and in 10 minutes at 0.01 to 0.1
µmol/L.57 When complete disassembly of microtubules is
induced by nocodazole, the microtubule reassembly starts just 2 minutes
after addition of paclitaxel.37 The antimigratory effect
of paclitaxel can most likely be attributed to the observed changes in
the cytoplasmic arrangements of ß-tubulin and smooth muscle
-actin, which affect cell shape, size, and
motility.6 8 28 29 58 Changes of the cytoplasmic
organization of vimentin might also influence cell shape and size.
Controversial results were described by others with various cell types,
such as fibroblasts,6 granulosa cells,30 or
epithelial cells.8 Immunofluorescence
stainings of haSMCs alone showed no clear evidence of a direct effect
of paclitaxel on the vimentin assembly. However, no defined
parameters or methods are available that characterize the
"turnover" of vimentin filaments, as described for actin
filaments or microtubules. Vimentin filaments are also found to
collocalize with microtubules and stabilize actin
structures.6 Thus, the observed alterations of the
vimentin distribution might be simply a consequence of changes in cell
shape and size induced by altered microtubules.
In rat VSMCs, the inhibition of cell migration occurred at
concentrations that were 10-fold lower than those needed to inhibit
cell proliferation.15 In the study by Sollott et
al,15 neointima formation in balloon-denuded
rat carotid arteries was significantly prevented by systemic
administration of paclitaxel, using plasma levels 100-fold below those
achieved in tumor patients.1 3
Both its rapid cellular uptake and long-lasting action over a broad concentration range as well as its efficacy in rat and human cells make paclitaxel a very promising candidate for local drug delivery to reduce the proliferative and migratory components that are involved in restenosis after angioplasty or stent implantation. In recent years, several catheter-based devices have been developed to deliver high doses of pharmacological or molecular agents locally to the site of angioplasty while keeping the risk of systemic side effects to a minimum.59
In preliminary animal experiments of this study using the microporous balloon for local paclitaxel delivery in the rabbit carotid artery, this compound also demonstrated its potential to inhibit neointima formation 4 weeks after intervention. No differences in the process of reendothelialization could be observed between paclitaxel-treated animals and control animals. Toxic or allergic side effects did not occur at any time point of the study. These results, however, only represent a very limited evaluation of paclitaxel therapy in vivo but substantiate in vitro findings in monocultures and cocultures with human vascular cells.60 Further experimental studies in larger animal models are required to evaluate the benefits and risks of local delivery of paclitaxel in coronary arteries.
|
Selected Abbreviations and Acronyms |
|---|
|
Received November 18, 1996; revision received January 16, 1997; accepted January 21, 1997.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Rowinsky EK, Donehower RC. Paclitaxel (Taxol). N Engl J Med. 1995;332:1004-1014.
2.
Manthey CL, Qureshi N, Stütz PL, Vogel
SN. Lipopolysaccharide antagonists block
Taxol induced signaling in murine macrophages. J
Exp Med. 1993;178:695-702.
3. Gelmon K. The taxoids: paclitaxel and docetaxel. Lancet. 1994;344:1267-1272.[Medline] [Order article via Infotrieve]
4. Schiff PB, Fant J, Horwitz SB. Promotion of microtubule assembly in vitro by Taxol. Nature. 1979;277:665-667.[Medline] [Order article via Infotrieve]
5. Donaldson KL, Goolsby GL, Kiener PA, Wahl AF. Activation of p34cdc2 coincident with Taxol-induced apoptosis. Cell Growth Differ. 1994;5:1041-1050.[Abstract]
6. Pletjushkina OJ, Ivanova OJ, Kaverina IN, Vasiliev JM. Taxol-treated fibroblasts acquire an epithelioid shape and a circular pattern of actin bundles. Exp Cell Res. 1994;212:201-208.[Medline] [Order article via Infotrieve]
7. Matsuoka H, Furusawa M, Tomoda H, Seo Y. Difference in cytotoxicity of paclitaxel against neoplastic and normal cells. Anticancer Res. 1994;14:163-168.[Medline] [Order article via Infotrieve]
8.
Gloushankova NA, Lyubimova AV, Tint IS, Feder HH,
Vasiliev JM, Gelfand IM. Role of the microtubular system in
morphological organization of normal and oncogene-transfected
epithelial cells. Proc Natl Acad Sci U S A. 1994;91:8597-8601.
9.
Schiff PB, Horwitz SB. Taxol stabilizes
microtubules in mouse fibroblast cells. Proc Natl Acad
Sci U S A. 1980;77:1561-1565.
10. Hämmerle H, Betz E, Herr D. Human endothelial cells are stimulated and vascular smooth muscle cells are inhibited in their proliferation and migration by heparins. Vasa. 1991;20:207-215.[Medline] [Order article via Infotrieve]
11.
Hanke H, Oberhoff M, Hanke S, Hassenstein S, Kamenz J,
Schmid KM, Betz E, Karsch KR. Inhibition of cellular
proliferation after experimental balloon angioplasty by
low-molecular-weight heparin. Circulation. 1992;85:1548-1556.
12. Schwartz RS, Holmes DR, Topol EJ. The restenosis paradigm revisited: an alternative proposal for cellular mechanisms. J Am Coll Cardiol. 1992;20:1284-1293.[Abstract]
13. Straubinger RM, Sharma A, Murray M, Mayhew E. Novel Taxol formulations: Taxol-containing liposomes. Monogr Natl Cancer Inst. 1993;15:69-78.
14.
Jordan MA, Toso RJ, Thrower D, Wilson L.
Mechanism of mitotic block and inhibition of cell proliferation by
Taxol at low concentrations. Proc Natl Acad Sci
U S A. 1993;90:9552-9556.
15. Sollott SJ, Cheng L, Pauly RR, Jenkins GM, Monticone RE, Kuzuya M, Froehlich JP, Crow MT, Lakatta EG, Rowinsky EK, Kinsella JL. Taxol inhibits neointimal smooth muscle cell accumulation after angioplasty in the rat. J Clin Invest. 1995;95:1869-1876.
16. Roth DR, Axel DI, Betz EL. In vitro model of the inner parts of a vessel wall with cultured human vascular cells. Coron Artery Dis. 1993;4:283-291.[Medline] [Order article via Infotrieve]
17. Axel DI, Brehm BR, Wolburg-Buchholz K, Betz EL, Köveker G, Karsch KR. Induction of cell-rich and lipid-rich plaques in a transfilter coculture system with human vascular cells. J Vasc Res. 1996;33:327-339.[Medline] [Order article via Infotrieve]
18. Jeppesen C, Nielsen PE. Photofootprinting of drug-binding sites on DNA using diazo- and azido-9-aminoacridine derivates. Eur J Biochem. 1989;182:437-444.[Medline] [Order article via Infotrieve]
19.
Gratzner HG. Monoclonal antibody to 5-bromo- and
5-iododeoxyuridine: a new reagent for detection of DNA
replication. Science. 1982;218:474-475.
20. Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. J Immunol Methods. 1986;89:271-277.[Medline] [Order article via Infotrieve]
21.
Gorczyca W, Gong J, Darzynkiewicz Z. Detection
of DNA strand breaks in individual apoptotic cells by the in
situ terminal deoxynucleotidyl transferase and nick
translation assays. Cancer Res. 1993;53:1945-1951.
22.
Hanke H, Strohschneider T, Oberhoff M, Betz E, Karsch
KR. Time course of smooth muscle cell proliferation in the
intima and media of arteries following experimental
angioplasty. Circ Res. 1990;67:651-659.
23. Lambert CR, Leone JE, Rowland SM. Local drug delivery catheters: functional comparison of porous and microporous designs. Coron Artery Dis. 1993;4:469-475.[Medline] [Order article via Infotrieve]
24.
Weidlinger FF, McLenachan JM, Cybulsky MI, Gordon JB,
Rennke HG, Hollenberg NK, Fallon JT, Ganz P, Cooke JP.
Persistent dysfunction of regenerated endothelium after
balloon angioplasty of rabbit iliac artery.
Circulation. 1990;81:1667-1679.
25.
Hassenstein S, Hanke H, Kamenz J, Oberhoff M, Hanke S,
Riessen R, Haase KK, Betz E, Karsch KR. Vascular injury and time
course of smooth muscle cell proliferation after experimental holmium
laser angioplasty. Circulation. 1992;86:1575-1583.
26.
Wallenstein S, Zucker CL, Fleiss JL. Some
statistical methods useful in circulation research. Circ
Res. 1980;47:1-9.
27. Majno G, Joris I. Apoptosis, oncosis and necrosis: an overview of cell death. Am J Pathol. 1995;146:3-15.[Abstract]
28.
Gotlieb AI, McBurnie-May L, Subrahmanyan L, Kalnins
VI. Distribution of microtubule organizing centers in migrating
sheets of endothelial cells. J Cell
Biol. 1981;91:589-594.
29.
Danowski B. Fibroblast
contractility and actin organization are stimulated by
microtubule inhibitors. J Cell Sci. 1989;93:255-266.
30. Herman B, Langevin MA, Albertini DF. The effects of Taxol on the organization of the cytoskeleton in cultured ovarian granulosa cells. Eur J Cell Biol. 1983;31:34-45.[Medline] [Order article via Infotrieve]
31. Bartoli MH, Boitard M, Fessi H, Beriel H, Devissaguet JP, Picot F, Puisieux F. In vitro and in vivo antitumoral activity of free and encapsulated Taxol. J Microencapsul. 1990;7:191-197.[Medline] [Order article via Infotrieve]
32. Crossin KL, Carney DH. Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis. Cell. 1981;23:61-71.[Medline] [Order article via Infotrieve]
33. Wahl AF, Donaldson KL, Fairchild C, Lee FYF, Foster SA, Demers GW, Galloway DA. Loss of normal p53 function confers sensitization to Taxol by increasing G2/M arrest and apoptosis. Nat Med. 1996;2:72-79.[Medline] [Order article via Infotrieve]
34. Donaldson KL, Goolsby GL, Wahl AF. Cytotoxicity of the anticancer agents cisplatin and Taxol during cell proliferation and the cell cycle. Int J Cancer. 1994;57:847-855.[Medline] [Order article via Infotrieve]
35.
Coomber BL, Gotlieb AI. In vitro
endothelial wound repair: interaction of cell migration
and proliferation.
Arteriosclerosis. 1990;10:215-222.
36. Donehower RC, Rowinsky EK. An overview of experience with Taxol (paclitaxel) in the USA. Cancer Treat Rev. 1993;19:63-78.
37.
McClain DA, Edelman GM. Density-dependent
stimulation and inhibition of cell growth by agents that disrupt
microtubules. Proc Natl Acad Sci U S A. 1980;77:2748-2752.
38. De Brabander M, Geuens G, Nuydens R, Willebrords R, Aerts F, De Mey J. Microtubule dynamics during the cell cycle: the effect of Taxol and nocodazole on the microtubule system of Pt K2 cells at different stages of the mitotic cycle. Int Rev Cytol. 1986;101:215-274.[Medline] [Order article via Infotrieve]
39.
Edelman G. Surface modulation in cell
recognition and cell growth. Science. 1976;192:218-226.
40. Manthey CL, Brandes ME, Perera PY, Vogel SN. Taxol increases steady-state levels of lipopolysaccharide-inducible genes and protein-tyrosine phosphorylation in murine macrophages. J Immunol. 1992;149:2459-2465.[Abstract]
41.
Otto AM, Ulrich MO, Zumbe A, Jimenez de Asua L.
Microtubule-disrupting agents affect two different events regulating
the initiation of DNA synthesis in Swiss 3T3 cells. Proc
Natl Acad Sci U S A. 1981;78:3063-3067.
42. Ding AH, Sanchez E, Nathan CF. Taxol shares the ability of bacterial lipopolysaccharide to induce tyrosine phosphorylation of microtubule-associated protein kinase. J Immunol. 1993;151:5596-5602.[Abstract]
43. Nishio K, Arioka H, Ishida T, Fukumoto H, Kurokawa H, Sata M, Ohata M, Saijo N. Enhanced interactions between tubulin and microtubule-associated protein 2 via inhibition of MAP kinase and CDC2 kinase by paclitaxel. Int J Cancer. 1995;63:688-693.[Medline] [Order article via Infotrieve]
44.
Rosette C, Karin M. Cytoskeletal control of gene
expression: depolymerization of microtubules
activates NF-kB. J Cell Biol. 1995;128:1111-1119.
45. Bhalla K, Ibrado AM, Torkina E, Tang C, Mahoney ME, Huang Y. Taxol induces internucleosomal DNA fragmentation associated with programmed cell death in human myeloid leukemia cells. Leukemia. 1993;7:563-568.[Medline] [Order article via Infotrieve]
46. Friedkin M, Legg A, Rozengurt E. Enhancement of DNA synthesis by colchicine in 3T3 mouse fibroblasts stimulated with growth factors. Exp Cell Res. 1980;129:23-30.[Medline] [Order article via Infotrieve]
47. Gradus-Pizlo I, Wilensky RL, March KL, Fineberg N, Michaels M, Sandusky GE, Hathaway DR. Local delivery of biodegradable microparticles containing colchicine or a colchicine analogue: effects on restenosis and implications for catheter-based drug delivery. J Am Coll Cardiol. 1995;26:1549-1557.[Abstract]
48.
March KL, Mohanraj S, Ho PPK, Wilensky RL, Hathaway
DR. Biodegradable microspheres containing a colchicine
analogue inhibit DNA synthesis in vascular smooth muscle cells.
Circulation. 1994;89:1929-1933.
49.
Weiss RH, Maduri M. The mitogenic
effect of thrombin in vascular smooth muscle cells is largely due to
basic fibroblast growth factor. J Biol Chem. 1993;268:5724-5727.
50. Stouffer GA, Owens GK. TGF-ß promotes proliferation of cultured SMC via both PDGF-AA-dependent and PDGF-AA-independent mechanisms. J Clin Invest. 1994;93:2048-2055.
51. Crossin KL, Carney DH. Microtubule stabilization by Taxol inhibits initiation of DNA synthesis by thrombin and by epidermal growth factor. Cell. 1981;27:341-350.[Medline] [Order article via Infotrieve]
52. Axel DI, Brehm BR, Wild DH, Köveker G, Karsch KR. Determination of platelet-derived growth factor in conditioned media of monocultures and cocultures of human vascular cells in a transfilter culture system. Eur Heart J. 1995;16(suppl):28. Abstract.
53. Axel DI, Brehm BR, Betz EL, Karsch KR. Formation of an in vitro atheroma using LDL and monocytes in a transfilter coculture system with human vascular cells. Circulation. 1993;88(suppl):I-323. Abstract.
54. Weber E, Hämmerle H, Vatti R, Berti G, Betz E. Co-cultivation of endothelial and smooth muscle cells on opposite sides of a porous membrane. Appl Pathol. 1986;4:246-252.[Medline] [Order article via Infotrieve]
55.
Long BH, Fairchild CR. Paclitaxel inhibits
progression of mitotic cells to G1 phase by interference
with spindle formation without affecting other microtubule functions
during anaphase and telophase. Cancer Res. 1994;54:4355-4361.
56. Derry B, Wilson L, Jordan MA. Substoichiometric binding of Taxol suppresses microtubule dynamics. Biochemistry. 1995;34:2203-2211.[Medline] [Order article via Infotrieve]
57.
Molè-Bajer J, Bajer AS. Action of Taxol on
mitosis: modification of microtubule arrangements and function of the
mitotic spindle in Haemanthus endosperm. J
Cell Biol. 1983;96:527-540.
58.
Kumar CC, Kim JH, Bushel P, Armstrong L, Catino
JJ. Activation of smooth muscle -actin promotor in
ras-transformed cells by treatments with antimitotic agents:
correlation with stimulation of SRF:SRE mediated gene transcription.
J Biochem. 1995;118:1285-1292.
59. Riessen R, Isner J. Prospects for site-specific delivery of pharmacologic and molecular therapies. J Am Coll Cardiol. 1994;23:1234-1244.[Abstract]
60. Kunert W, Küttner A, Herdeg C, Horch B, Göggelmann C, Axel DI, Oberhoff M, Karsch KR. Paclitaxel inhibits development of restenosis following experimental balloon angioplasty in the rabbit carotid artery. Eur Heart J. 1996;17(suppl):386. Abstract.
This article has been cited by other articles:
 |
|
 |
|
  M. Unverdorben, C. Vallbracht, B. Cremers, H. Heuer, C. Hengstenberg, C. Maikowski, G. S. Werner, D. Antoni, F. X. Kleber, W. Bocksch, et al. Paclitaxel-Coated Balloon Catheter Versus Paclitaxel-Coated Stent for the Treatment of Coronary In-Stent Restenosis Circulation, June 16, 2009; 119(23): 2986 - 2994. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Mahmud, J. A. Ormiston, M. A. Turco, J. J. Popma, N. J. Weissman, C. D. O'Shaughnessy, T. Mann, J. J. Hall, T. F. McGarry, L. A. Cannon, et al. TAXUS Liberte Attenuates the Risk of Restenosis in Patients With Medically Treated Diabetes Mellitus: Results From the TAXUS ATLAS Program J. Am. Coll. Cardiol. Intv., March 1, 2009; 2(3): 240 - 252. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. K. Pendyala, J. Li, T. Shinke, S. Geva, X. Yin, J. P. Chen, S. B. King III, K. A. Robinson, N. A.F. Chronos, and D. Hou Endothelium-Dependent Vasomotor Dysfunction in Pig Coronary Arteries With Paclitaxel-Eluting Stents Is Associated With Inflammation and Oxidative Stress J. Am. Coll. Cardiol. Intv., March 1, 2009; 2(3): 253 - 262. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Chiu, D. T. Ko, P. C. Austin, E. A. Cohen, J. L. Velianou, R. Goeree, G. Blackhouse, and J. V. Tu Paclitaxel Versus Sirolimus Stents in Diabetic and Nondiabetic Patients Circ Cardiovasc Qual Outcomes, March 1, 2009; 2(2): 96 - 107. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A Ostovan, R. Mollazadeh, J. Kojuri, and M. Mirabadi Experience with Paclitaxel-Eluting Infinnium Coronary Stents Asian Cardiovasc Thorac Ann, December 1, 2008; 16(6): 454 - 458. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Hamilos, J. Sarma, M. Ostojic, T. Cuisset, G. Sarno, N. Melikian, A. Ntalianis, O. Muller, E. Barbato, B. Beleslin, et al. Interference of Drug-Eluting Stents With Endothelium-Dependent Coronary Vasomotion: Evidence for Device-Specific Responses Circ Cardiovasc Intervent, December 1, 2008; 1(3): 193 - 200. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-K. Choung, S.-E. Jin, M. J. Lee, C.-K. Kim, and J.-M. Hwang Slow-Releasing Paclitaxel in Polytetrafluoroethylene/Polylactide-co-glycolide Laminate Delays Adjustment after Strabismus Surgery in Rabbit Model Invest. Ophthalmol. Vis. Sci., December 1, 2008; 49(12): 5340 - 5345. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Werk, S. Langner, B. Reinkensmeier, H.-F. Boettcher, G. Tepe, U. Dietz, N. Hosten, B. Hamm, U. Speck, and J. Ricke Inhibition of Restenosis in Femoropopliteal Arteries: Paclitaxel-Coated Versus Uncoated Balloon: Femoral Paclitaxel Randomized Pilot Trial Circulation, September 23, 2008; 118(13): 1358 - 1365. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Cyrus, H. Zhang, J. S. Allen, T. A. Williams, G. Hu, S. D. Caruthers, S. A. Wickline, and G. M. Lanza Intramural Delivery of Rapamycin With {alpha}v{beta}3-Targeted Paramagnetic Nanoparticles Inhibits Stenosis After Balloon Injury Arterioscler. Thromb. Vasc. Biol., May 1, 2008; 28(5): 820 - 826. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Schachner, C. Steger, S. Heiss, N. Bonaros, W. Sterlacci, G. Laufer, and J. Bonatti Paclitaxel treatment reduces neointimal hyperplasia in cultured human saphenous veins Eur. J. Cardiothorac. Surg., December 1, 2007; 32(6): 906 - 911. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S W Park, S H Lee, C H Kim, G S Jeon, S J Hong, J G Yi, and H J Jeon Inhibition of pseudointimal hyperplasia in swine TIPS models: the efficacy of local delivery of paclitaxel using a perforated balloon catheter Br. J. Radiol., September 1, 2007; 80(957): 702 - 707. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Finn, G. Nakazawa, M. Joner, F. D. Kolodgie, E. K. Mont, H. K. Gold, and R. Virmani Vascular Responses to Drug Eluting Stents: Importance of Delayed Healing Arterioscler. Thromb. Vasc. Biol., July 1, 2007; 27(7): 1500 - 1510. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Scheller, U. Speck, and M. Bohm Prevention of restenosis: is angioplasty the answer? Heart, May 1, 2007; 93(5): 539 - 541. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. J. Murphy, T. W. Johnson, M. H. Chamberlain, S. I. Rizvi, M. Wyatt, S. J. George, G. D. Angelini, K. R. Karsch, M. Oberhoff, and A. C. Newby Short- and long-term effects of cytochalasin D, paclitaxel and rapamycin on wall thickening in experimental porcine vein grafts Cardiovasc Res, February 1, 2007; 73(3): 607 - 617. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Scheller, C. Hehrlein, W. Bocksch, W. Rutsch, D. Haghi, U. Dietz, M. Bohm, and U. Speck Treatment of Coronary In-Stent Restenosis with a Paclitaxel-Coated Balloon Catheter N. Engl. J. Med., November 16, 2006; 355(20): 2113 - 2124. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. H. Lee, H. Y. Nam, T. Kwon, S. J. Kim, G. Y. Kwon, H. J. Jeon, H. J. Lim, W. K. Lee, J.-s. Park, J. Y. Ko, et al. Paclitaxel-coated expanded polytetrafluoroethylene haemodialysis grafts inhibit neointimal hyperplasia in porcine model of graft stenosis Nephrol. Dial. Transplant., September 1, 2006; 21(9): 2432 - 2438. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-J. Cho, T.-Y. Kim, H.-J. Cho, K.-W. Park, S.-Y. Zhang, J.-H. Kim, S.-H. Kim, J.-Y. Hahn, H.-J. Kang, Y.-B. Park, et al. The Effect of Stem Cell Mobilization by Granulocyte-Colony Stimulating Factor on Neointimal Hyperplasia and Endothelial Healing After Vascular Injury With Bare-Metal Versus Paclitaxel-Eluting Stents J. Am. Coll. Cardiol., July 18, 2006; 48(2): 366 - 374. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Patterson, S. Mapera, H.-H. Li, N. Madamanchi, E. Hilliard, R. Lineberger, R. Herrmann, and P. Charles Comparative Effects of Paclitaxel and Rapamycin on Smooth Muscle Migration and Survival: Role of Akt-Dependent Signaling Arterioscler. Thromb. Vasc. Biol., July 1, 2006; 26(7): 1473 - 1480. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Wessely, A. Schomig, and A. Kastrati Sirolimus and Paclitaxel on Polymer-Based Drug-Eluting Stents: Similar But Different J. Am. Coll. Cardiol., February 21, 2006; 47(4): 708 - 714. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Alvarez Jr and N. K. Kapur Drug Eluting Stent Technology: A Paradigm Shift in the Treatment and Prevention of Restenosis Journal of Pharmacy Practice, December 1, 2005; 18(6): 461 - 478. [Abstract] [PDF]
|
|
|
|
|
|
J.W. Fischer Dexamethasone: Effects on neointimal hyperplasia and vessel integrity Cardiovasc Res, December 1, 2005; 68(3): 350 - 352. [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. Nawarskas and L. A. Osborn Paclitaxel-eluting stents in coronary artery disease Am. J. Health Syst. Pharm., November 1, 2005; 62(21): 2241 - 2251. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. Granada, D. Ensenat, A. N. Keswani, G. L. Kaluza, A. E. Raizner, X.-m. Liu, K. J. Peyton, M. A. Azam, H. Wang, and W. Durante Single Perivascular Delivery of Mitomycin C Stimulates p21 Expression and Inhibits Neointima Formation in Rat Arteries Arterioscler. Thromb. Vasc. Biol., November 1, 2005; 25(11): 2343 - 2348. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A H Gershlick Drug eluting stents in 2005 Heart, June 1, 2005; 91(suppl_3): iii24 - iii31. [Full Text] [PDF]
|
|
|
|
|
|
M. A. Costa and D. I. Simon Molecular Basis of Restenosis and Drug-Eluting Stents Circulation, May 3, 2005; 111(17): 2257 - 2273. [Full Text] [PDF]
|
|
|
|
|
|
J. W. Moses, R. Mehran, E. Nikolsky, J. M. Lasala, W. Corey, G. Albin, C. Hirsch, M. B. Leon, M. E. Russell, S. G. Ellis, et al. Outcomes with the paclitaxel-eluting stent in patients with acute coronary syndromes: Analysis from the TAXUS-IV trial J. Am. Coll. Cardiol., April 19, 2005; 45(8): 1165 - 1171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. B. Hermiller, A. Raizner, L. Cannon, P. A. Gurbel, M. A. Kutcher, S. C. Wong, M. E. Russell, S. G. Ellis, R. Mehran, G. W. Stone, et al. Outcomes with the polymer-based paclitaxel-eluting TAXUS stent in patients with diabetes mellitus: The TAXUS-IV trial J. Am. Coll. Cardiol., April 19, 2005; 45(8): 1172 - 1179. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Lansky, R. A. Costa, M. Mooney, M. G. Midei, H. K. Lui, W. Strickland, R. Mehran, M. B. Leon, M. E. Russell, S. G. Ellis, et al. Gender-based outcomes after paclitaxel-eluting stent implantation in patients with coronary artery disease J. Am. Coll. Cardiol., April 19, 2005; 45(8): 1180 - 1185. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. K. Prasad, K. R. Resmi, L. K. Krishnan, and R. Vaishnav Survival of Endothelial Cells in vitro on Paclitaxel-loaded Coronary Stents J Biomater Appl, April 1, 2005; 19(4): 271 - 286. [Abstract] [PDF]
|
|
|
|
|
|
K. Peppel, L. Zhang, E. S. Orman, P.-O. Hagen, A. Amalfitano, L. Brian, and N. J. Freedman Activation of vascular smooth muscle cells by TNF and PDGF: overlapping and complementary signal transduction mechanisms Cardiovasc Res, February 15, 2005; 65(3): 674 - 682. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. S. Schwartz, N. A. Chronos, and R. Virmani Preclinical restenosis models and drug-eluting stents: Still important, still much to learn J. Am. Coll. Cardiol., October 6, 2004; 44(7): 1373 - 1385. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Scheller, U. Speck, C. Abramjuk, U. Bernhardt, M. Bohm, and G. Nickenig Paclitaxel Balloon Coating, a Novel Method for Prevention and Therapy of Restenosis Circulation, August 17, 2004; 110(7): 810 - 814. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Vogt, A. Stein, G. Rettemeier, N. Krott, R. Hoffmann, J. v. Dahl, A.-K. Bosserhoff, W. Michaeli, P. Hanrath, C. Weber, et al. Long-term assessment of a novel biodegradable paclitaxel-eluting coronary polylactide stent Eur. Heart J., August 1, 2004; 25(15): 1330 - 1340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R Sindermann, V. Verin, J. W Hopewell, H. P. Rodemann, and J. H Hendry Biological aspects of radiation and drug-eluting stents for the prevention of restenosis Cardiovasc Res, July 1, 2004; 63(1): 22 - 30. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Lansky, R. A. Costa, G. S. Mintz, Y. Tsuchiya, M. Midei, D. A. Cox, C. O'Shaughnessy, R. A. Applegate, L. A. Cannon, M. Mooney, et al. Non-Polymer-Based Paclitaxel-Coated Coronary Stents for the Treatment of Patients With De Novo Coronary Lesions: Angiographic Follow-Up of the DELIVER Clinical Trial Circulation, April 27, 2004; 109(16): 1948 - 1954. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X.-L. Zheng, Y. Gui, G. Du, M. A. Frohman, and D.-Q. Peng Calphostin-C Induction of Vascular Smooth Muscle Cell Apoptosis Proceeds through Phospholipase D and Microtubule Inhibition J. Biol. Chem., February 20, 2004; 279(8): 7112 - 7118. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Gershlick, I. De Scheerder, B. Chevalier, A. Stephens-Lloyd, E. Camenzind, C. Vrints, N. Reifart, L. Missault, J.-J. Goy, J. A. Brinker, et al. Inhibition of Restenosis With a Paclitaxel-Eluting, Polymer-Free Coronary Stent: The European evaLUation of pacliTaxel Eluting Stent (ELUTES) Trial Circulation, February 3, 2004; 109(4): 487 - 493. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tanabe, P. W. Serruys, M. Degertekin, G. Guagliumi, E. Grube, C. Chan, T. Munzel, J. Belardi, W. Ruzyllo, L. Bilodeau, et al. Chronic Arterial Responses to Polymer-Controlled Paclitaxel-Eluting Stents: Comparison With Bare Metal Stents by Serial Intravascular Ultrasound Analyses: Data From the Randomized TAXUS-II Trial Circulation, January 20, 2004; 109(2): 196 - 200. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. W. Stone, S. G. Ellis, D. A. Cox, J. Hermiller, C. O'Shaughnessy, J. T. Mann, M. Turco, R. Caputo, P. Bergin, J. Greenberg, et al. A Polymer-Based, Paclitaxel-Eluting Stent in Patients with Coronary Artery Disease N. Engl. J. Med., January 15, 2004; 350(3): 221 - 231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Scheller, U. Speck, B. Romeike, A. Schmitt, M. Sovak, M. Bohm, and H.-P. Stoll Contrast media as carriers for local drug delivery: Successful inhibition of neointimal proliferation in the porcine coronary stent model Eur. Heart J., August 1, 2003; 24(15): 1462 - 1467. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Park, W. H. Shim, D. S. Ho, A. E. Raizner, S.-W. Park, M.-K. Hong, C. W. Lee, D. Choi, Y. Jang, R. Lam, et al. A Paclitaxel-Eluting Stent for the Prevention of Coronary Restenosis N. Engl. J. Med., April 17, 2003; 348(16): 1537 - 1545. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-K. Hong, G. S. Mintz, C. W. Lee, J.-M. Song, K.-H. Han, D.-H. Kang, J.-K. Song, J.-J. Kim, N. J. Weissman, N. E. Fearnot, et al. Paclitaxel Coating Reduces In-Stent Intimal Hyperplasia in Human Coronary Arteries: A Serial Volumetric Intravascular Ultrasound Analysis From the ASian Paclitaxel-Eluting Stent Clinical Trial (ASPECT) Circulation, February 4, 2003; 107(4): 517 - 520. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tanabe, P. W. Serruys, E. Grube, P. C. Smits, G. Selbach, W. J. van der Giessen, M. Staberock, P. de Feyter, R. Muller, E. Regar, et al. TAXUS III Trial: In-Stent Restenosis Treated With Stent-Based Delivery of Paclitaxel Incorporated in a Slow-Release Polymer Formulation Circulation, February 4, 2003; 107(4): 559 - 564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Grube, S. Silber, K. E. Hauptmann, R. Mueller, L. Buellesfeld, U. Gerckens, and M. E. Russell TAXUS I: Six- and Twelve-Month Results From a Randomized, Double-Blind Trial on a Slow-Release Paclitaxel-Eluting Stent for De Novo Coronary Lesions Circulation, January 7, 2003; 107(1): 38 - 42. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. M. Lanza, X. Yu, P. M. Winter, D. R. Abendschein, K. K. Karukstis, M. J. Scott, L. K. Chinen, R. W. Fuhrhop, D. E. Scherrer, and S. A. Wickline Targeted Antiproliferative Drug Delivery to Vascular Smooth Muscle Cells With a Magnetic Resonance Imaging Nanoparticle Contrast Agent: Implications for Rational Therapy of Restenosis Circulation, November 26, 2002; 106(22): 2842 - 2847. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. N. Babapulle and M. J. Eisenberg Coated Stents for the Prevention of Restenosis: Part I Circulation, November 19, 2002; 106(21): 2734 - 2740. [Full Text] [PDF]
|
|
|
|
 |
|
 D. E. Ingber Mechanical Signaling and the Cellular Response to Extracellular Matrix in Angiogenesis and Cardiovascular Physiology Circ. Res., November 15, 2002; 91(10): 877 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Kataoka, E. Grube, Y. Honda, Y. Morino, S.-H. Hur, H. N. Bonneau, A. Colombo, C. Di Mario, G. Guagliumi, K. E. Hauptmann, et al. 7-Hexanoyltaxol-Eluting Stent for Prevention of Neointimal Growth: An Intravascular Ultrasound Analysis From the Study to COmpare REstenosis rate between QueST and QuaDS-QP2 (SCORE) Circulation, October 1, 2002; 106(14): 1788 - 1793. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yasuda, T. Noguchi, M. Gohda, T. Arai, N. Tsutsui, Y. Nakayama, T. Matsuda, and H. Nonogi Local delivery of low-dose docetaxel, a novel microtubule polymerizing agent, reduces neointimal hyperplasia in a balloon-injured rabbit iliac artery model Cardiovasc Res, February 1, 2002; 53(2): 481 - 486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Jorgensen, H. Kelbaek, S. Helqvist, G. V. H. Jensen, K. Saunamaki, J. Kastrup, O. Havndrup, H. Bundgaard, J. Kyst Madsen, M. Christiansen, et al. Predictors of coronary in-stent restenosis: importance of angiotensin-converting enzyme gene polymorphism and treatment with angiotensin-converting enzyme inhibitors J. Am. Coll. Cardiol., November 1, 2001; 38(5): 1434 - 1439. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E Regar, G Sianos, and P W Serruys Stent development and local drug delivery Br. Med. Bull., October 1, 2001; 59(1): 227 - 248. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Work, A. R. McPhaden, N. J. Pyne, S. Pyne, R. M. Wadsworth, and C. L. Wainwright Short-Term Local Delivery of an Inhibitor of Ras Farnesyltransferase Prevents Neointima Formation In Vivo After Porcine Coronary Balloon Angioplasty Circulation, September 25, 2001; 104(13): 1538 - 1543. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F Liistro and A Colombo Late acute thrombosis after paclitaxel eluting stent implantation Heart, September 1, 2001; 86(3): 262 - 264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Honda, E. Grube, L. M. de la Fuente, P. G. Yock, S. H. Stertzer, and P. J. Fitzgerald Novel Drug-Delivery Stent: Intravascular Ultrasound Observations From the First Human Experience With the QP2-Eluting Polymer Stent System Circulation, July 24, 2001; 104(4): 380 - 383. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Farb, P. F. Heller, S. Shroff, L. Cheng, F. D. Kolodgie, A. J. Carter, D. S. Scott, J. Froehlich, and R. Virmani Pathological Analysis of Local Delivery of Paclitaxel Via a Polymer-Coated Stent Circulation, July 24, 2001; 104(4): 473 - 479. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Boehm and E. G. Nabel Cell Cycle and Cell Migration : New Pieces to the Puzzle Circulation, June 19, 2001; 103(24): 2879 - 2881. [Full Text] [PDF]
|
|
|
|
|
|
A. W. Heldman, L. Cheng, G. M. Jenkins, P. F. Heller, D.-W. Kim, M. Ware Jr, C. Nater, R. H. Hruban, B. Rezai, B. S. Abella, et al. Paclitaxel Stent Coating Inhibits Neointimal Hyperplasia at 4 Weeks in a Porcine Model of Coronary Restenosis Circulation, May 8, 2001; 103(18): 2289 - 2295. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. I Axel, A. Frigge, J. Dittmann, H. Runge, I. Spyridopoulos, R. Riessen, R. Viebahn, and K. R Karsch All-trans retinoic acid regulates proliferation, migration, differentiation, and extracellular matrix turnover of human arterial smooth muscle cells Cardiovasc Res, March 1, 2001; 49(4): 851 - 862. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B.R Brehm, S.C Wolf, D Bertsch, M Klaussner, S Wesselborg, S Schuler, and K Schulze-Osthoff Effects of nebivolol on proliferation and apoptosis of human coronary artery smooth muscle and endothelial cells Cardiovasc Res, February 1, 2001; 49(2): 430 - 439. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Riessen, M. Fenchel, H. Chen, D. I. Axel, K. R. Karsch, and J. Lawler Cartilage Oligomeric Matrix Protein (Thrombospondin-5) Is Expressed by Human Vascular Smooth Muscle Cells Arterioscler. Thromb. Vasc. Biol., January 1, 2001; 21(1): 47 - 54. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. E. Drachman, E. R. Edelman, P. Seifert, A. R. Groothuis, D. A. Bornstein, K. R. Kamath, M. Palasis, D. Yang, S. H. Nott, and C. Rogers Neointimal thickening after stent delivery of paclitaxel: change in composition and arrest of growth over six months J. Am. Coll. Cardiol., December 1, 2000; 36(7): 2325 - 2332. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K R KARSCH and A C NEWBY Stent magic! The genie has escaped from the bottle Heart, November 1, 2000; 84(5): 469 - 470. [Full Text]
|
|
|
|
 |
|
 J. Al Suwaidi, P. B. Berger, and D. R. Holmes Jr Coronary Artery Stents JAMA, October 11, 2000; 284(14): 1828 - 1836. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Hou, P. I. Rogers, P. M. Toleikis, W. Hunter, and K. L. March Intrapericardial Paclitaxel Delivery Inhibits Neointimal Proliferation and Promotes Arterial Enlargement After Porcine Coronary Overstretch Circulation, September 26, 2000; 102(13): 1575 - 1581. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Herdeg, M. Oberhoff, A. Baumbach, A. Blattner, D. I. Axel, S. Schroder, H. Heinle, and K. R. Karsch Local paclitaxel delivery for the prevention of restenosis: biological effects and efficacy in vivo J. Am. Coll. Cardiol., June 1, 2000; 35(7): 1969 - 1976. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. J. Creel, M. A. Lovich, and E. R. Edelman Arterial Paclitaxel Distribution and Deposition Circ. Res., April 28, 2000; 86(8): 879 - 884. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Chandrasekar and J.-F. Tanguay Platelets and restenosis J. Am. Coll. Cardiol., March 1, 2000; 35(3): 555 - 562. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. G. Lehmann, J. J. Popma, J. A. Werner, A. J. Lansky, and R. L. Wilensky Vascular remodeling and the local delivery of cytochalasin B after coronary angioplasty in humans J. Am. Coll. Cardiol., March 1, 2000; 35(3): 583 - 591. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Liistro, G. Stankovic, C. Di Mario, T. Takagi, A. Chieffo, S. Moshiri, M. Montorfano, M. Carlino, C. Briguori, P. Pagnotta, et al. First Clinical Experience With a Paclitaxel Derivate-Eluting Polymer Stent System Implantation for In-Stent Restenosis: Immediate and Long-Term Clinical and Angiographic Outcome Circulation, April 23, 2002; 105(16): 1883 - 1886. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||