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Annals of Biomedical Engineering, Vol. 32, No. 4, April 2004 (2004) pp. 537543

The Effects of a Shear Flow on the Uptake of LDL and Acetylated LDL
by an EC Monoculture and an ECSMC Coculture
KOICHI NIWA, TATSUNORI KADO, JIRO SAKAI, and TAKESHI KARINO
Laboratory of Biofluid Dynamics, Research Institute for Electronic Science, Hokkaido University, North 12, West 6, North District,
Sapporo 060-0812, Japan
(Received 30 July 2003; accepted 6 November 2003)

an arterial wall where ECs are forming only the inner surface of a thick wall consisting of ECs, internal and external
elastic laminas, and layers of smooth muscle cells (SMC),
fibroblasts, and various matrix fibers. Therefore, we prepared ECSMC cocultures which we considered a model
closer to an arterial wall by seeding ECs directly over SMCs
and coculturing them, and tested whether they behaved in
the same manner as EC monocultures in the uptake of LDL
and acetylated LDL (Ac-LDL) in both the presence and
absence of a laminar shear flow.

AbstractTo elucidate the mechanisms of localized genesis and

development of atherosclerosis and anastomotic intimal hyperplasia in man, a coculture of bovine aortic endothelial cells (ECs)
and smooth muscle cells (SMCs) was prepared, and the effects of a
shear flow on the uptake of lipoproteins by the cells was studied by
incubating the ECSMC coculture as well as an EC monoculture
with a culture medium containing either DiI-LDL or DiI-Ac-LDL
and subjecting to a laminar shear flow. It was found that in both
the presence and absence of a shear flow that imposed the ECs
an area mean shear stress of 13.3 dynes/cm2 , the uptake of LDL
by an ECSMC coculture was much greater than that by an EC
monoculture, whereas that of Ac-LDL was almost the same. The
uptake of LDL by an EC monoculture increased slightly by being
exposed to a shear flow, whereas that by an ECSMC coculture
did not. In contrast to this, the uptake of Ac-LDL by both an EC
monoculture and an ECSMC coculture decreased drastically by a
shear flow, suggesting that the action of a shear flow on the uptake
of Ac-LDL by vascular cells is very different from that of LDL.

MATERIALS AND METHODS

Materials
Bovine aortic endothelial cells (ECs) and smooth muscle
cells (SMCs) were purchased from Cell Systems Inc. (WA,
USA) and Cell Applications Inc. (CA, USA), respectively.
Ascorbic acid, fibronectin, and Iscovs modified Dulbeccos
medium (IMDM) were obtained from Sigma-Aldrich
Co. (MO, USA). Low-density lipoproteins (LDL) and
acetylated-LDL (Ac-LDL) labeled with 1,10 -dioctadecyl3,3,30 ,30 -tetramethyl-indocarbocyanine perchlorate (DiI)
were from Biomedical Technologies Inc. (MA, USA).

KeywordsEndothelial cell, Smooth muscle cell, Coculture,

Shear stress, LDL, Ac-LDL, Mass transfer.

INTRODUCTION
It is suspected that wall shear stresses play an important
role in the pathogenesis and localization of atherosclerosis
and anastomotic intimal hyperplasia in man.1 Thus the effects of a shear flow and flow-induced shear stresses on the
morphology and functions of vascular endothelial cells including the uptake of low-density lipoproteins (LDL) have
been studied by many investigators by using a monolayer
of cultured vascular endothelial cells (ECs).26 The results
obtained on the effects of a shear stress on the uptake and
internalization of substances showed that imposition of a
shear stress to ECs increases fluid-phase endocytosis2 and
uptake of LDL by the cells.3 However, it is questionable
whether a monolayer of ECs can be used as a model of

Culture of ECs and SMCs and Preparation

of an ECSMC Coculture
ECs and SMCs were cultured in IMDM containing fetal
calf serum (FCS) at 20% by volume, penicillin (100 IU/ml)
and streptomycin (100 g/ml) in culture dishes in a humidified atmosphere of 5% CO2 and 95% air at 37 C. Cells at
passages of 712 were used for experiments.
Preparation of an ECSMC coculture was carried out
as follows. SMCs were seeded in 35-mm diameter culture
dishes at a density of 5 104 /cm2 . To enhance the production of collagen by SMCs, ascorbic acid was added to
culture medium at a concentration of 50 g/ml throughout the entire duration of culture. Under these conditions,
SMCs covered the entire surface of the culture dish within
2 days. After SMCs were cultured for indicated duration in

Address correspondence to Dr. Koichi Niwa, Laboratory of Biofluid

Dynamics, Research Institute for Electronic Science, Hokkaido University,
North 12, West 6, North District, Sapporo 060-0812, Japan. Electronic
mail: niwa@bfd.es.hokudai.ac.jp

537
C 2004 Biomedical Engineering Society
0090-6964/04/0400-0537/1

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each experiment, ECs were seeded directly over SMCs at a

density of 5 104 /cm2 , and then the cells were cocultured
for indicated duration in each experiment. EC monoculture
was also prepared by seeding ECs in culture dishes at a
density of 5 104 /cm2 . For the investigation of lipoprotein
uptake, three kinds of ECSMC cocultures, i.e., ECSMC
coculture of 10, 14, and 18 days, were prepared by culturing
SMCs for 4, 8, and 12 days, respectively, and then seeding
ECs directly over SMCs and coculturing them for additional
6 days. The culture medium was renewed every 23 days.
Experimental Setup and Procedures
To study the effects of a laminar shear flow (shear stress)
on the uptake of lipoproteins by the cells in an EC monoculture and an ECSMC coculture, we constructed a rotatingdisk shearing apparatus similar to that used in the study of
Ono et al.7 It consisted of a power supply, a DC motor, a
32-mm diameter rotating disk made of stainless steel and
attached to the shaft of the motor, and a Plexiglas housing
which served as a spacer between the motor and the rotating
disk and also a cap for the cell culture dish. By rotating the
disk immersed in a cell culture medium at a constant velocity, the ECs forming a monolayer in an EC monoculture or
an ECSMC coculture prepared on the bottom surface of a
35-mm diameter cell culture dish were exposed to a laminar
shear flow which imposed the ECs a shear stress which varied linearly from zero at the center of the cell culture dish
(which corresponded to the center of the rotating disk) to a
maximum at a location which corresponded to the periphery
of the rotating disk. The values of shear stresses imposed
on ECs were evaluated using the following formula:
= r / h
where is wall shear stress, is the viscosity of the culture medium, r is the radial distance from the center of the
rotating disk, is the angular velocity of rotation of the
rotating disk, and h is the clearance between the surface of
an EC monolayer and the rotating disk. The conditions set
in the present study were = 0.73 mPa s, h = 0.5 mm,
velocity of rotation = 820 rpm. Thus, the estimated shear
stress imposed on ECs varied from 0 at the center of the
cell culture dish to 20 dynes/cm2 at the periphery. Since the
area mean wall shear stress is given by 2/3 of the maximum
value at the periphery of the rotating disk, we expressed the
shear stress as 13.3 dynes/cm2 in the present study.
Experiments were carried out by filling 2 ml of IMDM
containing FCS at 20% by volume, penicillin (100 IU/ml),
streptomycin (100 g/ml), and either DiI-LDL or DiI-AcLDL at a concentration of 5 g/ml in a 35-mm diameter
cell culture dish in which an EC monolayer or an ECSMC
coculture was prepared, and subjecting the ECs forming a
monolayer in either an EC monoculture or an ECSMC coculture to a laminar shear flow by rotating the disk which imposed the ECs an area mean shear stress of 13.3 dynes/cm2

for 2 h in a CO2 incubator kept at 37 C. The amount of

DiI-LDL and DiI-Ac-LDL taken up by the cells was assessed as described below. The results were compared with
those obtained without the flow (control) to find the effect
of a shear flow (shear stress).
Observation of the Growth Pattern of ECs
in an ECSMC Coculture
To find out how much ECs covered the surface of SMCs,
ECs were stained with Ac-LDL labeled with DiI (DiI-AcLDL) which has been reported to be taken up specifically by
ECs.8 This was done as follows. An ECSMC coculture was
washed with PBS twice and incubated in culture medium
containing DiI-Ac-LDL at a concentration of 5 g/ml for
2 h in a CO2 incubator. Then the cells were washed with
PBS, immersed in Hanks balanced salt solution (HBSS),
and observed by a fluorescent microscope with standard
rhodamine excitation/emission filters.
Assessment of the Amount of Lipoproteins Taken
Up by the Cells
Uptake of LDL and Ac-LDL by cells was determined
according to the methods of Murakami et al.9 After finishing the flow experiment, the cell culture dish was removed
from the rotating-disk shearing apparatus, and the cells were
washed four times with cold PBS and solubilized with 2 ml
of PBS containing 0.2% Triton X-100. After centrifuging
cell lysates to remove cell debris, the fluorescence intensity
of supernatants was measured with a spectrofluorometer
(FP-750; Jasco, Japan) at excitation and emission wavelengths of 514 and 550 nm, respectively. The fluorescence
intensity was converted to the amount of lipoproteins using
a calibration curve of DiI-LDL and DiI-Ac-LDL, and the
amount of lipoproteins taken up by the cells was expressed
as the weight of lipoproteins taken up per dish per hour.
Statistical Analysis of Experimental Data
Results were expressed as a mean SD. Significance of
difference in the means of groups was evaluated by Students t-test. The minimum level of significance was set at
p < 0.05.
RESULTS
Growth Pattern of an ECSMC Coculture
Figure 1 shows the photographs of an ECSMC coculture taken at 1, 3, and 5 days after directly seeding ECs
over SMCs in a cell culture dish. As it can be seen from the
figure, it was not possible to distinguish ECs from SMCs
under a phase-contrast microscope. However, under a fluorescent microscope, ECs could be observed clearly as red
spots since they took up DiI-Ac-LDL. As evident from the
figure, the surface area occupied by ECs increased with

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539

FIGURE 1. Microscopic photographs of an ECSMC coculture at various times after seeding ECs over SMCs. ECs were seeded
directly over SMCs 4 days after seeding SMCs. The pictures shown in the upper and lower panels were taken respectively under a
phase contrast microscope and a fluorescent microscope following incubation of the ECSMC coculture with a medium containing
DiI-Ac-LDL. Original magnification: 200.

increasing the duration of culture, and whole area was covered with a confluent monolayer of ECs within 5 days after
seeding ECs. The thickness of the ECSMC coculture prepared in this manner was approximately 78 m.

SMC coculture of 10 days an area mean shear stress of

13.3 dynes/cm2 on the uptake of LDL by the cells in the
EC monoculture and the ECSMC coculture. It was found
that the uptake of LDL by an EC monoculture increased by
approximately 30% by the exposure of the cells to a shear

Uptake of Lipoproteins
Figure 2 shows the results of LDL uptake by EC monoculture and three kinds of ECSMC cocultures that were
prepared by culturing SMCs for 4, 8, and 12 days, respectively, and then seeding ECs directly over the SMCs and
coculturing them for 6 days. As evident from the figure, in
all the cases, the ECSMC coculture took up several times
more LDL than the EC monoculture did, indicating that the
LDL taken up by ECs was transferred to SMCs lying underneath. It appeared that the amount of LDL taken up by an
ECSMC coculture increased with increasing the duration
of culture of SMCs (increasing the number of SMCs) in the
ECSMC coculture. Not like the case of LDL, the uptake
of Ac-LDL did not increase by the presence of SMCs, and
the amount of Ac-LDL taken up by the ECSMC coculture
was almost the same as that taken up by an EC monoculture,
indicating that it was taken up only by ECs (Fig. 3).
Effects of a Shear Flow (Shear Stress)
on Lipoprotein Uptake
Figure 4 shows the effects of a laminar shear flow
which imposed the ECs in an EC monoculture and an EC

FIGURE 2. Uptake of DiI-LDL by an EC monoculture and three

kinds of ECSMC cocultures. ECSMC cocultures were prepared by seeding ECs directly over SMCs after culturing SMCs
for 4 or 8 or 12 days, and then coculturing them for 6 days.
The uptake of DiI-LDL by the cells were tested by incubating
the cells with a medium containing 5 g/ml of DiI-LDL for 2 h at
37 C in a CO2 incubator. The results are expressed as a mean +
SD (n = 39).

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FIGURE 3. Uptake of DiI-Ac-LDL by an EC monoculture and

three kinds of ECSMC cocultures. ECSMC cocultures were
prepared in the same manner as described in Fig. 2. The uptake
of DiI-Ac-LDL by the cells were tested by incubating the cells
with a medium containing 5 g/ml of DiI-Ac-LDL for 2 h at 37 C
in a CO2 incubator. The results are expressed as a mean + SD
(n = 39).

flow (shear stress). The effect of a shear flow on the uptake

of LDL by an ECSMC coculture was obscured by a drastic
increase in the amount of LDL taken up by the ECSMC
coculture even in the absence of a shear flow and by the large
values of standard deviation from mean values. Statistically
speaking, there was no significant difference between the
values of the amount of LDL taken up by the cells in EC
SMC cocultures exposed and not exposed to a shear flow,

FIGURE 5. Effects of a shear flow (shear stress) on the uptake

of DiI-Ac-LDL by an EC monoculture and ECSMC 10-dayscocultures. The effects of a shear flow on the uptake of DiI-AcLDL by the cells were tested by exposing the cells to an area
mean shear stress of 13.3 dynes/cm2 while incubating the cells
with a medium containing 5 g/ml of DiI-Ac-LDL for 2 h at 37 C
in a CO2 incubator. The results are expressed as a mean + SD
(n = 35).

indicating that the effect of a shear flow is minimal in this

case.
Figure 5 shows the effects of a shear flow on the uptake of
Ac-LDL by an EC monoculture and an ECSMC coculture.
As evident from the figure, the uptake of Ac-LDL by both
the EC monoculture and the ECSMC coculture decreased
drastically by the exposure of the cells to a shear flow (shear
stress), suggesting that the action of a shear flow (shear
stress) on the uptake of Ac-LDL by ECs is very different
from that of LDL.
DISCUSSION
The Merit of Using ECSMC Cocultures Prepared
by Direct Seeding of ECs Over SMCs

FIGURE 4. Effects of a shear flow (shear stress) on the uptake of DiI-LDL by an EC monoculture and ECSMC 10-dayscocultures. The effects of a shear flow on the uptake of DiI-LDL
by the cells were tested by exposing the cells to an area mean
shear stress of 13.3 dynes/cm2 while incubating the cells with
a medium containing 5 g/ml of DiI-LDL for 2 h at 37 C in a
CO2 incubator. The results are expressed as a mean + SD
(n = 35).

To study the effects of a shear flow (shear stress) on various functions of endothelial cells (ECs) such as syntheses
of biochemicals and uptake of lipids, it is desirable to use
real arteries and veins by carrying out animal experiments.
However, it is difficult to have many animals and prepare
vessels for experiments and control experimental conditions
same as those physiological ones in vivo. Thus, most of the
studies have been carried out using cultured ECs and in
vitro. These studies brought about some information useful for elucidating the role of hemodynamic factors in the
pathogenesis and localization of atherosclerosis and anastomotic intimal hyperplasia. The disadvantage of using an EC
monoculture is lack of the interaction of ECs with smooth
muscle cells (SMCs) which underlie the ECs in a real vessel. In fact, there are some reports that suggest that ECs and

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SMCs interact with each other and affect each others functions such as production of certain biochemicals,1012 proliferation of themselves,13,14 and uptake of lipoproteins.15,16
Most of the experiments carried out to investigate the interactions between ECs and SMCs employed coculture systems in which ECs and SMCs were seeded and grown separately on both sides of a microporous membrane so as to
make it possible to analyze various functions of the two
different types of cells separately.1014 In real arteries and
veins, ECs are isolated from SMCs which are in the media
by a membrane called internal elastic lamina in the same
manner as the case of ECSMC coculture system mentioned
above. However, the membrane is elastic and has numerous
pores whose diameters range from 2 to 7 m,17 enabling a
direct contact between the ECs coating the luminal surface
of the vessel in a monolayer and the SMCs forming the media of the vessel. Considering these facts, the coculture system described above may not be regarded as a physiological
model of an arterial wall since it lacks a direct contact between ECs and SMCs necessary to interact with each other.
Therefore, to assure a direct contact and a direct interaction
of ECs with SMCs and solve the problem, we constructed
a model of an arterial wall by seeding ECs directly over
SMCs and coculturing them until the ECs formed a confluent monolayer which covered the entire surface of the
SMCs.
The Uptake of Lipoproteins by an EC Monoculture
and an ECSMC Coculture
In our present study, we prepared three kinds of EC
SMC cocultures by culturing SMCs for different periods
and then seeding ECs directly over the SMCs and coculturing them. Using these cocultures, we studied the uptake
of LDL and Ac-LDL, and compared the results with those
obtained with EC monocultures.
It has been shown that LDL is taken up by ECs via two
pathways, that is, LDL receptor-mediated endocytosis and
nonspecific fluid phase endocytosis.18,19 It is considered
that Ac-LDL is also taken up by ECs via two pathways, that
is, scavenger receptor-mediated endocytosis 20 and nonspecific fluid phase endocytosis. Our results showed that the
uptake of Ac-LDL was much greater than that of LDL in
EC monocultures. Since the amount of LDL and Ac-LDL
added to the medium was the same, it is likely that the
amount of LDL and Ac-LDL taken up by ECs via nonspecific fluid phase endocytosis was also the same. Therefore,
it was considered that the uptake of a large amount of AcLDL by an EC monoculture was carried out via scavenger
receptor-mediated endocytosis. A similar observation was
reported by Sanan.21 He showed that the uptake of Ac-LDL
by confluent BAECs in culture was four times greater than
that of LDL.
It was found that the uptake of LDL by ECSMC coculture was much greater than that by EC monocultures, and

541

it appeared to increase with increasing the duration of culture of SMCs. It was considered that this was caused by the
increase in the number of SMCs since it was also observed
that the thickness of the ECSMC coculture increased with
increasing the duration of culture of SMCs prior to seeding
ECs.
It has been reported that transfer of LDL from plasma
to the wall of an aorta takes place largely independent of
LDL receptor-mediated endocytosis of ECs.22,23 Although
we could not determine the transport pathway of LDL from
ECs to SMCs in the present study, it is likely that LDL was
carried by vesicles through ECs and exocytosed to SMCs.15
To confirm the transport pathways of LDL in this coculture,
it is necessary to study further the ultrastructure of the junctions of the ECs and SMCs.
In the case of the uptake of Ac-LDL, since it has been
reported that SMCs could take up Ac-LDL via scavenger
receptor-mediated endocytosis,24 we expected that the uptake of Ac-LDL by ECSMC cocultures increase with increasing the duration of culture of SMCs hence the number
of the SMC in the coculture. However, the actual results
were completely different from what we expected. The
amount of AC-LDL taken up by ECSMC cocultures was
almost the same as that taken up by EC monocultures despite of the presence of SMCs in different numbers. The
results suggest that Ac-LDL was taken up only by ECs
even in ECSMC cocultures, or even if it was taken up
also by SMCs, the amount was negligibly small. It has been
reported that in hypercholesterolemic rabbits, scavenger receptors are expressed highly in SMCs located in the intima
of a balloon-injured portion of the aorta but not in SMCs
located in the media of uninjured portion of the aorta.24 It
has been also shown by several in vitro experiments that the
expression level of scavenger receptors on SMCs was regulated by some stimulants such as inflammatory cytokines24
and oxidative stresses.25 Thus, it was considered that the
expression level of scavenger receptors on the SMCs of
ECSMC cocultures used in our present study was very
small since the SMCs were not activated by any stimulus.
The Effects of a Shear Flow (Shear Stress)
on the Uptake of Lipoproteins
In the present study we used a rotating-disk shearing
apparatus to impose a shear flow on ECs. In this apparatus, since the estimated value of the shear stress imposed
on ECs varied from 0 at the center of the culture dish to
20 dynes/cm2 at the periphery, it was not possible to find
out the effect of a shear stress of a certain fixed value. However, since we could obtain a shear flow that imposed the
ECs an area mean shear stress of 13.3 dynes/cm2 which is
close to the mean wall shear stress prevailing in vivo with
this apparatus, we used this just to see if there is any effect
of a physiological level of shear flow on the uptake of LDL
and Ac-LDL by the cells in an ECSMC coculture.

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Our results showed that the uptake of LDL by EC monocultures increased by the exposure of the cells to a shear
flow (shear stress). This is in agreement with the results
reported by Davies et al.2 and Sprague et al.3 In contrast
to this, the uptake of Ac-LDL decreased by the imposition
of a shear flow (shear stress). Similar results were obtained
with ECSMC cocultures. It has been reported that ECs
possess at least two types of scavenger receptors and AcLDL are taken up by ECs via scavenger receptor-mediated
endocytosis.20,26 However, precise roles of these receptors
are not known yet. Although we cannot explain the mechanism of the flow-induced decrease in Ac-LDL uptake by
both EC monocultures and ECSMC cocultures at this moment, the present results indicate that the action of a shear
flow (shear stress) on the uptake of Ac-LDL is different
from that on the uptake of LDL.
In the present paper, we used both a shear flow and a
shear stress to describe the effects of a flow on the uptake
of lipoproteins by an EC monoculture and an ECSMC coculture since we were not sure whether it was a flow itself
or a flow-induced shear stress which affected the uptake of
lipoproteins by the cells. It is considered that a flow itself
will affect the contact time of lipoprotein molecules with
ECs, while a shear stress will affect the structure and function of ECs. It is possible that in the case of the uptake of
LDL, although the contact time of LDL with ECs is shortened by the presence of a flow, the uptake and intracellular
transport of LDL by ECs are augmented through the activation of pinocytosis and receptor-mediated internalization
by flow-induced shear stresses as shown by Davies et al.2
and Sprague et al.,3 resulting in an increase in the uptake
of LDL by both an EC monoculture and an ECSMC coculture. The fact that the uptake of LDL increased greatly
by the presence of SMCs underneath of ECs indicates that
SMCs are actively involved in the transport of LDL taken up
by ECs. In the case of Ac-LDL, it is likely that in addition
to shortening of contact time of Ac-LDL molecules with
ECs, the expression of scavenger receptors in ECs was not
enhanced by the shear stress, resulting in a drastic reduction
in the uptake of AC-LDL by both an EC monoculture and
an ECSMC coculture. This is only our speculation, and
it is necessary to clarify which of the flow itself and the
flow-induced shear stress affects the uptake of lipoproteins.

a condition of a slow flow and a low wall shear stress. Although it is not likely that Ac-LDL is produced in vivo,
there is a report that shows that both Ac-LDL and oxidized
LDL (Ox-LDL), which is supposed to play an important
role in atherogenesis, are taken up by ECs by binding to the
scavenger receptor SREC.20 These modified lipoproteins
are also taken up by SMCs by binding to a class-A scavenger receptor.25 Therefore, we considered that the uptake
of Ac-LDL by vascular cells may at least in part represent
the uptake of Ox-LDL. The results obtained with Ac-LDL
suggests that it is very likely that the uptake of Ox-LDL
by ECs in the arterial endothelium and its accumulation in
the subendothelial space occurs preferentially in regions of
slow flow and low wall shear stress, inducing the adhesion
to the endothelium, invasion into the subendothelial space,
and accumulation there of monocytes (macrophages) that
take up Ox-LDL and become foam cells in such regions,
leading eventually to localized genesis and development of
atherosclerotic lesions and intimal hyperplasia at such sites.
It should be pointed out, however, that in our present study,
the ECs in both monoculture and coculture with SMCs were
exposed to a shear flow only for 2 h. This might be too short
to assess the chronic effects of shear flow on various functions of vascular endothelial cells and smooth muscle cells.
To confirm the physiological significance of flow- or shear
stress-induced changes of the uptake of modified LDL, further studies including animal experiments are required.
In summary, we constructed a model of an arterial wall
by seeding ECs directly over SMCs and coculturing them,
and tested whether it behaved in the same manner as an EC
monoculture did in the uptake of lipoproteins. We found that
in an ECSMC coculture, the uptake of LDL was greatly
increased by the presence of SMCs, whereas the uptake
of Ac-LDL remained almost the same as that by an EC
monoculture, indicating that Ac-LDL was taken up only by
ECs. It was also found that a shear flow (a shear stress) imposes opposite effects on ECSMC cocultures in the uptake
of LDL and Ac-LDL. Our simple model of an arterial wall
would be useful to investigate the effects of various physical
and fluid mechanical factors on the uptake of lipoproteins
by an arterial wall in vitro.

Implication of the Results for Localized Development

of Vascular Diseases

This work was supported by a Grant-in-Aid for Scientific

Research no. 15300150 from Japan Society for the Promotion of Science and a Grant-in-Aid for Young Scientists (B)
no. 15780184 from The Ministry of Education, Culture,
Sports, Science and Technology of Japan.

It has been reported that slow flows and the resultant low
wall shear stresses are directly related to the localization of
atherosclerotic lesions and anastomotic intimal hyperplasia
in man.1,2729 In our present study, it was found that the uptake of Ac-LDL, which is a chemically modified LDL, by
an EC monoculture and an ECSMC coculture was greatly
suppressed by the imposition of a shear flow (shear stress).
In other words, the uptake of Ac-LDL was favored under

ACKNOWLEDGMENTS

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