465

Blood Flow and in Vivo Apparent Viscosity

in Working and Non-Working Skeletal
Muscle of the Dog after High and Low
Molecular Weight Dextran
L. GUSTAFSSON, L. APPELGREN, AND H.E. MYRVOLD

SUMMARY We studied the effect of high and low molecular weight dextran on blood flow and in vivo
apparent viscosity in the vasodilated vascular bed of working and non-working skeletal muscle. In 12
mongrel dogs, the calf muscle of one hindlimb was isolated. Vasodilation was induced either by sciatic
stimulation setting the muscle at rhythmic work or by intraarterial infusion of papaverine. Blood flow
was measured electromagnetic'ally at different perfusion pressures. In vivo apparent viscosity was
calculated by comparing pressure-flow relationships for blood and a reference solution. Viscosity in
vitro was determined in a cone-plate viscometer. A hyperviscous state was induced by intravenous
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infusion of high molecular weight dextran (HMWD). Hemodilution subsequently was produced by
administration of low molecular weight dextran (LMWD). After HMWD, blood flow decreased to 30%
of control values in the non-working group and to 45% of control values in the working group. After
subsequent infusion of LMWD, blood flow returned to 60% of control values in the non-working group
and to 70% of control values in the working group. In vivo apparent viscosity increased to values 250%
above control in the non-working group and to 120% above control in the working group following
HMWD. After subsequent infusion of LMWD in vivo, apparent viscosity decreased, but remained at
values 65% above control in the non-working group and 45% above control in the working group. Thus,
the flow impairment induced by HMWD was less pronounced in the working muscle, indicating a flow-
preserving effect of rhythmic muscle contractions in this state of disturbed blood rheology. In contrast,
the flow-improving effect of LMWD by hemodilution was more pronounced in the non-working muscle.
Circ Res 48: 465-469, 1981

VISCOSITY of whole blood and plasma is in-
creased after tissue injury, especially at low shear
rates (Gelin, 1956). A change is seen in plasma
proteins with an increased fibrinogen-albumin ratio
resulting in an increased plasma viscosity, de-
creased suspension stability, and occurrence of red
cell aggregation. The flow disturbance after trauma
thus includes two different alterations of the rheo-
logical properties of blood: aggregation of formed
elements and increase of plasma viscosity. Abnor-
mal rheological behavior with increased plasma vis-
cosity and cell aggregation also are sometimes en-
countered in certain forms of plasma cell dyscrasias
(Wells 1970, Somer 1975). It has been suggested
that infusion of high molecular weight dextran
(HMWD) to some extent mimics the hyperviscous
state following extensive trauma (Gelin 1956) and,
because of their molecular weight-dependent influ-
ence upon red cell aggregation, dextrans have been
used in model experiments for evaluation of the role
of cell aggregation (Djojosugito et al., 1970; Dick-
mans et al., 1971). Infusion of HMWD produces red
cell aggregation and increased plasma viscosity.
Low molecular weight dextran (LMWD), on the
other hand, corrects the hyperviscous state induced
by HMWD (Gelin 1956, 1962; Shoemaker et al.,
1965; Litwin et al., 1965; Gelin et al. 1968; Appelgren
and Lewis, 1970) and trauma (Gelin, 1956, 1962).
Blood flow and blood viscosity are further depen-
dent on hematocrit, and there is a decrease of flow
at increased hematocrits. In a previous study we
found that rhythmic muscle contractions facilitated
flow at increased hematocrits (Gustafsson et al.,
1980). The purpose of the present investigation was
to study the influence of rhythmic muscle contrac-
tions on in vivo apparent viscosity and flow during
a state of red cell aggregation and increased plasma
viscosity induced by HMWD and packed red cells
and, furthermore, to evaluate the effect of subse-
quent hemodilution with LMWD.

From the Department of Surgery I and II and Department of Anaes-
thesiology III, University of Goteborg, Goteborg, Sweden.
This study was supported by grants from the Swedish Medical Re-
search Council (B 78-17X-O066O-14B), from the Faculty of Medicine,
University of Goteborg,fromthe Goteborg Medical Society, and from the
Tore Nilson Foundation.
Address for reprints: Dr. L. Gustafsson, Department of Surgery I,
Laboratory of Biorheology, Sahlgrenska Sjukhuset, S-413 45 Goteborg,
Sweden.
Received May 28, 1980; accepted for publication October 16, 1980.
Methods
In 12 mongrel dogs (body weight 15-22 kg) an-
esthesia was induced and maintained with intrave-
nous thiopental sodium. The dogs were intubated
endotracheally and ventilated artificially with air
using a volume controlled respirator.
The flow measurements were performed in the
isolated calf muscle of one hindlimb of the dog (Fig.
 466 CIRCULATION RESEARCH
A.Fern.. Dex-Tyr
VFem. ' Perfusion
Papaverine
'rn> Flow probe
Adjustable screw clamp
Sciatic nerve
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FIGURE 1 The isolated calf muscle of the dog with
provisions for blood flow measurements, perfusion pres-
sure recording, sciatic nerve stimulation, and papaver-
ine infusion.
1), the surgical preparation being similar to the one
described in a previous study (Gustafsson et al.,
1980). The thigh muscles were divided above the
knee joint, as were all vessels except the popliteal
artery and vein. The skin was loosened around the
calf in order to exclude cutaneous blood flow, after
which the skin was repositioned, thus protecting
the preparation. The paw was excluded by a screw
clamp placed just above the ankle joint. The super-
ficial femoral artery was isolated and all side
branches were divided except two which were used
for pressure recording and drug infusion. Wide-bore
siliconized T-tubes were introduced in the femoral
artery and vein for intermittent extracorporeal per-
fusion of the isolated limb with oxygenated body-
warm dextran-Tyrode's solution (6% dextran 40 in
Ringer's solution) from a pressure bottle connected
to the arterial T-tube. The dextran-Tyrode's solu-
tion was Newtonian and served as a reference so-
lution. The viscosity of the solution was 2.2 cP and
osmolality was 300-315 mOsm/kg.
The perfusion pressure was measured with Sta-
tham pressure transducers, model P23, via arterial
and vein catheters and was recorded continuously
on a Grass polygraph, model 7C. Another side
branch of the artery was used for infusion of papav-
erine. To obtain different perfusion pressures, an
adjustable screw clamp was placed on the artery
proximal to the pressure catheter. Blood flow was
measured with an electromagnetic flow meter. (Ny-
VOL. 48, No. 4, APRIL 1981
cotron model 372), with the probe placed on the
artery proximal to the screw clamp. Zero flow was
obtained by occluding the artery distal to the flow
probe. Calibration of the flow probe was done in
vivo in each experiment for blood and the dextran-
Tyrode's solution.
In six dogs flow measurements were performed
on the limb during muscle stimulation via the cut
distal end of the sciatic nerve (rectangular pulses of
3-5 V and 3-5 msec duration at a rate of 4-6
impulses per second) from a Grass stimulator. In six
dogs flow measurements were performed on the
resting muscles when the vascular bed had been
dilated by continuous intraarterial infusion of pa-
paverine (80 mg/min).
A hyperviscous state was induced by intravenous
injection of HMWD (mean molecular weight Mw 1
million) 1.0 g/kg body weight in a 15% solution
together with 200 ml packed homologous red cells
(Hct 0.90). Hemodilution subsequently was pro-
duced by administration of LMWD (mean molec-
ular weight Mw 40.000) 1.5 g/kg body weight in a
10% solution.
Pressure-flow curves were constructed by step-
wise adjustments of the screw clamp. The pressure-
flow relationships in the working and vasodilated
non-working calf muscles were established for nor-
mal blood, after infusion of HMWD and after sub-
sequent infusion of LMWD for perfusion pressures
between 0 and 70 mm Hg. By adjusting the screw
clamp in distinct steps, blood flow, expressed in ml/
min per 100 g, was obtained as a function of perfu-
sion pressure. The screw clamp was adjusted from
maximal flow down to complete closure and re-
opened again in about 20 distinct steps altogether,
maintaining the perfusion pressure constant for 5-
10 sec at each step. The closing-reopening proce-
dure was repeated three to four times, and the
values of flow and perfusion pressures obtained
were plotted on a pressure-flow diagram. Pressure-
flow curves also were recorded for the reference
solution. During perfusion with the reference solu-
tion, the proximal shanks of the T-tubes were
clamped. The arterial T-tube was connected to a
pressure-bottle containing the dextran-Tyrode's so-
lution which was drained from the muscle via the
venous T-tube. P v then was adjusted by the level of
the outlet shank and efforts were made to obtain
the same venous pressure for the reference solution
as for blood at the same arterial driving pressure.
Thereby the distending pressure (Pa + Pv)/2 was
about the same at identical perfusion pressures for
blood and the reference solution.
Provisional values for apparent or effective blood
viscosity in vivo at different perfusion pressures
were calculated by comparing the pressure-flow
curves for blood and for the reference solution,
assuming the blood and reference solution viscosi-
ties in the calf muscle to be inversely proportional
to the recorded flows at a certain perfusion pressure.
Viscosities in vitro of whole blood and plasma
 DEXTRANS AND IN VIVO BLOOD RHEOLOGY/Gustafsson et al.
were determined in a Wells-Brookfield synchroelec-
tric cone-plate viscometer, model LVT, using 1-ml
samples of heparinized blood and plasma. Deter-
minations were made at shear rates of 23, 46, 115
and 230/sec. Hematocrits were measured in micro-
hematocrit tubes. For statistical analysis the Wil-
coxon rank sum test was used (Colton, 1974).
Results
Infusion of HMWD and packed red cells induced
only a small change in hematocrit from 0.44 to 0.43
in the working group, whereas hematocrit remained
unchanged at 0.48 in the nonworking group. After
LMWD hematocrit decreased to 0.33 in the working
group and to 0.35 in the non-working group.
After infusion of HMWD and packed red cells,
whole blood viscosity in vitro increased by 120% in
both groups of dogs. After subsequent infusion of
LMWD, whole blood viscosity decreased to values
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30% above control. Plasma viscosity increased by
270% after HMWD and decreased to values 140%
above control after subsequent infusion of LMWD.
The results from the in vitro viscosity measure-
ments are shown in Figure 2.
The results of the pressure-flow measurements
are shown in Fig. 3. At corresponding perfusion
pressures and at normal hematocrits, flow was
higher in the pharmacologically vasodilated resting
0 23 id
Sheaf rate
VISCOSITY
Normal
. HMWO
--LMVD
Ptasmo
23 46 115
Shear rate s"'
FIGURE 2 In vitro viscosity data for whole blood and
plasma for normal blood ( ) , after HMWD ()
and after LMWD (- ). Spread of data is given
in four points at shear rates of 23, 46, 115, and 230/sec
(mean SEM).
467
OmJ mm KtOg
150
NoifTXjl
--- HMWD
- - LMWO
P o - P v mmHg
FIGURE 3 Pressure-flow relationships in the working
and vasodilated non-working skeletal muscle for normal
blood ( ;, after HMWD (- - - -), and after LMWD
(- -) Spread of data is given in three represent-
ative points at perfusion pressures of 25, 50, and 70 mm
Hg (mean SEM). The pressure-flow curves for the
reference solution, a dextran-Tyrode's solution, are in-
dicated by ( ).
muscles than in the rhythmically contracting mus-
cles. After infusion of HMWD skeletal muscle,
blood flow decreased in both groups. In the vaso-
dilated non-working muscles, blood flow decreased
by 70% with the most pronounced decrease at lower
perfusion pressures. In the rhythmically contracting
muscles blood flow decreased by 55%, i.e., the flow
decrease was less pronounced in the rhythmically
contracting muscles than in the resting muscles (P
< 0.05). After subsequent infusion of LMWD, blood
flow increased, but to only 60% of initial control
values in the non-working group and to 70% of
initial values in the working group. Note that, ex-
pressed as a flow improvement from the HMWD to
the LMWD state, the flow increase was more pro-
nounced in the non-working group than in the
working group after LMWD.
Calculated apparent viscosities in vivo as a func-
tion of perfusion pressure are shown in Figure 4.
After infusion of HMWD and packed red cells, in
vivo apparent viscosity increased to values 240%
above control at high perfusion pressures (70 mm
Hg) and to 280% above control at lower perfusion
pressures (25 mm Hg) in the vasodilated nonwork-
ing muscles. In the rhythmically contracting mus-
cles in vivo, apparent viscosity increased to values
120% above initial values at high perfusion pres-
sures and to 140% above initial at lower perfusion
pressures. After subsequent infusion of LMWD,
230
apparent viscosity in vivo decreased, although not
to initial control values but to values 65% above
initial values in the non-working group and to 45%
above initial values in the rhythmic contracting
group. Thus, the reduction of in vivo apparent
viscosity after LMWD was relatively more pro-
nounced in the non-working group.
 468
VISCOSITY
IN VIVO
VISCOSITY
K> Non-working 10- Vforking
9 9
8 Hcl
8 HMWD
7 LMWD
7 "48
6 6
5- 5-
4
3 44
2 .-Tyr.
1 by the occurrence of inertial pressure losses during
50 100
PoPyrnmHg
FIGURE 4 Average relationships between perfusion
pressure and calculated apparent viscosity in vivo in the
working and vasodilated non-working skeletal muscle
for normal blood ( ) , after HMWD () and after
LMWD ( -). The apparent viscosity in vivo was
calculated by comparing the flow values for blood and
the reference solution at identical perfusion pressure.
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Discussion
In the present investigation, pressure-flow rela-
tionships and viscosity changes in vitro and in vivo
were studied during a hyperviscous state induced
by HMWD and packed red cells and after subse-
quent infusion of LMWD. The amount 1.0 g/kg of
HMWD given has been shown to cause disturbance
of flow and flow distribution and pronounced red
cell aggregation as evidenced by vital microscopy
(Litwin et al., 1965; Gelin et al., 1968).
The flow and viscosity changes were analyzed in
two different states of the skeletal muscle vascular
bed: the vasodilated resting muscle and the rhyth-
mically contracting muscle. By inducing a pro-
nounced vasodilation, vessel geometry adjustments
because of smooth muscle activity were avoided.
Under normal conditions, blood flow in the papav-
erine-dilated nonworking muscles exceeded the flow
in the exercise-dilated muscles, indicating that the
twitching movements of the muscles provide a me-
chanical hindrance to flow. Furthermore, an addi-
tional vasodilating effect probably is obtained phar-
macologically compared to metabolically induced
vasodilation (Gaehtgens et al., 1975).
Infusion of HMWD and packed red cells resulted
in a decreased flow and increased in vivo apparent
viscosity in both resting and exercising muscles.
However, the flow decrease was less marked in the
rhythmically contracting muscle. Subsequent infu-
sion of LMWD reduced in vivo apparent viscosity
and increased flow in both groups, although the
pre-experimental control values were not reached.
The apparent viscosity in vivo was obtained by
perfusing the limb with a reference solution of
known viscosity (Djojosugito et al., 1970). From
comparison of flow values for blood and dextran-
Tyrode's solution at identical perfusion pressures,
apparent viscosity in vivo could be calculated. The
concept of determining viscosity in vivo is of im-
portance since a considerable discrepancy has been
CIRCULATION RESEARCH VOL. 48, No. 4, APRIL 1981
found to exist between viscosity as measured in
vitro and in vivo. Thus, in vivo apparent viscosity
- Normo
has been shown to be considerably lower than that
in vitro (Whittaker and Winton, 1933, Levy and
Share, 1953, Djojosugito et al., 1970). The difference
Hcl has been attributed mainly to the Fahraeus-Lind-
43
qvist effect in small vessels (Fahraeus and Lind-
qvist, 1931). However, Benis et al. (1970, 1973)
demonstrated that the discrepancy can be explained
50 100 flow of cell-free reference solution in larger vessels
nmHg
outside the microcirculation proper. In the present
study, the calculated in vivo apparent viscosity was
lower than in vitro whole blood viscosity for normal
blood and around 2.2 cP at high perfusion pressures.
This is in accordance with other studies of this sort,
e.g., Benis et al. (1973) who found normal blood to
exhibit an effective relative viscosity of about 2.
After HMWD, in vivo apparent viscosity increased
relatively more than whole blood viscosity in vitro
at least at lower perfusion pressures. This has been
observed earlier and might be due to microvessel
obstruction by red cell aggregates (Djojosugito et
al., 1970). It is, however, important to have in mind
that HMWD alters the fluidity of blood in two
ways: aggregation of red cells and increase in plasma
viscosity. Their relative contribution to the flow
decrease has been studied in vivo by Dickmans et
al. (1971) using isolated intestinal segments and by
Hint (1964) using the isolated rabbit ear. They
compared dextran-blood solutions with different
molecular weights at identical plasma viscosities
and hematocrits for their effect on blood flow at
different perfusion pressures and found only small
effects of red cell aggregation, at least at normal
perfusion pressures. However, a considerable flow
reduction was observed when plasma viscosity was
increased (Hint, 1964). In the present study, in vivo
apparent viscosity values in working muscle vary
almost in direct proportion to changes in plasma
viscosity, suggesting a greater importance of plasma
viscosity than red cell aggregation. The contribu-
tion of increased plasma viscosity is evident after
infusion of LMWD when in vivo apparent viscosity
is still well above control values despite an almost
normal in vitro whole blood viscosity.
When comparing the pressure-flow relationships
after HMWD and packed red cells in the resting
and exercising muscles, we found that the flow
decrease was less pronounced in the exercising mus-
cles, indicating a flow-preserving effect of the
rhythmic muscle contractions during this state of
disturbed rheology. A similar flow-preserving effect
has been shown to exist after acutely induced poly-
cythemia (Gustafsson et al., 1980). A high hemato-
crit was found to be more deleterious to the oxygen
availability in the resting muscle than in the exer-
cising muscle. In the present study, the rhythmi-
cally contracting muscles are also more tolerant to
the hyperviscous state induced by HMWD. The
mechanism for the flow-preserving effect during
 DEXTRANS AND IN VIVO BLOOD RHEOLOGY/Gustafsson et al.
hyperviscosity might be that the muscular contrac-
tions facilitate the passage of red cell aggregates by
squeezing them through the precapillary and cap-
illary network. The rhythmic muscle contractions
might also increase the flow velocity in the postcap-
illary venules where flow and shear rates are pro-
portionally low and viscosity proportionally high,
especially after induced red cell aggregation and at
low perfusion pressures.
LMWD caused hemodilution and almost re-
stored whole blood viscosity in vitro. After LMWD,
the flow increase was somewhat more pronounced
in the resting than in the exercising muscle. This
might indicate that the rhythmic muscle contrac-
tions act mainly on cellular elements which become
less important after hemodilution.
The results from the present study indicate that
there is a flow-preserving effect of rhythmic muscle
contractions during hyperviscosity induced by high
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molecular weight dextran. In vivo apparent viscos-
ity is probably dependent on both red cell aggre-
gation and the plasma viscosity. Their relative con-
tributions cannot be estimated however, from the
present study. No quantitative relationships exist
between in vitro whole blood or plasma viscosity
with in vivo apparent viscosity, but these param-
eters vary in the same direction in vivo and in vitro.
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L Gustafsson, L Appelgren and H E Myrvold

Circ Res. 1981;48:465-469

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doi: 10.1161/01.RES.48.4.465
Circulation Research is published by the American Heart Association, 7272 Greenville Avenue, Dallas, TX 75231
Copyright 1981 American Heart Association, Inc. All rights reserved.
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