Eur J Appl Physiol

DOI 10.1007/s00421-016-3349-3

ORIGINAL ARTICLE

Musclespecific acute changes inpassive stiffness ofhuman

triceps surae afterstretching
KosukeHirata1 EriMiyamotoMikami1,2 HiroakiKanehisa1 NaokazuMiyamoto1

Received: 6 December 2015 / Accepted: 24 February 2016

Springer-Verlag Berlin Heidelberg 2016

Abstract
Purpose It remains unclear whether the acute effect of
stretching on passive muscle stiffness differs among the
synergists. We examined the muscle stiffness responses of
the medial (MG) and lateral gastrocnemii (LG), and soleus
(Sol) during passive dorsiflexion before and after a static
stretching by using ultrasound shear wave elastography.
Methods Before and after a 5-min static stretching by
passive dorsiflexion, shear modulus of the triceps surae
and the Achilles tendon (AT) during passive dorsiflexion
in the knee extended position were measured in 12 healthy
subjects.
Results Before the static stretching, shear modulus was
the greatest in MG and smallest in Sol. The stretching
induced significant reductions in shear modulus of MG, but
not in shear modulus of LG and Sol. The slack angle was
observed at more plantar flexed position in the following
order: AT, MG, LG, and Sol. After the stretching, the slack
angles of each muscle and AT were significantly shifted to
more dorsiflexed positions with a similar extent. When considering the shift in slack angle, the change in MG shear
modulus became smaller.
Conclusion The present study indicates that passive
muscle stiffness differs among the triceps surae, and that
the acute effect of a static stretching is observed only in
the stiff muscle. However, a large part of the reduction of

Communicated by Olivier Seynnes.

* Naokazu Miyamoto
miyamoto@nifsk.ac.jp
1

National Institute ofFitness andSports inKanoya,

1 Shiromizu, Kanoya, Kagoshima 8912311, Japan

Japan Society forthe Promotion ofScience, Tokyo, Japan

passive muscle stiffness at a given joint angle could be due

to an increase in the slack length.
Keywords Ultrasound shear wave elastography Shear
modulus Gastrocnemius Soleus Ankle joint
Abbreviations
ANOVA Analysis of variance
AT Achilles tendon
EMG Electromyography
LG Lateral gastrocnemius
MG Medial gastrocnemius
MTU Muscletendon unit
MVC Maximal voluntary contraction
RMS Root mean square
ROM Range of motion
Sol Soleus

Introduction
Pre-exercise stretching is usually performed for decreasing passive muscle stiffness. Understanding the effect
of stretching on muscle stiffness has long been a topic of
interest for not only athletes and coaches but also clinicians and researchers. In human experiments, passive muscle stiffness has been widely evaluated from the relationship between joint angle and passive joint torque (Gajdosik
2001; Magnusson 1998; Magnusson etal. 1997). However,
the joint torque is not a measure specific to the responses
of individual muscles, because the joint torque results from
a composite of contractile (i.e., synergists and antagonists)
and non-contractile tissues such as tendon, skin, ligament and articular structures. Based on the change in joint
torque, therefore, it is difficult to determine alteration in the

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stiffness of individual muscles. Because of this limitation,

it remains unknown whether the acute effect of stretching
on passive muscle stiffness differs among the synergists.
The development of imaging techniques such as ultrasonography allows us to evaluate underlying architectures
of the muscletendon unit (MTU) in humans invivo. Using
B-mode ultrasonography, previous studies have examined
changes in muscle fascicle length and behavior of tendinous tissues during passive lengthening of the MTU (Abellaneda etal. 2009; Blazevich etal. 2012; Herbert etal.
2002; Morse etal. 2008). However, the conclusions of previous studies using B-mode ultrasonography on the effect
of stretching on muscle stiffness have not reached a consensus. For example, the muscle stiffness of the medial gastrocnemius (MG) has been reported to be reduced (Morse
etal. 2008; Nakamura etal. 2011) or not be affected (Kato
etal. 2010; Konrad and Tilp 2014b). As one of the reasons for the inconsistent conclusions over studies, methodological issue for evaluating the muscle stiffness may be
involved. In addition to the fact that the passive torque is
related to the viscoelastic properties of the entire musculoarticular complex, B-mode ultrasonography per se can provide information only about architecture such as muscle
tendon junction displacement and fascicle length, not about
mechanical properties such as stiffness. Thus, other methods are required to more directly and properly assess the
passive stiffness of individual muscles in human invivo.
Additionally, previous studies which examined passive
lengthening of the plantar flexors (i.e., passive dorsiflexion) have very often investigated only MG (Abellaneda
etal. 2009; Blazevich etal. 2012; Konrad and Tilp 2014b;
Morse etal. 2008; Nakamura etal. 2011), perhaps because
of the difficulty to properly quantify the elongation of other
muscles such as the lateral gastrocnemius (LG) and soleus
(Sol) by using B-mode ultrasonography. Muscle stiffness
is influenced by the muscle fiber type composition; type
I muscle fibers exhibit greater passive stiffness (Kovanen
etal. 1984b; Mutungi and Ranatunga 1996, 1998). In the
human triceps surae, muscle fiber type composition has
been shown to be different among the synergists (Johnson
etal. 1973). Therefore, it is hypothesized that the responses
in passive muscle stiffness to passive ankle dorsiflexion
and their changes induced by an acute bout of stretching is
greater in Sol composed of 70100% type I muscle fibers
than in MG and LG composed of >50% type II muscle fibers (Johnson etal. 1973).
One of the methods to resolve the aforementioned concerns is to use ultrasound shear wave elastography. This is
a recently developed imaging technique that can quantify
localized tissue stiffness along the principal axis of the
probe, based on the propagation speed of remotely induced
shear wave (Bercoff etal. 2004; Palmeri etal. 2008). Muscle shear modulus measured by ultrasound shear wave

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Eur J Appl Physiol

elastography is shown to be highly correlated with Youngs

modulus obtained via traditional materials testing (Eby
etal. 2013). By the use of this technique, the present study
aimed to test the hypotheses mentioned above by measuring the muscle stiffness responses of MG, LG, and Sol during passive dorsiflexion before and after an acute stretching
intervention.

Materials andmethods
Subjects
Twelve healthy subjects (eight men and four women)
participated in the present study (168.87.5cm,
61.78.9kg, 20.42.9years; meanSD). None of the
subjects reported any ongoing neuromuscular diseases or
musculoskeletal injuries specific to the ankle or knee joints.
Before participation, all subjects were fully informed of the
purpose and experimental procedures. They were asked to
refrain from strenuous exercise 48h before testing. Each
subject completed an informed consent form. The Ethics
Committee on Human Research of National Institute of
Fitness and Sports in Kanoya approved the investigation,
and all of the experimental procedures were performed in
accordance with the Declaration of Helsinki.
Experimental setup andprotocol
Subjects lay prone on a dynamometer (CON-TREX MJ,
PHYSIOMED, Germany) bed with their right knees fully
extended. The right foot was tightly fixed to the dynamometers footplate. The rotation axes of the ankle and the footplate were aligned as closely as possible. The ankle joint
angle was passively dorsiflexed from 50 plantar flexion
[neutral position (i.e., the sole of the foot at right angles to the
tibia axis) defined as 0, with larger values for dorsiflexion]
to the end ROM which was defined as the angle that the subjects felt pain, at an angular velocity of 1/s. The very slow
angular velocity was used to obtain a better resolution for the
muscle stiffness via ultrasound shear wave elastography and
to avoid or minimize the stretch reflex (Hirata etal. 2015;
Hug etal. 2013; Morse etal. 2008), which could stiffen the
muscles. The angular displacement of the ankle joint was
measured with an electronic goniometer (SG110/A, Biometrics, UK) fixed to the ankle joint with double-sided adhesive
tape. In this study, all reported angle measurements refer to
the ankle joint angle assessed with the goniometer, not the
angle of the dynamometer footplate. Throughout the passive
lengthening, the subjects were requested to relax completely
and to not resist the movement of the footplate. In order to
familiarize the subjects to the procedure and to ensure that
they were as relaxed as possible, a familiarization session of

Eur J Appl Physiol

two cycles was performed followed by the start of the testing

session. This familiarization session also has a role to avoid
a conditioning effect of passive lengthening on the stiffness
of muscle and tendon (Hirata etal. 2015; Konrad and Tilp
2014a, b). Therefore, the measurements were performed during the 3rd cycle and later (PRE). Then, immediately after a
5-min static stretching (see below), the same passive lengthening procedure was applied (POST) to the pre-determined
end ROM.
Static stretching maneuver
Static stretching was administered to the right lower leg
of the subject. The posture of the subject and setup were
similar to those for the muscle stiffness measurement as
mentioned above. The ankle joint angle was held at the end
ROM of dorsiflexion for 5min. Throughout the stretching,
the subjects were requested to relax completely and not
offer any voluntary resistance.
Ultrasound shear wave elastography
Two ultrasound shear wave elastography systems (Aixplorer Ver. 6 and Ver. 7, Supersonic Imagine, France) with
a 50mm linear array probe (SL15-4, Supersonic Imagine,
France) with musculoskeletal preset were used to quantify
the passive stiffness of each muscle and the Achilles tendon
(AT). Each ultrasound probe was placed over the bellies of
MG and LG, over the medial or lateral aspect of Sol distal
to muscle-tendon junction of MG or LG, or over AT, along
the fascicle direction. The optimal probe location was determined for subject-by-subject to identify several fascicles
without interruption across the B-mode image in a certain
plane and get good-looking images. The determination of
probe location was performed prior to the placement of electromyography (EMG) electrodes. Since four targets (three
muscles and one tendon) were investigated with two systems
in the present study, the passive lengthening as mentioned
above was repeated over the same ROM for assessing the
other targets. The order of testing for MG, LG, Sol, and AT
within the two passive lengthening was randomly assigned
and counterbalanced across subjects. Care was taken not to
press and deform the muscles while scanning.
EMG
In order to ensure that the lengthening procedures were
conducted under passive condition, surface EMG signals during the prescribed tasks were obtained from MG,
LG, and Sol. After shaving, rubbing with sandpaper, and
cleaning with alcohol, pre-amplified (gain: 500, bandpass filtering: 5450Hz) bipolar active surface electrodes
(electrode shape: parallel-bar, size: 1mm width8mm

length, inter-electrode distance: 12mm; FA-DL-141,

4-assist, Japan) were placed next to each ultrasound position of MG, LG, and Sol, along the fascicle direction of
each muscle. The reference electrode was placed over the
left medial malleolus. The torque, joint angle, and EMG
data were simultaneously stored on a personal computer
for later analyses using a 16 bit analogue-to-digital converter (PowerLab/16SP, ADInstrument, Australia) with a
sampling frequency of 1kHz. Also, these data were manually synchronized with elastography recording (sampling
frequency=1Hz).
Validity andrepeatability ofshear wave elastography
measurements
The validity of measurement of shear modulus by using the
ultrasound shear wave elastography has been described in
detail in previous reports (Miyamoto etal. 2015b; Yoshitake etal. 2014). Briefly, the validity was evaluated in tissue-mimicking phantoms which were calibrated by a conventional stressstrain test. The Youngs modulus measured
by shear wave elastography was highly correlated with
measured values of Youngs modulus obtained via traditional materials testing (r>0.99), with the regression line
(y=1.01x5.45) close to the identity line. The inter-trial
absolute difference was 0.3kPa. The repeatability during
the passive dorsiflexion was evaluated in three subjects
for each muscle. The intra-class correlation coefficients
(1, 2) were 0.990.01, 1.000.00, and 0.970.2 for
MG, LG, and Sol, respectively. Absolute differences were
1.20.1, 0.70.3, and 0.80.3kPa for MG, LG, and
Sol, respectively. Additionally, previous studies have suggested that shear modulus measured by ultrasound shear
wave elastography could be considerably affected by pennation angle (Gennisson etal. 2010; Masetti etal. 2012).
However, more recently we have shown that the effect of
pennation angle on the measured shear modulus was negligibly small (<1.3%) (Miyamoto etal. 2015a).
Data analysis
The passive torque was corrected for the gravity effect of
the footplate. All passive lengthening procedures used for
measurements of muscle stiffness were performed up to the
end ROM. However, data from 50 plantarflexion up to 25
dorsiflexion were used for the analyses, as all subjects could
reach this ankle angle before and after a 5-min static stretching. For EMG data, the root mean square values (RMSEMG) were computed over 0.5s period at each joint angle
for each muscle. Then, the RMS-EMG value of each muscle
was normalized to that obtained during isometric maximal
voluntary contraction (MVC) which performed at 0 of the
ankle joint. For elastographic data, the data processing was

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Eur J Appl Physiol

performed by using software of the ultrasound scanner for

every 1 of the ankle joint. Actually, the software provides
the Young modulus by multiplying the shear modulus by the
constant 3, based on the assumption of an isotropic material,
which is obviously not true for the muscle. Hence, we calculated shear modulus by dividing the obtained Youngs modulus by 3. Depending on the subject and target, a circular area
as large as possible with exclusion of aponeurosis and subcutaneous adipose tissue (Fig.1c) was chosen as the region
of interest for the shear modulus calculation. No value in the
circular region reached the saturation limit of shear modulus
for the elastography system (267kPa) except for AT. Based
on the shear modulus-ankle joint relationships, the slack
angles of each muscle were determined as the first increase
above the variation in shear modulus at the plantarflexed
position (Fig.1b). This determination of the slack angle was
visually conducted (Hirata etal. 2015; Lacourpaille etal.
2014) by three experimenters who were blinded to subject
identification, target, and time (i.e., PRE and POST) information. The mean value of the three experimenters was used
for further analyses. Due to the saturation limit of shear
modulus for the elastography system, the shear modulus of
AT were saturated at about 25 (i.e., 25 of plantar flexion). Thus, shear modulus measurements of AT were used
only for the determination of the tendons slack angle.
Statistics
Statistical analyses were performed for the values at every
5 of the ankle joint angle. For the RMS-EMG and shear
modulus data, separate three-way analyses of variance
(ANOVAs) [time (i.e., PRE, POST)muscleangle]
with repeated measures were used. Two-way ANOVAs with
repeated measures were used for the torque (timeangle)
and slack angle [timetarget (i.e., MG, LG, Sol, AT)].
When appropriate, additional analyses (Tukey and paired
t-tests with Bonferroni correction) were performed. For
maximal ROM data, a paired t test was performed. Pearsons product-moment correlation coefficient (r) was determined to assess the correlation between changes in passive
torque and shear modulus. The significance level for all
comparisons was set at P<0.05. Data are reported as means
and SD. The statistical analyses were performed by using
statistical software (SPSS Statistics 21, IBM Japan, Japan).

Results
ROM, passive torque andEMG
After 5min static stretching, ROM was significantly increased from 28.94.6 to 31.25.8
(ROM =2.32.0) (P =0.003). Passive plantar

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Fig.1Typical example of responses in passive plantar flexion torque

(a) and shear modulus (b) of the medial (MG; circles) and lateral gastrocnemius (LG; triangles), soleus (Sol; diamonds), and Achilles tendon (AT; squares) during passive lengthening before a static stretching.
Arrows in shear modulus-joint angle curves indicate the slack angles
of MG, LG, Sol, and AT. The bottom images (c) are typical examples
of shear modulus measurements of MG obtained at 40, 0, and 20
of dorsiflexion angle. The colored region represents the shear modulus map with the scale to the right of the figure. Circle is the region of
interest for determination of shear modulus (color figure online)

flexion torque increased exponentially with an increase in

the ankle joint angle (Fig.1a). The passive torque was
reduced after a 5-min static stretching, which reached
significance at 1025 (P<0.05, Fig.2); the reduction
at 25 was 6.98.2Nm. The EMG activities during
the passive lengthening were relatively small (<1.0%
MVC) at plantar flexed positions whereas they were
slightly increased toward the end ROM (<3.2% MVC)
for each muscle, despite the subjects being asked to
relaxed during the passive lengthening. For RMS-EMG
data, a three-way ANOVA revealed that there was only a
significant main effect of angle (P=0.014) without any
interactions.

Eur J Appl Physiol

Fig.2Changes in passive plantar flexion torque during passive

lengthening before (PRE; filled) and after (POST; open) a 5-min
static stretching. *Significantly different between PRE and POST
(P<0.05)

Fig.3Changes in shear modulus of the medial (MG; circles) and lateral gastrocnemius (LG; triangles), and soleus (Sol; diamonds) during passive lengthening before (PRE; filled) and after (POST; open)
a 5-min static stretching. *Significantly different between PRE and
POST in MG (P<0.05)

Shear modulus
Slack angle
Figure 1b represents examples of ultrasound images
and of the shear modulus-ankle joint angle relationship, showing the characteristic curvilinear relationships
with shear modulus increasing steeply toward the end
ROM, especially in MG. Figure3 shows pooled data of
shear modulus of each muscle during passive lengthening before and after the static stretching. A three-way
ANOVA showed a significant timemuscleangle
interaction (P =0.022). Follow-up two-way ANOVAs
revealed that a significant timeangle interaction was
observed only for MG shear modulus whereas there was
only a significant main effect of Angle (P<0.001) without main effect of time or timeangle interaction for
LG and Sol. According to further analyses, the shear
modulus of MG after the static stretching was significantly smaller than that before the stretching at 1025
of the ankle joint angle (Fig.3); the reduction at 25 was
29.417.2kPa (20.38.9% of PRE value). Additionally, Tukey post hoc tests demonstrated that shear modulus was significantly greater in MG than in LG and Sol
above 5 and greater in LG than Sol above 5 both
before and after the stretching (P<0.001).
Correlation analysis was further performed to examine
a potential association between changes in passive torque
and shear modulus of MG. The relative change [i.e., (POST
valuePRE value)/PRE value] in passive torque was significantly correlated to that in shear modulus of MG across
subjects (r=0.620, P=0.031).

As observed in Fig.1b, the slack angle differed among the

three muscles. Table1 shows the slack angles of the triceps surae and AT before and after the static stretching. A
two-way ANOVA showed significant main effects of time
(P =0.001) and target (P<0.001) without timetarget
interaction. The mean difference in slack angle between PRE
and POST across Target was 1.8, showing that the slack
angles of each muscle and AT were significantly shifted
toward more dorsiflexed positions after the static stretching.
A Tukey post hoc test revealed that the slack angle was significantly smaller (i.e., more plantar flexed position) in the
following order: AT<MG<LG<Sol (P<0.05).
Effect ofslack angle onshear modulus
Figure 4 shows the relationships between shear modulus
and ankle angle from slack for each muscle. Separate twoway ANOVAs for each muscle showed that a significant
time angle interaction (P =0.003) was observed only
for MG whereas there was only a significant main effect of
angle without main effect of time or timeangle interaction for LG and Sol. Follow-up analyses demonstrated that
shear modulus of MG after the static stretching was significantly smaller than that before the stretching at 30 and 35
from the slack angle (P =0.045 and P =0.008, respectively) although the magnitude of the reduction in shear
modulus after the stretching was considerably smaller

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Eur J Appl Physiol

Table1Slack angles of the medial and lateral gastrocnemius, soleus,

and Achilles tendon before and after a static stretching
Dorsiflexion angle at slack ()

MG
LG
Sol
AT

Before

After*

19.14.9
14.05.8
4.12.0

17.25.0
12.55.5
2.07.3

44.82.3

43.32.8

MG medial gastrocnemius, LG lateral gastrocnemius, Sol soleus, AT

achilles tendon
* Significant main effect of time without timetarget interaction
(P<0.05). Values are meanSD

Fig.4Relationships between shear modulus and ankle angle from

slack for the medial (MG; circles) and lateral gastrocnemius (LG;
triangles), and soleus (Sol; diamonds) before (PRE; filled) and after
(POST; open) a 5-min static stretching. *Significantly different
between PRE and POST in MG (P<0.05)

(4.6kPa; 6.8% of PRE) compared with that based on the

shear modulus-joint angle relationship (Fig.3).

Discussion
The major findings of the present study were that (1)
shear modulus during passive lengthening of the triceps
surae was the greatest in MG and smallest in Sol, (2) the
effect of a static stretching on shear modulus was significant only in MG, and (3) the magnitude of the reduction
of MG shear modulus after the static stretching became
smaller when considering the shift in slack angle induced
by the stretching. Shear modulus measured by ultrasound
shear wave elastography has been shown to be highly correlated with the material stiffness (Youngs modulus) of a

13

muscle measured by a conventional stressstrain test (Eby

etal. 2013). Thus, the present findings suggest that passive
muscle stiffness differs among the triceps surae, and that
the acute effect of a static stretching is observed only in the
stiff muscle (i.e., MG). However, a large part of the reduction of passive muscle stiffness at a given joint angle could
be due to an increase in the slack length.
Intermuscle differences inpassive stiffness
Previous invitro studies have shown that type I muscle
fibers have greater passive stiffness compared with type II
muscle fibers (Mutungi and Ranatunga 1996, 1998), possibly due to differences in titin isoform within each fiber
(Wang etal. 1991) and/or in content of collagen which
composes intramuscular connective such as the perimysium and endomysium (Kovanen etal. 1984a; Zimmerman etal. 1993). At the start of this study, therefore, it was
expected that, during passive lengthening of the triceps
surae, Sol composed of 70100% type I muscle fibers
exhibits greater muscle stiffness than MG and LG composed of >50% type II muscle fibers (Johnson etal. 1973).
However, contrary to the expectation, muscle stiffness at
a given ankle joint angle was greater in MG and LG than
Sol. This result suggests that factors other than muscle fiber
type composition should be involved to determine the intermuscle differences in passive stiffness among the triceps
surae. Kawakami etal. (2000) have reported that the sarcomere length of MG is longer than that of Sol at a given
ankle joint angle in the knee extended position. Additionally, the slack angle was observed at more plantar flexed
position in the following order: AT, MG, LG, and Sol. Taking these observations into account together with the finding that passive force generally develops at or near the optimal length (Davis etal. 2003; Friden and Lieber 2002), it is
possible that the passive stiffness of the individual muscles
of the triceps surae is strongly influenced by architectural
characteristics such as sarcomere length and corresponding
slack length.
Theoretically, passive stretching elicits the stretch
reflex, which would stiffen the lengthened muscles.
In this study, the rate of lengthening was set at a sufficiently slow angular velocity (1/s) to avoid or minimize
the stretch reflex. Nevertheless, the EMG activities during the lengthening procedure were negligible at plantar
flexed positions (<1% MVC) whereas the EMG activities were slightly increased toward the end ROM for each
muscle (<~3% MVC). Thus, the reflexive muscle activities would likely have contributed significantly to the
muscle stiffness during passive lengthening. However,
the present result that regarding RMS-EMG only a significant main effect of Angle was observed without any
interactions indicates that the muscle activities during the

Eur J Appl Physiol

lengthening procedure were not different among the three

muscles and between before and after the static stretching.
Thus, it is unlikely that the muscle activities during passive lengthening are responsible for the observed differences in passive stiffness between muscles and between
pre- and post-stretching.
Acute effect ofstatic stretching onMG passive stiffness
The stretching maneuver adopted here increased ROM and
decreased passive torque at a given ankle joint angle. These
results indicate that the stretching maneuver could be effective to increase the flexibility of the entire plantar flexor
muscle group. However, after the static stretching, passive
muscle stiffness was significantly reduced only in MG. In
other words, the static stretching could not alter the passive muscle stiffness of LG and Sol. Freitas etal. (2015)
have revealed that MG passive stiffness measured by ultrasound shear wave elastography was reduced only when sufficient mechanical stress was imposed on the muscle during
stretching. During the static stretching in the present study,
the ankle joint angle was held at the end ROM of dorsiflexion, where shear modulus of LG and Sol was less than half
of that of MG (Fig.3). Therefore, it is reasonable that the
stretching effect was observed only in MG.
The muscle stiffness of MG was significantly reduced
by 20% at 25 after the static stretching (Fig.3). The
significance remained even when considering the shift
in slack angle induced by the static stretching (Fig.4).
Thixotropic property of a muscle has been proposed as
one of the possible mechanisms for the reduced muscle
stiffness after the static stretching (Rassier etal. 2005).
However, it is well known that muscle stiffness of this
nature can be reduced by relatively small amounts of
movement (Campbell and Lakie 1998). Moreover,
Morse etal. (2008) have subjected the plantar flexors
including MG to a number of quick stretches that would
be expected to abolish the short range elastic component or possibly change the conformation of titin. They
concluded that thixotropic properties of a muscle are
unlikely to be the mechanism leading to increased flexibility as a result of static stretching. Another mechanism
is decreased stiffness of intramuscular connective tissues
(Morse etal. 2008). As mentioned above, the connective tissue, particularly the perimysium, is considered to
be a major extracellular contributor to passive stiffness
(Purslow 1989). The ultrasound shear wave elastography scanner used in the present study can quantify localized tissue stiffness and we have taken care to exclude
the outside connective tissues such as epimysium, fascia,
and aponeurosis from the analyses of shear wave elastographic data. Gajdosik (2001) suggested that lengthening
deformation of the connective tissues within the muscle

belly (e.g., perimysium and endomysium) could influence

passive stiffness. Thus, the reduction in passive muscle
stiffness of MG after static stretching observed in the present study could be due to the deformation of the perimysium and/or endomysium, not to that of the epimysium.
However, the extent of the substantial reduction of MG
passive stiffness was considerably smaller (7%) when
considering the shift in slack angles after the stretching (Fig.4). Additionally, in the present study, the slack
angle was significantly observed at more plantar flexed
position in the following order: AT<MG<LG<Sol.
After the stretching, the slack angles of each muscle and
AT were significantly shifted toward more dorsiflexed
positions with a similar extent. Taking these observations
into account together with the fact that the triceps surae
and AT are serially connected, the 2 shift of the slack
angle in AT necessarily induces the same degree of the
slack angles in the triceps surae. Therefore, it is possible
that the reduction of MG muscle stiffness at the dorsiflexed positions after the stretching is attributed mainly
to the shift of slack angle of AT.
A limitation of the present study is that passive lengthening and stretching were performed only in the knee
extended position, in spite of the fact that MG and LG are
bi-articular muscles crossing for the knee and ankle joints.
Thus, it is unclear whether the differences in passive muscle stiffness and slack angle among triceps surae (MG or
LG vs. Sol) in the knee extended position can be observed
in the knee flexed position. Another limitation is that shear
modulus measurement was performed at only one region
for each muscle. Especially, Sol has a complex multipennate arrangement of the muscle fibers (Agur etal. 2003;
Hodgson etal. 2006). Thus, it is unclear whether the current findings hold true for other regions (e.g., proximal
vs. distal, medial vs. lateral) within each muscle. Further
investigation is warranted to clarify these.
In conclusion, the present study has shown that passive
muscle stiffness during passive lengthening of the triceps
surae was the greatest in MG and smallest in Sol. Additionally, after a 5-min static stretching of the triceps surae,
muscle stiffness was reduced only in MG. These findings
suggest that the acute effect of a static stretching on passive
muscle stiffness is muscle-specific and occurs only in the
stiff muscle. However, a large part of the reduction of passive muscle stiffness at a given joint angle could be due to
an increase in the slack length.
Acknowledgments This work was supported by JSPS KAKENHI
Grant Number 25702038.
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict
of interest.

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