European Journal of Orthodontics 12 (1990) 370-379 © 1990 European Orthodontic Society

Myotomy of the lateral pterygoid muscle and

condylar cartilage growth
Robert J. Hinton
Department of Anatomy, Baylor College of Dentistry, Dallas, Texas, U.S.A.

SUMMARY The influence of lateral pterygoid muscle activity on growth of the condylar cartilage
of the mandible remains controversial. As a case in point, two recent studies involving total
resection (myectomy) and sectioning (myotomy) of this muscle have come to contradictory
conclusions. In the study reported here, it was found that young growing rats in which the lateral
pterygoid muscle was bilaterally cut experienced significantly retarded mitotic activity and bone
formation relative to their sham-operated counterparts. Changes in the shape and topographical
distribution of cartilage over the condylar head, as well as the distribution of labelled cells, were
also noted in myotomy animals at 4 days following surgery.
Considerable resorption at the insertion of the muscle on the mandibular ramus was apparent at
14 days after surgery. However, the myotomy surgery was accompanied by a significant post-
operative loss of body weight and incisal supra-eruption indicative of feeding difficulties, thereby
complicating interpretation of the results.

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Due largely to the work of Petrovic and associ-
ates (Charlier et al, 1969; Petrovic, 1972; Petro-
vic et al., 1975; Petrovic and Stutzmann, 1977),
the functional activity of the lateral pterygoid
muscle is widely regarded as an important media-
tor of growth at the condylar cartilage of the
mandible. However, most studies have employed
indirect means (e.g. orthodontic appliances) of
presumably altering the activity of this muscle,
and only rarely (McNamara, 1980) has verifica-
tion of this altered activity been obtained.
Recently, it has been shown that chronic stimula-
tion of the lateral pterygoid muscle directly via
in-dwelling electrodes produces an augmen-
tation in the thickness of the condylar cartilage
and an increase in the overall length of the
mandible (Kantomaa and Ronning, 1982).
Petrovic and associates (Petrovic et al., 1975)
have reported that reduction or elimination of
lateral pterygoid functional activity following
total resection (myectomy) of the muscle signifi-
cantly reduces the number of prechondroblasts
undergoing mitosis in the cartilage, as well as
overall mandibular length. However, subsequent
studies have concluded that growth of the condy-
lar cartilage, as measured by fluorescent markers
in the subchondral bone, is not inhibited by
cutting (myotomy) of the muscle (Goret-Nicaise
etai, 1983; Awn et al., 1987). The purpose of this
study was to investigate the short-term effect of
bilateral myotomy of the lateral pterygoid
muscle on the growth of the condylar cartilage,
with an emphasis on the precise nature of the
metabolic and morphologic changes effected in
the cartilage.
Materials and methods
In 26 day-old male Sprague-Dawley rats, the
fascia overlying the posterior fibres of the tem-
poralis muscle was incised and the muscle fibres
carefully elevated to expose the superior aspect
of the infratemporal region. The condylar neck
was then grasped with forceps and pulled later-
ally, thereby straightening the course of the
lateral pterygoid muscle and enabling it to be cut
by microscissors thrust downward into the fossa.
This straightening of the muscle usually resulted
in it being cut near its midpoint or closer to its
origin, thereby minimizing trauma near the
condyle that might result in fibrosis and limi-
tation of movement. This procedure was per-
formed bilaterally on all myotomy group ani-
mals. A second group, in which the
infratemporal region was exposed surgically and
the condylar neck pulled laterally, but with no
LATERAL PTERYGOID MYOTOMY
cutting of the muscle, served as a sham surgery
group.
Twenty-eight of the animals were killed 4 days
following surgery by intravenous injections of
sodium pentobarbital; the other 24 were killed 14
days following surgery. The degree to which the
lateral pterygoid muscle had been damaged was
determined by visual inspection and recorded at
sacrifice. Two hours prior to death, six to eight
animals from each group were injected intrave-
nously (femoral vein) with 1 j*Ci/g body weight
of [45Ca]-chloride in water (New England Nuc-
lear, specific activity = 21.9 mCi/mg). One hour
prior to death, another six to eight animals from
each group were injected intravenously with 2
fiCi/g body weight of [3H]-thymidine (New Eng-
land Nuclear, specific activity = 6.7 Ci/mmol).
At sacrifice, both condyles were removed; one
was randomly chosen for biochemical assay, and
the other for histology and radioautography. In
the condyle designated for analysis of [3H]-
thymidine incorporation, the articular disc was
carefully detached so as not to damage or strip
any part of the prechondroblastic zone of the
cartilage. The condylar cartilage was then
severed from the ramus at the cartilage-bone
interface, dissected free of excess soft tissue, and
frozen at — 20°C for later biochemical assay.
Histological examination of the condylar tissue
thus removed for analysis confirmed that the
sample to be assayed contained little bone,
ensuring that the radioactivity derived largely
from labelled cells in the prechondroblastic zone.
Condyles reserved for biochemical analysis to
determine [3H]-thymidine incorporation were
first homogenized in ice-cold 10 percent trichlor-
acetic acid. (TCA) containing 1 mM thymidine.
The homogenates were placed on ice for 30
minutes, then centrifuged at 3000 g and 4°C for
15 minutes. The precipitates were washed twice
with ice-cold 10 per cent TCA without thymi-
dine, then hydrolysed for 20 minutes in 2 ml of 10
per cent TCA at 90°C. Following hydrolysis, the
tubes were kept on ice for 15 minutes, then
centrifuged at 3000 g for 10 minutes at 4°C. An
aliquot (500 y\) of the supernatant was added to
10 ml of ACS (Aqueous Counting Scintillant,
Amersham) and assayed for radioactivity in a
Beckman LS 7500 Scintillation counter. A
further aliquot was utilized to estimate DNA
content using the diphenylamine method (Bur-
ton, 1956). Calf thymus DNA (Sigma) served as
standard, and DNA content was read at 600 nm.
371
The results were expressed as dpm//ig DNA in
the acid insoluble precipitate.
The mandibular condyles designated for
biochemical analysis of [45Ca]-uptake were
removed, weighed, and then homogenized in 2
ml ice-cold 0.15 NaCl containing 0.003 M
NaHCO 3 at pH 7.4. Following centrifugation at
3000 g for 60 minutes at 4°C, the sediment was
stirred in 5 ml 0.1 M CaCl2 in 0.05 M Tris-HCl
buffer, pH 7.4 at 25°C for 30 minutes, then
centrifuged at 3000 g for 30 minutes. The
sediment was washed twice in 10 ml lots of 5 mM
Tris-HCl, pH 7.4 and centrifuged at 3000 £ for 30
minutes. Following washing, the sediment was
stirred in 10 ml 0.5 M HC1 for 2 hours at 25°C.
After centrifugation at 3000 g for 30 minutes, an
aliquot (750 /il) of the supernatant was added to
10 ml of ACS (Aqueous Counting Scintillant,
Amersham) and assayed for radioactivity in a
Beckman LS7500 scintillation counter. The
results were expressed as cpm/mg tissue.
In condyles reserved for radioautographic and
histological analysis, the disc and ramus were left
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intact; following routine histological processing
and paraffin embedding, they were sectioned in
either the coronal or sagittal plane at 5 microns.
Sections selected from the mid-condylar region
were then dipped in Kodak nuclear track emul-
sion (NTB II), dried, and placed in a light-tight
box at 4°C for 6 weeks. The exposed slides were
then developed in Kodak D-19 for 6 minutes,
rinsed in distilled water, and placed in Kodak
rapid fixer for 4 minutes. Following a wash in
distilled water, the slides were lightly stained in
haematoxylin and eosin. In several animals from
each group, representative sections were traced
to assess the localization of the radioactive label.
A cell was considered labelled if five or more
grains lay over the nucleus. Utilizing a Nikon
Optiphot microscope fitted with a Leitz drawing
tube, the location of each labelled cell along the
contour of the condyle was plotted under dark-
field illumination at a magnification of approxi-
mately x 80.
Results
Post-surgical growth
By 4 days following surgery, the incisors in the
myotomy group were supra-erupted in virtually
every case. In addition, the lateral pterygoid
muscle was found to be cut over one-third to half
172: R. J. HINTON

Table 1 Percentage body weight gain/lossf. Mean + SD. characterized the muscle in the sham surgery
group. By 14 days following surgery, the muscle
Time after surgery was found at death to be cut over half or more of
14 days
its cross-section in all but one myotomy animal.
4 days 7 days
Incisal supraeruption was still present in most
Sham surgery 30.6±4.9 70.4 ±6.5 135.3+12.2 myotomy animals, although to a lesser extent
Myotomy -15.3 + 8.2* 7.4+13.3* 27.3+15.0* than at 4 days.
(7) (6) (6) Considerable intergroup differences in weight
gain were present at both death intervals (Table
• Mean is different from sham surgery group at 1). At 4 days post-surgery, myotomy group
t Calculated as a percentage of body weight immediately
prior to surgery. animals had lost on average 15 per cent of their
pre-surgical body weight, whereas the sham
surgery group had experienced approximately a
its height (i.e. through the superior head and well 31 per cent gain in body weight since surgery.
into the inferior head) in half of the animals and Inspection of weight gain at 7 days post-surgery
cut nearly through in the other half. Upon indicated that the beginnings of weight gain by
examination at death, the lateral pterygoid the myotomy group animals had begun by this
muscle in the myotomy group appeared quite time interval. By 14 days post-surgery, the
pale in comparison to the reddish colour which myotomy animals had gained weight noticeably,

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Figure 1 (a,b) Coronal sections of representative condyles

from 4-day sham surgery (a) and myotomy (b) groups. In
each case, the medial aspect is to the right. Note the greatly
attentuated thickness of cartilage on the lateral aspect in the
myotomy animal, as contrasted with the more even distribu-
tion in the sham surgery animal. This myotomy animal, while
typical, does not by any means represent the most extreme
skewing of cartilage thickness observed in this group. (H & E.
Bar represents 400 //m.)
Figure 2 (a,b) Higher power view of same condyle as in Fig.
lb, showing lateral (a) and medial (b) aspects of condylar
cartilage in myotomy animal. A major part of the difference
in cartilage thickness appears to be due to the greatly
diminished size of the layer containing differentiated, but not
yet hypertrophied chondrocytes (arrows) on the lateral side.
(H & E. Bar represents 200 //m.)
LATERAL PTERYGO1D MYOTOMY 373

but at a significantly slower rate than the sham
surgery group.
Morphology and radioautography
4 Days after surgery. At 4 days following surgery
the cartilage in the myotomy group was consis-
tently different from sham surgery animals in its
morphology in coronal (frontal) section and to
a lesser degree in sagittal section. In coronal
section, the sham surgery condyles exhibited a
relatively even thickness of cartilage extending in
a sickle-shape from the medial to the lateral poles
of the condylar head (Fig. la). In contrast, the
myotomy group condyles were characterized by
a strongly asymmetrical distribution of the carti-
lage, with very thin cartilage laterally giving way
to a considerably thicker cartilage medially,
often peaking in thickness in the medial one-
third of the contour (Fig. lb). The reduced
thickness on the lateral aspect of the cartilage
appeared to be largely due to a greatly attenuated
chondroblastic region (Fig. 2a,b). This atypical

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Figure 3 (a-f) Drawings of the distribution of [3H]-thymidine labelled cells in three animals from each group killed 4 days
following surgery. Coronal sections are depicted, with each dot representing the position of one labelled cell. In each instance, the
medial aspect is to the right. Thin irregular line delineates the approximate location of the cartilage-bone interface, (a-c) Sham
surgery, (d-f) Myotomy.
374
distribution of cartilage was a common, but not
universal finding in the myotomy sample, being
present in seven of nine animals for which
coronal sections were available. Inspection of
plots of the distribution of labelled cells showed
that the preponderance of labelled prechondro-
blastic cells in the myotomy animals were found
in the medial half of the cartilage, with very few
to none laterally. By comparison, the distribu-
tion in the sham surgery animals was much more
uniform (Fig. 3a-f). In both groups, uptake of
the [3H]-thymidine label was confined almost
exclusively, as would be expected, to the pre-
chondroblastic layer of the cartilage and to the
subchondral bone deep to the cartilage, with an
occasional labelled cell in the articular disc or
fibrous articular layer of the cartilage. Resorp-
tion at the insertion of the lateral pterygoid
muscle on the condylar neck appeared to be
accentuated in some myotomy animals relative
to the sham surgery group.
In sagittal section at 4 days, the condylar
(b),
Figure 4 (a,b) Superior aspect of condylar cartilage in
representative 4-day sham surgery (4a) and myotomy (b)
animals. Anterior is to theright.(Sagittal section, H & E. Bar
represents 400 ^m.)
R. J. HINTON
cartilage in the sham surgery group was crescent-
shaped, gradually increasing in thickness from
anterior to posterior, attaining its greatest thick-
ness at the posterosuperior and posterior aspects.
The cartilage in the animals which had under-
gone myotomy 4 days earlier was considerably
thinner over the anterior and superior aspects of
its contour than in the sham surgery group (Fig.
4a,b), often constituting a zone only a few cell
layers thick. In addition, in some myotomy
animals the cartilage appeared truncated on its
posterior aspect, with the tissue in this region
resembling bone more than cartilage (Fig. 5b); in
sham surgery animals, a cartilaginous tissue of
varying thickness was present in this region (Fig.
5a). Although the appearance and thickness of
cartilage in the posterior region of the condyle
varied in the sham surgery group from medial to
lateral as well as between animals, nothing
comparable to the appearance described above
in certain myotomy animals was observed in any
of the sham surgery condyles. However, this
finding was not typical of all myotomy animals.
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14 Days after surgery. In coronal section, the
condylar cartilage in myotomy animals showed
little of the tissue asymmetry that was so typical
of the 4-day myotomy animals (Fig. 6a),
although the cartilage thickness on the lateral
aspect was still attenuated in some animals
relative to sham surgery animals. However, the
labelled prechondroblastic cells were still largely
found in the medial half of the cartilage as noted
in the 4-day myotomy animals. The most striking
aspect of the 14-day myotomy animals was the
considerable resorption that had taken place on
the medial side of the ramus at the insertion of
the lateral pterygoid muscle. Although some
resorption was characteristic of this area in sham
surgery animals, the resorption in the myotomy
animals was so extensive as to undermine the
medial extent of the condylar cartilage in many
cases (Fig. 6b). This phenomenon resulted in the
condyle head appearing 'bent' medially and the
ramus thinned compared to sham surgery ani-
mals (Fig. 6c). In sagittal section, this extreme
resorption resulted in a very characteristic scal-
loping of the condylar neck in the more medial
sections with a normal appearance laterally. The
appearance of the cartilage in 14-day myotomy
animals was in general more similar to that of the
other (sham) group, although some myotomy
animals exhibited greatly thinned cartilage with a
LATERAL PTERYGOID MYOTOMY
(i
Figure 5 (a,b) Posterosuperior and posterior aspect of condylar cartilage in representative sham surgery (a) and myotomy (b)
animals. Anterior is to the left. (Sagittal section, H & E. Bar represents 400 ^m.)
reduced number and thickness of hypertrophic
cells.
Biochemical analysis
Two-way analysis of variance with group and
time interval as factors indicated significant
effects for both group (F=35.7, P^O.001 and
time (F=11.3, PsSO.01) for [3H]-thymidine
incorporation, but the interaction of the two
factors was not significant. A similar result was
obtained for [45Ca]-uptake: both group (F= 23.5,
P^0.001) and time effects (F=7.3, /><0.05)
were significant, but not their interaction.
[3H]-thymidine incorporation (dpm/^g DNA)
was significantly reduced in the myotomy sample
compared to the sham surgery group by 4 days
following surgery, and continued to be so 14 days
following surgery (Table 2). [45Ca]-uptake (cpm/
mg tissue) was similarly affected at both post-
surgical intervals. Values for both parameters,
even though corrected for differences in tissue
weight or DNA content, were in the myotomy
375
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group routinely one-half to one-third the corres-
ponding figure in the sham surgery group. [3H]-
thymidine incorporation, but not [45Ca]-uptake,
declined significantly over time within each
group.
Although labelling indices were not calculated
due to small sample sizes, the sections taken for
[3H]-thymidine radioautography supported the
results obtained from the biochemical assay of
[3H]-thymidine incorporation. As Fig. 3a-f
show, the primary difference between the myo-
tomy animals and the sham surgery group lay in
the greatly reduced number of labelled cells in the
prechondroblastic layer of the cartilage, with
more similar labelling in the subchondral bone.
Discussion
The biochemical analyses of [3H]-thymidine in-
corporation and [45Ca]-uptake support the ear-
lier findings of Petrovic and co-workers using a
rat myectomy model. They found that the
376 R. J. HINTON

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Figure 6 (a) Coronal section of condyle in 14-day myotomy animal. Note resorption at the insertion of the lateral pterygoid
muscle (LPM), 'undermining' the condylar cartilage. (H & E. Bar represents 400 fim.) (b) Higher-power view of insertion of
lateral pterygoid muscle on condylar neck in myotomy animal, showing extensive resorption (arrows), (c) Similar view of
insertion of lateral pterygoid muscle in a sham surgery animal. (Bar represents 400 /im for b and c.)
LATERAL PTERYGOID MYOTOMY
Table 2 [3H]-thymidine incorporation and [45Ca]-uptake in
sham surgery and myotomy groups. Mean + SD.
[3H]-thymidine
incorporation [45Ca]-uptake
(dpm//jg DNA) (cpm/mg tissue)
4 days after surgery
Sham surgery 440.8 + 75.2 786.1 + 172.6
(8) (6)
Myotomy 212.9 + 68.4** 434.9+120.9**
(6)
(7)
14 days after surgery
Sham surgery 305.2+139.0 569.5 + 169.2
(7) (5)
Myotomy 111.8 + 94.3** 309.8+115.6*
(7) (5)
* Mean is different from the sham surgery mean at
fZ0.05.
** Mean is different from the sham surgery mean at
number of [3H]-thymidine labelled prechondro-
blasts in the condylar cartilage decreased signifi-
cantly at 2-4 weeks following total resection of
the lateral pterygoid muscle; in addition, the
overall length of the mandible was significantly
shortened (Petrovic and Stutzmann, 1972; Stutz-
mann and Petrovic, 1974; Petrovic et al., 1975).
The results of my biochemical assays contrast
with the data obtained by Goret-Nicaise and
colleagues (Goret-Nicaise et al., 1983; Awn et al.,
1987) using a rat myotomy model. Using fluores-
cent markers in undecalcified bone sections to
measure the rate of bone deposition deep to the
condylar cartilage, they concluded that unila-
teral cutting of the lateral pterygoid muscle
resulted in no significant difference in the growth
rate of the condylar cartilage. Unfortunately,
rats of several different ages were utilized and/or
killed at differing ages, and it is not clear whether
the statistical analyses were performed between
age-matched samples or rather between the
entire experimental and the entire control
groups. In addition, their use of a unilateral
myotomy model leaves open the possibility that
the action of the intact muscle on the contra-
lateral side could affect the movements taking
place at the operated joint. The use of a bilateral
myotomy procedure in the present study tends to
mitigate against this possibility. My data, most
pertinently the [45Ca]-uptakefigures,would sug-
gest that a diminution in at least the amount of
mineralization occurs at 14 days following myo-
377
tomy, the only time interval which overlaps with
those studied by Awn et al. (1987). However, the
data of Awn and co-workers permitted the
examination of the rate of mineralization, so the
two sets of results are not strictly comparable. On
the other hand, Goret-Nicaise et al. (1983),
employing a similar model in older rats, noted
that myotomy of the lateral pterygoid muscle
appeared to produce 'complex deformations of
the condyle', including resorption at its anterior
border and increased bone growth at its pos-
terior border. Although their results were con-
fined to sagittal sections of the condyle, they bear
some similarity to the skewing of cartilage
thickness and resorption-associated deforma-
tions observed in my study.
Any interpretation of these results cannot
overlook the appreciable, often extreme, loss of
body weight which accompanied bilateral myo-
tomy of the lateral pterygoid muscle. Body
weight differences among the experimental
groups have not been reported in previous
studies, yet it appears likely that they occurred,
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probably to a significant degree. Since the lateral
pterygoid muscle is known to be strongly active
during incision in the rat (Weijs and Dantuma,
1975), it is hardly surprising that myotomy of
this muscle would greatly interfere with incision,
as evidenced by the supra-eruption of the inci-
sors. Supra-eruption of the incisors has also been
noted following total resection of the lateral
pterygoid muscle (Petrovic et al., 1982). In
animals provisioned with standard rat pellets,
the inability to incise would severely interfere
with feeding, so that it is not unexpected that a
significant loss of body weight occurred in the
myotomy sample in this study.
However, perhaps the most probable explana-
tion for. the sequelae of the myotomy surgery in
this study is that they represent the consequences
of hypofunction of the mandibular articulation.
It is well-established that motion is necessary for
the appearance (Hall, 1968) and maintenance
(Hall, 1972; Glineberg et al., 1982; McNamara,
1980; Hinton, 1985) of secondary cartilage. In
the case of the condylar cartilage, enhanced
protrusion of the lower jaw prompted either by
an intra-oral appliance (Petrovic, 1972; McNa-
mara and Carlson, 1979) or by electrical stimula-
tion of the lateral pterygoid muscle (Kantomaa
and Ronning, 1982) has also been found to
increase proliferation and/or thickness of the
condylar cartilage, whereas forced retraction of
378
the mandible (Petrovic et al, 1975; Asano, 1986)
or even amputation of the incisor teeth to
discourage incision (Hinton and Carlson, 1986;
Hinton, 1988) has the opposite effect. What is
not clear is whether the myotomy affects func-
tional movements of the mandible more through
its surgical sequelae (i.e. scarring) or through the
elimination of lateral pterygoid muscle pull with
the associated diminished movement or protru-
sive capability. Although some scarring may
have been present in the myotomy group ani-
mals, the extent of incisal supra-eruption in the
myotomy animals appeared no worse at 14 days
than at 4 days; in addition, in the last week before
sacrifice, the myotomy animals increased 20 per
cent in body weight compared to 38.1 per cent in
sham surgery animals—a lower, but not insub-
stantial, rate of weight gain. Thus, any scarring
that was present was presumably not severe
enough to greatly hamper the food intake capa-
bilities of the myotomy animals in the second
week following surgery.
The reason for the changes in the topographi-
cal distribution of cartilage and of [3H]-thymi-
dine labelled cells over the condylar head in
myotomy animals, which have apparently not
been previously reported, is unclear. It is possible
that they may reflect an altered pattern of
loading occasioned by the damage to the lateral
pterygoid muscle or perhaps an impairment of
transverse movements with an associated de-
crease in functional stimulus. However, data
which might permit evaluation of these possibili-
ties (e.g. histological sections which would illus-
trate the orientation of the condyle to the
mandibular fossa or data on mandibular move-
ments following surgery) were not obtained as
part of the study.
Address for correspondence
Dr Robert J. Hinton
Department of Anatomy
Baylor College of Dentistry
3302 Gaston Avenue
Dallas
TX 75246
U.S.A.
Acknowledgements
This study was supported by NIH grant
R. J. HINTON
DE06982. Thanks are due to Trina Morgan and
Sheryl Lumbley for their technical assistance.
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