European Journal of Orthodontic* 12 (1990) 51-76 O 1990 European Orthodontic Society

The structure of the periodontal ligament: an update

B. K. B. Berkovitz
Department of Anatomy, King's College London, England

Introduction ligament collagen is comprised of type III colla-

gen (Butler et al., 1975). The amount of type III
The functions of the periodontal ligament are of
collagen in adult connective tissues is normally
interest to a number of specialists. Its ability to
low, but in foetal (and granulation) tissues it is
remodel and allow for tooth movement is of
present in significant quantities (Chung and
interest to the orthodontist. Its ability to gener-
Miller, 1974; Epstein, 1974; Shuttleworth and
ate forces sufficient to move teeth is of interest to
Forrest, 1975; Barnes et al., 1976; Pierard
those concerned with the mechanism of tooth
and Lapiere, 1976). Immunofluorescence studies
eruption. Its ability to withstand the consider-
appear to demonstrate that type IN collagen is
able forces of mastication is of interest to those
not restricted to any particular zone of the
studying the mechanism of tooth support. Its
periodontal ligament, but is dispersed through-
ability to generate proprioceptive information
out (Wang et al., 1980; Takita et al., 1987).
important in the control of mastication is of
Furthermore, there is evidence that more type III
interest to neurophysiologists. However, study

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collagen may be present in erupting teeth corn-
of the periodontal ligament should not be limited
to those involved only in dental research. The
periodontal ligament has properties which make
it a particularly useful model to investigate
problems associated with soft fibrous connective
tissues, and connective tissue biologists are gra-
dually becoming aware of it. The periodontal
ligament has much in common with soft connec-
tive tissues elsewhere in the body. In addition, it
has other, somewhat unusual, features. An
appreciation of its many characteristics may
provide the basis for a greater understanding of
its biological behaviour. One factor to be borne
in mind when reviewing research on the perio-
dontal ligament is that most investigators have
used animal material.
Like all soft fibrous connective tissues, the
periodontal ligament consists of a fibrous stroma
in a gel of ground substance containing cells,
blood vessels and nerves. The fibrous stroma
consists primarily of collagen (with very small
amounts of oxytalan) and the cells are mainly
fibroblasts (Fig. 1).

Collagen in the periodontal ligament

Biochemically, the major type of collagen found
in the periodontal ligament is type I collagen.
This type of collagen also forms the major
protein of skin, bone, tendon and dentine.
Additionally, about 20 per cent of periodontal
Figure 1 Photomicrograph of decalcified longitudinal sec-
tion of periodontal ligament showing the general appearance
of the tissue. The bulk of the cells are fibroblasts, many of
which appear fusiform. A layer of osteoblasts lines the
surface of the alveolar bone (A) and a layer of cementoblasts
lines the surface of the cement (B). A blood vessel is arrowed.
52
pared to erupted (Takita et al., 1987). It is not
clear whether the two types of collagen are in
some way separated or linked within the micro-
structure of the ligament. There is evidence from
other sites that the two types of collagen mol-
ecule have the capacity to form crosslinked
copolymers. The molecule of both types of
collagen can form mixed fibrils with infinitely
varying proportions of the two components
(Henkel and Glanville, 1982). Pathological con-
ditions can arise which are characterized by an
alteration in the ratio of types I and III collagen
(for example, Penttinen et al., 1975; Pope et al.,
1975). The functional significance of type III
collagen within the penodontal ligament is not
known, although it might relate to properties
such as collagen turnover, the determination of
collagen fibril diameters or the provision for
some degree of mobility.
Recently, the presence of type XII collagen (a
non-fibril linking form) has been demonstrated
in the periodontal ligament, although its func-
tional significance is not known (Dublet et al.,
1988).
Although it was once claimed that the major-
ity of collagen within the periodontal ligament
was randomly orientated to produce the appear-
ance of an indifferent fibre plexus (Shackleford,
1971, 1973; Svejda and Skach, 1973), this view is
no longer held, the indifferent fibre plexus being
regarded as probably an artefact related to the
method of preparation (Sloan et al., 1976).
Collagen is now generally accepted as being
arranged in the form of principal bundles, having
different orientations in different parts of the
ligament (i.e. crestal, horizontal, oblique, apical,
interradicular). The collagen fibre bundles form
branching networks which course around the
neurovascular bundles (Fig. 2). Close to cemen-
tum the bundles are 3-10 /im in diameter, while
near the alveolar wall they are less numerous but
thicker, with a diameter of 10-20 /im (Zwarych
and Quigley, 1965; Shackleford, 1971; Sloan,
1982). The majority of the remaining bundles in
the periodontal ligament are 1 -4 /im in diameter.
Due to their frequent branching and anastomos-
ing, it is not possible to trace an intact network of
bundles across the periodontal space. The
networks have a complex three-dimensional ar-
rangement with overlapping of the bundles in
adjacent layers (Sloan, 1978a, 1979, 1982). It is
believed by some that this complex arrangement
ensures that, irrespective of the direction of an
B. K. B. BERKOVITZ
applied force, some collagen bundles are always
placed in tension.
When viewed in the scanning electron micro-
scope, the principal collagen fibres do not neces-
sarily run a straight course from tooth to alveolar
bone but may appear wavy (Fig. 3). When
viewed between crossed polars, the presence of
alternating light and dark bands is also inter-
preted as evidence of the wavy nature of the
collagen (Gathercole and Keller, 1982). The
model most widely used for studying the biologi-
cal significance of such waves in collagen is
tendon. Here, the collagen is folded in the form
of a zigzag which is so regular that it has a
quantifiable periodicity and a quantifiable angu-
lar deflection from the fibre axis (Diamant et al.,
1972; Nicholls^a/., 1984). In rat-tail tendon, the
periodicity is about 100 /im and the angular
deflection from the fibre axis is between 10°-15°
(depending on age) (Diamant et al., 1972). This
regular waviness has been termed a 'crimp' and
may confer special mechanical properties on the
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tissue. In a typical stress/strain curve for tendon
there is an early, small, easily extensible, non-
linear toe region interpreted as resulting from
straightening of the crimp (Gathercole and
Keller, 1982). When loads are applied to teeth,
one observes evidence of a biphasic response
(Parfitt, 1960; Picton et al., 1974; Wills et al.,
1978; Moxham and Berkovitz, 1983) and some
believe the first elastic phase is related to straigh-
tening out of the principal collagen fibre bundles.
However, care must be taken in interpreting the
possible biological significance of waviness of
periodontal ligament collagen. It is not clear
whether it is homologous with the crimping seen,
for example, in rat-tail tendon. In the periodon-
tal ligament, the only data concerning periodicity
reports a value of about 16 /im while the angular
deflection is in excess of 20° (Gathercole and
Keller, 1982). There is also little information as
to whether the periodontal fibres are straigh-
tened sufficiently during loading to allow for any
tension to be transmitted to the alveolar bone
(Gabel, 1956). Furthermore, could the waviness
be an artefact associated with the preparation of
the material?
There is evidence for a relationship between
collagen fibril diameters and the mechanical
properties of a connective tissue (Parry et al.,
1978; Flint et al., 1984). One connective tissue
that has proved particularly useful in investigat-
ing this relationship is the rabbit hind limb
STRUCTURE OF THE PERIODONTAL LIGAMENT
Figure 2 Scanning electron micrograph of decalcified transverse section of periodontal ligament showing collagen fibres
arranged as a complex branching network passing from the tooth (A) to alveolar bone (B). Blood vessels are arrowed.
tendon of flexor digitorum profundus. This
tendon is subjected to tensional forces through-
out its length but there are, in addition, compres-
sive forces within a localized area adjacent to the
bony prominences of the calcaneum and talus
where the tendon passes forwards into the sole of
the hind limb. Mcrrilees and Flint (1980) have
determined the collagen fibril diameters in areas
of tension and compression. In areas under
tension, the distribution was found to be bimodal
with major peaks at about 30 nm and 150 nm,
and with many large fibrils present. In areas
subjected to compression, however, there was a
unimodal (although slightly skewed) distribution
with a peak at about 30 nm and considerably
fewer of the large size fibrils. Flint et al. (1984)
also found a direct relationship between collagen
fibril diameter distribution and functional load-
ing in skin. Although the periodontal ligament
exhibits obvious differences both structurally
and functionally when compared with rabbit
hind limb tendon (and skin), in the absence of
any other data it appeared worthwhile similarly
53
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to investigate collagen fibril diameters. It was
found that periodontal collagen fibril diameters
show a sharp, unimodal distribution (Fig. 4) with
a peak around 45 nm (Berkovitz et al., 1981). If
the size and distribution of collagen fibril dia-
meters are in any way indicative of function, then
the periodontal ligament may be subjected to
compression during loading.
Unlike most mammalian teeth, procumbent
sheep incisors appear to be supported primarily
by the gingival connective tissue of the lower
dental pad and not by the periodontal ligament
(Spence, 1978). Shore et al. (1988) found that the
distribution of collagen fibril diameters was
bimodal in the lower dental pad with a popula-
tion standard deviation significantly larger than
the unimodal distribution of fibril diameters
found in the periodontal ligament. These authors
concluded that their results were consistent with
the view that the sheep incisor is supported
primarily by tension within the fibres of the lower
dental pad.
Merrilees and Flint (1980) also reported differ-

Figure 3 Scanning electron micrograph of transverse sec-
tion of periodontal ligament showing wavy appearance of
collagen fibres. (Courtesy of Dr P. Sloan.)
Figure 4 Transmission electron micrograph showing a transversely sectioned collagen bundle in the periodontal ligament. The
collagenfibrildiameters have a unimodal distribution with a mean of about 45 nm. The ground substance occupies about 65 per
cent of the volume of the collagen bundle. A = part of fibroblast.
B. K. B. BERKOVITZ
ences in the axial periodicity of the collagen in
rabbit hind limb tendon of flexor digitorum
profundus. In areas subjected to tension, the
periodicity was approximately 62 nm; in areas of
compression, approximately 54 nm. For the
periodontal ligament, the axial periodicity has
been reported as approximately 57 nm (Mox-
ham, 1985).
One property of certain types of collagen is
that, with age and possibly function, there is a
considerable increase in fibril diameters up to
maturity (Torp et al., 1975; Flint et al., 1984).
The factors controlling the growth of collagen
fibrils are poorly understood (Gathercole and
Keller, 1982; Flint et al., 1984), although reac-
tions with the ground substance and function are
clearly important considerations. Apart from a
very small increase in periodontal collagen fibril
diameters between unerupted and erupted rat
molars, no further increase with age has been
observed (Berkovitz et al., 1984). A comprehen-
sive study by Luder et al. (1988) similarly
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reported no increase in collagen fibril diameters
for human periodontal ligament.
Many authors, utilizing both biochemical and
autoradiographical techniques, have reported
that there is a continuous and rapid turnover of
periodontal collagen. Indeed, turnover of pro-
tein in this site is one of the most rapid in the
STRUCTURE OF THE PERIODONTAL LIGAMENT
whole body, the half-life of the collagen being in
the order of days (for example, Skougaard et al.,
1970; Rippin, 1976, 1978; Beertsen and Everts,
1977; Sodek, 1977, 1978; Sodek et al., 1977;
Orlowski, 1978; Perera and Tonge, 1981a; Van
den Bos and Tonino, 1984; Imberman et al.,
1986; Taverne et al., 1986; Sodek and Ferrier,
1988). Difficulties are encountered in determin-
ing turnover rates for collagen, for example due
to the presence of intracellular degradation of
newly synthesized collagen and due to possible
re-utilization of protein (for example, Laurent,
1987). This, together with differences related to
the type of method used for measurement (for
example, autoradiography versus biochemical
techniques), to the tooth type, age and site of
sampling, and to the nature of the species used
may help account for values for turnover times of
periodontal collagen ranging from about 2 days
to 45 days.
Despite differences in procollagen processing,
types I and III collagen are believed to be
metabolized at similar rates during the rapid
turnover of collagen within the periodontal
ligament (Sodek, 1978; Sodek and Limeback,
1979). Limeback el al. (1978) and Limeback and
Sodek (1979) have shown that periodontal fibro-
blasts in vitro synthesize both type I and type III
collagen in the approximate ratios seen in vivo.
However, the difference in metabolic rate
demonstrated in vivo between periodontal and
skin fibroblasts (Sodek, 1976, 1977) were not
reproduced in vitro (Limeback et al., 1978). Also
the amount of type III collagen produced de-
creased on subculturing (Limeback and Sodek,
1979).
The significance of this high rate of turnover in
the periodontal ligament is not understood,
although it may be associated with the need for
rapid adaptation of the tissue. It may be related
to the necessity for tooth movement. However,
there does not appear to be a consistent relation-
ship between turnover rates of collagen and
eruption rates. Thus, although turnover rates
appear to be increased when eruption is reacti-
vated following the extraction of opposing teeth
(Rippin, 1976,1978; Kanoza e/a/., 1980), there is
no corresponding increase in turnover rates
when eruption rates are doubled in the rat incisor
(Van den Bos and Tonino, 1984). Furthermore,
there are no quantitative differences in intracellu-
lar organelles associated with protein synthesis in
periodontal fibroblasts of erupting rat teeth
55
compared with those prevented from erupting
(Shore et al., 1985) or between erupting and
erupted rat molars (Berkovitz et al., 1984).
Alternatively, the high turnover rate may be
related to masticatory forces. Arguing against
this view, however, is the observation that turn-
over rates appear to vary little even after a rodent
incisor tooth has been removed from occlusion
(Beertsen and Everts, 1977; Shore et al, 1982;
Van den Bos and Tonino, 1984) or pinned (Shore
et al., 1985). Indeed, there is some evidence that
turnover increases in hypofunctional rat molars
(Rippin, 1976, 1978; Kanoza et al., 1980).
Two further observations relating to turnover
of collagen are noteworthy. First, when turnover
is increased following reactivation of eruption
caused by extraction of opposing teeth, the effect
is seen for both type I and type III collagen
(Kanoza et al., 1980). The initial phase of
hypofunction is accompanied by an increase in
the volume density of phagocytosed collagen in
the periodontal ligament fibroblasts (Beertsen,
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1987). Second, in an organ culture system where
blocks of mouse alveolar bone containing all
three molars are maintained, it has been reported
that the application of orthodontic loads results
in a significant increase in the proportion of type
HI collagen (Duncan et al., 1984).
The precise location of the zone of shear within
the periodontal ligament which allows a tooth to
erupt with respect to the alveolar bone has not
been fully established (for review, see Sloan,
1982). Based on histological evidence, Sicher
(1942) had proposed the existence of an interme-
diate zone or plexus in the middle of the ligament
where remodelling occurred between the ends of
bone-related and tooth related collagen fibres.
Primarily evident only in longitudinal sections of
continuously growing incisors, the presence of
such an intermediate plexus has been reinter-
preted by Sloan (1978b) as representing merely
an optical effect explained entirely by the ar-
rangement of the middle layer collagen into
sheets rather than bundles.
The absence of any localized zone within the
middle of the ligament showing a higher rate of
turnover compared with adjacent zones has been
demonstrated by many workers. Autoradio-
graphic studies generally indicate a rapid and
uniform uptake (and subsequent uniform loss) of
label across the entire width of the periodontal
ligament (Stallard, 1963; Carneiro and Fava de
Moraes, 1965; Thomas, 1967; Magnusson, 1968;
56 B. K. B. BERKOVITZ

Rippin, 1976, 1978; Beertsen and Everts, 1977;
Perera and Tonge, 1981a). However, some evi-
dence suggests that turnover may be faster
adjacent to the alveolar bone (Stallard, 1963;
Crumley, 1964; Anderson, 1967; Melcher and
Correia, 1971; Kanoza et al., 1980). Collagen in
different regions along the length of the tooth
may have different turnover rates. For example,
Rippin (1976) and Perera and Tonge (1981a)
reported that average half-lives for collagen in
the crestal and apical regions of the ligament
were approximately 6.5 and 2.5 days respectively
in erupting and in young rat molars. In adult rats
these values were approximately 11 and 7 days
respectively (Rippin, 1978).
Based on studies relating to general morpho-
logy, autoradiography and ultrastructural quan-
tification of intracellular organelles, Beertsen et
al. (1974), Beertsen and Everts (1977) and Beert-
sen et al. (1979, 1982) place the zone of shear
associated with the continuously growing rodent
incisor in the mid-region of the ligament, the
nective tissue and wound healing tissue, it has
been suggested that the high turnover rate may
be the result of mechanical stresses derived from
intermittent occlusal forces that cause micro-
trauma in the periodontal ligament (Limeback et
al., 1978).
There appears to be a significant difference in
the development of collagen fibres of the perio-
dontal ligament according to whether the teeth
have, or do not have, deciduous predecessors. In
the case of deciduous teeth or permanent molars,
the principal oblique fibre groups are evident
before the tooth has erupted into the oral cavity
(Fig. 5) (for example, Bernick, 1960; Trott, 1962;
Levy and Bernick, 1968; Magnusson, 1968;
Atkinson, 1972). However, Grant and Bernick
(1972), Grant et al. (1972) and Bernick and
Grant (1982) reported that, for the permanent
dentition of monkeys, only the presumptive

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inner tooth-related part of the ligament moving
with the tooth, the outer bone-related part
remaining stationary. A similar view was
expressed by Melcher and Correia (1971) follow-
ing observations on reactivated eruption in rat
molars. Berkovitz et al. (1980), however, provide
some evidence that the zone of shear may be
situated closer to the tooth. These authors
removed the proliferating basal region of the rat
incisor and found that, as the tooth fragment
continued to erupt, almost the entire width of the
periodontal ligament was left intact immediately
behind in the vacated socket.
The properties of collagenous tissues partly
depend on the nature of the intermolecular
crosslinks of the collagen. The major reducible
crosslink of periodontal ligament collagen is
dehydrodihydroxylsinonorleucine (Pearson et
al., 1975). In this, it resembles granulation and
embryonic tissues (Forrest et al., 1972; Bailey et
al., 1973; Barnes et al., 1976). Although the
proportion of reducible crosslinks relative to the
total collagen content of some tissues decreases
with age (Bailey and Shimokamoki, 1971), no
such change was observed in bovine molar
periodontal ligament both before and after erup- Figure 5 Photomicrograph of decalcified longitudinal sec-
tion (Pearson et al., 1975). This may reflect the tion of a permanent molar tooth just emerging into the oral
high rate of collagen turnover in the ligament, cavity. The periodontal ligament is composed of well-formed
such that the normal process of maturation to principal fibres that are orientated in a superior oblique
direction from the cementum of the root of the tooth to the
non-reducible crosslinks is not completed. alveolar bone. (Courtesy of Dr D. A. Grant and the Journal
Because of the similarities with embryonic con- of Periodontology.)
STRUCTURE OF THE PERIODONTAL LIGAMENT
dentogingival fibres are prominent during the
eruptive phase of tooth development, the
remainder of the periodontal ligament contain-
ing only loosely structured collagenous elements
which do not span the periodontal space (Fig. 6).
These authors relate the difference to the chrono-
logical variation in the sequence of deposition of
alveolar bone. These findings are of significance
if it is believed that collagen plays a role in the
mechanism of tooth eruption (for review, see
Moxham and Berkovitz, 1982a).
Oxytalan fibres in the periodontal ligament
Oxytalan fibres are pre-elastin type fibres (for
example, Fullmer et al., 1974; Bradamante and
Svajger, 1977; Shuttleworth and Smalley, 1983)
and are characterized by their ability to stain
following pretreatment with an oxidizing agent
(Fullmer, 1960; Rannie, 1963). There is little
information about the biochemical composition
or turnover rate of oxytalan fibres. In the
Figure 6 Photomicrograph of decalcified longitudinal sec-
tion of a permanent premolar (A) just emerging into the oral
cavity. The bulk of the periodontal ligament lacks significant
numbers of principal fibres passing from the root of the tooth
to alveolar bone.
57
periodontal ligament, they tend to lie parallel to
the root surface (Fullmer et al., 1974). Oxytalan
fibres form a three-dimensional meshwork,
extending from the cement to the peripheral
periodontal blood vessels (Simpson, 1967; Sims,
1975, 1976). They have not been shown to pass
from tooth to bone. The meshwork exhibits a
predominantly apico-occlusal orientation, with a
laterally interconnecting system of fine fibrils.
While also noting the close relationship between
oxytalan fibres and blood vessels, Bowling and
Rygh (1988) could not observe an extensive
attachment of the fibres to the cement in the
crestal region. At the light microscope level,
oxytalan fibres can range in diameter from 0.5-
2.5 urn (Simpson, 1967; Beertsen et al., 1974).
Ultrastructurally, they can be readily dis-
tinguished from collagen as they consist of a
parallel assemblage of circular microfibrils about
12 nm in diameter, with variable amounts of
amorphous interfibrillar material and show no
banding (Fig. 7). They can be distinguished from
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Figure 7 Transmission electron micrograph showing a
longitudinally sectioned oxytalan fibre (A) comprised of a
parallel assemblage of micro fibrils, and some collagen fibrils
(B).

elastin by their lack of an amorphous core.
Ultrastructurally, oxytalan fibres cut transver-
sely vary in outline from being irregular to ovoid
(Fig. 8). Mean values for the major and minor
axes of oxytalan fibres at the ultrastructural level
vary from about 1.5-0.6 /im and 0.5-0.2 pm
according to site and possibly species (Sims,
1984; Shore et al., 1984). These data suggest that
light microscopy techniques artefactually
thicken the fibres. Quantitatively, oxytalan fibres
are quoted as occupying approximately 0.3 per
cent of the volume of the extracellular matrix in
the rat (Shore et al., 1984) and about 3 per cent in
humans (Jonas and Riede, 1980).
Various functions have been ascribed to oxy-
talan in the periodontal ligament:
1. It has been suggested that oxytalan fibres play
a role in tooth support, increasing the rigidity
of the periodontal ligament (Rannie, 1963;
Fullmer et al., 1974; Edmunds et al., 1979;
Jonas and Riede, 1980). This view is based on
two observations. First oxytalan is said to be
closely related to the collagen fibre bundles.
Secondly, increased amounts of oxytalan are
said to be found in the ligaments of teeth
subjected to increased functional demand.
Whereas with orthodontic loading Jonas and
Riede (1980) reported fibroplasia, fibre leng-
Figure 8 Transmission electron micrograph showing a transversely sectioned oxytalan fibre (A) surrounded by transversely
sectioned collagen fibrils.
B. K. B. BERKOVITZ
thening and narrowing of oxytalan fibre
diameter in humans, Bowling and Rygh
(1988) found no change over a similar time
scale using rat molars.
2. It has been postulated that, because oxytalan
fibres are found close to periodontal fibro-
blasts, they may serve as guides for cell
migration during tooth eruption (Beertsen et
al., 1974).
3. Due to the close relationship of oxytalan
fibres to the periodontal blood vessels and
putative nerve endings, it has been suggested
that the fibres act as part of a mechanorecep-
tive system which modulates the behaviour of
the vessels within the ligament, either directly
or by the production of a more general neural
response (Sims, 1973, 1977, 1983).
As yet, there is little experimental evidence
indicating the function of oxytalan fibres. Few
changes have been observed in the fibres when
the functional state of the ligament is altered
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(Berkovitz et al., 1984; Shore et al., 1984). It is
also not clear why the fibres do not mature to
elastin as in other connective tissues. Whether
this reflects the high rate of turnover, or the
possible lack of tension within the ligament is not
known (Shore et al., 1984).
STRUCTURE OF THE PERIODONTAL LIGAMENT
Ground substance in the periodontal ligament
Although the ground substance has not been the
subject of a great deal of research in the past, its
importance in the functioning of connective
tissues is now widely accepted (for reviews, see
Pearson, 1982; Shuttleworth and Smalley, 1983).
As mentioned for collagen, the volume of the
extracellular matrix occupied by ground sub-
stance has also been investigated in the tension
and pressure zones of the rabbit hind limb
tendon of flexor digitorum profundus (Merrilees
and Flint, 1980). It has been reported that there
was considerably more ground substance within
collagen fibre bundles in areas subjected to
compression compared with areas subjected to
tension (50% and 27% respectively). Berkovitz et
al. (1981) calculated from electron micrographs
that the fibre bundles in the periodontal ligament
contain large volumes of ground substance
(approximately 65%—see Fig. 4). This may
indicate that the ligament is adapted to resist
tooth displacement by compressional systems.
However, this interpretation based upon the
volume of ground substance would appear to
conflict with the results of Gillard et al. (1977)
and Pearson (1982) relating to the type of ground
substance. Gillard et al. (1977) found that the
predominant glycosaminoglycan in connective
tissues under tension is dermatan sulphate,
whereas chondroitin sulphate predominates in
tissues under compression. Flint el al. (1984) also
report a progressive and significant increase in
the proportion of dermatan sulphate in skin up
to maturity. Pearson (1982) reported that derma-
tan sulphate is the major component of the
sulphated galactosaminoglycans in bovine perio-
dontal ligament and that the proteoglycans are
proteodermatan sulphate and a proteoglycan
containing dermatan sulphate-chondroitin sul-
phate hybrids. Indeed, a fully sulphated, authen-
tic chondroitin sulphate was not detected. Some
changes were observed in the ground substance
during the process of tooth eruption but their
significance is not fully understood.
As for collagen, the turnover of structural
non-collagenous protein in the ligament is also
rapid (Baumhammers and Stallard, 1968; Sodek,
1977, 1978;Orlowski, 1978). Indeed, it is claimed
to be about five times faster than collagen (Van
den Bos and Tonino, 1984).
Much interest has recently been shown in the
complex glycoprotein, fibronectin (for example,
59
Mosher, 1984; Hynes, 1985). It is thought to be
important in many basic connective tissue func-
tions. It may promote attachment of cells to the
substratum, especially to collagen fibrils (Jones
el al., 1986). As cells preferentially adhere to
fibronectin, it may be involved in cell migration
and orientation. Considering these functions,
together with the high rate of turnover in the
ligament, it is not surprising that fibronectin may
have considerable biological significance within
the periodontal ligament.
Immunofluorescent techniques at the light
microscope level have revealed that fibronectin is
uniformly distributed throughout the periodon-
tal ligament in both the erupting and fully
erupted state (Connor et al., 1984; Takita et al.,
1987). Ultrastructural studies have localized it
over collagen fibres and at certain sites at the cell
collagen interface (Pitaru et al., 1987). As loss of
fibronectin has been observed during the termi-
nal maturation of many extracellular connective
tissue matrices (Lindner et al., 1975; Dessau et
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al., 1978; Hassall et al., 1978), its continued
presence within the periodontal ligament may be
indicative of a failure of the ligament to 'mature'.
Connor et al. (1983) have shown that 99 per cent
of periodontal ligament cells in vitro synthesize
both fibronectin and type I collagen, while the
remaining 1 per cent of cells synthesize only
fibronectin.
Periodontal fibroblasts
The periodontal ligament generally appears to be
a very cellular tissue, approximately 50 per cent
of its volume (excluding blood vessels) being
occupied by cells (Beertsen and Everts, 1977;
Shore and Berkovitz, 1979; Shore et al., 1984).
However, species differences do occur as the
cellularity for sheep periodontal ligament is
considerably lower (Berkovitz, 1985). There is
also some evidence of a decrease in cellularity
with age (for example, Klingsberg and Butcher,
1960; Grant and Bernick, 1972; Toto et al.,
1975).
Determination of cell shape is difficult in
routine histological sections because the fibro-
blast cytoplasm takes up little stain. The shape is
deduced mainly from the nucleus which stains
more intensely. In the human, fibroblasts have
been described as ovoid or flattened in appear-
ance (Fullmer, 1967). Examination of the liga-
ment suggests that fibroblasts are often fusiform
and elongated (for example, Garant and Cho,
60
1979) (see Fig. 1). From a scanning electron
microscope study, Roberts and Chamberlain
(1978) concluded that the cells were pleomor-
phic, but identified four general types. However,
care must be taken before accepting this classifi-
cation as it is conceivable that artefacts resulting
from specimen preparation could have been
introduced. It is possible that cells could have the
same basic shape but that a lack of preferential
orientation may give an appearance of pleomor-
phism. Ultimately, the shape can only be deter-
mined by three-dimensional reconstruction tech-
niques.
In the rat incisor, periodontal fibroblasts have
a preferential orientation in the inner, cement-
related part of the ligament. While having an
elongated outline in longitudinal sections, the
cells also have a similar shape in transverse
sections of the tooth (Fig. 9). Visualization of the
cells in different planes led Shore and Berkovitz
(1979) to conclude that the cells were flattened
discs, having a mean diameter of approximately
30 fj.m. The outline was irregular due to the
presence of cytoplasmic processes.
The shape of periodontal fibroblasts may have
relevance when considering the cell motility/
contractility hypothesis of tooth eruption (for
example, Beertsen et al., 1974; Melcher and
Beertsen, 1977). There is evidence from autora-
diographical studies that periodontal fibroblasts
move occlusally at a rate equal to that of
eruption. Furthermore, if the eruption rate of the
rodent incisor is increased by cutting the tooth
free of occlusion (i.e. rendering it unimpeded),
there is a concomitant increase in the rate of
migration of cells in the cement-related part of
the periodontal ligament (Beertsen, 1975; Beert-
sen and Everts, 1977). Although providing evi-
dence of a shift in the position of periodontal
fibroblasts, such work does not in itself indicate
whether the cells are moving actively to generate
a force of eruption or are merely being trans-
ported passively with the ligament, the eruptive
force being generated by another mechanism. If
periodontal fibroblasts, by a combination of
migration and contraction, generate 'fractional
forces' (Stopak and Harris, 1982), their morpho-
logy might be expected to resemble that of
fibroblasts migrating and generating traction in
vitro. However, such cells in vitro are not
rounded in outline but appear to be highly
elongated (for example, Hay, 1982; Bellows et
al., 1982a and b).
B. K. B. BERKOVITZ
When viewed at the ultrastructural level, the
periodontal fibroblast exhibits the features one
might expect of a cell actively involved in the
synthesis and secretion of protein. It may be
regarded as a non-regulated secretory cell, in that
its secretion rate is maintained at a relatively
constant level. The cell has a prominent nucleus
which generally shows at least one nucleolus, and
occupies approximately 25 per cent of the cell by
volume (Beertsen and Everts, 1977; Yamasaki et
al., 1987). In aged mice, more than 17 percent of
cells were seen to be multinucleated (Cho and
Garant, 1984a). The cytoplasmic organelles
abundant in periodontal fibroblasts are rough
endoplasmic reticulum, mitochondria, Golgi
complex and vesicles (Figs. 10 and 11). The
rough endoplasmic reticulum occupies between
5-10 per cent of the volume of the cytoplasm
(Beertsen and Everts, 1977; Berkovitz et al.,
1984; Yamasaki et al., 1987). There is evidence
that fibroblasts produce collagen in excess of
their normal requirements, the excess being
broken down intracellularly (Bienkowski el al.,
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1978). To date, such degradation has been shown
for type I (but not type III) collagen and 10-40
per cent of newly synthesized procollagen may be
degraded intracellularly before processing into
collagen fibrils (Bienkowski, 1983). The reason
for excess production is unknown although any
increased demand for collagen secretion could
presumably rapidly be met by a simple reduction
in the rate of this intracellular degradation.
The synthetic pathway for collagen in perio-
dontal fibroblasts is similar to that in other
tissues (for example, Cho and Garant, 1981a;
Weinstock, 1981). Thus, the presence of labelled
proline can be demonstrated in the rough endo-
plasmic reticulum within 5 minutes after injec-
tion. By 20 minutes after injection, radioactivity
becomes concentrated in the Golgi apparatus.
Labelled secretory granules believed to contain
the procollagen molecules pass to their secretory
sites in fibroblast processes by 30 minutes and
deposit their labelled procollagen into the
matrix. By 4 hours, the majority of silver grains
are located over collagen fibrils extracellularly.
Sodek (1976) calculated that the conversion of
new collagen into periodontal fibres was highly
efficient (nearly 100%) compared with, say,
gingival fibres where about 50 per cent of new
collagen was degraded. Furthermore, Guis et al.
(1973) interpreted the small quantities of salt-
soluble collagen extractable from the periodon-
STRUCTURE OF THE PERIODONTAL LIGAMENT

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Figure 9a Photomicrograph of decalcified longitudinal section of rat incisor periodontal ligament. Fibroblasts in the inner,
cement-related part of the ligament (A) arc preferentially orientated and appear fusiform. B = tooth. C = blood vessel in outer,
alveolar bone-related part of ligament. D = alveolar bone.
Figure 9b Photomicrograph of decalcified transverse section of rat incisor periodontal ligament. Fibroblasts in the inner,
cement-related part of the ligament (A) appear fusiform. B = blood vessel in outer, alveolar bone-related part of ligament.
C = alveolar bone.

tal ligament and representing the most recently network is essential for the organized transport
synthesized procollagen molecules, as indicating of collagen precursors from the rough endoplas-
that the collagen molecules mature more rapidly mic reticulum to the Golgi apparatus, and for the
than in other tissues. That an intact microtubular eventual transport and exocytosis of collagen
62 B. K. B. BERKOVITZ

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lim
^
Figure 10 Transmission electron micrograph of periodontal fibroblast. Note the intracellular organelles associated with
protein synthesis, particularly the rough endoplasmic reticulum.

Figure 11 Transmission electron micrograph of periodontal fibroblast showing microfilament bundle ( Q lying beneath the cell
membrane. Microtubules (arrowed) .are present throughout the cytoplasm. A = lysosome. B = vesicles associated with cell
membrane.
STRUCTURE OF THE PERIODONTAL LIGAMENT
secretory granules, was demonstrated by Cho
and Garant (1981b) who showed collagen secre-
tion was completely inhibited following the
administration of colchicine, a drug known to
disrupt microtubules.
Limeback et al. (1983) analysed the amount
and types of collagen synthesized by clones of
periodontal ligament fibroblasts in vitro. Their
results indicate that primary cultures contain a
heterogeneous mixture of cells that not only
synthesize different levels of collagen but also
produce different ratios of type I and type III
(and even some type V) collagen. It is not yet
clear whether these findings reflect the in vivo
situation.
To balance the rapid synthesis and secretion of
collagen (and of ground substance) into the
extracellular matrix, there must be an equivalent
rapid breakdown (for reviews, see Carmichael,
1982; Werb, 1982; Krane, 1985; Laurent, 1987).
The nature and control of this degradative
process is poorly understood. Whereas at one
time it was thought that collagen degradation
was an extracellular process, the apparent pres-
ence of intracellular collagen profiles in fibro-
blasts associated with sites of rapid collagen
breakdown, such as the periodontal ligament
(Fig. 12) (for example, Ten Cate, 1972; Beertsen
etal., 1974,1978; Eley and Harrison, 1975; Shore
and Berkovitz, 1979), cranial sutures and wound
repair (for example, Ten Cate and Freeman,
1974), has been interpreted as indicating that, in
healthy tissues, all collagen degradation may be
intracellular. Where degradation is occurring
slowly, the scarcity of profiles and the small
sampling area imposed by electron microscopy
make detection of intracellular profiles unlikely
(Ten Cate and Deporter, 1975).
Ten Cate et al. (1976) have provided evidence
for a temporal sequence in the intracellular
digestion of collagen. In the first stage, a well-
defined collagen fibril(s) is phagocytosed by the
fibroblast to form a phagosome consisting of the
fibril in a clear matrix, surrounded by a unit
membrane. Primary lysosomes fuse with the
phagosome to form a phagolysosome in which
there is a gradual increase in electron density of
the matrix. Further enzymatic degeneration of
the fibril results in a progressive loss of its
characteristic banded structure.
It might be arged that the intracellular appear-
ance of collagen profiles results from oblique
sectioning across surface invaginations. Alterna-
63
tively, the profiles could be internally polymer-
ized fibrils which have never been secreted into
the extracellular space. Indeed, as already men-
tioned, biochemical studies have demonstrated
that fibroblasts in vitro can degrade considerable
amounts of newly synthesized procollagen
within the cell before secretion (Bienkowski et
al., 1978; Bienkowski, 1983). However, it might
be expected that this collagen would not show
the normal banded appearance of native colla-
gen fibrils.
Attempts have been made to clarify the nature
of intracellular collagen profiles by the injection
of the electron-dense marker thorium dioxide
(Ten Cate et al., 1976) and by histochemical
techniques for the localization of alkaline and
acid phosphatases (Deporter and Ten Cate,
1973; Ten Cate and Syrbu, 1974; Garant, 1976;
Beertsen et al., 1978; Weinstock, 1981; Yajima,
1986). The identification of these substances
within the profiles was regarded by the authors as
evidence that profiles were related to intracellu-
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lar lysosomes and that the collagen had once
been in the extracellular environment. Ultima-
tely, however, it would seem that the intracellular
nature of the collagen can only be resolved by
serial sectioning, and studies using this method
have been undertaken by Svoboda et al. (1979a
and b) and Melcher and Chan (1981). These
authors examined periodontal fibroblasts cul-
tured on tendon collagen in vitro and gingival
fibroblasts in vivo. Their observations clearly
showed that a number of the collagen profiles
were wholly intracellular. That such collagen did
not represent a degradative process involving
newly synthesized collagen was shown in the in
vitro study, as the intracellular collagen had the
distinctive, large diameter of the tendon collagen
on which the cells were cultured.
One question concerning collagen degradation
is whether periodontal fibroblasts need to secret
a collagenase (which is activated extracellularly)
to cleave fibrils extracellularly as a prerequisite to
subsequent phagocytosis. At present, the physio-
logical role of collagenases is still unclear. Elec-
tronmicroscopic studies of collagen degradation
in vivo and in vitro appear to illustrate that there
is no major enzymatic lysis of collagen fibrils
extracellularly during physiological resorption
(Svoboda et al., 1979a and b; Rose et al., 1980;
Melcher and Chan, 1981; Yajima, 1986). Colla-
gen fibrils can be engulfed at one end by cellular
processes, or the cell membrane can envelop the
m B. K. B. BERKOVITZ

I/an

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Figure 12 Penodontal fibroblast containing an intracellular collagen profile (arrowed) sectioned longitudinally (a) and
transversely (b).

middle region of a fibril(s) while the ends remain
'functional'. The enclosed part of the fibril
(coated by ruthenium red-positive material—
Melcherand Chan, 1981) is interiorized in whole
or in part, eventually fusing with primary lyso-
somes to form phagolysosomes. Thus, it is
possible that part of a fibril may be undergoing
degradation within a fibroblast while the rest of
the fibril may be 'functioning' extracellularly. It
is not known what happens to the deficiency in
the fibril. Is function restored along the length of
the fibril by synthesis and replacement of a new
segment? If this is so, does the new replacement
fibril segment immediately attain the same dia-
meter as that of the older, existing fibril and is it
comprised of the same type of collagen? If
replacement did occur but was not very efficient,
this could perhaps account for the excessive
synthesis of collagen mentioned earlier.
Further evidence that intracellular collagen
STRUCTURE OF THE PERIODONTAL LIGAMENT
fibrils had once been in the extracellular com-
partment has been deduced from examination of
the periodontal ligaments of scorbutic guinea-
pigs where the synthetic phase of collagen is
inhibited. Trie continuing presence of intracellu-
lar collagen under these conditions (Ten Cate et
al., 1976) has been explained on the basis of
phagocytosis by the fibroblast. Additional evi-
dence that collagen-containing profiles are endo-
cytic rather than exocytic comes indirectly from
autoradiographic studies. At early time intervals
after 3H-proline injection (i.e. 30 min), collagen
containing vacuoles were unlabelled; only elon-
gated secretory granules were labelled (Wein-
stock, 1981). If the collagen containing vacuoles
participate in collagen secretion (rather than
resorption), one would have expected them to be
labelled during the time of active collagen fibril
formation.
Even if collagen profiles are intracellular and
are the result of phagocytosis, the question
remains as to whether there are sufficient profiles
to account for collagen degradation occurring
entirely intracellularly. Calculations based on
published turnover times for collagen (assuming
this to be for a single population of collagen) and
comparing the number of collagen fibrils intra-
cellularly with that extracellularly, led Berkovitz
and Shore (1982) to conclude that, if intracellular
degradation occurred within a time span of
between about 25-50 minutes, then degradation
could be accounted for by an entirely intracellu-
lar process. Unfortunately, no information is yet
available on the temporal sequence of this pro-
cess.
Among other intracellular organelles functio-
nally important within fibroblasts are microfila-
ments and microtubules. A network of microfila-
ments with a diameter of approximately 6 nm is
present throughout the cytoplasm of the fibro-
blast and may be gathered together into recog-
nizable bundles or stress fibres beneath the cell
membrane (Fig. 11). Microfilaments are present
within many diverse cell types and are involved in
endocytosis, exocytosis and cell locomotion and
motility (for example, Allison, 1973; Willingham
et al., 1981). Microtubules are present as a system
of randomly orientated, non-branching cylinders
with an approximate diameter of 22 nm (Fig. 11).
They play a role in intracellular transport and in
the formation and/or maintenance of cell shape
(for example, Allison, 1973). Indeed, disruption
of microtubules by the drug colchicine leads to a
65
change in cell shape and to an inhibition in
collagen synthesis by periodontal fibroblasts
(Garant and Cho, 1979; Bellows et al., 1982a).
Despite its interference with collagen synthesis,
colchicine had no effect on the number of
intracellular collagen profiles, suggesting that
collagen phagocytosis is microtubule indepen-
dent (Beertsen et al., 1984).
The centriole of a cell comprises the centriole,
basal body and ciliary shaft. Solitary cilia (Fig.
13) have been described as lying within cell
imaginations in mouse incisor periodontal fibro-
blasts where they are described as being preferen-
tially located in the occlusally-directed part of
the cytoplasm in purported migrating fibroblasts
(Beertsen et al., 1979), and as containing no more
than nine tubule doublets (as opposed to the
normal 9 + 2 configuration). The significance of
cilia in fibroblasts is not known.
A structural feature evident in periodontal
fibroblasts but said to be absent in other adult
connective tissues is the juntional complex
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(Shore et al., 1981). The most common type is the
simplified desmosome, characterized by an
increased density of the cell membrane and
underlying cytoplasm, and by the presence of a
narrow intercellular space of about 15 nm (Fig.
14). It differs from the typical desmosome by the
absence of bundles of inserting tonofilaments
and of a dense intercellular line. Gap junctions
are also occasionally present.
The hypothesis that fibroblasts may generate
the force associated with tooth eruption has
previously been mentioned. Melcher and Beert-
sen (1977) have suggested that the presence of
microtubules and microfilaments in cells pro-
vides a structural basis for a motile system. The
extension of cell processes and attachment to
collagen fibrils or other cells, and the subsequent
withdrawal of these processes, could draw the
collagen fibrils closer together which, because of
the alignment of the fibres, could result in
movement of the tooth in an occlusal direction.
Some evidence in support of this view has been
derived from in vitro studies in which monkey
periodontal ligament fibroblasts were incorpor-
ated into 3-dimensional collagen gels and
resulted in contraction of the gel (Bellows et al.,
1981). Bellows et al. (1982a) also demonstrated
that sufficient tension was developed to bring
together fragments of tooth and bone initially
placed some distance apart within the gel; this
contraction was inhibited by colcemid and cyto-
66 B. K. B. BERKOVITZ

1 »'«•?»» ^

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Figure 13 Transmission electron micrograph of a solitary cilium (arrowed) lying within an apparent invagination of the
periodontal fibroblast cell membrane.

Figure 14 Transmission electron micrograph showing simplified desmosome (arrow) between opposing cell membranes of two
periodontal fibroblasts.

chalasin D, suggesting that microtubules and has been carried out by Bellows et al. (1982a and
microfilaments were involved. b). The cells appear spindle-shaped. Ultrastruc-
The most detailed study of the morphology of turally, they are similar to myofibroblasts, cells
periodontal fibroblasts during gel contraction which are seen during wound contraction and
STRUCTURE OF THE PERIODONTAL LIGAMENT
which share properties (as the name suggests) of
both muscle cells and fibroblasts (for example,
Gabbiani, 1979). They possess thick cell coats,
considerable amounts of microfilamentous
material dispersed throughout the cytoplasm,
numerous structures resembling gap junctions,
occasional crenulated nuclei and only small
amounts of rough endoplasmic reticulum (Fig.
15). In exhibiting these features, contracting
fibroblasts in vitro differ from periodontal fibro-
blasts in vivo which have an irregular disc-shape,
a cytoplasm containing considerable amounts of
rough endoplasmic reticulum, and microfila-
mentous material primarily in the form of stress
fibres beneath the cell membrane. Gap junctions
are infrequent in vivo where the more common
type of intercellular contact is the simplified
desmosome (Shore et al., 1981). Significantly, as
contraction of the collagen gel in vitro ceased, the
morphology of the cultured fibroblasts changed
to resemble periodontal ligament fibroblasts in
vivo; the cells assumed a more rounded morpho-
logy, exhibited endoplasmic reticulum and
showed few gap junctions but more desmosome-
like contacts. This change in morphology of
fibroblasts during gel contraction presumably is
related to a change in function, knowledge of
Figure 15 Transmission electron micrograph ofpart of a periodontal fibroblast cultured on a collagen gel and visualized during
the contraction stage. The cytoplasm is filled with microfilamentous material and there is a paucity of rough endoplasmic
reticulum. Material kindly supplied by Dr C. A. Shuttleworth.
67
which might enhance our understanding of the
role of periodontal fibroblasts in vivo.
The ultrastructure of periodontal fibroblasts
has been analysed in teeth exhibiting different
eruptive behaviours. The aim was to determine
whether there were any morphological charac-
teristics which might be consistent with the view
that the cells do exhibit a high degree of motility
and/or the ability to generate tension, and
whether these characteristics vary with the pat-
tern of eruption. For example, there may be
differing energy requirements (and thus differ-
ences in mitochondrial volume) and variations in
the amounts of microfilaments and microtu-
bules. However, on comparing periodontal liga-
mentfibroblastsin (a) erupting and fully-erupted
rat molars (Berkovitz et al., 1984), (b) normal
incisors and unimpeded incisors erupting about
twice as fast (Beertsen and Everts, 1977; Shore et
al., 1982) and (c) normal and immobilized rat
incisors (Shore et al., 1985), few differences were
evident and none that appeared consistent with
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the view that fibroblasts generate the eruptive
force.
Two distinct connective tissues exist around
the continuously growing rat mandibular inci-
sor. The periodontal ligament proper lies on the
0-5
68
lingual aspect of the tooth and, being attached to
tooth and bone, could translate a fractional force
generated by fibroblasts to the tooth. The ena-
mel-related connective tissue on the labial aspect
has no such attachment to the tooth or bone and
its fibroblasts have never been implicated in
generating an eruptive force by traction. When
quantitative comparisons of various cellular par-
ameters are made, it is evident that the two cell
types are virtually indistinguishable (Berkovitz,
1989). If structure is related to function, the two
cell types would not appear to have disparate
functions.
The anti-microtubular drug colchicine is
known to significantly retard tooth eruption in
the continuously growing rodent incisor (Berko-
vitz 1972; Chiba et al., 1980; Beertsen et al.,
1984). In an ultrastructural study, Beertsen et al.
(1984) found a direct relationship between the
eruption rate and the extent of microtubular
disruption following the administration of differ-
ent amounts of colchicine. Although this might
appear to lend support to the cell motility theory
of tooth eruption, the observations from the
preceding two paragraphs should be borne in
mind as should the more generalized effects of
colchicine which themselves might contribute to
the change in eruption (Berkovitz, 1976).
Garant and Cho (1979) and Cho and Garant
(1985) have demonstrated that certain organelles
within fibroblasts of the transseptal region of the
gingiva may be polarized. This they interpret as
being related to migration of the cell, such that
the Golgi complex is situated between the nuc-
leus and the leading (distal) process. These
authors also believe that the leading edge is the
main site where collagen is secreted (Cho and
Garant, 1984b) and that there may be a func-
tional link between the processes of migration
and secretion. Perhaps, also, the possible move-
ment of the cell through the matrix helps to align
the collagen. Beertsen et al. (1979) also reported
that fibroblasts within the ligament of the rat
incisor were polarized with respect to the cen-
triole which lay in the occlusally-directed part of
the cytoplasm, while in the erupted molar liga-
ment the Golgi region was usually situated in
that pole of the cell directed towards the alveolar
wall and the occlusal plane. These authors
interpreted the polarity as evidence of unidirec-
tional movement or unidirectional deposition or
phagocytosis of collagen.
As osteoblasts and cementoblasts of the perio-
B. K. B. BERKOVITZ
dontal ligament become incorporated into alveo-
lar bone and cellular cementum, replacement
cells must be provided within the ligament to
permit osteogenesis and cementogenesis to conti-
nue. Periodontal fibroblasts are also generated
throughout life. It is not known whether peri-
dontal fibroblasts, cementoblasts and osteo-
blasts all arise from a common precursor or
whether each cell type has its own specific
precursor cell. Although progenitor cells can be
identified by their ability to incorporate tritiated
thymidine, little is known about their origin and
life cycle.
Studies have shown that between approxima-
tely 0.5-3 per cent of cells within the periodontal
ligament are initially labelled following an injec-
tion of tritiated thymidine (Jensen and Toto,
1968; Toto and Kwan, 1970; Roberts and Jee,
1974; McCulloch and Melcher, 1983c). Such
variation may be related to diurnal periodicity
(Roberts, 1975a) or to age, location within the
ligament or individual variation (Gould et a!.,
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1982). With age, there is an overall reduction in
the labelling index (Jensen and Toto, 1968; Toto
and Borg, 1968; Toto et al., 1975; McCulloch
and Melcher, 1983c).
Dividing cells are located predominantly para-
vascularly and migrate towards the bone and
cement surfaces (McCulloch and Melcher,
1983b). As there is no increase in the number of
cells within the periodontal ligament with age,
the periodontal ligament can be regarded as a
slowly renewing tissue (Perera and Tonge,
1981b; McCulloch and Melcher, 1983b).
Homeostatic mechanisms must exist within the
tissue whereby cell generation is in equilibrium
with cell death and/or cell migration (McCulloch
and Melcher, 1983c). Evidence of some cell death
has been presented by Schellens et al. (1982) and
McCulloch and Melcher (1983a). In the normal
physiological state, few labelled cells appear to
be lost from the periodontal ligament by incor-
poration into bone or cement (McCulloch and
Melcher, 1983b).
The relatively low labelling index seen in the
normal physiological state can be significantly
increased following various experimental pro-
cedures. However, the heterogeneity of the pat-
terns of cell proliferation elicited by orthodontic
forces (for example, Roberts and Chase, 1981),
endocrine factors (Roberts, 1975b), trauma
(Gould et al., 1977, 1980) and electrical stimula-
tion (Davidovitch et al., 1980) suggests that the
STRUCTURE OF THE PERIODONTAL LIGAMENT
progenitor cells are a mixed population with
regard to the induction of proliferation (Roberts
et al., 1982). For example, in contrast to the
primarily paravascular proliferation of trauma-
tized periodontal ligament (Gould et al., 1977,
1980), cells entering S phase after orthodontic
stimulation are widely distributed throughout
the tissue and migrate mainly towards alveolar
bone to form osteoblasts (Roberts and Chase,
1981). Thus, orthodontic pressure and injury
may recruit progenitors that are distinctly differ-
ent cell types (osteogenic versus fibroblastic), or
the proliferating progenitors may represent indi-
vidual components of the same sequence.
With continuous infusion of thymidine using
osmotic minipumps, the labelling index rises
from about 1 per cent at the beginning to
between 30 per cent and 50 per cent by the end of
40-60 days (Gould et al., 1982; McCulloch and
Melcher, 1983a). This has been interpreted as
indicating that the 50 per cent or more of
unlabelled cells have either lost the capacity for
mitosis or have cycle times greater than 60 days
(McCulloch and Melcher, 1983a). The cell cycle
time for many of the proliferating cells associated
with orthodontically stimulated teeth has been
calculated to be about 32 hours (Roberts, 1975a).
However, there is a small population of labelled
cells whose label is not initially diluted, indicat-
ing they are blocked in the G 2 phase of the cell
cycle.
Gould et al. (1980) and Gould (1983) have
studied the morphology of progenitor cells in the
periodontal ligament. Whilst it may be expected
that such progenitor cells would have a relatively
undifferentiated appearance (for example, small
size, scarcity of intracellular organelles and a
high nuclear/cytoplasmic ratio), some cells tak-
ing up tritiated thymidine have been shown to be
relatively differentiated, even containing intra-
cellular collagen profiles (Gould et al., 1980;
Gould, 1983). Roberts et al. (1982) have used
nuclear size as a marker to distinguish different
populations of progenitor cells. In orthodonti-
cally stimulated teeth, they reported that osteo-
blast precursors were kinetically separable into
two distinct groups. Preosteoblasts, the imme-
diate proliferating precursors of osteoblasts, had
large nuclei (> 170 /im3) and were derived from
relatively undifferentiated fibroblast-like cells
which had smaller nuclei ( < 80 /im3).
In attempting to correlate structure and func-
tion, the periodontal ligament has been com-
69
pared with other 'adult' soft connective tissues.
While these two types of tissue have a number of
features in common, there are also properties
present within the ligament which are more
characteristic of'foetal'connective tissues. These
properties include rate of turnover, type of
collagen, nature of collagen crosslinks, collagen
fibril diameter, nature of ground substance, the
degree of cellularity and the presence of intercel-
lular contacts (Moxham et al., 1984). An appre-
ciation of this concept may help to provide a
clearer understanding of the relationship
between the ligament's structure and function.
The vasculature of the periodontal ligament
Apart from providing nutrition, the vasculature
of the periodontal ligament has been implicated
in the mechanisms of tooth eruption and tooth
support (for reviews, see Moxham and Berko-
vitz, 1982a and b). It is often stated that, for a
fibrous connective tissue, the periodontal liga-
ment is highly vascular. However, few data are
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available to compare the density of vessels in the
ligament against those in other fibrous tissues.
The periodontal vasculature has recently been
reviewed by Ed wall (1982).
The vessels of the ligament are thin walled and
tend to run parallel to the long axis of the root.
Values for the volume density of blood vessels in
the ligament vary according to tooth type and
site, from about 7 per cent in the adult mouse
molar (Gould et al., 1977; McCulloch and
Melcher, 1983c), up to about 50 per cent near the
growing base of the rat incisor (Moxham et al.,
1985). There is also evidence that vascularity can
vary between the eruptive and post-eruptive
phases of tooth development (Moxham et al.,
1987).
There are few studies concerned with measur-
ing tissue fluid pressures within the periodontal
ligament and the subject is fraught with difficul-
ties. Nevertheless, such information is important
when considering the possible role of the vascula-
ture and tissue fluid pressures in the tooth
eruptive and support mechanisms. Lamb and
Van Hassel (1972) and Palcanis (1973) have
reported tissue fluid pressures of about 10 mm
Hg. However, Walker et al. (1978) report the
pressures to be about 2 mm Hg below atmos-
pheric pressure.
Information concerning functional aspects of
the vascularity requires physiological-type
experiments to determine properties such as rates
70
Figure 16 Transmission electron micrograph of part of the wall of a periodontal ligament capillary showing fenestrations.
of blood flow. However, because of anatomical
considerations, reliable techniques for use with
respect to the ligament have yet to be developed.
Little direct information can be deduced from
structural observations on fixed material. How-
ever, observations showing the presence of fenes-
trated capillaries may have functional signifi-
cance.
Fenestrated capillaries are found in organs
engaged in the production or absorption of fluids
or where there is a need for a more direct
exchange with the blood stream (for example,
endocrine glands, the gastrointestinal tract, the
kidneys—Majno, 1965). Indeed, physiological
studies show that the permeability characteristics
of fenestrated capillaries differ markedly from
those of continuous capillaries (Dresel el al.,
1966; Eliassen et al., 1973; Haraldsson et al.,
1982). Brief reports indicate that fenestrations
are present in vessels of the periodontal ligament
(Fig. 16) (Corpron et al., 1976; Frank et al.,
1976). There are no other reports of fenestrations
in dense fibrous connective tissues. Studies
involving ultrastructural quantification of the
distribution of fenestrations in the periodontal
ligament of the rat dentition have been under-
taken by Moxham et al. (1985, 1987), involving
the continuously growing incisor and the fully
erupted and erupting molar of rats. Significantly
more fenestrations were found: per mm3 of tissue
in the erupting molar than in the erupted molar;
near the base of the incisor than more occlusally;
and in the molar (either erupting or erupted)
than in the incisor. Such observations may
provide evidence of a link between fenestrated
vessels and eruption. However, the authors
indicated that the differences in the distribution
B. K. B. BERKOVITZ
of fenestrations could be an effect of eruption
rather than its cause and that the precise func-
tions of fenestrations were still uncertain.
Acknowledgements
I am grateful to Mr L Kelberman for photo-
Downloaded from by guest on June 25, 2015
graphic assistance and to Miss M Cheesewright
for typing the manuscript.
Address for correspondence
D r B K B Berkovitz
Department of Anatomy
King's College London
Strand
London WC2R 2LS
England
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