Journal of Oral Rehabilitation 2000 27; 587–594
The effect of oestrogen deficiency on the alveolar bone
resorption caused by traumatic occlusion
S. KAWAMOTO & E. NAGAOKA Second Department of Prosthetic Dentistry, Kagoshima University Dental School,
Kagoshima City, Japan
SUMMARY This study, using 132 female rats, was
designed to investigate whether oestrogen loss fa-
cilitates alveolar bone alterations induced by trau-
matic occlusion. Rats were ovariectomized (OVX)
or underwent sham-operation (Sham). Seven days
after surgery, half of the rats in each group were
subjected to experimental traumatic occlusion
(trauma), and the other half were left untreated.
Thus, there were four groups: OVX + trauma,
Sham+ trauma, OVX, and Sham. Rats in each
group were killed 1, 3, 5, 7, or 10 days after the
introduction of occlusal trauma. The resected
Introduction
The integrity of the tissues supporting the teeth is
essential for proper mastication. Periodontal diseases,
although initiated mainly by bacterial irritants, are also
fostered by other factors, including abnormal mechani-
cal stress. The first description of trauma from occlu-
sion, by Karolyi (1901), was followed by numerous
investigations. In animal studies, the possible role of
excessive biting forces in the pathogenesis of periodon-
tal disease was clarified (Glickman & Smulow, 1965;
Kenney, 1971; Lindhe & Svanberg, 1974). Addition-
ally, it has been reported that the clinical signs of
occlusal trauma are related to the severity of periodon-
titis (Pihlstrom et al., 1986; Jin & Cao, 1992). The
impairment of osseous support by extended periodon-
tal diseases is not readily restored and complicates
efforts to preserve the natural tooth. With continued
advances in effective procedures for periodontal disease
prevention and treatment with preservation of the
supporting bone, it is increasingly important to identify
© 2000 Blackwell Science Ltd
mandibles were processed without decalcification,
and histomorphometric measurements were per-
formed in the alveolar bone adjacent to the peri-
odontal ligament of the first molar. The statistical
assessment of the time- and group-specific differ-
ences by analysis of variance revealed significant
differences between the OVX +trauma and
Sham+ trauma groups in the resorption parame-
ters, but not in the formation parameters. The re-
sults show that the alveolar bone dynamics induced
by traumatic occlusion are enhanced by oestrogen
deficiency.
the dynamics of alveolar bone in response to various
factors.
Much information is available concerning the effects
of systemic agents on skeletal bone metabolism, partic-
ularly with respect to the status of osteoporosis. In
human post-menopausal subjects, ovarian hormone
deficiency gives rise to an imbalance of calcium
homeostasis leading to osteopenia (Heaney, Recker &
Saville, 1978), and the same is true in an ovariec-
tomized rat model of oestrogen deficiency (Kalu,
Hardin & Cockerham, 1984). For further characteriza-
tion of the aetiologic mechanism of oestrogen defi-
ciency-induced osteoporosis, quantitative analyses
using bone histomorphometry have been extensively
performed (Wronski et al., 1985; Turner, Evans & Wak-
ley, 1993; Dempster et al., 1995; Yamaura et al., 1996).
These studies established that oestrogen deprivation
induces net loss of bone volume due to increased bone
resorption and inadequate compensation by bone for-
mation. The effects of generalized skeletal osteopenia
on the jaw bones have been the focus of increased
587
588 S. KAWAMOTO & E. NAGAOKA
interest in dentistry. Several studies suggested an effect
of oestrogen loss on oral bones in human subjects
(Wowern & Stoltze, 1979; Wowern, 1981; Kribbs,
1983; Nishimura, Hosokawa & Atwood, 1992), but
there have been only a few detailed analyses using
animal models (Miller et al., 1991; Elovic, Hipp &
Hayes, 1995).
We hypothesized that the sensitivity of the alveolar
bone tissue to local factors is modified by systemic
regulators of bone mineral metabolism. In the present
study, our objective was to examine whether oestrogen
loss enhances the alveolar bone changes generated by
abnormal occlusal force.
Materials and methods
Female Wistar rats, aged 43 weeks, were purchased
from Kyudo Inc.* for this study. The animal use proto-
col was reviewed and approved by the Animal Experi-
ment Committee, Kagoshima University Dental School.
During the experimental period, the rats were kept at
22 °C with a 12 h/12 h light – dark cycle and received
20 g per day of a commercially available standard diet
containing 1·19% calcium, 1·13% phosphorus, and 40
IU of vitamin D3. They were given tap water ad libitum.
After acclimatization for 1 week, they were divided
randomly into four treatment groups with/without the
experimental traumatic occlusion introduced 7 days af-
ter ovariectomy (OVX) or sham surgery (Sham), as
follows: group 1 (OVX +trauma), group 2 (Sham +
trauma), group 3 (OVX) and group 4 (Sham). At the
surgery, each rat was anaesthetized with an i.p. injec-
tion of sodium pentobarbital at 62·5 mg/kg. The bilat-
eral ovariectomy in groups 1 and 3, and the sham
surgery in the other two groups, were performed via
the dorsal approach. Additionally, experimental trau-
matic occlusion was applied in groups 1 and 2: metal
was bonded to the occlusal surface of the maxillary left
molars with dental adhesive material† to produce trau-
matic occlusion at 7 days after surgery. To confine the
vertical dimensions, all metal was trimmed to 1-mm
thickness before bonding. Groups 3 and 4 served as
non-trauma control groups. All rats were injected in-
traperitonially with Calcein‡ and tetracycline hy-
drochloride§ 3 days and 1 day, respectively, prior to
* Kumamoto, Japan.
†
Super Bond C&B, Sun Medical Co., Japan.
‡
Nacalai Tesque Inc., Kyoto, Japan.
§
Achromycin; Lederle Ltd., Tokyo, Japan.
© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587–594
sacrifice. Four to seven rodents in each group were
killed with an overdose of sodium pentobarbital at
each of 1, 3, 5, 7, and 10 days after the introduction of
the traumatic occlusion. In addition to these rats, as
baseline controls, five rats in each of groups 3 and 4
were killed on the day of introduction of occlusal
trauma (on day 0). Blood samples were collected by
cardiac puncture immediately after death.
For confirmation of successful ovariectomy, the
serum level of estradiol (E2) was measured using a
radioimmunoassay.
Histomorphometry
The mandible was removed and bisected at the midline
suture. The left half was processed without decalcifica-
tion for quantitative analysis of bone histomorphomet-
ric parameters. Samples were fixed with 10% formalin
in phosphate buffer, then dehydrated by immersion in
a graded series of ethanol solutions and defatted in
acetone for 3 days each. The specimens were infiltrated
with the embedding medium¶. The embedded samples
were sliced in the bucco-lingual direction at the first
molar and ground to 25-mm thickness with a cutting
and grinding system**. The prepared sections including
the apex of the mid-mesial root of the first molar were
first observed with fluorescence microscopy, and then
stained with methylene blue and basic fuchsin for light
microscopy.
Histomorphometric measurements were performed
on the alveolar bone surface adjacent to the periodon-
tal ligament (PDL). The microscopic images generated
by Nikon OPTIPHOTO and BIOPHOTO were displayed
on a colour monitor through a TV camera, and all
parameters were quantified using semiautomatic image
analysis software†† on an NEC personal computer (PC-
9801DX). The perimeter measurements were obtained
in rectangular fields, at magnification of 340 ×, in
sequence from the buccal crest to the lingual crest
along the alveolar wall, as illustrated in Fig. 1. Accord-
ing to the method standardized by Parfitt et al. (1987),
the mineralized surface (MS) and mineral apposition
rate (MAR), derived from the fluorescence, were em-
ployed as kinetic parameters. The osteoclast surface
(Oc.S), eroded surface (ES), and osteoid surface (OS),
¶
Rigolac, Oken-shoji, Tokyo, Japan.
** Exact BS3000 and MG4000, Exact, Germany.
††
COSMOZONE 1S, Nikon, Japan.
 EFFECT OF OESTROGEN DEFICIENCY ON ALVEOLAR BONE RESORPTION
as static parameters, were measured and expressed as
percentages of the measured bone surface (BS) on
stained sections.
Statistical analysis
The values of serum E2 were evaluated by the Mann–
Whitney U-test. Data obtained from bone histo-
morphometry are reported as the mean value and
standard error of measurement for each group. For
assessment of the group-specific differences, all values
obtained were evaluated statistically using analysis of
variance (ANOVA), followed by comparison of the val-
ues at each time point by the Mann – Whitney U-test. A
P value of less than 0·05 was considered to indicate a
significant difference. Analyses were performed with a
STATVIEW 4.02J program on a Macintosh computer.
Results
E2 assay
No time-specific differences of the serum E2 level after
surgery were detected in any of the four groups. The
values obtained at each time point in the OVX groups
(groups 1 and 3) were therefore summed and com-
pared with those in the Sham groups (groups 2 and 4).
In these two pairs of groups, the mean serum E2 levels
were 1·418 and 6·987 pg/mL, respectively (PB0·05).
Fig. 1. Schematic drawing of measured areas on a bucco-lingual
section at the mid-mesial root of the first molar. Each rectangular
measured field is 0·45 mm2.
© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587 – 594
589
The E2 levels remained lower in the OVX groups than
in the Sham groups throughout the experimental
period.
Histomorphometry
Figure 2 shows the alterations in the static parameters
of the histomorphometry after introduction of occlusal
trauma, Table 1 the values of the histomorphometric
parameters at each time point, and Table 2 the results
of the tests of significance of differences at each time
point and of group-specific differences tested by ANOVA.
Static parameters
In the non-trauma groups (groups 3 and 4), moderate
sequential changes in each parameter were seen during
the experimental period, although there were no sig-
nificant difference between these two groups.
In both the trauma groups (groups 1 and 2), in
contrast, time-dependent changes of each parameter
were clearly demonstrated. ES/BS and Oc.S/BS, as re-
sorption parameters, showed transient increase and
then gradual decrease to the baseline level. Oc.S/BS
reached a peak on day 1 of traumatic occlusion and
then decreased gradually. The values of Oc.S/BS on
day 1 in groups 1 and 2 were increased to 431 and
347% of those of the baseline controls, respectively.
ES/BS increased and remained elevated until day 7,
then declined to the baseline. The ES/BS values on
days 1–7 in groups 1 and 2 ranged from 189 to 244%
and from 121 to 164% of those of the baseline con-
trols, respectively. On the other hand, OS/BS, as a
formation parameter, decreased transiently during the
period. The minimum values of OS/BS relative to the
baseline controls were 60% in group 1 and 35% in
group 2. The changes of each parameter in group 1
were generally similar to those in group 2. Significant
differences between the two groups were detected for
Oc.S/BS, but not for other indices (ANOVA).
Statistical comparison of groups 2 and 4 revealed
significant differences between them for Oc.S/BS, ES/
BS and OS/BS.
Kinetic parameters
The values of MS were depressed transiently in the
trauma groups (groups 1 and 2), but not in the non-
590 S. KAWAMOTO & E. NAGAOKA
Fig. 2. Alterations of the static parameters. (A) Osteoclast surface
(Oc.S/BS). Significant differences were found between groups 1
and 2, 1 and 3, and 2 and 4 (by ANOVA). (B) Eroded surface
(ES/BS). Significant differences were found between groups 1
and 3, and 2 and 4 (by ANOVA). (C) Osteoid surface (OS/BS). No
significant differences were detected.
© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587–594
trauma groups (groups 3 and 4). ANOVA revealed a
significant difference between MS in the non-trauma
groups, but not in the trauma groups. No significant
changes of MAR were seen during the experimental
period in any group.
Discussion
Oestrogen deficiency, manifested as a generalized
skeletal osteopenia, was noted in the OVX groups
(groups 1 and 3) within 7 days after surgery. This
finding is consistent with the data reported by Ismail et
al. (1988), and it provided confirmation that the circu-
lating levels of oestrogen were already reduced before
the introduction of experimental occlusal trauma in
the OVX group.
For the examination of the influence of occlusion
status on tooth-supporting tissue, experimental studies
in animals have been extensively performed. In some
studies, experimental traumatic occlusion was simu-
lated by raising the vertical dimension of occlusion
(Bhaskar & Orban, 1955; Wentz, Jarabak & Orban,
1957; Safavi et al., 1974; Jorgensen, 1980). The de-
struction of the alveolar bone proper by traumatic
occlusion was assessed by radiographic or histologic
methods. However, there have been no quantitative
analyses of occlusal trauma in bone tissues. King, Keel-
ing & Wronski (1991) employed histomorphometry to
demonstrate the time course of changes in alveolar
bone induced by orthodontic tooth movement. We also
designed the present study to quantify histomorpho-
metrically the bone changes caused by traumatic occlu-
sion. Moreover, we examined whether the alterations
were intensified by systemic and metabolic factors.
Previous studies have demonstrated that the alveolar
bone adjacent to the PDL undergoes a remodelling
sequence. In the rat, it was confirmed that the molar
teeth migrate spontaneously, corresponding to bone
modelling and remodelling in the tooth socket (Baron,
1973). Using dynamic histomorphometric methods, Vi-
gnery & Baron (1980) demonstrated coordinated activ-
ities of bone formation and resorption in the tooth
socket during molar drift. In our study, similarly, both
bone formation and resorption activities were noted in
the tooth sockets of untreated rats.
In the present study, sequential changes of bone
resorption and formation were identified after the ap-
plication of experimental traumatic occlusion. It has
 EFFECT OF OESTROGEN DEFICIENCY ON ALVEOLAR BONE RESORPTION 591

Table 1. Histomorphometric parameters before and after introduction of traumatic occlusion

Day 1. OVX+trauma 2. Sham+trauma 3. OVX 4. Sham

Oc.S/BS (%) 0 – – 1·560 (1·127)† 1·260 (0·287)‡

1 6·124 (1·449) 4·374 (1·020) 1·685 (0·600) 1·054 (0·402)
3 5·354 (1·049) 2·900 (0·889) 1·668 (0·714) 1·412 (0·425)
5 4·441 (1·228) 1·947 (0·672) 1·596 (0·251) 0·853 (0·299)
7 3·571 (0·668) 1·583 (0·368) 1·996 (0·689) 0·664 (0·114)
10 1·116 (0·593) 2·120 (0·858) 1·120 (0·716) 0·580 (0·089)

ES/BS (%) 0 – – 12·132 (4·228)† 16·434 (3·556)‡

1 23·841 (3·849) 23·968 (4·799) 7·567 (2·625) 7·812 (0·844)
3 29·634 (2·311) 19·993 (1·602) 14·621 (3·789) 13·614 (2·633)
5 22·933 (0·463) 26·983 (5·088) 11·968 (2·174) 11·402 (1·147)
7 24·627 (2·569) 23·798 (4·583) 13·647 (2·581) 12·079 (1·964)
10 12·453 (2·825) 16·740 (2·617) 13·846 (4·751) 9·534 (2·977)
OS/BS(%) 0 – – 14·574 (7·495)† 16·801 (4·455)‡
1 12·552 (1·850) 6·025 (4·249) 17·895 (4·922) 13·692 (5·993)
3 8·783 (2·667) 6·522 (1·940) 10·622 (6·765) 16·318 (2·952)
5 17·673 (2·447) 11·587 (3·307) 15·811 (2·542) 17·861 (1·629)
7 12·400 (1·796) 11·105 (3·651) 17·139 (7·302) 17·885 (2·351
10 15·378 (4·267) 17·760 (4·223) 13·372 (6·263) 12·815 (2·545)
MS (%) 0 – – 5·128 (1·022)† 4·046 (0·441)‡
1 * * 5·389 (0·577) 3·640 (0·378)
3 5375 (1·230) 5·098 (1·232) 5·468 (0·519) 3·913 (1·435)
5 3·605 (0·939) 3·095 (1·133) 5·119 (0·729) 3·380 (0·635)
7 4·081 (1·139) 3·488 (0·631) 5·793 (1·106) 3·860 (0·777)
10 6·278 (1·774) 3·547 (1·152) 5·515 (0·491) 3·530 (0·060)
MAR (mm/day) 0 – – 8·021 (0·240)† 8·152 (0·474)‡
1 * * 8·467 (0·297) 8·094 (0·199)
3 8·776 (0·196) 8·034 (0·444) 8·249 (0·311) 8·210 (0·208)
5 8·709 (0·196) 8·648 (0·324) 8·324 (0·327) 8·181 (0·289)
7 8·533 (0·817) 8·458 (0·224) 8·884 (0·126) 8·147 (0·055)
10 8·026 (0·029) 8·158 (0·543) 8·247 (0·350) 8·320 (0·090)

The measured value of each parameter is expressed as the mean and standard error for the mean (in parentheses). Four to seven rats
were killed per timepoint in each group.
* Not determined; † baseline control of the OVX groups (groups 1 and 3); ‡ baseline control of the Sham groups (groups 2 and 4); –,
not applicable.

been reported that transient accelerated resorption and
suppressed formation parallel to the bone dynamics
were found at pressure sites in orthodontically-treated
rats (King et al., 1991). In that study, persistent force
produced by an orthodontical appliance brought about
a sequence of cellular activities resulting in enhanced
resorption and suppressed formation at pressure sites,
and the curves for both resorption and formation were
attenuated as a result of the tooth relocation. We
observed findings similar to these in our study, even
though the mechanical stress was applied in a different
manner. It is probable that the intermittent depressive
stress produced by traumatic occlusion is weakened by
intrusion of the tooth and/or elongation of the con-
tralateral molars. The degree of elongation was re-
ported to be 26 mm/day for molars (Ohshima et al.,
1991), and 750 mm/day for incisors (McKee & War-
shawsky, 1984).
Regarding the formation activities, the transient re-
duction of OS/BS reported here was similar to that in
orthodontically pressed sites. The values of OS/BS re-
turned to the baseline on day 10 after the application
of traumatic occlusion. Bridges, King & Mohammed
(1988) reported that the bone density lost due to tooth
movement was recovered within about 2 weeks in
adult rats. In our study, it remained unclear whether
bone formation was elevated in response to the bone
loss and whether the bone loss was restored.
The kinetic indices, MS and MAR, did not show
significant differences in the trauma groups. The values

© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587 – 594
592 S. KAWAMOTO & E. NAGAOKA

Table 2. Differences (P values) between the groups before and after introduction of traumatic occlusion (by the
Mann–Whitney U-test)

Day Groups 1–2 Groups 1–3 Groups 1–4 Groups 2–3 Groups 2–4 Groups 3–4

Oc.S/BS (%) 0 – – – – – NS
1 NS NS B0·05 NS B0·05 NS
3 NS B0·05 B0·05 NS NS NS
5 NS NS B0·05 NS NS NS
7 B0·05 NS B0·05 NS NS NS
10 NS NS NS NS NS NS

ANOVA† B0·05 B0·05 * * B0·05 NS

ES/BS (%) 0 – – – – – NS
1 NS B0·05 B0·05 B0·05 B0·05 NS
3 B0·05 B0·05 B0·05 NS B0·05 NS
5 NS B0·05 B0·05 NS B0·05 NS
7 NS B0·05 B0·05 NS NS NS
10 NS NS NS NS NS NS

ANOVA† NS B0·05 * * B0·05 NS

OS/BS (%) 0 – – – – – NS
1 NS NS NS NS NS NS
3 NS NS NS NS B0·05 NS
5 NS NS NS NS NS NS
7 NS NS NS NS NS NS
10 NS NS NS NS NS NS

ANOVA† NS NS * * B0·05 NS
MS (%) 0 – – – – – NS
1 * * * * B 0·05
3 NS NS NS NS NS NS
5 NS NS NS NS NS NS
7 NS NS NS NS NS NS
10 NS NS NS NS NS NS

ANOVA† NS NS * * NS B0·05
MAR (mm/day) 0 – – – – – NS
1 * * * * * NS
3 NS NS B0·05 NS NS NS
5 NS NS NS NS NS NS
7 NS NS NS NS NS B0·05
10 NS NS B0·05 NS NS NS

ANOVA† NS NS * * NS NS

* Not determined; NS, not significant; –, not applicable.

†
Group-specific differences tested by ANOVA.

of MS indicate the accumulation of surface mineralized
during the last 3 days prior to sacrifice. We speculate
that the measured sites were subjected to irregular
pressure and tension induced by jiggling force, and that
therefore the effect of oestrogen loss, observed in the
non-trauma groups, was masked. No significant differ-
ences of MAR were detected among the four groups.
This result is consistent with the observation made by
Yamaura et al. (1996) that the mineralizing function of
osteoblasts is not facilitated by OVX.
In this study, we examined whether systemic agents
acted synergistically on the alterations of bone kinetics
produced by local factors. Several investigations have
demonstrated, during molar tooth drifting, the effects
on bone turnover of metabolic and hormonal factors,
such as bisphosphonate (Hardt, 1988), prostaglandins
(Lasfargues & Saffer, 1992), 1,25(OH)2D3 (Takano-Ya-
mamoto, Kawakami & Yamashiro, 1992), and parathy-
roid extracts (Roberts, 1975). However, in our study in
bucco-lingual sections, OVX alone produced no signifi-

© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587–594

 EFFECT OF OESTROGEN DEFICIENCY ON ALVEOLAR BONE RESORPTION
cant alteration of bone kinetics compared with sham
operation. A possible reason for this discrepancy is the
difference in the measured surfaces of sections. The
sequences of bone remodelling are more marked in the
mesio-distal or horizontal sections than in the bucco-
lingual sections of the tooth socket, because molars
drift distal to the mandible. Liu & Baylink (1984)
reported that in calcium-deprived rats, the osteoclast
activity was not significantly changed on bucco-lingual
sections obtained at the molar socket.
It is well known that surgical termination of the
ovarian function causes the alteration of cellular dy-
namics and results in systemic bone loss (Wronski et
al., 1985; Wronski et al., 1989). Examination of the
changes of bone cell populations in the PDL of parathy-
roidectomized rats revealed effects of a systemic agent
on the osteoclast progenitor pool (Vignery & Baron,
1978). It was reported that a significant increase in the
number of granulocyte-macrophage colony forming
units, representing osteoclast progenitors, was ob-
served as early as 7 days after OVX (Nirupama, Shevde
& Pike, 1996). Accordingly, in the present study, we
speculate that the recruitment of osteoclasts and their
precursors in PDL was increased within 7 days post-
OVX, and as a result, a large number of osteoclasts
were activated in response to the hyperfunction of the
periodontium. This speculation seems reasonable as an
explanation for the observed findings that OVX en-
hanced the sensitivity of alveolar bone to the resorp-
tive action of the excessive occlusal forces.
In conclusion, the results of this study show that the
resorptive activity produced by localized abnormal me-
chanical stress on alveolar bone is enhanced by oestro-
gen deficiency that is manifested as a generalized
skeletal osteopenia.
Acknowledgments
This investigation was partly supported by the Grant-
in-Aid from the Ministry of Education, Science and
Culture, Japan (No. 07672119).
References
BARON, R. (1973) Remaniement de la lame cribriforme et des
fibres desmondotales au cours de ra migration physiologique.
Journal de Biolobie Buccale, 1, 151.
BHASKAR, S. & ORBAN, B. (1955) Experimental occlusal trauma.
Journal of Periodontology, 26, 270.
© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587 – 594
593
BRIDGES, T., KING, G. & MOHAMMED, A. (1988) The effect of age
on tooth movement and mineral density in the alveolar tissues
of the rat. American Journal of Orthodontics, 93, 245.
DEMPSTER, D.W., BIRCHMAN, R., XU, R., LINDSAY, R. & SHEN, V.
(1995) Temporal changes in cancellous bone structure of rats
immediately after ovariectomy. Bone, 16, 157.
ELOVIC, R.P., HIPP, J.A. & HAYES, W.C. (1995) Maxillary molar
extraction causes increased bone loss in the mandible of
ovariectomized rats. Journal of Bone and Mineral Research, 10,
1087.
GLICKMAN, I. & SMULOW, J.B. (1965) Effect of excessive occlusal
forces upon the pathway of gingival inflammation in humans.
Journal of Periodontology, 36, 141.
HARDT, A.B. (1988) Bisphosphonate effects on alveolar bone
during rat molar drifting. Journal of Dental Research, 67, 1430.
HEANEY, R.P., RECKER, R.R. & SAVILLE, P.D. (1978) Menopausal
changes in calcium balance performance. Journal of Laboratory
and Clinical Medicine, 92, 953.
ISMAIL, F., EPSTEIN, S., FALLON, M.D., THOMAS, S.B. & REINHARDT,
T.A. (1988) Serum bone gla protein and the vitamin D en-
docrine system in the oophorectomized rat. Endocrinology, 122,
624.
JIN, L.J. & CAO, C.F. (1992) Clinical diagnosis of trauma from
occlusion and its relation with severity of periodontitis. Journal
of Clinical Periodontology, 19, 92.
JORGENSEN, E.B. (1980) A 3-month study in monkeys of occlusal
dysfunction and stress. Scandinavian Journal of Dental Research,
88, 171.
KALU, D.N., HARDIN, R.R. & COCKERHAM, R. (1984) Evaluation of
the pathogenesis of skeletal changes in ovariectomized rats.
Endocrinology, 115, 507.
KAROLYI, M. (1901) Beobachtungen uber pyorrhea alveolaris.
O8 sterreichisch-Ungarische Vierteljahresschrift für Zahnheilkunde, 17,
279.
KENNEY, E.B. (1971) A histopathologic study of incisal dysfunc-
tion and gingival inflammation in the rhesus monkey. Journal
of Periodontology, 42, 3.
KING, G.J., KEELING, S.D. & WRONSKI, T.J. (1991) Histomorpho-
metric study of alveolar bone turnover in orthodontic tooth
movement. Bone, 12, 401.
KRIBBS, P.J. (1983) Oral findings in osteoporosis. Part2. Relation-
ship between residual ridge and alveolar bone resorption and
generalized skeletal osteopenia. Journal of Prosthetic Dentistry,
50, 719.
LASFARGUES, J.J. & SAFFER, J.L. (1992) Effect of prostaglandin
inhibition on the bone activities associated with the sponta-
neous drift of molar teeth in the rat. Anatomical Record, 234,
310.
LINDHE, J. & SVANBERG, G. (1974) Influence of trauma from
occlusion on progression of experimental periodontitis in the
beagle dog. Journal of Clinical Periodontology, 1, 3.
LIU, C.C. & BAYLINK, D.J. (1984) Differential response in alveolar
bone osteoclasts residing at two different bone sites. Calcified
Tissue International, 36, 182.
MCKEE, M.D. & WARSHAWSKY, H. (1984) In vivo experimentation
on rat incisor enamel organs through a surgical window.
Anatomical Record, 210, 693.
594 S. KAWAMOTO & E. NAGAOKA
MILLER, S.C., BOWMAN, B.M., MILLER, M.A. & BAGI, C.M. (1991)
Calcium absorption and osseous organ-, tissue-, and envelope-
specific changes following ovariectomy in rats. Bone, 12, 439.
NIRUPAMA, K., SHEVDE, N. & PIKE, J.W. (1996) Estrogen modu-
lates the recruitment of myelopoietic cell progenitors in rat
through a stromal cell-independent mechanism involving
apoptosis. Blood, 87, 2683.
NISHIMURA, I., HOSOKAWA, R. & ATWOOD, D.A. (1992) The knife-
edge tendency in mandibular residual ridges in women. Jour-
nal of Prosthetic Dentistry, 67, 820.
OHSHIMA, S., KOMATSU, K., YAMANE, A. & CHIBA, M. (1991)
Prolonged effects of hypofunction on the mechanical strength
of the periodontal ligament in rat mandibular molars. Archives
of Oral Biology, 36, 905.
PARFITT, A.M., DREZNER, M.K., GLORIEUX, F.H., KANIS, J.A., MAL-
LUCHE, H. & MEUNIER, P.J. (1987) Bone histomorphometry:
standardization of nomenclature, symbols, and units. Journal of
Bone and Mineral Research, 2, 595.
PIHLSTROM, B.L., ANDERSON, K.A., AEPPLI, D. & SCHAFFER, E.M.
(1986) Association between signs of trauma from occlusion
and periodontitis. Journal of Periodontology, 57, 1.
ROBERTS, W.E. (1975) Cell population dynamics of periodontal
ligament stimulated with parathyroid extract. American Journal
of Anatomy, 143, 363.
SAFAVI, H., RUBEN, M.P., MAFLA, E.R. & BLOOM, A.A. (1974)
Periodontal traumatism produced by sustained increase in oc-
clusal vertical dimension: a histopathological study. Journal of
Periodontology, 45, 207.
TAKANO-YAMAMOTO, T., KAWAKAMI, M. & YAMASHIRO, T. (1992)
Effect of age on the rate of tooth movement in combination
with local use of 1,25 (OH) 2D3 and mechanical force in the
rat. Journal of Dental Research, 71, 1487.
TURNER, R.T., EVANS, G.L. & WAKLEY, G.K. (1993) Mechanism of
action of estrogen on cancellous bone balance in tibiae of
© 2000 Blackwell Science Ltd, Journal of Oral Rehabilitation 27; 587–594
ovariectomized growing rats: inhibition of indices of formation
and resorption. Journal of Bone and Mineral Research, 8, 359.
VIGNERY, A. & BARON, R. (1978) Effect of parathyroid hormone
on the osteoclastic pool, bone resorption and formation in rat
alveolar bone. Calcified Tissue Research, 26, 23.
VIGNERY, A. & BARON, R. (1980) Dynamic histomorphometry of
alveolar bone remodeling in the adult rat. Anatomical Record,
196, 192.
WENTZ, F.M., JARABAK, J. & ORBAN, B. (1957) Experimental
occlusal trauma imitating cuspal interferences. Journal of Peri-
odontology, 29, 117.
WOWERN, N.V. (1981) Micrographic and histomorphometric in-
dices of mandibles for diagnosis of osteopenia. Scandinavian
Journal of Dental Research, 23, 47.
WOWERN, N.V. & STOLTZE, K. (1979) Comparative bone morpho-
metric analysis of mandibles and 2nd metacarpals. Scandinavian
Journal of Dental Research, 2, 358.
WRONSKI, T.J., DANN, L.M., SCOTT, K.S. & CINTRON, M. (1989)
Long-term effects of ovariectomy and aging on the rat skele-
ton. Calcified Tissue International, 45, 360.
WRONSKI, T.J., LOWRY, P.L., WALSH, C.C. & IGNASZEWSKI, L.A.
(1985) Skeletal alterations in ovariectomized rats. Calcified Tis-
sue International, 37, 324.
YAMAURA, M., NAKAMURA, T., TSURUKAMI, H., HIJIOKA, A.,
NARUSAWA, K. & OHNISHI, H. (1996) Local bone turnover in the
metaphysis of the proximal tibia and the lumber vertebra
during the early periods after ovariectomy in rats. Calcified
Tissue International, 58, 52.
Correspondence: Shin-ichiro Kawamoto, Second Department of
Prosthetic Dentistry, Kagoshima University Dental School, 8-35-1
Sakuragaoka, Kagoshima City 890, Japan. E-mail: kwmt-
s@dentb.hal.kagoshima-u.ac.jp