ORIGINAL ARTICLE

Alteration of tooth movement by

reveromycin A in osteoprotegerin-
deficient mice
Chisato Minamoto,a Ken Miyazawa,a Masako Tabuchi,a Miyuki Hirano,a Manami Mizuno,a Mamoru Yoshizako,a
Yasuyoshi Torii,a Yuichirou Asano,a Takuma Sato,a Makoto Kawatani,b Hiroyuki Osada,b Hatsuhiko Maeda,c
and Shigemi Gotoa
Nagoya and Saitama, Japan

Introduction: Osteoprotegerin-deficient mice develop severe high-turnover osteoporosis with porous low-
density trabecular bone from an age-related increase in osteoclast activity and are useful alveolar bone
models of osteoporosis or frail periodontal tissue. Bisphosphonate (BP), a first-line drug for osteoporosis, is
bone-avid, causing side effects such as brittle and fragile bones and jaw osteonecrosis after tooth extraction.
In orthodontics, active movement is precisely controlled by temporarily suppressing and resuming movement.
BP impedes such control because of its long half-life of several years in bone. Therefore, we investigated the
novel osteoclast-specific inhibitor reveromycin A (RMA), which has a short half-life in bone. We hypothesized
that tooth movement could be precisely controlled through temporary discontinuation and re-administration of
RMA. Methods: Osteoprotegerin-deficient mice and wild-type mice were developed as tooth movement
models under constant orthodontic force. A constant orthodontic force of 10 g was induced using a nickel-
titanium closed coil spring to move the maxillary first molar for 14 days. We administered BP (1.25 mg/kg) or
RMA (1.0 mg/kg) continuously and then discontinued it to reveal how the subsequent movement of teeth and
surrounding alveolar bone was affected. Results: Continuous BP or RMA administration suppressed osteoclast
activity and preserved alveolar bone around the roots, apparently normalizing bone metabolism. Tooth move-
ment remained suppressed after BP discontinuation but resumed at a higher rate after discontinuation of
RMA. Conclusions: RMA appears useful for controlling orthodontic tooth movement because it can be sup-
pressed and resumed through administration and discontinuation, respectively. (Am J Orthod Dentofacial
Orthop 2020;157:680-9)

I
n bone tissue, bone resorption by osteoclasts and
bone formation by osteoblasts occurs continually
without interruption. Similarly, in orthodontic tooth
movement, bone is resorbed by osteoclasts at the site
where compression is applied while bone is formed by
osteoblasts at the tension site, so the alveolar bone is
continuously remodeled.1-4 Osteoclasts are the only
a
Department of Orthodontics, School of Dentistry, Aichi Gakuin University, Na-
goya, Japan.
b differentiation and function of osteoclasts by strongly
Chemical Biology Research Group, RIKEN, Saitama, Japan.
c
Department of Oral Pathology, School of Dentistry, Aichi Gakuin University, Na-
goya, Japan.
All authors have completed and submitted the ICMJE Form for Disclosure of Po-
tential Conflicts of Interest, and none were reported.
This study was partially supported by JSPS KAKENHI (No. JP 18K09870).
Address correspondence to: Ken Miyazawa, Department of Orthodontics, School
of Dentistry, Aichi Gakuin University, 2-11 Suemori-dori, Chikusa-ku, Nagoya,
464-8651, Japan; e-mail, miyaken@dpc.agu.ac.jp.
Submitted, November 2018; revised and accepted, April 2019.
0889-5406/$36.00
Ó 2020 by the American Association of Orthodontists. All rights reserved.
https://doi.org/10.1016/j.ajodo.2019.04.037
cells capable of destroying and resorbing calcified
bone tissue, and their differentiation, maturation, and
function are strictly regulated by the receptor activator
of NF-kB ligand (RANKL) present on the membrane of
osteoblasts or bone-marrow stromal cells.5 Upon the
recognition of RANKL, osteoclasts and osteoclast pro-
genitors differentiate into mature osteoclasts. Osteopro-
tegerin (OPG) synthesized by osteoblasts is a member of
the tumor necrosis factor receptor superfamily and func-
tions as a decoy receptor for RANKL. OPG inhibits the
suppressing the interaction between RANKL and
RANK. Overexpression of OPG in mice causes severe os-
teopetrosis through the suppression of bone resorp-
tion.6-9 In contrast, bone resorption in
osteoprotegerin-deficient (OPG KO) mice is increased
because of the upregulated production of osteoclasts.
In OPG KO mice, despite having normal bone tissue at
birth, bone loss becomes apparent in trabecular bone
at 1 week of age, and in cortical bone, mainly

680
Minamoto et al
consisting of cancellous bone, at 4 weeks of age.
Because of enhanced osteoclastic activity during
development, mature OPG KO mice have severe high-
turnover osteoporosis with porous and low-density
trabecular bone compared with wild-type (WT)
mice.10,11 Therefore, OPG KO mice are an instrumental
animal model to simulate the condition of alveolar
bone in patients with osteoporosis or fragile periodontal
tissue.
Increasingly often in recent years, young individuals
who are still developing, as well as older adults and
elderly individuals, now undergo orthodontic proced-
ures for prosthetic pretreatment or prevention of peri-
odontal disease. However, many adult patients who
request orthodontic treatment have fragile periodontal
tissue or show signs of systemic osteoporosis.
Osteoporosis is classified into low-turnover osteopo-
rosis, caused by aging-related downregulation of bone
formation, and high-turnover osteoporosis, caused by
upregulation of bone resorption due, for example, to
menopause.12,13 Because bisphosphonate (BP) is the
drug of choice for osteoporosis, the number of ortho-
dontic patients on BP is expected to increase. However,
BP remains in bone for many years after administration
and is associated with side effects such as dense and
fragile bones14-16 and jaw osteonecrosis after tooth
extraction.17-19
In orthodontic treatment, during active movement,
precise control is also necessary by temporarily suppress-
ing and then resuming the movement. BP is a hydrolysis-
resistant PP1 derivative that has a high affinity for bone
and inhibits osteoclastic bone resorption.20 BP is used
clinically for the treatment of osteoporosis. However,
because of the long half-life of BP, it remains in bone
for several years after administration,21 which would
likely interrupt the precision of orthodontic tooth move-
ment. We, therefore, focused on the osteoclast-specific
inhibitor reveromycin A (RMA), an acid polyketide
recently isolated from actinomycetes.22 RMA is not
readily taken up by most cells, but active osteoclasts,
which dissolve bone by secreting acid, can take up
RMA in acidic environments.23 RMA has a high potential
to inhibit bone resorption by inducing osteoclast
apoptosis via the suppression of isoleucyl-tRNA synthe-
tase in osteoclasts. In addition, RMA has an extremely
short half-life and is taken up readily by osteoclasts
that are actively resorbing bone but not by osteoclast
progenitors or osteoclasts not actively resorbing it (inac-
tive osteoclasts). After uptake, RMA induces apoptosis in
the osteoclasts, thereby inhibiting bone resorption.20 By
taking advantage of these properties, we hypothesize
that orthodontic tooth movement can be precisely
American Journal of Orthodontics and Dentofacial Orthopedics
681
controlled through repeated cycles of administration,
discontinuation, and resumption of RMA.
In this study, using OPG KO mice with high-turnover
osteoporosis, we developed an animal model of experi-
mental tooth movement under constant orthodontic
force to investigate the movement of teeth and the sur-
rounding alveolar bone. To verify our hypothesis that
tooth movement is suppressed by BP even after discon-
tinuation because of its long half-life and bone avidity
but is resumed after discontinuation of RMA because
of its short half-life, we administered BP or RMA contin-
uously and then discontinued their administration to
reveal their effects on tooth movement and the sur-
rounding alveolar bone.
MATERIAL AND METHODS
Eight-week-old male OPG KO mice (n 5 24, experi-
mental group) and WT C57BL/6J mice (n 5 24, control
group) were used in this study. All mice were purchased
from CLEA Japan (Tokyo, Japan) and were reared in the
Animal Facility of Aichi Gakuin University School of
Dentistry under the same environmental conditions
(room temperature, 22 6 2
C; humidity, 50 6 10%;
and 12-h light/dark cycle). Animals were fed CE-2 solid
diet (CE-2; CLEA Japan, Tokyo, Japan), and tap water
was provided ad libitum. This study was approved by
the Institutional Animal Care and Use Committee of
our institution and was conducted following the Institu-
tional Animal Care and Use Committee policies and pro-
cedures. This report complies with the Animals in
Research: Reporting In Vivo Experiments guidelines.
To move teeth experimentally, general anesthesia
was induced using inhalational diethyl ether in 8-
week-old OPG KO or WT mice. A nickel-titanium closed
coil spring with a force of 10 g was placed on the maxil-
lary incisors and left first molar (M1) to move the molar
mesially for 14 days in experimental animals (loaded
side). The right M1 was used as control (unloaded side;
Figs 1, A and B). Experimental reagents were RMA
3Na salt and alendronate sodium hydrate (Teiroc; Teijin
Pharma, Tokyo, Japan).
Mice were divided into 3 groups (n 5 4 each) for the
continuous administration of physiological saline (sa-
line) for 14 days (SA group), RMA (1.0 mg/kg body
weight) for the first 7 days followed by saline for the re-
maining 7 days (RMA1/ group), or RMA for the entire
14 days (RMA1 group). Saline or RMA was administered
intraperitoneally twice daily starting 3 days before the
placement of the 10 g of force with nickel-titanium
closed coil spring. BP (1.25 mg/kg body weight) was
administered intraperitoneally to 3 groups of animals
(SA, BP1/, and BP1), but the administration was
May 2020  Vol 157  Issue 5
682 Minamoto et al

Fig 1. A, Schematic diagram showing the placement of a closed coil spring and the direction of tooth
movement. B, Closed coil spring mounted on M1. C, Time schedule in experimental tooth movement
(n 5 4, each group). BP administration intraperitoneally once daily started 5 days before initiation of
experimental tooth movement. RMA administration intraperitoneally twice daily starting 3 days before
the initiation of experimental tooth movement. BP1, bisphosphonate administration group; BP1/, bi-
sphosphonate discontinuation group; RMA1, RMA administration group; RMA1/, RMA discontinu-
ation group; SA, saline administration group. D, Site of remaining alveolar bone height measurement.
a, M1 total alveolar bone height; b, M2 total alveolar bone height; a0 , M1 remaining alveolar bone height;
b0 , M2 remaining alveolar bone height. E, Site of bone density measurement. M1M, M1 mesial root;
M1DP, M1 distopalatal root; M1DB, M1 distobuccal root.

May 2020  Vol 157  Issue 5 American Journal of Orthodontics and Dentofacial Orthopedics
Minamoto et al 683

Fig 2. A, Micro-CT images showing experimental tooth movement after 14 days of BP or RMA admin-
istration. BP1, bisphosphonate administration group; BP1/, bisphosphonate discontinuation group;
RMA1, RMA administration group; RMA1/, RMA discontinuation group; SA, saline administration
group. B, Distance, (mm) between M1-M2 in the BP administration. C, Distance, (mm) between M1-
M2 in the RMA administration. Statistical significance: n.s., not significant. *P \0.05; **P \0.01.

once daily starting 5 days before placement of the 10 g
of force with nickel-titanium closed coil spring (Fig 1, C).
Regarding the dose of RMA and BP, this experiment was
conducted with reference to studies by Tanaka et al24
and Shoji et al,25 respectively. These studies demon-
strated that RMA and BP are both effective in reducing
tooth movement.
Microcomputed tomography (CT) was performed us-
ing a 3-dimensional micro x-ray CT device developed for
use in small laboratory animals (R_mCT; Rigaku, Tokyo,
Japan), with tube voltage and current set at 90 kV and
88 mA, respectively, run time at 2 min, and pixel size
at 20 3 20 3 20 mm. Under inhalational anesthesia, an-
imals underwent imaging 7 days after initiation of
experimental tooth movement, and the maxillary bone
was excised and scanned after 14 days of experimental
tooth movement (Figs 1, C and 2, A). Movement dis-
tance (Figs 2, B and C) was measured using TRI/3D-

American Journal of Orthodontics and Dentofacial Orthopedics May 2020  Vol 157  Issue 5
684 Minamoto et al

Fig 3. A, Interradicular septa in the alveolar crest after 14 days of BP or RMA administration in exper-
imental tooth movement. B, Remaining alveolar bone height in the BP administration. C, remaining
alveolar bone height in the RMA administration. D, Hematoxylin and eosin staining of periodontal tissue
(alveolar bone surrounding the M1 root) taken after 14 days of BP or RMA administration in experi-
mental tooth movement. E, Bone density in the BP administration (BV/TV). F, Bone density in the
RMA administration (BV/TV). BP1, bisphosphonate administration group; BP1/, bisphosphonate
discontinuation group; BP, non-bisphosphonate administration group; RMA1, RMA administration
group; RMA1/, RMA discontinuation group; RMA, non-RMA administration group. Statistical signif-
icance: n.s., not significant. *P \0.05; **P \0.01.

BON software (Ratoc System Engineering Co, Ltd, To- the occlusal view with the narrowest gap between M1
kyo, Japan) to determine how far the teeth had moved. and M2 was observed. To observe the state of the alve-
The images were rotated and adjusted to ensure that olar crest of the left M1 interradicular septum, we

May 2020  Vol 157  Issue 5 American Journal of Orthodontics and Dentofacial Orthopedics
Minamoto et al
observed the roots of M1 and the surrounding alveolar
bone in a sagittal view at 14 days after the appliance
was inserted (Fig 3, A).
The remaining alveolar bone height was measured as
described by Mizuno et al26 with minor modifications.
Total alveolar bone height was defined as the height
from the cementoenamel junction line to the root apex
line. The ratio of the remaining alveolar bone height be-
tween M1 and M2 was calculated by the average of M1
and M2 (remaining alveolar bone height/total alveolar
bone height). In addition, the ratio of the remaining
alveolar bone height between the left M1 and M2 was
compared with that in the right maxilla (Figs 1, D and
3, A-C).
After 14 days of experimental tooth movement, the
maxillary bone was excised, fixed with 10% neutral buff-
ered formalin solution, and decalcified in 10% ethylene-
diaminetetraacetic acid (pH 7.2) at 4
C for 4 weeks.
Then, the jaw bone was embedded in paraffin using
the conventional method and was cut into 5-mm serial
horizontal sections. The section located at one-third of
the dental root (between the furcation site and apex)
was examined and subsequently stained with hematox-
ylin and eosin. Periodontal tissue was examined under a
light microscope. Bone volume was measured in the in-
terradicular septum as described previously by Sprogar
et al,27 with minor modifications. The borders of the tis-
sue area examined were defined by the maxillary first
molar mesial root, the maxillary first molar distopalatal
root, and the maxillary first molar distobuccal root,
and expressed as a percentage of the bone volume in
the interradicular septum vs the total volume of the
area (Fig 1, E).
Blood was collected from OPG KO and WT mice un-
der diethyl ether anesthesia. Serum alkaline phospha-
tase (ALP) activity was measured using the Liquitech
ALP kit (Roche Diagnostics KK, Tokyo, Japan), and
the blood concentration of tartrate-resistant acid phos-
phatase was measured using an enzyme-linked immu-
nosorbent assay kit (Immunodiagnostic Systems,
Ontario, Canada).
Statistical analysis
Statistical analysis was performed using GraphPad
Prism software (version 6, GraphPad Software, San
Diego, Calif). Normality tests for each group were per-
formed using the Shapiro-Wilk test. Multiple compari-
sons of the mean and standard deviation of the
experimental data were made using Kruskal-Wallis and
Mann-Whitney U tests with Bonferroni correction. In
all analyses, a probability of P\0.05 was considered sig-
nificant.
American Journal of Orthodontics and Dentofacial Orthopedics
685
RESULTS
Representative micro-CT images were taken after
14 days of experimental tooth movement, as shown
in Figure 2, A. After 7 and 14 days of experimental
tooth movement, the movement distance in the SA
group was significantly greater in OPG KO mice
than in WT mice (Figs 2, A-C). In OPG KO mice, the
movement distance in the BP1/ and BP1 groups
after 7 and 14 days of experimental tooth movement
was significantly shorter than that in the SA group,
with values similar to the BP1/ and BP1 groups af-
ter 7 days and the BP1/ group after 14 days seen in
the SA group of WT mice. After 14 days, tooth move-
ment in the BP1/ group of OPG KO mice was
significantly shorter than that in the SA group of
WT mice. There was no significant difference in the
distance between the BP1/ and BP1 groups (Figs
2, A and B).
In contrast, after 7 days of experimental tooth move-
ment in OPG KO mice, the movement distance tended to
be suppressed in both the RMA1/ and RMA1 groups
compared with the SA group, but the 3 groups did not
significantly differ. After 14 days of experimental tooth
movement in OPG KO mice, movement distance was
significantly shorter in the RMA1 group than in the
SA group, with the values getting closer to those in the
SA group of WT mice. In OPG KO mice, movement dis-
tance in the RMA1/ group ranged between the dis-
tance observed in the RMA1 and SA groups (Figs 2, A
and C).
After 14 days of experimental tooth movement, the
SA group of OPG KO mice developed osteoporosis at
the M1 root furcation site and significant alveolar
bone resorption around the M1 distopalatal root and be-
tween M1 and M2 teeth on the loaded side, compared
with the SA group of WT mice (Figs 3, A-C). Among
OPG KO mice, bone resorption at the M1 root furcation
site, around the M1 distopalatal root, and between M1
and M2 teeth were significantly suppressed in the
BP1/ and BP1 groups compared with the SA group
(Figs 3, A and B).
OPG KO mice showed a widening of the periodontal
ligament space at the tension site and narrowing of the
space at the compression site on the loaded side after
14 days of experimental tooth movement. In addition,
alveolar bone resorption was significant, and trabecular
bone was sparse compared with the unloaded side (Fig 3,
D). Unlike the SA group, bone resorption in the interra-
dicular septa as well as trabecular bone loss was sup-
pressed in the BP1/ and BP1 groups of OPG KO
mice, with no significant difference between the 2
groups (Figs 3, D and E).
May 2020  Vol 157  Issue 5
686 Minamoto et al

Fig 4. A, Serum TRAP levels after 14 days of BP administration in experimental tooth movement. B,
Serum ALP levels after 14 days of BP administration in experimental tooth movement. BP1, bi-
sphosphonate administration group; BP1/, bisphosphonate discontinuation group; BP, non-
bisphosphonate administration group. C, Blood TRAP levels after 14 days of RMA administration in
experimental tooth movement. D, Serum ALP levels after 14 days of RMA administration in experi-
mental tooth movement. RMA1, RMA administration group; RMA1/, RMA discontinuation group;
RMA, non-RMA administration group. Statistical significance: n.s., not significant. *P \0.05;
**P \0.01.

In OPG KO mice, bone resorption in alveolar bone at
the M1 root furcation site, near the M1 distopalatal root,
and between M1 and M2 teeth was suppressed in the
RMA1 group, compared with the SA group. In the
RMA1/ group, bone resorption at these locations
was similar to that observed in the SA group (Figs 3, D
and F).
After 14 days of experimental tooth movement on
the loaded side in OPG KO mice, bone resorption in
the interradicular septa and trabecular bone loss were
suppressed, and alveolar bone was preserved in the
RMA1 group, compared with the SA group. In contrast,
bone resorption in the interradicular septa did not differ
significantly between the RMA1/ and SA groups (Figs
3, D and F).
In the SA group, the serum tartrate-resistant acid
phosphatase (TRAP) level was significantly higher in
OPG KO mice than in WT mice. In OPG KO mice, the
TRAP level was significantly lower in the BP1/ and
BP1 groups than in the SA group, with no significant
difference between the former 2 groups (Fig 4, A).
In addition, serum ALP level in the SA group was
significantly higher in OPG KO mice than in WT mice.
In OPG KO mice, serum ALP level was significantly lower
in the BP1/ and BP1 groups than in the SA group,
with no significant difference between the former 2
groups (Fig 4, B).
In OPG KO mice, serum TRAP level in the
RMA1 group was significantly lower than that in the
SA group, with values being closer to those in the SA

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Minamoto et al
group of WT mice. TRAP level in the RMA1/ group
tended to remain between the levels in the SA and
RMA1 groups, with no significant intergroup difference
(Fig 4, C). In addition, in OPG KO mice, the ALP level in
the RMA1 group was significantly lower than that in the
SA group, whereas the ALP level in the RMA1/ group
remained between levels observed in the SA and
RMA1 groups (Fig 4, D).
DISCUSSION
Among the methods developed to facilitate tooth
movement, the Waldo method, in which an elastic
band is placed between M1 and M2,28 is commonly
used to move teeth in mice and rats. Using this method,
Shoji et al25 showed that BP administration suppressed
alveolar bone resorption and normalized tooth move-
ment in OPG KO mice. However, because of the long
half-life of BP in bone, Yabumoto et al29 used RMA
with a relatively short half-life and obtained results
almost identical to those with BP administration. In a
study using a nickel-titanium coil spring in OPG KO
mice, Tanaka et al24 reported that continuous adminis-
tration of RMA suppressed tooth movement and alveolar
bone resorption. To expand on their findings, we used
the same method to simulate actual orthodontic treat-
ment in the present study. Experimental tooth move-
ment was continued even after discontinuation of the
drugs to verify our hypothesis that tooth movement re-
sumes after discontinuation of RMA but remains sup-
pressed after discontinuation of BP. In this way, we
could simulate actual clinical settings in orthodontic
treatment, with continuous administration of RMA in
cases requiring no tooth movement and discontinuation
of RMA in cases requiring tooth movement. The findings
of this study will contribute to determining whether the
movement of teeth with the administration of drugs can
be planned with precision.
After continuous administration of BP for 14 days,
M1 tooth movement in OPG KO mice reduced signifi-
cantly to a level similar to normal tooth movement in
WT mice. In addition, alveolar bone resorption in the in-
terradicular septa or between M1 and M2 teeth was sup-
pressed significantly, thereby maintaining the trabecular
bone. Moreover, the TRAP level in OPG KO mice was
significantly lower in the BP1 group than in the SA
group, with significant downregulation in systemic oste-
oclastic activity in the former group. Administration of
BP reduced serum ALP level, which is usually extremely
high in OPG KO mice, suppressing overactive osteo-
blasts. Previously, Fujimura et al30 induced experimental
tooth movement using 10 g of force with nickel-
titanium coil springs in 8-week-old WT C57BL/6 mice
American Journal of Orthodontics and Dentofacial Orthopedics
687
and administered BP locally for 12 consecutive days to
reduce tooth movement, the number of osteoclasts,
and root resorption associated with orthodontic tooth
movement. In addition, using the Waldo method in 7-
week-old male rats, Kim et al31 induced experimental
tooth movement and prevented backward movement
by continuously administering BP via the tail vein.
Even though the species, injection site, and concentra-
tion of BP in these examined animals differed from those
used in our study, their findings still support ours that
continuous administration of BP suppresses tooth
movement.
The present findings also revealed that even after
discontinuation of BP, the effects were similar to those
observed after the 14-day administration of BP, presum-
ably because of the deposition of BP with its long half-
life in the bone matrix. In a study using nickel-titanium
coil springs in 12-week-old female rats, Kaipatur et al32
suppressed tooth movement by administering BP for
12 weeks even though the administration was discontin-
ued before commencing experimental tooth movement.
Although the timing of experimental tooth movement
and BP administration in their study was different
from ours, the findings are in agreement that tooth
movement is inhibited even after discontinuation of
BP. It is therefore likely that the administration of BP
will interfere with orthodontic tooth movement even af-
ter its discontinuation.
As to the effect of RMA administration on bone
metabolism turnover in OPG KO mice, a significant
decrease in TRAP and ALP levels, which are usually
high as seen in the SA group, suggests that RMA normal-
izes the activity of osteoclasts and osteoblasts systemi-
cally. In addition, continuous administration of RMA
for 14 days in experimental tooth movement suppressed
trabecular bone loss and alveolar bone resorption in the
interradicular septa and between M1 and M2 teeth in
OPG KO mice. Furthermore, the movement distance
was also significantly reduced to a level similar to that
observed in WT mice, suggesting that tooth movement
was normalized to some degree in OPG KO mice. In
contrast, the TRAP level in the RMA1/ group was be-
tween that in the SA and RMA1 groups with no signif-
icant intergroup difference, whereas serum ALP level in
the RMA1/ group was between that in the SA and
RMA1 groups with significant intergroup differences.
In addition, in the RMA1/ group, trabecular bone
loss and alveolar bone resorption in the interradicular
septa and between M1 and M2 teeth were significant af-
ter 14 days of RMA administration, and the extent of
tooth movement was somewhere between those in the
SA and RMA1 groups. These findings reflect the phys-
ical and pharmacological properties of RMA, including
May 2020  Vol 157  Issue 5
688
specificity toward active osteoclasts, a short half-life,
and drug action only when circulating in the blood. In
other words, the above findings show that in the
RMA1 groups, metabolic bone turnover was suppressed
during RMA administration but was resumed after its
discontinuation.
Because of the differences in doses and durations of
preorthodontic administration, the efficacies of BP and
RMA cannot be compared. However, this study has
shown that continuous administration of BP or RMA
significantly suppressed tooth movement while prevent-
ing alveolar bone resorption and preserving trabecular
bone in OPG KO mice; nonetheless, tooth movement
resumed after discontinuation of RMA but not BP, which
was deposited in the bone matrix for an extended period.
In summary, the findings of this study suggest that
owing to its fast metabolic rate and short half-life in
the body,23 RMA can be used to suppress tooth move-
ment, and its discontinuation permits resumption of
movement during orthodontic treatment.
The limitations of this study include a small sample
size. It was difficult to breed OPG KO mice, and there-
fore, it was difficult to achieve the desired number. The
sample size was determined from past studies by Tanaka
et al24 and Yabumoto et al.29 For WT mice, the number
was set according to the sample size of OPG KO mice.
Further studies with a larger sample size are necessary
to evaluate the possible associations revealed here.
CONCLUSIONS
In OPG KO mice, continuous administration of BP or
RMA suppressed elevated osteoclast activity, thereby
preserving alveolar bones around the roots, indicating
a normalization of bone metabolism. Significant reduc-
tion in tooth movement and alveolar bone resorption
around the roots were continuously observed even after
BP administration was discontinued. In contrast, tooth
movement was observed when RMA administration
was discontinued, and increases in tooth movement dis-
tance and alveolar bone resorption were observed on
experimental day 14. These results suggest that RMA is
a useful orthodontic agent that suppresses tooth move-
ment as required by continuation and discontinuation of
administration.
ACKNOWLEDGMENTS
The authors thank T. Nogawa (RIKEN) and A. Okano
(RIKEN) for the preparation of Reveromycin A.
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May 2020  Vol 157  Issue 5