h-
S. Soma1, S. Matsumoto1, Y. Higuchi3,
T. Takano-Yamamoto4, K. Yamashita4,
K. Kurisu2, and M. Iwamoto2*
'Ogo Dental Clinic, 2-19-16, Tsukamoto, Yodogawa, Osaka
532-0026, Japan; 2Department of Oral Anatomy and
Developmental Biology, Faculty of Dentistry, Osaka
University, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan;
4Department of Orthodontics, Faculty of Dentistry,
Okayama University, 2-5-1, Shikada, Okayama, Okayama,
700-7825, Japan; and 3Chugai Pharmaceutical Company,
2-1-9, Kyobashi, Chuo-ku, Tokyo, 104, Japan;
*corresponding author, mal(dent.osaka-u.ac.jp
J Dent Res 79(9):1717-1724, 2000
ABSTRACT
We previously reported that whereas systemic
continuous infusion of parathyroid hormone
(PTH) accelerated orthodontic tooth movement,
systemic but intermittent injection of PTH did not
increase the rate of tooth movement. Analysis of
these data suggested that continuous
administration of PTH could be applicable for
orthodontic therapy. In the present study, we
investigated whether local and chronic application
of PTH(I-34) would accelerate orthodontic tooth
movement. To increase the residence time of PTH
in the injected area, we used methylcellulose
(MC) gel (2% W/v) for a slow-release formulation
of PTH. MC gel containing PTH (PTH-MC)
continuously released biologically active PTH into
the acceptor medium for more than 72 hrs in vitro.
When male rats received a local injection of PTH-
MC into the subperiosteum in the mesio-palatal
region of the maxillary first molar (M') every
other day, M' movement, which was mesially
drawn by an orthodontic coil spring attached to
the maxillary incisors, was accelerated in a dose-
dependent manner. PTH-MC injection at 1 pg/400
g body weight caused a 1.6-fold increase in the
rate of tooth movement. The acceleration of tooth
movement by PTH-MC injection was marked on
days 6, 9, and 12. Local injection of PTH
dissolved in saline without MC did not
significantly accelerate tooth movement on day 6
or later. Histological examination revealed active
osteoclastic bone resorption and a widened
periodontal space on the compression side of the
periodontal tissue in the PTH-MC-injected rats.
These results suggest that local injection of PTH
in a slow-release formulation is applicable to
orthodontic therapy.
KEY WORDS: PTH, local injection, methylcellulose,
slow release, orthodontic tooth movement.
Received May 19, 1999; Last revision February 8, 2000;
Accepted February 22, 2000
Local and Chronic Application
of PTH Accelerates Tooth
Movement in Rats
INTRODUCTION
Development of methods for accelerating orthodontic tooth movement
could shorten the period of active orthodontic treatment. Since the rate
of tooth movement depends on the activities of bone resorption and bone
remodeling on the compression side of the periodontal tissue, it would be
expected that activation of osteoclastic bone resorption and bone
remodeling would be important for accelerating orthodontic tooth
movement. Previous studies have demonstrated that administration of bone-
resorbing factors such as prostaglandin E,, prostaglandin E2, and
1,25(OH)2-vitamin D3 in combination with mechanical force led to more
rapid bone turnover and hence faster orthodontic tooth movement
(Yamasaki et al., 1980, 1984; Chao et al., 1988; Collins and Sinclair, 1988;
Lee, 1990; Takano-Yamamoto et al., 1992a,b).
We have also reported that continuous systemic infusion of PTH was
effective in accelerating orthodontic tooth movement (Soma et al., 1999).
PTH infusion enhanced osteoclastic bone resorption in the compressed
periodontal tissue without causing an increase in bone loss in the other areas
of the periodontal tissue. In contrast, intermittent injection of PTH at the
same dose did not significantly affect the experimental tooth movement.
These findings suggest that continuous administration is necessary for the
acceleration of tooth movement by the infusion of PTH. An early in vivo
study demonstrated that direct administration of PTH into the bone marrow
of long bones induced osteoclast formation in the injected area (Takano-
Yamamoto and Rodan, 1990), suggesting that PTH may increase bone
resorption activity in the injected area in vivo. Taken together, the above
findings suggest that the local and chronic administration of PTH into the
periodontal tissue may be effective for acceleration of orthodontic tooth
movement when the retention time of PTH in the injected area is increased
by the loading of PTH in a slow-release formulation.
Slow-release preparations of gonadotropin-releasing hormone agonists
and antagonists are clinically used in the therapy for steroid-dependent
cancers, precocious puberty, or estrogen-dependent disorders (Filicori and
Flamigni, 1988; Geisthoevel et al., 1989; Oostdijk et al., 1990).
Biodegradable materials as drug carriers have been applied to the induction of
osseous regeneration in bone defects (Kenley et al., 1994), continuous
administration of antibiotics into the periodontal sulcus in periodontitis
(Okuda et al., 1992), or ophthalmic instillation therapy for glaucoma (Lee and
Lee, 1993; Kumar and Himmelstein, 1995). Since drugs carried in
biodegradable materials continuously diffuse into the surrounding tissues for a
long time, the injected drugs may reside at a considerable concentration
around the injected area and give their pharmacological effects continuously
around the injection area for a certain period. Methylcellulose (MC) is a
highly hydrophilic, non-toxic, polymer, and its derivatives can be easily
mixed with aqueous solutions. The local injection of lidocaine carried in MC
gel (MCcomR, Orion-Farmos, Turku, Finland) is used clinically in long-
lasting nerve block therapy. Among various biodegradable materials, MC gel
1717
1718 Soma et a/. J Dent Res 79(9) 2000
formulations containing hydrophilic drugs are thoughit to be
A suitable for parente-al drug delivery (Paavola et al., 1995).
Therefore, it would be expected that MC gels would be
applicable to the slow release of PTH. The followinig two

I-_
I
- -r:f-d
Il I-
requirements are important for the design of a MC gel for the
slow release of PTII: (I) The MC gel should release PTH
slowly, and (2) PTH released from the MC gel should retain its
biological activity.
The purpose of the present study was to investigate whether
chronic local administration of PTH accelerates orthodontic
tooth movement. We first examined the pharmacokinetics of
B PTH release from MC gels and the bioactivity of the released
PTH by using an in vitro drug-release system and chondrocyte
culture system, respectively. Then we examined the effects of
E 200 local injection of PTH dissolved in MC gel on the rate of
c
0
.2
E 150 I experimental tooth movemenit in rats.

a)
a,
E 100
MATERIALS & METHODS
0
.9 In vitro Drug Release Experiment
c)U 50
0- Ca Preparation oJ PTH-ccontaining Methi1Icellulose Gels
-O . . . [li
. . . . ,r-L-1
. .
/b)r in vitro Drug Release Experimnent
r 12 24 36 48 60 72 We prepared a MC solutioni by adding 4 g of MC powder (Wako
Time of incubation (h) Purc Chemical Industry, Osaka, Japan) to 100 mL of'salinie at 90 to
100°C with constant stirring. After the MC had dissolved
completely, the solution was cooled and kept at 4°C. Synthictic
C 80 r
human PTH(1-34) (Peptide InstitLtC Inc., Mino, Japan) dissolved
in 0.9%0 saline at 2 p.g/p.L was mixed with thc same volumIeC of MC
a,)
solution, resulting in a final concentrationi of 2.0%'S (\\/) PTH in
ICD
60 1- the MC solution.
0) Drug Release Ex.perinents
.- cn 40 l- We used a two-compartment in vilro systcm (Paavola et al., 1995)
C to examine the release of PTEH from the MC gel (Fig. IA). Brifcly,
20 F a 200- L quantity of MC solution containinig PTI I was placcd in a
donor compartment having a membrane filter bottom whose
0 effective diffusion area was 0.79 cm3. The donor compar-timicit
CDE 12 24 36 48 60 72 containing the MC gel was immeised in an acceptor compartimieint
Time of incubation (h) filled with 500 F.L of Dulbecco's modificd Eagle's mediulIl
(DMEM) containing 0.1P00 boviic scrum albuminll (BSA). Then the
752 system was stiiTed at 37C. The acceptor mediumii was harvested
every 12 hrs, and 500 FiL of nlCw mcdium was added to the
6 acceptor compartment afte'r each harvcstinig. The concentr-ation of

oE Fi ure 1. Drug release experiments. (A) Schematic diagram of drug-

4 release system. PTH-containing gels lal were placed in a donor
o- compartment having a membrane filter bottom (b). The donor
m com partment was inserted into an acceptor compartment (c). The
C X
0o whole system was placed on a rotator (d). Medium (e) in the acceptor
2 compartment was replaced every 1 2 hrs. (B) The amount of PTH
C,2 released from the MC gel into the medium was measured by
radioimmunoassay. (C) Cumulative release of PTH from MC gel into
the acceptor medium was plotted. The Y-axis presents the amount of
0 PTH in the acceptor medium/amount of PTH originally mixed in MC
o . ) N) m1 -* gel x 100. (D) Biological activity of PTH in the acceptor medium.
a cn Confluent cultures of chondrocytes were treated with PTH-containing
o 2r3 :r 3 3 acceptor medium collected at the indicated times or with the same
-0CD C
--i
:E
L:
c -I E E. amount of freshly prepared PTH (Fresh PTH) and labeled with [35S]-
D B 3 3 sulfate as described in "MATERIALS & METHODS". Histograms show
the amount of [35S]-sulfate incorporation, which represents the
control 10-8 M PTH 10-7 M PTH systhesis of sulfated glucosaminoglycan. The data shown are presented
as the mean ± SD af five cultures.
J Dent Res 79(9) 2000 Tooth Movement with PTH Infusion 1719
PTII in the harvested mediLlIl was rneasuLi-ed by a
radiolilmIurloassay (IRMA kit, ImmullotopiCs Inc., San Clemente, A
CA). Al'ter the radioinilmulnoassay, the concentration of' PTH r\2
II
released fiomil the MC' gel was adjusted to 10' M or 10' M by 1
dilution of tIle PTI I with DMEM containinn 0.1I0(, BSA, and we
uscd this diluted solutioll to examine the biological activity.
microsyringe
Bioas,sav vi PTH Ac tivitY
It is known that PTIH at 10) M to 10-7 M stimulates sulfated
glycosaminoglycaii synthiesis in chonidrocytes (Koike et oi., 1990).
Theref ore. Ws e uscd a chonidi ocyte culile-C system to examine
wlihethie- PTII relcased iromii the MC -cl i-etainied its biological
activity. C'honidiocytes wxcic isolated f-om i ib cartilages of fouir- B
weck-old New Zealand whlite rabbits (Nippon Dobutsu Co., Osaka,
Japan) as describcd prcviously (Kato ct u/., 1987). The cells were
seedced at a dcnsity o' 1 .5 x 104 cells/6-iimm well and maintained in
0.1 m L of' D[vMEM containinig 1l0'", i'etal bovine seruLIm (FBS). L
When the cells had become conilueilt, they were thetn niCibated for
24 hli-s in 0. 1 iiiL DMEM suppIleimieited with 0.3'%, FBS and
various concentrations of PTH. (ne iC'i of' [3 S]-sulfate was added
20 hl-s befoie the end of' the inCubation. We deteriniied sulf'ated- 'U
glucosamil noglycal syntilesis by mcasuLilng the incolporatioll of
135S I-sul'atc into mater-ials precipitated with cetylpyridinium 'a '1.

chloi-ide after protcase digestion (Kato eit al, 1980).

Animal Experiments
c
J
P,e)raiuwion ol PTH I/c6 tion Gets fi,n Local hIjICctiOl7
PTII dissolveci in saline at 0,2 or 2.0(Lg/I_ltwas mixcd with the
same volumile of MC solution (4%,). Thus, the 1iiial concenitr-atioln of
PTI I in the MC gel (2'"o /\) was 0.1 or 1.0()g /_L, respectively.
To compare the ei'lects of the gel formulation with those of saliic
as a vxclicie, we also pi epared PTII dissolved in saline at 1.0 }:,
Vug/jl_L for local inhjection. LS m
0()rhodoniic Tootli M ivmcnicot
Oirtilodonitic tooth movement was perlormed accordinig to the
metIlod of 131-udvik and Rygh (1993), as described in detail in a
D
prcvious report (Soma cl al, 1999). Blriefly, the maxillar-y fir-st
ii olao- (Ml) on the righit side (tooth movemilent side) was moved
mesially by a force magnitude of 30 g foi 12 days, effected by an
orthio(lonitic closed-coil spring (Sentalloy 509-21, Tomy
International C'o, Tokyo, lapan) ligated between the maxillary
incisor-s and M' (Figs. 2A, 2B).
Au/miiail Ivl)xerilnents
Filty-six male Wistar rats. each wx eighing fiomli 350 g to 380 g (20 Figure 2. Orthodontic tooth movement. (A,B) The right maxillary first
wks old: Orincital Kobo C'o., Tokyo, Japan), WerIC assigned to the molar (Ml ) was moved mesially by a closed-coil spring ligated between
followin(g 7 treatment groups, each group consistilng of' 8 rats: incisors (Is) and Mt. PTH was injected into the subperiosteum in the
(I )eontol group treated with only ortilodonitic fol-ce, (2) local mesio-palatal region of M1 ('h (C) The distance between Mi and M2
iljection group treated with orthiodonitic ioIbce and local injection of was measured with calipers. (D) Histological examination was
vehicle dissolved in MC' gel, (3) local injectioll group treated with
performed in the area indicated by the rectangle.
orthodonitic foi-ce and local iljectioni of 0. ig PTH dissolved in MC
gel, (4) local ilnjection group treated with olihodolntic torce and local mircosyriiige (1701 LT, I( jLL, lamilton Co., Reno, NV, USA), into
inlectioni of' jg PTH dissolved in MC' gel, (5) local injectioln grotip the subperiosteum in the mcsio-palatal region of the right MI iln local
ti-eated withi orthlodonitic foirce and local injection of I jig PTH injectioni groups cinder general anesthicsia (Fig. 2A). The palatal
dissolvecd in 0.9'Y, saliie. (6) systemic injection group treated with mticosa was so thin that wc could confirimi that the tip ol thc needle
orthiodonitic for-ce and systemic injectioin of I jig PTIFI dissolved in had been placed propeily in the target airca ulider the periosteuLi.
MC' gel, and (7) local iijectioni group treated with only local iljectioni Since the rats wcrc anesthietized maniy times duLrilIg thc cxperimental
of I f_g PTII dissolved in MC` gel (without orthiodonltic force). Each peiiod, we used cthcr for general anesthiesia. Because ethcr diffuses
,cl with or without PTI I wxas injected cveiy othel day, by meanis of a so rapidly throughl thc lunIgs, therc would be littic prccipitation of
1 720 Soma et a/. J Dent Res 79(9) 2000
ether, even if the animals were daily exposed to ether, when ether is
A compared with ketamine, barbiturate derivatives, or other injectable
anesthetics. The systemic injection group received a subcutaneous
a,b injection of 1 FL of MC gel containing 1 jig PTH in the dorso-
1.0 1 cervical region. To prevent the injection site from becoming infected,
N
we gave each rat a daily intramuscular injection of 30 mg/kg BW of
kanamycin sulfate (Meiji Pharmaceutical Co., Tokyo, Japan). A
C 0.8 previous study (Yamamoto and Rodan, 1990) showed that PTH
C
o E
0.6 1I I
I 1 I injection into bone marrow induces osteoclasts in vivo within 30 hrs.
Thus, in the present study, orthodontic tooth movement was initiated
24 hrs after the first injection of PTH. An impression of the maxilla
was taken, while the rats were under ether anesthesia, for the
nV-0.4
.0 preparation of plaster dental casts every third day (Fig. 4). For taking
precise impressions of maxillae, we used a putty and paste technique
C 0.2 utilizing a hydrophilic vinyl polysiloxane impression material
.
(Exaflex, putty and injection type, GC Corp., Tokyo, Japan), the
U U
I _ -
_I I I* -
| I | | |I I distortion ratio of which was less than 0.3%. Rigid stock trays were
PTH prepared for rat maxillae. The tray loaded with the putty was seated
cotl 0 0.1 1.0 1.0 1.0
(1 g/400 g BW) on the maxilla. Then, the injectable paste was loaded in the putty tray,
MC - - +
and some paste was syringed into the maxillary dentition. The tray
kbcal was seated at the maxilla and kept in the mouth until fully set. After
injection local local local systemic the impression had been taken, the coil spring was adjusted to
generate a drawing force of 30 g. We evaluated the amount of
orthodontic tooth movement by measuring the interproximal distance
between M1 and the second molar on the cast under a microscope at a
B magnification of 40x with calipers having an accuracy of 0.05 mm
(Fig. 2C). Measurement of the distance between MI and M2 of each
animal was performed three times by three well-trained measurers,
who had not been informed of which group was being graded. Intra-
measurer error was not greater than 0.05 mm. Almost identical results
X 0.8 were obtained among the measurers. On day 12 of the tooth
CU movement, all animals were killed by excessive inhalation of ether.
2 0.6 For the histological examination, the maxilla on both tooth-
movement side (right side) and control side (left side) was dissected,
E_ fixed in 4% paraformaldehyde, decalcified in 4% formic acid, and
E 0.4 embedded in paraffin. Specimens were cut into serial mesio-distal
.- sections of 8-,um thickness and stained with hematoxylin and eosin.
CU 0.2 Since orthodontic tooth movement is related to osteoclastic bone
C resorption on the compression side of the periodontal tissue,
0 0.04 histological examinations were focused on the periodontal tissue on
i5I I0 3 6 9 12
the mesial side of the disto-buccal root of M1 (Fig. 2D). We and other
researchers have reported that this area showed osteoclastic bone
Days of tooth movement resorption in response to mechanical compression when MI was
moved mesially by orthodontic force (Waldo and Rothblatt, 1954;
Figure 3. Effects of local injection of PTH on orthodontic tooth movement. Takano-Yamamoto et al., 1992; Soma et al., 1999).
(A) The right maxillary first molar (M1) was moved as described in Fig. 2. The animal protocols used in the present study were approved
During tooth movement, rats in each group were treated with PTH as by the Animal Care Committee at Osaka University Dental School.
described in "MATERIALS & METHODS". Local injection groups were
treated with subperiosteal injection of vehicle or PTH in the mesio-palatal
region of M1. The systemic injection group received a subcutaneous Statistical Analysis
injection of 1.0 jig PTH-containing MC gel in the dorso-cervical area. The Statistical differences of measured data of tooth movement in
distance between M1 and M2 was measured on day 12 of tooth groups were evaluated by one-factor ANOVA. Scheffe's F test was
movement. The data shown are presented as the mean ± SD for 8 rats. ap
<0.01 compared with control group. 6p < 0.01 compared with local performed to evaluate differences between each pair of groups. All
injection group treated with vehicle dissolved in MC gel. F value = 14.52; results were expressed as the mean ± SD.
degree of freedom between groups = 5. (B) Time course of the effects of
local injection of PTH on orthodontic tooth movement. Each animal
received a local injection of MC gel (vehicle in MC,-), 1.0 ,ug PTH- RESULTS
containing MC gel (PTH in MC,O), or 1.0,ug PTH-containing saline (PTH
in saline, ). The distance between M1 and M2 was measured on the PTH Release from MC Gel Formations in vitro (Fig. 1)
indicated days of tooth movement. The data shown are presented as the As shown in Fig. IA, PTH-containing MC gel was placed in
mean ± SD for 8 rats.ap <0.05 and bp < 0.01 compared with vehicle in
MC. Cp <0.01 compared with PTH in saline. the donor compartment, which was then immersed in the
acceptor medium. The medium in the acceptor compartment
J Dent Res 79(9) 2000 Tooth Movement with PTH Infusion 1721
was collected every 12 hrs, and the quantity of PTH released
from the gel into the medium was measured by
radioimmunoassay. In the first 12-hour incubation, about
40% of the PTH in the MC gel was released, and the
remainder was gradually released until 72 hrs of incubation
(Fig. I B). The cumulative amount of PTH reached a plateau
by 72 hrs of incubation (Fig. IC). The amount of cumulative
PTH released from the MC gel during the experiment did
not exceed 80% of the amount of PTH in the MC gels. This
is probably due to non-specific adsorption of PTH to the
plastic wells (donor and acceptor compartment wells) and
filters, although we added 0.l1% BSA to the MC gel
formulation of PTH. As shown in Fig. ID, the PTH-
containing medium collected from the acceptor compartment
stimulated sulfated-glucosaminoglycan synthesis in
chondrocytes in a dose-dependent manner. The biological
activity of PTH released from the MC gel was almost the
same as that of freshly prepared PTH. Analysis of these data
indicates that the MC gel continuously released biologically
active PTH for three days. Therefore, in the following
animal experiments, we injected PTH-containing MC gel
every other day.
Effects of Local Injection of PTH on the Rate
of Orthodontic Tooth Movement (Figs. 3, 4)
M' had moved mesially by 0.54 + 0.08 mm in the control
group on day 12 of tooth movement after the application of
orthodontic force as described in "MATERIALS &
METHODS". M' movement in the local injection group
treated with MC gel only was comparable with that in the
control group. Local injection of PTH dissolved in the MC gel
dose-dependently accelerated Ml movement. Local injection
of 1.0 Vwg PTH contained in the MC gel caused an
approximately 1.6-fold increase in M1 movement when
compared with local injection of MC gel only (Figs. 3A, 4).
Neither local injection of PTH dissolved in saline nor systemic
injection of PTH dissolved in MC gel increased M' movement
on day 12. As shown in Fig. 3B, the tooth movement was
markedly accelerated on day 6 and later. Although the local
injection of PTH dissolved in saline appeared to accelerate
tooth movement on day 3, M' movement in this group on day
12 was not different from that for the local injection group
treated with vehicle dissolved in MC gel. Time courses of MI
movement in the control group, local injection group treated
with 0.1 p.g PTH dissolved in MC, and systemic injection
group treated with 1.0 Vig PTH were almost the same as the
time course for the local injection group treated with MC gel
only (data not shown). Throughout the experimental period, no
signs of inflammation were recognized in the palatal mucosa,
where the gels had been injected subperiosteally.

Figure 4. Photographs of the dental casts obtained from the maxillae

on day 12 of tooth movement. (A) Control group. (B) Local injection
group treated with vehicle dissolved in MC gel. (C) Local injection
group treated with 0.1 .tg PTH dissolved in MC gel. (D) Local injection
group treated with 1.0 ig PTH dissolved in MC gel. (E) Local injection
group treated with 1 .0 pLg PTH dissolved in saline. (F) Systemic
injection group treated with 1.0 jig PTH dissolved in MC gel. (G) Local
injection group treated with 1.0 tig PTH dissolved in MC gel (without
orthodontic forcel. Note that the interproximal space between M1 and
M2 was widest in the rat treated with the local injection of 1.0 p.g PTH
dissolved in MC gel (D).
1722 Soma et al. J Dent Res 79(9) 2000
Histological Changes in the Compressed
Periodontal Tissue (Fig. 5)
Histological examiinationi revealed that the applicationl of'
orthodontic forcc on M' in the local iijectioni group treated
with vehicle dissolved in MC gel caused narrowing of the
periodonital space anid activation of osteoclastic bone
resorption in the coilpr-essed periodontal tissUC (Fig. 5A). The
local injection group treated with 1.0 jg PTII in MC gcl
showed widening of the periodontal space (Fig. SC), wheni
compared with the local injction' group trcated witi velhicle
dissolved in the gel. Local injcction of' PTH-containing MC
gel did not activate osteoclastic bone r-esor-ptionI by itself
unless orthodontic force was silultaeICously applied on M'
(Fig. SE). PTH injection did not enilhaice ostcoclastic bone
resorption on the side opposite the treated MN in these thice
groups (Figs. 5B, SD, SF).

DISCUSSION
The present results showed that the local injectioni of PTI at I
jig/400 g BW evcry othel- day (0.25 jig1I 00 g BW/every other
day 0.125 jig/100 g BW/day) caused an iicrease in the rate
of tooth movemeint. We previously repor-ted that the rate of
tooth movemilenit was iicr-eased by systeimiic inLusioni of PTII at
4 jig/100 g BW/day (Soma cu a!., 1999). The requiL-ed dose of
PTH in local adminiistr-ationi was 1/32 of that used in systemilic
itfusioni for accclerating ortliodonitic tooth moverment
Therefore, local in jection of PTI I appears to have an advantage
in reducilng the dose of PTI. A preVioUs study showcd that
systemic inftusioni of PTH at 4 jig/l100 g BW/day did not
decrease the bone volumIec (llock and (Gera. 1992). Conisisteint
with that report, we observed carlice- that systemic TITH
infusioni at 4 ig/ 100 g BW/day for 12 days did not cause cither
an iicrease in the sermI1 calciulIl level or a decrease in bone
mineral density during orthiodonitic tooth moveimicit (Soma et
al., 1999). Thus, local admiiiiistiationi of P'T1I appears to
accelerate orthodonitic tooth movemIecilt without SystCeilic bone
loss and appears applicable for orthiodonitic thcrapy without any
undesirable side-effects on systeimiic bone metabolisimi.
The irats treated witli local iiiectioji of MC gel containiiig
PTH showed a wider periodonital space on the comilpressioIn side
of the periodontal tissuc when compar-ed with raits injected witli
MC gel only (Figs. SA, SC), thus iindicatinig that bone
resorption in the compressed periodontal tissue was enhanced
by the PTH injection. Siice the PTI I injection did nlot causc thc
Figure 5. Histological changes in the periodontal tissue on the
compression side on day 12 of tooth movement. Sections show the
mesial side of the disto-buccal root of M'. (Refer to Fig. 2D for
orientation.) (A) MI on tooth movement side in a rat treated with local
injection of vehicle dissolved in MC gel and orthodontic force. (B) Ml
on control side in a rat treated with local injection of vehicle dissolved
in MC gel and orthodontic force. (C) MI on tooth movement side in a
rat treated with local injection of 1.0 jig PIH dissolved in MC gel and
orthodontic force. (D) M' on control side in a rat treated with local
injection of 1 .0 p.g PTH dissolved in MC gel and orthodontic force. (E)
MI on tooth movement side in a rat treated only with local injection of
1.0 p. PTH dissolved in MC gel (without orthodontic force). (F) MI on
control side in a rat treated only with local injection of 1 .0 [ig PTH
dissolved in MC gel (without orthodontic force) R, disto-buccal root of
M1; arrowhead, osteoclast. Note widened periodontal space and
extensive osteoclastic bone resorption in the compressed periodontal
tissue of M' in the rats treated with PTH dissolved in MC gel and
orthodontic force in combination (C).
J Dent Res 79(9) 2000 Tooth Movement with PTH Infusion
appearance of osteoclasts in non-compressed periodontal tissue
in the absence of orthodontic force (Fig. 5E), it is likely that
local injection of PTH at a certain dose enhanced osteoclastic
bone resorption only with concomitant mechanical compressive
force. A previous study demonstrated that interleukin-1, 6 and
tumor necrosis factor-co, all potent bone-resorbing cytokines,
were rapidly increased in concentration in periodontal tissue
when orthodontic force was applied to teeth (Uematsu et al.,
1996). PTH was also shown to stimulate osteoblasts to produce
bone-resorptive factors such as interleukin-6 and leukemia
inhibitory factor (Lowik et al., 1989; Greenfield et al., 1993;
Pollock et al., 1996). Moreover, an earlier in vitro study
indicated that PTH and interleukin-lo and 1,3 synergistically
enhanced osteoclastic bone resorption (Dewhirst et al., 1987).
In view of these reports, the histological findings in the present
study strongly suggested that PTH injected into the
subperiosteum enhances production of bone-resorbing
cytokines induced by mechanical force in the compressed
periodontal tissue. Thus, application of local injection of PTH
and mechanical force in combination appears to induce
effective bone resorption for orthodontic tooth movement.
The drug-release experiments showed that the
concentration of PTH in the donor medium was significantly
higher than that in the acceptor medium harvested until 48 hrs
of incubation, in good agreement with data from a previous
study (Paavola et al., 1995). Thus, MC gel was predicted to
release PTH continuously into the surrounding tissue for 48
hrs. In the animal experiments, PTH dissolved in saline did not
accelerate orthodontic tooth movement. When PTH and
calcitonin were administered intravenously or subcutaneously,
their concentrations in the blood peaked at about 15 min and
returned to normal levels in several hours (Patton et al., 1994).
Therefore, PTH dissolved in saline was likely to have been
transferred rapidly from the subperiosteal tissue to the
surrounding vessels and degraded in a short time. Many
previous studies showed that intermittent systemic injection of
PTH stimulated bone-forming activity without affecting
osteoclastic bone resorption (Hock et al., 1988, 1989; Liu and
Kalu, 1990; Liu et al., 1991; Hock and Gera, 1992; Ibbotson et
al., 1992; Schmidt et al., 1995), whereas continuous infusion of
PTH was found to induce both bone formation and resorption
without a net decrease in the bone mass (Malluche et al., 1982;
Podbesek et al., 1983; Kitazawa et al., 1991; Hock and Gera,
1992; Jerome et al., 1992). We assume that PTH locally
injected in a MC gel formulation resides at a certain
concentration around the injected tissue, imitating a state of
continuous infusion of PTH, and thereby activates bone
resorption in the compressed periodontal tissue.
For ideal application of locally injected PTH to orthodontic
treatment, PTH by one injection should be present continuously
over a month in the target area. Therefore, it is key that suitable
biodegradable materials be developed for the continuous release
of PTH at the desired rate for a long period of time. It is also
important that PTH carriers not have a tendency to spread into a
larger area from the injection site in a short period of time. In
some cases, orthodontists should consider not only the desired
tooth movement of specific teeth but also the reciprocal effects
on anchor teeth. If anterior teeth are retracted, reciprocal effects
are exerted on posterior teeth during treatment with edgewise
appliances. For minimal displacement of anchor teeth, PTH
should be administered locally at the teeth whose movement is
1723
desired. If PTH administration is applied to orthodontic patients,
local injection would seem to be more suitable than systemic
administration. Application of local injection of PTH may have
the potential not only to shorten the treatnent time but also to
improve the treatment results. On this point, we are now
examining several injectable drug carriers that release PTH
continuously in situ during orthodontic tooth movement.
ACKNOWLEDGMENTS
This work was supported by a Research Grant from the
Ministry of Education, Science and Culture of Japan and by
support from Chugai Pharmaceutical Co., Ltd.
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