Effects of Doxycycline and Corticotomy on
Interleukin-1 ββββ Expression during Orthodontic
Tooth Movement
Won Hee Lim
The Graduate School
Yonsei University
Department of Dental Science
Effects of Doxycycline and Corticotomy on
Interleukin-1 ββββ Expression during Orthodontic
Tooth Movement
A Dissertation
Submitted to the Department of Dental Science
and the Graduate School of Yonsei University
in partial fulfillment of the
requirements for the degree of
Doctor of Philosophy of Dental Science
Won Hee Lim
June 2007
감사의 글
모든 순간 마다 보이지 않는 세심함으로 인도해 주신 하나님께 감사를 드
립니다. 본 논문이 완성되기까지 부족한 저를 항상 격려해 주시고 사랑으로
이끌어 주신 백형선 교수님께 깊은 감사를 드립니다. 그리고, 어려웠던 유학
시절부터 깊은 사랑을 베풀어 주신 김종관 교수님께 말도 다 할 수 없는 감
사를 드립니다. 삼년의 시간 동안 분에 넘치는 사랑을 주신 전윤식 과장님께
는 어떻게 말로 표현할 수 없는 감사를 올립니다. 학창 시절부터 지도 교수
님으로 사랑과 격려를 주신 이승일 교수님께도 깊은 감사를 드립니다. 세심
한 지도를 해주신 황충주 과장님께도 깊은 감사를 올립니다.
끊임 없는 이해와 사랑을 베풀어 주신 양가 어머님께 감사드리고 마지막으
로 사랑하는 남편에게 사랑과 고마움의 마음을 담아 전합니다.
모든 분께 진심으로 감사 드립니다.
2007년 6월
저자 씀
Table of Contents
Abstracts (English) ·····································································iii
I. Introduction···························································································1
II. Materials and Methods
A. Animals·····················································································4
B. Experimental Procedures ·························································4
C. Real Time RT-PCR ···································································5
D. Tooth Movement ·······································································8
E. Statistical Analysis ··································································10
III. Results ·······························································································11
IV. Discussion·························································································22
V. Conclusion ·························································································26
References······························································································27
Figure Legends ······················································································32
Abstract (Korean)···················································································34
i
List of Figures and Tables
Table 1. Genes analyzed by quantitative TaqMan PCR·································· 8
Figure 1. Schematic drawing of the rat model (A) orthodontic appliance placed (B)
orthodontic appliance placed after corticotomy·········································6
Figure 2. Landmarks on lateral cephalometric radiographs. Oc, most posterior point of
squama occipitalis; Pa, most superior point of parietal bone; T, most inferior
point of tympanic bone; Na, most anterior point of nasal bone ···················9
Figure 3. IL-1β mRNA expression (Means and SEM) in the no doxycycline + appliance
group (NDA)·····························································································12
Figure 4. IL-1β mRNA expression (Means and SEM) in the doxycycline + appliance
group (DA) ·······························································································13
Figure 5. IL-1β mRNA expression (Means and SEM) on pressure side for the DA and
NDA groups······························································································14
Figure 6. IL-1β mRNA expression (Means and SEM) on control side for the DA and
NDA groups····························································································15
Figure 7. mRNA levels (relative to control) for IL-1β in the corticotomy + appliance
group (CA) ·······························································································17
Figure 8. mRNA levels (relative to control) for IL-1β per group on pressure side in
CA·············································································································18
Figure 9. mRNA levels (relative to control) for IL-1β per group on control side in
CA·············································································································19
Figure 10. Tooth movement in the DA and NDA groups ··········································20
Figure 11. Tooth movement in the CA and A group··················································21
ii
Abstract
Effects of Doxycycline and Corticotomy on Interleukin-1β
Expression during Orthodontic Tooth Movement
Backgrounds: Cytokines are thought to play an important role in bone
remodeling during tooth movement. The aim of this study was to investigate the effect
of systemic administration of doxycycline and corticotomy on expression of Interleukin-
1β(IL-1β) during orthodontic tooth movement.
Methods: Real time reverse transcription polymerase chain reaction was
performed to measure messenger RNA (mRNA) expression of IL-1β at day 7 and 14
following application of orthodontic force to the maxillary first molar in 36 nine-week-old
male Wistar rats. Doxycycline was administrated throughout the study in 9 animals
(doxycycline + appliance group:DA) while other 9 animals served as treatment controls
(no doxycycline + appliance group:NDA). In 9 of these animals corticotomy was
performed after insertion of orthodontic appliances (corticotomy + appliance group:CA)
while the remaining 9 animals received only the orthodontic appliance (appliance
group:A). The animals were euthanized day 7 and 14 after appliance insertion and
gingival tissue samples were obtained for analysis.
iii
Results: A similar expression of IL-1β was shown at day 7 following application
of force in both DA and NDA groups. IL-1β expression at day 14 was significantly
decreased compared to that at day 7 for both DA and NDA groups. The increased
expression of IL-1β was detected on CA group seven days after the application of force
and these differences were statistically significant between these CA and A groups
fourteen days after experiment.
Conclusion: The results suggest that expression of IL-1β during orthodontic
tooth movement were not affected by administration of doxycycline, but the levels of
cytokine both day 7 and day 14 were increased in corticotomy groups. It may support
the hypothesis that osteoclast recruitment stimulated by orthodontic force was not
influenced by doxycycline, but regional acceleratory phenomenon after performance of
corticotomy increases bone remodeling during orthodontic tooth movement.
KEY WORDS: Doxycycline, Corticotomy, Tooth movement, IL-1β Expression
iv
1
Effects of Doxycycline and corticotomy on Interleukin-1β
Expression during Orthodontic Tooth Movement
Won Hee Lim, D.D.S., M.S.
Department of Dental Science, Graduate School, Yonsei University
(Directed by Prof. Hyoung Seon Baik, D.D.S., M.S.D., PhD.)
I. Introduction
Pressure onto the periodontal ligament (PDL) space induced by orthodontic forces
results in vascular compression, which leads to cellular activation and the release of
pro-inflammatory molecules such as prostaglandins and cytokines.1 Prostaglandin E2
(PGE2) and interleukin-1β (IL-1β) levels in human gingival crevicular fluid (GCF) has
been shown to be elevated during orthodontic tooth movement.2 Thus it appears that
these inflammatory mediators may regulate biological processes related to alveolar
bone remodeling for initiation of tooth movement, although the mechanism is not yet
fully understood.1,3
PGE has been known to be closely involved in bone resorption.4 Yamasake et al. has
shown that the rate of tooth movement was almost double in sites receiving PGE1
injections compared to vehicle-control.5 Conversely, anti-inflammatory drugs may
reduce the rate of bone resorption by interfering with biologic mechanisms induced by
2
PGs.6,7 IL-1β, a major physiologic form of IL-1, is a known potent cytokine involved in
initiation of bone resorption.8 Interestingly, IL-1β may induce elevation of PGE
synthesis in a dose- and time-related order.9 Inversely, administration of IL-1β and/or
PGE significantly enhanced the rate of tooth movement.10 These findings suggest that
elevation of these mediator levels may serve as a marker for increased local bone
remodeling.11
It has been shown that systemic and local administration of pharmaceutical agents may
affect tooth movement.12-14 As the use of antibiotics increases, this may cause
variations in normal bone turnover in orthodontic patients.15 Doxycycline modulates
alveolar bone metabolism, most likely due to an inhibitory effect on bone resorption
including inhibition of several matrix metalloproteinases.16 In still other studies,
doxycycline has been shown to affect osteoclast function.17 Tetracyclines have also
been shown to reduce expression of cytokines involved in bone resorption including IL-
1 and TNF-α.18 Minocycline added to gingival fibroblast cultures inhibited synthesis of
PGE2.19 Moreover, it has been shown that minocycline may prevent a decrease of bone
mineral density in ovariectomized rats.20
It has been shown that healing activity was dramatically increased adjacent to the
injury site in osseous tissue.21 This biologic process, regional acceleratory
phenomenon (RAP), potentiates healing by localized burst of hard and soft tissue
remodeling and involves increase in bone turnover and decrease in regional bone
density.22 The characteristics of RAP during initial phase appears an enhanced porosity
in cortical bone due to increase in osteoclastic activity.23 Interestingly, tooth movement
3
was enhanced in rats with osteoporosis.24 These findings suggest that localized
transient osteoporosis, which means increased mobilization of calcium, decreased
bone density and increased bone turnover, result in rapid tooth movement after RAP
invoked by corticotomy.25 In still other studies, there has been strong indirect evidence
that the physiologic events associated with RAP result in corticotomy-facilitated tooth
movement.26 PGE2 has also been shown to facilitate RAP in the repair of experimental
fractures.22 However, an assessment of PGE2 and IL-1β after corticotomy have not
been made to see the RAP.
Administration of doxycycline and corticotomy during orthodontic tooth movement
might add interacting factors for the study of osteoclast.1,13,18 The aim of this study was
to investigate whether systemic administration of doxycline and corticotomy induced
different gene transcription of IL-1β in the gingival tissues involved in orthodontic tooth
movement.
4
II. Material and Methods
A. Animals
Thirty six nine-week-old, male Wistar rats, approximate weight of 280 g, from Yonsei
Univerisity vivarium were used. All animals were kept in stainless-steel cages in air-
conditioning and subject to standard 12-hour light/dark cycle. They were fed with a
pellet diet (8811M0001, Extrusion, Superfeed Co., LTD, Gangwon, Korea) and tap
water ad libitum. They were checked everyday in regard to their health status.
Doxycycline was administrated throughout the study in 9 animals (doxycycline +
appliance group:DA) while other 9 animals served as treatment controls (no
doxycycline + appliance group:NDA). In 9 of these animals corticotomy was performed
after insertion of orthodontic appliances (corticotomy + appliance group:CA) while the
remaining 9 animals received only the orthodontic appliance (appliance group:A). From
each group, the animals were euthanized day 7 and day 14.
B. Experimental Procedures
A constant force of 20 g was generated using a closed coil spring (Sentalloy, 0.009 x
0.036, Tomy Incorporated, Tokyo, Japan) to move the maxillary first molar mesially. The
springs were attached to the maxillary left first molar and the maxillary incisors using a
stainless steel ligature wires. Light-curing bonding
Material (3M Unitek, Nonrovia, CA, USA) was applied on perforations produced by
diamond bur along the mesio-lingual and disto-lingual line angle of the maxillary first
molar and the distal sides of the incisors to ensure maximum retention of the coil spring
5
(Figure 1A). Reactivation was not done during the study period. Animals scheduled for
antibiotic treatment received 5 mg/kg bodyweight/day.doxycycline (Dentistar, Hana
Pharmaceutical company, Hwa Sung, Kyonggido, Korea) added to tap water. For CA
group, the vertical corticotomy cuts without flap reflection were performed through the
cortical layer of the bone using fine fissure bur (Figure 1B). A GluStitch (Salvin Dental
Specialties, Inc., NC, USA) was applied to improve tissue adhesion around the wound
area on the rats receiving corticotomy.
The weight of the animals was recorded on the day of the appliance insertion and
before death. All procedures were performed using general anesthesia induced by
subcutaneous injection of Zoletil (Tiletamine 125 ml, Zolazepam 125 ml; 0.04 ml Virbac,
060516 carros, France) and Rompun (Xylazire hydroxychloride 23.32 mg/ml; 0.01 ml
Bayer AG, 51368 Leverkusen, German). At the end of each experimental period the
animals were euthanized using an overdose of an anesthetic. The gingiva around the
maxillary first molar was excised, frozen in liquid nitrogen and kept at -80°C until use.
Real time reverse transcription polymerase chain reaction (real time RT-PCR) was
used to quantify IL-1β as reported elsewhere.27,28
C. Real Time RT-PCR
1. RNA extraction
TRI zol reagent (Molecular Research Center, Inc., Cincinnati, OH, USA)was
used to extract total RNA from each sample
2. cDNA synthesis
One µg total RNA was reverse transcribed into cDNA using a Promega’s
6
A
B
Figure 1. Schematic drawing of the rat model (A) orthodontic appliance placed (B)
orthodontic appliance placed after corticotomy
7
Reverse Transcription system (Promega Corp., Madison, WI, USA). Reverse
transcriptase was added in the mixture containing RT POX buffer, 2.5 mM MgCl2,
Oligo (dT) primer, dNTP Mix and Rnasin. The reagents were incubated at 42°C
for 15 min, and then heated to 95°C for 5 min.
3. Real time RT-PCR
Real time RT-PCR was performed in an ABI Prism 7000 Sequence Detection
System (Applied Biosystems, Foster City, CA, USA). Primers and probes of IL-
1β and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) are shown in
Table I. PCR amplifications were performed in a total volume of 25 µl,
containing 50ng cDNA sample, 1X TaqMan Universal PCR Master Mix, 300 nM
of each primer and 250 nM TaqMan probe following the manufacturer’s
protocol. Initial thermal cycling conditions were 2 min at 50°C and then 10 min
at 95°C. Cycle conditions for IL-1β and GAPDH were 15 sec at 95°C and 1 min
at 60°C, and RNAs for IL-1β and GAPDH were amplified by 40 cycles. For
PCR, oligonucleotide hybridization probe was labeled with 6-
carboxyfluorescein as reporter fluorescence and 6-carbocy-tetramethy-
rhodamine as quencher fluorescence.
4. Quantification using the Standard Curve Method
For quantification of results, cDNA fragments of the known sequences were
constructed as standards. The amount of GAPDH mRNA and mRNA levels of
cytokine were measured using this Standard Curve method. The results were
adjusted by the amount of GAPDH mRNA. Levels of these mRNA in each
sample were expressed as the ratio versus that of control. Every sample was
measured in triplicate.
8
Table 1. Genes analyzed by quantitative TaqMan PCR
Gene Primer/probe Sequence
IL-1β Forward primer
Reverse primer
Probe
CAA CCA ACA AGT GAT ATT CTC CAT G
GAT CCA CAC TCT CCA GCT GCA
CTG TGT AAT GAA AGA CGG CAC ACC CAC C
TNF-α Forward primer
Reverse primer
Probe
CAT CTT CTC AAA ATT CGA GTG ACA A
TGG GAG TAG ACA AGG TAC AAC CC
CAC GTC GTA GCA AAC CAC CAA GTG GA
Sequences for the primers and probes are shown in 5’to 3’ orientation; probes contain
a FAM on the 5’ end and a TAMRA on the 3’ end.
D. Tooth Movement
Each animal was placed in a specially made craniostat for standardized radiography. A
wooden wedge was placed into each ear to ensure position. An intraoral radiographic
apparatus (IMS 2000 Inermecs Co., LTD, Seoul, Korea) was used with dental film
(Insight Kodak Co., Rochester, NY, USA). The amount of tooth movement was
measured on lateral cranial radiographic images by one masked examiner. Horizontal
and vertical reference lines were used for cephalometric measurement. The horizontal
reference plane was defined as the most anterior point of the nasal bone and the most
posterior point of the squama occipitalis, and the vertical reference plane was as the
most superior point of the parietal bone and the most inferior point of the tympanic
9
bone.29 The two parallel lines to the parietal-tympanic plane were used to measure the
amount of mesial movement of the maxillary first molar (Figure 2). One line was drawn
on the most anterior point of the anterior border of the maxillary second molar crown,
and the other on the most posterior point of the posterior border of the maxillary first
molar crown.29 The distance between the maxillary first and second molar digitized with
a film scanner (PowerLook 1100, UMAX, Hyo Sung Electric Light LTD, Seoul, Korea)
was measured using cephalometric image analysis (V-Ceph. Cybermed Inc, Seoul,
Korea).30 The distance was measured thrice to statistically evaluate the reproducibility
of the examiner. The examiner exhibited high reproducibility.
Figure 2. Landmarks on lateral cephalometric radiographs. Oc, most posterior point of
squama occipitalis; Pa, most superior point of parietal bone; T, most inferior point of
tympanic bone; Na, most anterior point of nasal bone.
10
E. Statistical Analysis
Data were expressed as means ± standard error of the mean and analyzed by
nonparametric standard techniques such as Wilcoxon and Mann-Whitney tests as
appropriate. A p-value <0.05 was required for statistical significant.
11
III. Results
The orthodontically treated animals drank and ate less during the first 3-4 days of study
but rebounded to normal food and water ingestion and did not seem to be bothered by
the orthodontic appliance, hence body weight increased over the experimental period.
Four animals were excluded from the analysis due to appliance displacement.
In the NDA group, a statistically significant difference in mRNA levels for IL-1β was
found between the pressure side (left side) and the contralateral control side(right side).
mRNA expression of IL-1β on the pressure side was greater at 7 days compared with
at 14 days, while that on the control side showed no difference between at 7 and 14
days (Figure 3). The similar results were observed in the DA group (Figure 4).
Comparing the NDA group to the DA group, no difference was found in IL-1β mRNA
expression on the pressure side day 7 and 14. On day 14, IL-1β mRNA expression was
statistically significantly decreased on both groups (Figure 5). On control side, there
was no difference between the groups as well. On the other hand, IL-1β mRNA
expression on control side of DA group revealed statistically significant increase from
day 7 to day 14 (Figure 6).
12
Figure 3. IL-1β mRNA expression (Means and SEM) in the no doxycycline + appliance
group (NDA). * P < 0.05
13
Figure 4. IL-1β mRNA expression (Means and SEM) in the doxycycline + appliance
group (DA). * P < 0.05
14
Figure 5. IL-1β mRNA expression (Means and SEM) on pressure side for the DA and
NDA groups. * P < 0.05
15
Figure 6. IL-1β mRNA expression (Means and SEM) on control side for the DA and
NDA groups. * P < 0.05
16
In the CA group, a statistically significant difference in IL-1β mRNA expression was
found between the pressure side and the control side at day 7 and 14. IL-1β mRNA
expression on the pressure side was greater at day 7 compared to day 14 (Figure 7).
Comparing the A group to the CA group, no significant difference was found in IL-1β
mRNA expression on the pressure side day 7. On the other hand, IL-1β mRNA
expression in the CA group showed a statistically significant expression on day 14
(Figure 8). On the other hand, IL-1β mRNA expression on control side of CA group
revealed statistically significant difference on day 7 compared to that in the A group
(Figure 9).
No significant difference in tooth movement was found between the DA and NDA groups
day 7 and day 14 (Figure10). However, a statistically significant increase in tooth
movement was found between the A and CA groups (Figure 11).
17
Figure 7. mRNA levels (relative to control) for IL-1β in the corticotomy + appliance
group (CA). * P <0.05
18
Figure 8. mRNA levels (relative to control) for IL-1β per group on pressure side in CA.
* P <0.05
19
Figure 9. mRNA levels (relative to control) for IL-1β per group on control side in CA.
* P <0.05
20
Figure 10. Tooth movement in the DA and NDA groups.
21
Figure 11. Tooth movement in the CA and A group. * P <0.05
22
IV. Discussion
Tooth movement has, as would be expected, an impact on the magnitude of
periodontal tissue response.1 This influence can be modulated by the administration of
doxycycline, probably because of the differences in alveolar bone response.19,31-32
Tetracycline has been known to act as a collagenase inhibitor, and influence the
recruitment and function of osteoclasts.33-43 Additionally, the absolute volume of the
alveolar bone was significantly increased in rats received doxycycline under
orthodontic force.19 These results elicited evaluation of the biologic process on cellular
levels related to tooth movement after administration of doxycycline.
There was a statistically significant decrease in IL-1β mRNA expression from day 7 to
day 14 for both DA and NDA groups. It coincides with the study in which the maximum
levels in IL-1β and IL-6 were detected on day 3 following application of orthodontic
force and then decreased on day 7 and 10 for return to the baseline levels.44
There was no difference of IL-1β mRNA expression between the NDA and the DA
groups at any time point. It was demonstrated that odontoclast and osteoclast
recruitment stimulated by orthodontic force was not affected by doxycycline treatment
on day 7.19 Therefore, it can be speculated that doxycycline treatment may not affect a
biologic process related to tooth movement for short time period. However, care should
be taken on administration of doxycycline for long time period because IL-1β mRNA
expression on control sides in the DA group was statistically significant increased from
day 7 to day 14.
23
IL-1β mRNA expression on pressure side at day 14 is significantly greater in the CA
group compared to the A group. This trend could be explained as a reflection of the
regional acceleratory phenomenon (RAP) where healing activity was dramatically
increased adjacent to the injury site after surgical wounding of the osseous tissue.21,22 It
was reported that evidence of RAP was first observed 10 days after mucoperisteal
surgery in rats.45 It may be interpreted that IL-1β mRNA expression in the CA group
may be affected on day 14 in this study.
IL-1β mRNA expression on control side at day 7 is significantly greater in the CA group
compared to the A group. A plausible explanation may be found in a previous study that
tissue reactions to mechanical force were not localized to the loaded tooth.46 Moreover,
it appeared that tissue responses extended to the adjacent tooth, like the whole
hemimaxilla reacted to the orthodontic force applied to the first molar.
The primary aim of this study was to investigate the cytokines expression using RT-
PCR and the distance between the maxillary first and second molars was measured to
use as an indicator of the possible tooth movement. No statistically significant
difference in the rate of orthodontic tooth movement was found between DA and NDA
groups throughout the experiment. However, a statistically significant difference in the
rate of orthodontic tooth movement was found between CA and A groups on day 7,
although these tooth movement may not reflect the minor displacement due to
inaccuracy of the assessment method.
As antibiotics including tetracycline and erythromycine are frequently used for acne
24
treatment, administration of antibiotics may result in variations in normal bone turnover
during orthodontic treatment.15,47 Doxycycline was selected among tetracyclines for this
study, in part, because of the potent collagenase inhibitor, and because it has been
also known to decrease the total number of osteoclasts and prevent alveolar bone loss
following periodontal surgery in rats.19 The outcome of this study could be clinically
applied in that it is possible to treat patients undergoing short- term antibiotics therapy
because our data seem to indicate that the levels of cytokines, after orthodontic tooth
movement, were not affected during experimental periods. The rate of orthodontic tooth
movement was not affected by administration of doxycycline, either. A previous study
also showed that tetracycline has a beneficial effect on tooth movement by reducing
the amount of root resorption.19 A plausible explanation for decreased root resorption is
primarily by the inhibition of matrix metalloproteinases.14 In addition, tetracycline could
prevent gingival inflammation during tooth movement.34 Taken together, the orthodontic
treatment may be continued as usual for the short-term administration of doxycycline
during orthodontic treatment. However, effects of doxycyline during orthodontic
treatment have to be undertaken with larger number of samples before the clinical
application of this concept. Future research will be also focused on the influences of
cytokines on the individual activity after administration of doxycycline depending on
dose and duration.
The outcome of this study could provide the biologic process for fast rate of tooth
movement following application of orthodontic force combined by corticotomy. Our data
seem to indicate that the levels of cytokine, after orthodontic tooth movement followed
by corticotomy, were elevated for both on day 7 and on day 14. Especially, the
25
differences in the levels of cytokine on day 14 were statistically significant. The rate of
orthodontic tooth movement was also increased in corticotomy groups. Taken together,
the orthodontic treatment combined with corticotomy may be performed in cases where
reduction of orthodontic therapy treatment time is required.
The cytokines levels evaluated in human GCF did not provide the information of origin
in cytokines; at transcription, at translation or simply a product.3 For this reason, the
mRNA expression of these cytokines using RT-PCR was needed to investigate the
cellular induction of cytokines. RT-PCR has the advantage of quantification of the
cytokines, especially in cases where the available samples to be analyzed are too
small to quantify the cytokines at the protein level. 27 In addition, RT-PCR is thought to
be a very powerful and sensitive method, while the ELISA technique for cytokine
protein detection allows only a limited number of cytokines.27
26
V. Conclusion
Inflammatory cytokines expressed during orthodontic tooth movement can be
quantified on a cellular level using RT-PCR. This study evaluated the effect of
doxycycline and corticotomy on IL-1β, believed to play important roles in bone
remodeling following application of orthodontic forces. The results suggest that the
expression of IL-1β is not affected by doxycycline, but a statistically significant
expression of IL-1β was shown at day 14 in the CA groups.
These observations support the hypothesis that osteoclast recruitment stimulated by
orthodontic force is not influenced by doxycycline, and regional acceleratory
phenomenon after performance of corticotomy increases bone remodeling during
orthodontic tooth movement.
Moreover, our study has shown that RT-PCR appears powerful and sensitive method to
quantify mRNA expression in cases where the available samples too small to quantify
the cytokines at the protein level and cellular induction of cytokines with a very short
half-life needs to be assessed during orthodontic tooth movement.
27
References
1. Krishnan V, Davidovitch Z. Cellular, molecular, and tissue-level reactions to
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32
Figure Legends
Figure 1. Schematic drawing of the rat model (A) orthodontic appliance placed (B)
orthodontic appliance placed after corticotomy
Figure 2. Landmarks on lateral cephalometric radiographs. Oc, most posterior point of
squama occipitalis; Pa, most superior point of parietal bone; T, most inferior point of
tympanic bone; Na, most anterior point of nasal bone.
Figure 3. IL-1β mRNA expression in the no doxycycline + appliance group:NDA.
* P < 0.05
Figure 4. IL-1β mRNA expression in the doxycycline + appliance group: DA.
* P < 0.05
Figure 5. IL-1β mRNA expression on pressure side for the DA and NDA groups.
* P < 0.05
Figure 6. IL-1β mRNA expression on control side for the DA and NDA groups.
* P < 0.05
Figure 7. mRNA levels for IL-1β in the corticotomy + appliance group: CA.
* P <0.05
33
Figure 8. mRNA levels for IL-1β per group on pressure side in CA. * P <0.05
Figure 9. mRNA levels for IL-1β per group on control side in CA. * P <0.05
Figure 10. Tooth movement in the DA and NDA groups.
Figure 11. Tooth movement in the CA and A group. * P <0.05
34
국문국문국문국문 요약요약요약요약
독씨싸이클린독씨싸이클린독씨싸이클린독씨싸이클린 (Doxycycline)과과과과 코티카토미코티카토미코티카토미코티카토미 (Corticotomy)가가가가 교정적교정적교정적교정적 치아치아치아치아
이동시이동시이동시이동시 Interleukin-1ββββ(IL-1ββββ)에에에에 미치는미치는미치는미치는 영향영향영향영향
연세대학교 대학원 치의학과
(지도 백형선 교수님)
임원희
싸이토카인 (cytokines)은 교정 치료시 일어나는 골 개조에 중요한 역할을 하는 것
으로 알려져 있다. 이 실험의 목적은 치아 이동시 독시싸이클린 (doxycycline)의
투여와 corticotomy 시행시 Interleukin-1β(IL-1β) 표현의 변화를 보고자 함이다.
9주된 36마리의 수컷 위스타(Wistar) 쥐의 상악 제일구치에 교정력을 가한 후에
IL-1β messenger RNA(mRNA)의 표현을 보고자 real time reverse transcription
polymerase chain reaction을 행하였다. 9마리의 쥐에서는 독시싸이클린 투여 없이
치아 이동을 위한 교정 장치만 걸고, 다른 9마리의 쥐에서는 치아 교정 장치와 함
께 독시싸이클린을 투여하였다. 또 다른 9마리의 쥐에서는 교정 장치를 건 후,
corticotomy를 시행했고, 나머지 9마리 쥐는 교정 장치만 걸었다. 6마리의 쥐는 아
무 것도 시행하지 않았다. 쥐들은 교정 장치 삽입 7일 혹은 14일 후에 희생되었으
며 분석을 위해 상악 제일구치 부위의 잇몸 조직을 채취하였다.
35
독시싸이클린을 투여한 군과 투여하지 않은 군을 비교 시 7일째에는 비슷한 IL-1β
의 표현을 보였고, 14일에서는 두 그룹 모두에서 IL-1β의 양이 감소하였다.
Corticotomy를 시행한 군과 시행하지 않은 군을 비교 할 때 7일과 14일에서
corticotomy를 행하고 치아 이동을 한 군에서 IL-1β의 양이 크게 증가하였다.
결론적으로, 치아 이동 시 doxycycline의 투여는 IL-1β의 표현에 영향을 주지 않
았으나, corticotomy와 함께 치아 이동을 한 경우에는 IL-1β의 증가를 볼 수 있었
다.
핵심되는 말: 독시싸이클린 (doxycycline), 코티카토미 (corticotomy). 치아 이동, IL-
1β 표현 (expression)
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