Stimulatory effects of low-power laser irradiation on

bone regeneration in midpalatal suture during

expansion in the rat
Shiro Saito, DDS, ~ and Noriyoshi Shimizu, DDS, PhD b
Chiba, Japan

The purpose of this study was to investigate the effects of low-power laser irradiation on bone
regeneration during expansion of a midpalatal suture in rats. GaUium-aluminum-arsenide diode laser
100 mW irradiation was applied to the midpalatal suture during expansion carried out over 7 days
(3 or 10 minutes per day), 3 days (7 minutes per day for day 0-2 or 4-6), and 1 day (21
uninterrupted minutes on day 0). The bone regeneration in the midpalatal suture estimated by
histomorphometric method in the 7-day irradiation group showed significant acceleration at 1.2- to
1.4-fold compared with that in the nonirradiated rats, and this increased rate was irradiation dose-
dependent. Irradiation during the early period of expansion (days 0 to 2) was most effective,
whereas neither the later period (days 4 to 6) nor the one-time irradiation had any effect on bone
regeneration. These findings suggest that low-power laser irradiation can accelerate bone
regeneration in a midpalatal suture during rapid palatai expansion and that this effect is dependent
not only on the total laser irradiation dosage but also on the timing and frequency of irradiation. We
suggest laser therapy may be of therapeutic benefit in inhibiting relapse and shortening the
retention period through acceleration of bone regeneration in the midpalatal suture. (Am J Orthod
Dentofac Orthop 1997;111:525-32.)

Rapid palatal expansion ( R P E ) is the
preferred treatment approach to correct a con-
stricted maxillary dental arch. It is known, however,
that a long period of retention is necessary to
prevent early relapse of the expanded arch. ~'2 Al-
though the reason for the early relapse is not fully
clear, bone regeneration in the midpalatal suture
after expansion may affect the posttreatment re-
lapse. It would be potentially beneficial therefore to
accelerate bone formation in the midpalatal suture
after expansion to prevent relapse of arch width and
to abbreviate the retention period.
Various biostimulatory effects of low-power la-
ser irradiation have been reported since 1971, 3 and
the acceleration of bone formation by laser treat-
ment has been a focus of contemporary research.
Most of these reported studies were limited to
clinical 4.5 or qualitative studies that use in vivo
experimental models. ('-1~ The effects of laser irradi-
From Nihon University School ol Dentistry at Matsudo, Chiba, Japan.
This research was supported in part by a grant-in-aid fi)r scientific research
from the Ministry of Education, Science and Culture of Japan (06454586).
"Postgraduate student, Department of Orthodontics.
"Assistant protessor, Department of Orthodontics.
Reprint requests to: Dr. Shiro Saitt~, Dcpartmen| of Orthodontics, Nihon
University Sch~ol of Dentistry at Matsudo, 2-870-1, Sakaechu-nishi, Mat-
sudo city, Chiba, 271, Japan.
Copyright © 1997 by the American Association of Orthodontists.
0889-5406/97/$5.00 ~- 0 8/i/69698
ation on bone regeneration in the suture are not
known.
We investigated the stimulatory effects of vari-
ous low-power laser irradiation on bone regenera-
tion during expansion of midpalatal suture in rats.
These effects were evaluated with quantitative bone
histomorphometry and histologic examination.
MATERIALS AND METHODS
Expansion of Midpalatal Suture
A total of 76 male Wistar strain rats were used for this
experiment (56 rats for histomorphometric and 20 rats for
histologic examination). The rats were kept in separate
cages in a 12-hour light/dark environment at the constant
temperature of 23° C and provided with food and water ad
libitum. The health status of each rat was evaluated by
daily body weight monitoring for more than 1 week before
the start of the experiment (at 6 weeks old, the weight was
180 _+ 10 gm).
For the histomorphometric examination, 56 rats were
divided, according to the laser treatment protocol, into
seven groups of eight rats each (Fig. 1), as follows:
1. Intact group: Nontreated.
2. Nonirradiation control group: Midpalatal suture
was expanded but nonirradiated.
3. Two 7-day irradiation groups: Irradiation for either
(a) 3 minutes per day or (b) 10 minutes per day was

525
526 Saito and Shimizu
Bone labeling Calcein
1.
2. Nonirradiation conta'ol group
F-
3. 7-day irradiation group - ~
[_
[~
4. 3-day irradiation group
(7min/day) [--
5. Single irradiation group (21 rain, on day 0) ~ 7
• : Expansion of midpalatal suture
Fig. 1. Irradiation schedule in each group.
Fig. 2. Lateral view during laser irradiation of midpala-
tal suture in rat. Suture was expanded indirectly by
inserting metal ring between maxillary incisors. Irradia-
tion was performed by placing optical fiber (arrow) in
contact with tissue to prevent reflection of laser beam
onto tissue surface.
applied once daily from just after expansion (on
day 0) until day 6 (total dosage: 126 J and 420 J,
respectively).
4. Two 3-day irradiation groups: Irradiation for 7 min-
utes per day was repeated once daily on either (a)
days 0 to 2 or (b) days 4 to 6 (total dosage: 126 J).
5. Single irradiation group: Irradiation for 21 uninter-
rupted minutes (126 J) was performed just after the
expansion, on day 0.
After anesthetizing with an intraperitoneal injection of
sodium pentobarbital (50 mg/kg body weight), all rats
except the intact group were subjected to expansion of
midpalatal suture according to the method of Sawada and
Shimizu. 12 Expansion was performed by inserting a 1.5
mm thick circular metal ring composed of stainless steel
American Journal of Orthodontics and Dentofacial Orthopedics
May 1997
Tetr~ycline Calcein
+
Day 0 1 2 3 4 5 6
Intact group
•
3 min/day '~ V V V V V V
10 rain/day '~ V V V V V V P
on days 0-2 V V It,
on days 4-6 • V V V IP
ID
V : Laser irradiation
0.5 mm diameter wire (Sankin, Osaka, Japan) between the
maxillary incisors. The ring was retained by a 0.012-inch
diameter wire in holes that were drilled laterally in the
incisors just above the gingival papilla with a no. 1/4 round
bur. The thickness of the metal ring was determined on
the basis of the results of a preliminary study indicating
that 1.5 mm expansion between the incisors induced the
maximum expansion rate in the midpalatal suture without
continuous decrease of the animal's body weight.
Procedure of Laser Irradiation
As the low-power laser source, a gallium-aluminum-
arsenide (Ga-AI-As) diode laser device (Panalas 1000
prototype: Matsushita Industrial Equipment Inc.) was
used. The technical specifications of this laser device are
as follows: wave length: 830 nm; output power: 100 to 700
roW, variable. Continuous wave at 100 mW output power
(power density: 35.3 J/second/cm 2) was used. Diode laser
device has been often used for experimental and clinical
studies on bone repair. 5,9,1°
The laser beam was delivered by an optical fiber 0.6
mm in diameter, and irradiation was administered under
general anesthesia by placing the end of the optical fiber
tip in contact with the palatal mucosa at the midline and
the median point between the anterior edges of incisors
and incisive papilla (Fig. 2). This site was determined on
the basis of results of the preliminary study indicating that
the midpalatal suture showed parallel expansion that was
subject to quantitative analysis of new bone formation.
Bone Histomorphometry
For triple bone labeling, all rats were injected subcu-
taneously with calcein (Doiin Co.) at the dose of 8 mg/kg
on day 0 and day 6, and they were also injected with
oxytetracycline (Terramycin intramuscular solution,
Taito-Pfizer) at the dose of 50 mg/kg on day 3. The rats
were killed on day 7 with an intraperitoneal injection of a
American Journal of Orthodontics and Dent@ciat Orthopedics
Volume I 1 I, No, 5
lethal dose of sodium pentobarbital. The maxillae, includ-
ing the irradiation site were removed, fixed in ethyl
alcohol overnight, and prepared for histomorphometric
examination. The bone specimens were immersed in
Villanueva bone stain solution (5 mg/ml in 70% methanol;
Maruto Instrument Co.) for 72 hours. This Villanueva
bone staining method permits simultaneous visualization
and analysis of osteoid, mineralized bone and bone label-
ing of calcein and tetracycline. 13.~4 The specimens were
embedded in methyl methacrylate resin, using routine
methods. Each plastic-embedded bone specimen was cut
in the frontal plane at a right angle to the occlusal plane
of the upper molars. Five sections of approximately 100
Ixm thickness centering around the irradiation site were
made and then ground to 30 Ixm thickness.
The ground sections were photographed with a fluo-
rescent microscope (Olympus BHS, Olympus) under ul-
traviolet light, and a division scale was also photographed
at the same magnification for quantitative evaluation.
Each labeling line and the outline of the osteoid seams
were traced from the photographs. These traces were then
entered into a personal computer. The primary parame-
ters, which are mentioned later, were measured by using
image analysis software (Ultimage, Ver. 2.0, Graftek
France). Measurement for bone histomorphometry was
performed in areas measuring 500 Ixm horizontally × 600
~tm vertically located 200 Ixm under the surface of the
osseous palate because bone formation was irregular
adjacent to the oral and nasal cavities, making those areas
unsuitable for quantitative measurement. The error value
of the method was less than 8.8 × 10.3 mm 2 for the area
measurements and 0.4 Ixm for the linear measurements
assessed with measurements of each specimen five times.
The primary parameters measured for this experiment
were as follows:
1. Newly formed mineralized bone area: The area
covered with lines labeled on day 0 and day 6.
2. Osteoid area: The area of osteoid seams stained
orange by Villanueva bone stain.
3. Mineralized width on days 0-3: Average width
between lines labeled on day 0 and day 3.
4. Mineralized width on days 3-6: Average width
between lines labeled on day 3 and day 6.
The mean mineralized width was defined as the cross
width between two lines of 10 points that were placed at
intervals of 100 txm vertically along both sides of the
suture in the area of measurement.
As a secondary parameter derived from the mineral-
ized width, the mineral appositional rate (MAR), the
distance between labels (jxm) divided by labeling period
(days), was calculated separately for days 0-3 and days 3-6
as an indicator of the width of mineralized bone each day.
Histologic Examination
For histologic examination, 20 rats were divided into
three groups of five each: an intact group, a nonirradiation
Saito and Shimizu 527
control group and two 7-day irradiation groups (3 or 10
minutes per day). Treatment in each group was applied at
the time of bone histomorphometric examination. All rats
were fixed on day 7 and the maxillae were removed as
previously described. After demineralization in 10%
EDTA, they were embedded in paraffin and sectioned at
5 fxm. The sections were stained with hematoxylin eosin
and examined.
Statistical Analysis of Data
The values in each figure and table are presented as
the mean _+ SD for each group. Intergroup comparisons
of the mean value were conducted with Tuckey's t test for
the analysis of body weight change and Student's t test for
equal variance for other comparisons. In the analysis of
the effect of laser irradiation for 7 days, the average values
of multiple groups were compared by one-way analysis of
variance (ANOVA). A value ofp < 0.05 was considered a
significant difference.
RESULTS
Changes of Body Weight During the
Experimental Period
The body weights of the rats in the nonirradia-
tion control group and in all of the irradiation
groups decreased on day 1 but subsequently recov-
ered. This reduction was significant until day 3 (p <
0.05), but there was no significant difference there-
after. No statistically significant differences were
revealed by testing with Tuckey's t test among the
nonirradiation control and the various irradiation
groups (Fig. 3).
Effects of Laser Irradiation on Bone Regeneration
The ground sections stained by the Villanueva
bone staining method revealed clear triple bone
labeling along the bone edges of the midpalatal
suture and the osteoid inside of the bone edges (Fig.
4). The width between each labeling line was wider
in the laser irradiation groups compared with the
nonirradiation control group. The quantitative eval-
uation by histomorphometric method showed that
the newly formed mineralized bone area in the 7-day
irradiation groups (3 and 10 minutes/day) was sig-
nificantly increased (by 21.5% at 3 minutes/day; by
34.4% at 10 minutes/day) compared with the area in
the nonirradiation control group (p < 0.01) (Fig. 5,
A). This increase occurred in an irradiation dose-
dependent m a n n e r (p < 0.01, one-way A N O V A ) .
Laser irradiation significantly stimulated the M A R
during the experimental period (11.6 txm/day in the
3-minute/day group; 13.8 txm/day in the 10-minute/
day group) c o m p a r e d with that in the nonirradiation
control (8.2 ixm/day) (p < 0.01), and this effect also
528 Saito and Shimizu
240
220
200
180
I I I i I i l l
0 1 2 3 4 5 6 7 day
Intact group
x Nonirradiadoncontrolgroup
-----~ 3 min/day
10 min/day ~-" 7-dayirradiationgroup
on days0-2 ~ 3-dayirradiationgroup
on days4-6
• Singleirradiationgroup (21 min on day O)
Fig. 3. Changes of mean body weight during experi-
mental period. Mean body weight in intact group was
significantly higher on days 1 through 3 than that in any
expansion groups (p < 0.05), although there were no
significant differences between nonirradiation control
group and other laser irradiation groups (by Tuckey's t
test).
occurred in an irradiation dose-dependent manner
(p < 0.01, one-way ANOVA). Both the MAR
determined in the early period (days 0-3) and the
later period (days 3-6) of the experiment were
significantly increased compared with the corre-
sponding control periods. However, the MAR in the
early period was significantly greater than that in the
later period (days 0-3 MAR/days 0-7 MAR: 58.6%
in the 3-minute/day group; 58.0% in the 10-minute/
day group)(p < 0.05), whereas no significant differ-
ence was seen in the nonirradiation control group
(Fig. 5, B).
In the 3-day irradiation groups, laser irradiation
performed during the early period (days 0-2) signif-
icantly stimulated the newly formed mineralized
bone area and MAR compared with those in the
nonirradiation control group (t9 < 0.01), whereas
laser irradiation in the later period (days 4-6) did
not affect them (Fig. 6, A and B). This increase was
due to acceleration of bone regeneration only dur-
ing the earlier part of the experimental period and
American Journal of Orthodontics and Dentofacial Orthopedics
May 1997
A -- B
C D
Fig. 4. Photographs taken under fluorescent micro-
scope in each group examined. (A) intact group; (B)
nonirradiation control group; (C) 7-day irradiation group
(3 minutes/day); and (D) 7-day irradiation group (10
minutes/day). Upper and lower side in each image is
oriented toward nasal cavity and toward oral cavity,
respectively. Midpalatal suture (*) was expanded except
in intact group (A). Width between each labeling line
was wider in laser irradiation groups (C and D) com-
pared with nonirradiation control group {B). Measure-
ments of primary parameters were performed in 500 x
600 i~m area located 200 i~m deep to surface of
osseous palate (white frame). Bar = 100 t~m.
not to that during the later period (days 3-6). Single
irradiation did not affect the newly formed mineral-
ized bone area (Table I).
When the osteoid area of each irradiation
groups was compared with the control, there was no
significant difference in any irradiation group tested
(Table II).
Histologic Examination
Microscopic observation of the expanded mid-
palatal suture in the expansion groups revealed
extension of the transverse fibers and enlargement
of the capillaries in the suture, which may be due to
mechanical tension. No pathologic change such as
excessive intimation or scald was observed around
American Journal of Orthodontics and Dentofacial Orthopedics Saito and Shimizu 529
Volume 1 1 1 , N o . 5

A B
~ ~" 600 15 [ ] day 3-6 t
.~ ~ 500 [ ] day 0-3 t

s 400 10

" 300 I I ] °
0
200 e'.,

, 100
I I
z 0

e~, ej. %

% "oo

Fig. 5. A. Effect of laser irradiation (3 or 10 minutes/day) for 7 days on newly formed

mineralized bone area. Newly formed mineralized bone area in laser irradiation group was
significantly increased compared with that in nonirradiation group (1: P < 0.01). This effect was
irradiation dose dependent (one-way ANOVA, p < 0.01). B. Effect of laser irradiation (3 or 10
minutes/day) for 7 days on mineral appositional rate (MAR). MAR during experimental period
in irradiation group was significantly increased compared with that in nonirradiation control
group (1-:P < 0.01). This increase was irradiation dose dependent (one-way ANOVA, p < 0.01).
MARs in both earlier (days 0 to 3) and later (days 3 to 6) period were significantly increased
compared with corresponding period in control group; moreover, MAR in earlier period was
significantly higher than that in later period (*: p < 0.05).

the irradiation site of the suture in the 3- and
10-minute per day irradiation groups (Fig. 7).
DISCUSSION
In this study, we clearly demonstrated the stim-
ulatory effects of low-power Ga-A1-As diode laser
irradiation on bone regeneration in the midpalatal
suture during maxillary expansion by using a histo-
morphometric method. This method is a reliable
technique that is frequently used in quantitative
evaluation of bone remodeling in vivo. 14 The results
of our study indicate that the newly formed miner-
alized bone area and MAR were significantly in-
creased by the 7-day irradiation and that this in-
crease was dose dependent. The MAR activation
produced by the 7-day irradiation was more pro-
nounced in the earlier period. Furthermore, when
laser irradiation was applied only for the early 3
days, bone regeneration in the newly formed miner-
alized bone area was stimulated significantly com-
pared with the control, whereas irradiation applied
during the later 3 days had no effect on it. This
phenomenon was thus induced by early but not by
late stimulation of the bone regeneration.
Although tension in the expanding site should be
highest just after expansion and should gradually
decrease thereafter, the similarity of the bone for-
mation rates in the earlier and the later 3-day
periods after expansion (48% and 52%, respec-
tively) observed in the control group suggests that
the activity of bone regeneration in response to
tension is constant throughout the expansion period.
If it were to be applied clinically, laser irradiation
would be effective during the early rather than the
later stages of expansion. Although only the early
period irradiation stimulated bone regeneration,
sustained irradiation over a 7-day period accelerated
the MAR in both of these periods. This finding
suggests that, although laser application in the early
stage activates bone regeneration in the expanded
suture, the later laser treatment may play a role in
the maintenance of this bone regeneration activity.
We predict, in the absence of continued application
of laser treatment, the regenerative activity may
diminish. When we examined the effect of single
irradiation on day 0 for 21 minutes at the same total
dosage as the 7-day irradiation for 3 minutes or the
3-day irradiation for 7 minutes, it did not affect bone
530 Saito a n d S h i m i z u American Journal of Orthodontics and Dentofacial Orthopedics
May 1997

A B
~ 600 15 [ ] day 3-6
r~
.= .~ 500
,.Q [ ] day 0-3 t

~. 400 T ,0
.E 300 o
E
200 5
100
..=
Z 0 0

"Od

Fig. 6. A. Effect of laser irradiation (7 minutes/day) for 3 days on newly formed mineralized
bone area. Laser irradiation on days 0 to 2 significantly increased newly formed mineralized
bone area compared with that in nonirradiation control (t: P < 0.01), whereas irradiation on
days 4 to 6 did not have any effect on area. B. Effect of laser irradiation (7 minutes/day) for
3 days on MAR. Laser irradiation on days 0 to 2 significantly increased MAR compared with
that in nonirradiation control group (1-:P < 0.01), whereas irradiation on days 4 to 6 did not
affect MAR. MAR activation by laser irradiation was observed only in earlier period (days 0
to 3) with significant difference compared with corresponding period in control group (*:
p < 0.05).

Table I. Mean newly formed mineralized bone area in the Table II. Mean osteoid area in each experimental group.
single irradiation group. There was no significant difference There was no significant difference between the area in the
between the areas in the irradiation and nonirradiation control nonirradiation control group and that in any of the irradiation
groups (Student's t test) groups (Student's t test)

Mean newlyformed Experimental group Mean osteoid area (×10 3 mm 2)

mineralized bone area
~ l group (×10-3 mm2)
Nonirradiation control group 363.9 Z 50,7
Single irradiation group (21 347.9 + 39.67
minutes irradiation, on day 0)
regeneration. It is likely that bone regeneration is
dependent not only on the total dosage of laser
irradiation but also on the timing and the manner of
irradiation. Specifically, irradiation at the early stage at
the active site and repeated irradiation for a certain
period would be effective. These results are consistent
with those of earlier reports that multiple irradiation is
more effective than a single dose for acceleration of
bone formation 9 or fibroblast growth. 15
The mechanism by which laser irradiation pro-
motes bone formation is not fully understood.
Nonetheless, our results corroborate the results of
an in vitro study by Ozawa et al. 16 They reported
Intact group 136.35 -+ 15.18
Nonirradiation control group 144.72 -+ 43.59
7-day irradiation group (3 min/day) 150.83 -+ 43.13
7-day irradiation group (10 min/day) 148.85 -+ 38.75
3-day irradiation group (on days 0-2) 160.50 -4-22.66
3-day irradiation group (on days 4-6) 136.48 -~ 25.08
Single irradiation group (21 minutes 153.87 +- 26.23
irradiation, on day 0)
that the number of calcified bone nodules formed by
isolated rat calvarial osteoblasts was significantly
increased by low-power laser irradiation and con-
cluded that laser irradiation early in the cell culture
period is more effective in inducing nodule forma-
tion due to differentiation of osteoblasts and that
osteoblasts may have a specific optimal dosage of
irradiation for acceleration of differentiation.
Schneider et al. 17 reported the rapid generation of
myofibroblasts from fibroblasts when stimulated by
laser irradiation.
American Journal of Orthodontics and Dentofacial Orthopedics" Saito and Shimizu 531
Volume 111, No. 5

Because osteoblasts may be recruited along the

bone edges from undifferentiated precursor cells in
the expanding midpalatal suture, laser irradiation
could potentially stimulate the recruitment and/or
maturation of osteoblasts. This possible mechanism
is also supported by the results of several other
studies indicating that laser irradiation is effective
when applied at healing sites, such as those of bone
fracture,4"6 bone defect;s,7,8 tooth extraction,~,1° and
transplantation of bone morphogenetic protein. 11 In
contrast, laser irradiation is not effective when ap-
plied at inactive or normal tissue sites.
There have also been some studies of stimula-
tion of collagen synthesis, ~,~'~ including that by
Saperia et al., 2° which clearly demonstrated that the
mRNA level of type I collagen was increased by
laser irradiation during skin wound healing. As type
I collagen is a major matrix protein in bone, the
observation of laser stimulation of collagen level is
at least indirectly supportive of a stimulatory effect
of laser irradiation on bone formation.
In the irradiated groups, the osteoid area was
increased as well as bone regeneration. But this
increase was not significantly different from that in
the nonirradiation controls. We observed that the
osteoid formation was usually irregular and the
osteoid area was much smaller than the mineralized
bone area. It is possible that the lack of statistically
significant difference could be for these reasons.
We found that the body weight of the rats in the
irradiated groups was no different from that in the
nonirradiation control group, and no evidence of
disease was present at the site of irradiation. These
observations are not surprising as low-power laser
therapy is widely used with considerable efficiency in
the fields of orthopedic surgery,4 dermatology,21 and
pain control 22 without side effects.
In orthodontics, RPE is a common treatment
strategy, but the expanded arch easily relapses un- Fig. 7. Microscopic photographs in (A) intact group
less it is retained for a prolonged period. The rat, (B) nonirradiation control group rat, and (C) 7-day
possible causes of relapse may include unstable oral irradiation group (10 minute) rat. Extension of transverse
fibers, enlargement of the capillaries in suture, and
myofunction,23 regeneration of sutures connected to plump osteoblasts lining bone edge were observed in
other facial bones, 24 tension of palatal connective both expansion groups (B and C), and no pathologic
tissue, and alveolar bone remodeling. 25 We argue, change was observed in either nonirradiation control or
however, that one of the major cause of early irradiation groups. Bar = 50 i~m.
relapse after expansion could be the insufficient
bone regeneration at the midpalatal suture. In this expansion was found to be effective. This may be a
study, we demonstrated an increase of approxi- serendipitous and favorable protocol for clinical
mately 35% of the newly formed mineralized bone application of laser therapy: Several visits to the
area in the midpalatal suture with laser irradiation. orthodontic office for brief periods of daily irradia-
Although the single irradiation had no effect on tion in the early stages of expansion may be all that
bone formation, intermittent application of laser is needed to stimulate bone regeneration in the
over several time periods in the early stages of suture. Because the relationship between bone re-
532 Saito and Shimizu
generation in the midpalatal suture and the stability
of expansion was not within the scope of our study,
relapse potential after laser therapy during expan-
sion will be clarified in further studies.
The Ga-AI-As diode laser is known to be a
high-tissue penetration laser because hemoglobin
and water have a low coetficient of absorption for
it. 26 A previous study revealed that approximately
50% of the diode laser beam (60 mW) penetrates a
depth of 1.0 mm in human or bovine mandibular
cortical bone, 27 and it is likely that the diod~ laser
used in the current experiment may have had
enough energy to penetrate the midpalatal suture in
the rat.
CONCLUSION
In conclusion, low-power Ga-AI-As diode laser irra-
diation significantly stimulated bone regeneration in the
midpalatal suture during RPE in rats. Although further
studies are still required, the introduction of laser therapy
to RPE in orthodontic treatment seems feasible, and it
may be of great therapeutic benefit in inhibiting relapse or
shortening the retention period, or both.
We are indebted to Professor T. Iwasawa, Depart-
ment of Orthodontics, and Professor H. Yamamoto,
Department of Pathology, Nihon University School of
Dentistry at Matsudo, for their advice and critical com-
ments. We also thank Mr. A. Kaneda, senior engineer at
Matsushita Industrial Equipment, Inc., for providing the
laser device.
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