173

Original Article
173
Bull. Tokyo dent. Coll., Vol. 45, No. 3, pp. 173179, August 2004
INTRODUCTION
One of the key factors in preventing dental
caries has been recognized to be increasing
the resistance of teeth to acid by encouraging
the development of re-mineralization mecha-
nisms on the enamel surface. The daily topi-
cal application of a uoride, such as sodium
uoride (NaF), acidulated phosphate uo-
ride solution (APF), or stannous uoride
(SnF
2
), usually by the daily use of a uoride-
containing dentifrice, has been reported to
be of benet both in professional clinics and
self care. Pit-and-ssure sealants are also useful
COMPARISON OF SHORT-TERM IN VITRO FLUORIDE
RELEASE AND RECHARGE FROM FOUR DIFFERENT
TYPES OF PIT-AND-FISSURE SEALANTS
HIROSHI KOGA, ATSUSHI KAMEYAMA*, TAKASHI MATSUKUBO,
YOSHITO HIRAI* and YOSHINORI TAKAESU
Department of Epidemiology and Public Health, Tokyo Dental College,
1-2-2 Masago, Mihama-ku, Chiba 261-8502, Japan
* Third Department of Conservative Dentistry, Tokyo Dental College,
1-2-2 Masago, Mihama-ku, Chiba 261-8502, Japan
Received 18 October, 2004/Accepted for Publication 15 December, 2004
Abstract
Objective : The purpose of this in vitro study was to assess the effects of four commercial
uoride-containing pit-and-ssure sealants on caries prevention. Materials and Methods:
Four sealants containing uoride, Fuji III, Fuji III LC (GC Co., Tokyo), Teethmate F-1
(Kuraray Medical Co., Osaka) and Helioseal F (Vivadent Co., Liechtenstein) were used
to investigate uoride release and recharge. Disk-shaped specimens prepared from each
material were immersed in distilled water at a temperature of 37C. After seven days,
acidulated phosphate uoride solution (APF) was applied to each specimen, and it was
then again immersed in distilled water for 14 days. We then determined how much
uoride had been released into the immersing water. Fuji III LC was used with APF
solution to investigate the uoride uptake. Results : Fuji III had the highest uoride
release, and Fuji III LC had the highest uoride recharge. Helioseal F and Teethmate
F-1 had almost no uoride recharge. Fuji III LC/APF had a higher uoride uptake to
enamel than Fuji III LC. Conclusions : These results suggest that GIC-sealants in the oral
cavity can serve as a uoride reservoir and contribute to retaining a low uoride level in
oral uids, thereby preventing caries.
Key words: Fluoride-containing pit-and-ssure sealant Fluoride release
Fluoride recharge Fluoride uptakeCaries prevention
174 H. KOGA et al.
in protecting against acid attack and can pre-
vent caries in both primary and permanent
molars.
It has been more than 30 years since
Buonocore
3)
, and then Takeuchi and Kizu
15)
,
developed plastic-based pit-and-ssure seal-
ants in the 1960s. Since then, tremendous
improvements and developments in new
materials such as cyanoacrylate, bisphenol
A-glycidyl methacrylate (Bis-GMA), methyl
methacrylate (MMA), and polyurethane have
been achieved and used clinically. Glass-
ionomer-cement (GIC) based sealants were
developed in the 1970s, and it was discovered
that uoride, which happened to be con-
tained in these pit-and-ssure sealants, was
slowly released into the oral cavity and con-
tributed to both preventing dental caries and
their recurrence in surrounding regions
2,12)
.
Therefore, intentionally adding uoride to
resin pit-and-ssure sealants through physical
and chemical means has become widespread,
not only to prevent dental caries but to seal
the pits-and-ssures as well. However, clinical
studies establishing the preventive effects of
uoride contained in sealing materials have
not yet been published.
The purpose of this study was to evaluate
the clinical usefulness of representative
uoride-containing pit-and-ssure sealants
by examining their uoride release during
short periods and by testing their ability to
recharge by measuring the uoride uptake to
the enamel surface adhering to the sealing
materials in vitro.
MATERIALS AND METHODS
1. Fluoride-containing pit-and-ssure
sealants materials
The pit-and-ssure sealants containing
uoride we investigated are listed in Table 1.
They were Fuji III (FIII, GC Co., Tokyo, Japan),
Fiji III LC (FIII L, GC Co., Tokyo, Japan),
Teethmate F-1 (TF1, Kuraray Medical Co.,
Osaka, Japan), and Helioseal F (HSF,
Vivadent Co., Liechtenstein). These had dif-
ferent hardening mechanisms and contained
uoride that was either there unintentionally
or had been introduced intentionally.
2. Fluoride release from uoride-containing
sealant materials
The sealing materials were shaped into disks
with an acrylic ring (11-mm outer diameter,
9-mm inner diameter, and 2.5-mm depth)
according to the manufacturers directions.
The sealant material was placed into the
acrylic ring lying on a piece of acrylic, and
another piece of acrylic was placed on top. The
acrylic pieces were then squeezed together to
form a sealant disk. Only the FIII disks were
left for a xed time to allow chemical harden-
ing the other materials were polymerized and
hardened with light-irradiation (Jetlite 1000,
Table 1 Fluoride-containing pit-and-ssure sealants tested in this experiment
Trade name Manufacturer/main components Polymerization
Fuji III GC, Tokyo, Japan Chemical
(FIII) Powder: aluminum uorosilicate glass,
Liquid: polyacrylic acid and
poly basic carboxyl acid
Fuji III LC GC, Tokyo, Japan Light/Chemical curing
(FIII L) Powder: aluminum uorosilicate glass,
Liquid: polyacrylic acid and 2-hydroxyethyl
metacrylate (HEMA)
Teethmate F-1 Kuraray Medical, Osaka, Japan Light curing
(TF1) MDP, MF-MMA
Helioseal F Vivadent, Liechtenstein Light curing
(HSF) Fluorosilicate glass
175 FLUORIDE RELEASE AND RECHARGE FROM SEALANTS
J. Morita Co., USA) from the top and bottom
for 40sec. After hardening, both the top and
bottom acrylic were removed. Five disks were
shaped from each sealant material. Each pre-
pared disk was immersed in 5ml of distilled
water in a 10-ml plastic test tube and placed
without agitation in an incubator at 37C. The
5ml of distilled water used for immersion was
changed every 24hrs.
3. Fluoride recharge of applied APF from
sealants
The sealant disks were removed from the
test tubes after immersion for seven days,
rinsed with distilled water, and immersed in
acidulated phosphate uoride solution (APF:
9,050ppmF, pH3.8) in a 200-ml plastic beaker
for 4min. The disks were then removed from
APF solution, re-rinsed with distilled water,
and immersed in 5ml of distilled water in
a plastic test tube, enabling uoride to be
released. The immersing water was changed
every day and this procedure was repeated
over 14 days. The distilled water samples con-
taining uoride ions were kept in a refrigera-
tor at 4C until their uoride concentrations
could be determined.
4. Fluoride ion concentration measurements
The concentration of uoride ions released
from the sealants was analyzed with a com-
bination uoride ion-selective electrode
(Model 96-09BN, Orion Research Co., Cam-
bridge, MA) connected to an ion-analyzer
(EA920, Orion Research Co., Cambridge,
MA). To determine the uoride concentra-
tion, 0.1, 1.0, and 10ppm standard solutions
of uoride ions were prepared, and further
standard solutions of 0.05 and 0.02ppm uo-
ride ions were used for low uoride solutions
below 0.1ppm each time.
5. Fluoride uptake to bovine enamel treated
with FIII L and FIII L/APF application
Five bovine anterior teeth were polished
with a hearthstone powder and a polishing
brush for about one minute each and incised
between the tooth crown and root with a dia-
mond cutter. Each tooth crown was divided
into four specimens to obtain a total of 20
bovine enamel specimens. Five bovine enamel
specimens were randomly chosen, and lled
with the pit-and-ssure sealant, FIII L (light/
chemical curing), according to the manufac-
turers directions. Two groups consisting of
ve such bovine enamel specimens were pre-
pared. In the rst group, uoride uptake to
the enamel was accomplished with FIII L
alone in the second group, APF was applied
seven days later. Five bovine enamel samples
were evaluated as the control.
A bovine enamel specimen sealed with FIII
L was immersed in 5ml of 0.05M mono-
calcium phosphate saturated solution with
a pH of 7.4 adjusted with 5M potassium
hydroxide
10)
. This was used as articial saliva
in a plastic test tube. The immersion solution
was changed everyday. In the APF applica-
tion group, bovine enamel specimens were
immersed in the APF solution for four min-
utes seven days after immersion started,
rinsed with distilled water, and re-immersed
in the articial saliva. In both groups, immer-
sion in the articial saliva was maintained over
14 days.
6. Bovine enamel specimen sampling after
treatment with FIII L and FIII L/APF
applications and determination of
uoride uptake
After the immersion experiments, the pit-
and-ssure sealants were rmly yet carefully
removed from the bovine enamel specimens
of both groups with a spatula. After rinsing
with distilled water, a window (55mm) was
made on the enamel surface of the specimens
with dental wax, and the specimen was xed
to an acrylic rod (8-mm diameter, 11-cm
long). The enamel was sampled with acid
etching according to Ohkawas method
13)
.
The enamel specimens were immersed in
0.4ml of 0.5M perchloric acid in a plastic test
tube for 10sec and then neutralized by add-
ing 1.6ml of 0.5M sodium tricitrate. The
same enamel specimens were continuously
subjected to further acid treatment through
immersion for 20 and then 30sec. The uo-
ride concentrations in the sampling solutions
176
were analyzed with the uoride ion electrode
method using a buffer solution, 0.5M per-
chloric acid, and 0.5M sodium tricitrate (1: 4)
as adopted by Ohkawa
13)
. The calcium con-
centrations in the enamel sample solutions
were measured with atomic absorption spec-
trometry (ASS: Model 508, Hitachi). The
amount of enamel in each sample was calcu-
lated using a content of 36% calcium and 2.95
specic gravity for the values in bovine
enamel. The area of the enamel specimen was
determined with the tin foil method
13)
.
7. Statistical analysis
The means and standard deviations were cal-
culated for the groups and evaluated by analy-
sis of variance (Tukey Kramer test, StatView-J
5.0). A value of p0.05 was regarded as
signicant.
RESULTS
Figure 1 plots the cumulative release of
uoride ions from the four types of uoride-
containing pit-and-ssure sealants. APF solu-
tion was applied on the seventh day. The
cumulative release of uoride ions up to the
seventh day was the highest for FIII at
23745.2 (SD) g/cm
2
, FIII L and TF1 had
similar levels of approximately 53g/cm
2
,
and HSF had the lowest value of 3.72.8g/
cm
2
. As we can see in Fig. 2, the rate at which
uoride ions were released from FIII was
71.912.6g/cm
2
/day on the rst day, and it
rapidly decreased to 19.33.3g/cm
2
/day
on the fth day. The rates at which uoride
ions were released from FIII L and TF1 had
similar patterns. The initial uoride release
rates ranged from 17.218.4g/cm
2
/day, and
decreased to a third (4.04.7g/cm
2
/day) of
this on the fourth day, indicating that uoride
ions were released more slowly from FIII L
and TF1 than from FIII.
We estimated the release of uoride ions
from the four sealants after APF had been
applied on the seventh day and conrmed
that uoride was recharged in the FIII and
FIII L GIC-sealants, although this was not
observed in the TF1 or HSF resin sealants.
Moreover, uoride recharge was 1.5-fold
greater in FIII L (light/chemical sealant
74.19.4g/cm
2
) than in FIII (conventional
sealant, 58.79.5g/cm
2
), and the differ-
ence was statistically signicant (p0.05, see
Fig. 3). Both instances of uoride recharges
were retained for about one day and it then
took three days to reach the same release rate
as that before APF had been applied.
Figure 4 plots uoride concentrations in
enamel layers in the GI, GII, and control
groups. These concentrations for GI and GII
were signicantly higher than those of the
control groups (p0.05). The uoride con-
centrations in GI were very high, i.e.,
10,1004,570 (SD) ppm in the rst layer,
6,0703,430ppm in the second layer, and
3,3501,840ppm in the third layer. The
Fig. 1 Cumulative uoride-release from four different
pit-and-ssure sealants
Fig. 2 Fluoride-release from each uoride-containing
pit-and-ssure sealant in vitro
H. KOGA et al.
120
100
80
60
40
20
0
Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
FIII
HSF
FIII L
TF1
APF application
F
l
u
o
r
i
d
e

r
e
l
e
a
s
e

(

g
/
c
m
2
)
500
400
300
200
100
0
Days
1
FIII
HSF
FIII L
TF1
2
APF application
3 4 5 6 7 8 9 10 11 12 13 14
C
u
m
u
l
a
t
i
v
e

f
l
u
o
r
i
d
e

(

g
/
c
m
2
)
177
uoride concentration in GII was about
13,9002,390 (SD) ppm in the rst layer,
8,5503,260ppm in the second layer, and
3,8701,620ppm in the third layer. The
mean uoride uptake to the enamel surface
was slightly higher in GII than GI, but there
were no statistically signicant differences.
DISCUSSION
The uoride release rates and periods of
uoride-containing pit-and-ssure sealants
are clinically important in establishing to
what extent dental caries can be prevented
and whether they will recur in the boundary.
We evaluated the uoride released from four
types of uoride-containing pit-and-ssure
sealants in this study, uoride recharge
resulting from APF application, and uoride
uptake to enamel treated with FIII L/APF in
vitro, to evaluate their clinical signicance in
preventing caries.
Since conventional GIC (FIII) is extremely
sensitive to water, GIC that has been chemi-
cally polymerized after being used in llings is
detached if water and saliva contact it, sug-
gesting that the rate at which uoride ions are
released is increased due to it detaching. As
FIII L is light-hardening due to resin poly-
merization (HEMA), the degree to which it
detaches is less than that of FIII, resulting in
uoride being released more slowly. This
means that because FIII L is less sensitive to
water it retains much more AFP/FIII.
We conrmed that uoride is released
from glass ionomer/resin pit-and-ssure seal-
ants for long periods. We found a marked
recharge after applying APF to both the con-
ventional and autohardening GICs used for
lling materials
11)
. In the present study, the
concentration of uoride ions released from
a light/chemical GIC sealant (FIII L) decreased
to about a third immediately after APF was
applied, which agreed with these results.
Similar results when examining short-term
uoride ion release and uoride recharge
(APF, NaF, SnF
2
) in GIC materials have been
reported, but the reported recharge of uo-
ride ions from the application solutions used
in dental materials has been very low for NaF
within the neutral pH range and it has been
highly dependent on pH
4)
.
In the 1990s, Arends and Christoffersen
1)
and Featherstone
6)
proposed that dental caries
could be prevented on supercial layers of
enamel by inhibiting demineralization under
common ions such as calcium ions and phos-
phate ions and by promoting remineraliza-
tion under uoride ions on the surface. They
also stated that signicant remineralization
occurred even at very low concentrations
of uoride ions (0.030.05ppm) in the oral
cavity. Consequently, uoride in dental mate-
Fig. 4 Fluoride uptake (meanS.D.) to bovine enamel
with Fuji III LC (GI) and Fuji III LC/APF (GII)
Fig. 3 Comparison of uoride recharge (FR, mean
S.D.) after APF was applied for 4 min.
FR is dened as the difference in uoride release
between that at 7 days and that at 8 days. *p0.05
FLUORIDE RELEASE AND RECHARGE FROM SEALANTS
20,000
16,000
12,000
8,000
4,000
0
1st layer
FIII L/APF-treated group (GII)
FIII L-treated group (GI)
Control
2nd layer 3rd layer
F
l
u
o
r
i
d
e

c
o
n
c
e
n
t
r
a
t
i
o
n

(
p
p
m
)
100
*
*
FIII FIII L HSF TF1
80
60
40
20
0

F
R

(

g
/
c
m
2
)
178
rials that contributes to the release of uoride
ions is very important, both locally and
throughout the entire oral cavity. However, in
this study and many others, examining the
release of uoride ions from pit-and-ssure
sealants and uoride-containing materials
has been done in batch systems with distilled
water in vitro, resulting in limitations in evalu-
ating the dynamics of uoride ions emanat-
ing from these materials. Therefore, it is nec-
essary to examine the dynamics of uoride
ions under conditions that are the same as the
oral environment, for example, through study-
ing pH cycling and the in situ oral system
5)
.
Ripa
14)
suggested that there were limita-
tions with intentionally adding uoride to
dental materials and devices to prevent dental
caries from forming or enamel from deminer-
alizing. These limitations extended to delivery
of drugs (including uoride) to prevent oral
diseases (including those of soft and hard
tissues) because the clinico-epidemiological
ndings were inconclusive
14)
.
The preventive effects the glass-ionomer
pit-and-ssure sealant (FIII), were surveyed
clinico-epidemiologically for three years by
covering the permanent teeth of 91 children
aged 410 years without caries with FIII
9)
.
They found that the DMFT value decreased
to 66.5%. Furthermore, clinically reapplying
glass-ionomer would be sufciently simple
for the public health sector, and its effects
in preventing dental caries in children are
undeniable.
We rejected the null hypothesis that there is
little difference in uoride recharge after APF
treatment on the seventh day between seal-
ants (Fig. 3), because we found the uoride
recharge in the FIII and FIII L GIC sealants
to be much higher than that in the TF1 and
HSF resin sealants (p0.05). The uoride
recharge in FIII L was also signicantly higher
than that in FIII (p0.05). Therefore, we con-
rmed that GIC sealants serve as uoride
reservoirs in this experiment. In relation
to this, we also demonstrated that the acid-
resistance in enamel treated with GIC sealant
had been greatly improved
8)
.
We did not reject the null hypothesis that
uoride uptake to the enamel surface by
FIII L was almost the same as that with FIII L/
APF treatment on the seventh day, because
there were no signicant differences between
the two uoride concentrations. However,
the uoride concentration in each layer of
enamel with FIII L/APF treatment (GII)
tended to be slightly higher than that with
FIII treatment (GI), although both these uo-
ride levels were extremely high. Regarding
the uoride uptake in enamel with resin seal-
ants (MF-MMF co-polymerization polymer),
Tanaka et al. reported that the concentration
of total uoride was ca. 3,500ppm on a 10-m
supercial layer, that KOH-soluble uoride
(non-bound) was 2030%, and that bound
uoride was 7080% on layers up to 60m
16)
.
These results were signicantly higher than
those in the control group. Kawai et al.
obtained the same results for another com-
posite resin they investigated
7)
.
Our present study concurred with these
previous results and found that uoride-
recharge from pit-and-ssure sealants and
lling materials on the surface of teeth
occurred within a short time and that their
application to teeth increased acid resistance
on the surface.
In the future, we should run clinico-epide-
miological studies and in vivo experiments to
evaluate the effects of uoride-containing
sealants on preventing caries.
CONCLUSIONS
Of the four types of pit-and-ssure sealants,
conventional GIC (FIII) had the highest rate
of uoride release; a similar release pattern
was observed in GIC pit-and-ssure sealant
(FIII L) and resin sealant containing uoride
releasing polymer (TF1). The rate at which
uoride was released was reduced to about a
third of the initial level within a few days, and
release continued slowly thereafter. The uo-
ride recharge in FIII L when APF was applied
to the GIC sealant was signicantly higher
than in FIII (p0.05). These results suggest
that GIC-sealants in the oral cavity reservoirs
H. KOGA et al.
179
differ in their properties and that they can
contribute to low levels of uoride being
retained in the oral uid, thereby preventing
caries.
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Reprint requests to:
Dr. Hiroshi Koga
Department of Epidemiology and
Public Health,
Tokyo Dental College,
1-2-2 Masago, Mihama-ku,
Chiba 261-8502, Japan
FLUORIDE RELEASE AND RECHARGE FROM SEALANTS