Fluoride bioavailability in saliva and plaque.
ABSTRACT Different fluoride formulations may have different effects on caries prevention. It was the aim of this clinical study to assess the fluoride content, provided by NaF compared to amine fluoride, in saliva and plaque.
Eight trained volunteers brushed their teeth in the morning for 3 minutes with either NaF or amine fluoride, and saliva and 3-day-plaque-regrowth was collected at 5 time intervals during 6 hours after tooth brushing. The amount of collected saliva and plaque was measured, and the fluoride content was analysed using a fluoride sensitive electrode. All subjects repeated all study cycles 5 times, and 3 cycles per subject underwent statistical analysis using the Wilcoxon-Mann-Whitney test.
Immediately after brushing the fluoride concentration in saliva increased rapidly and dropped to the baseline level after 360 minutes. No difference was found between NaF and amine fluoride. All plaque fluoride levels were elevated after 30 minutes until 120 minutes after tooth brushing, and decreasing after 360 minutes to baseline. According to the highly individual profile of fluoride in saliva and plaque, both levels of bioavailability correlated for the first 30 minutes, and the fluoride content of saliva and plaque was back to baseline after 6 hours.
Fluoride levels in saliva and plaque are interindividually highly variable. However, no significant difference in bioavailability between NaF and amine fluoride, in saliva, or in plaque was found.
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ABSTRACT: The assessment of the fluoride kinetics in whole saliva as well as in the different salivary phases (supernatant saliva and sediment) is essential for the understanding of fluoride bioavailability. To assess the fluoride content, provided by sodium fluoride and amine fluoride, in the supernatant saliva and in salivary sediment. Seven trained volunteers were randomly attributed to 2 groups in a cross-over design and brushed their teeth in the morning for 3 min with a product containing either sodium fluoride or amine fluoride. Saliva was collected before, immediately after tooth brushing and 30, 120, and 360 min later and measured. The samples were centrifuged 10 min at 3024 × g. Fluoride content of the supernatant saliva and of the sediment was analysed using a fluoride sensitive electrode. All subjects repeated the study cycles 2 times, and statistical analyses were made using the nonparametric sign test for related samples, the Wilcoxon-Mann-Whitney-test for independent samples. There was a significant increase in fluoride immediately after tooth brushing in both groups in saliva and sediment. The distribution of fluoride between salivary sediment and supernatant saliva (ratio) varied considerably at the different collection times: decreased from 17.87 in baseline samples of saliva to 0.07 immediately and to 0.86 half an hour after tooth brushing in the sodium fluoride group and from 14.33 to 2.85 and to 3.09 in the amine fluoride group. Furthermore after 120 min and after 360 min after tooth brushing the ratio increased from 17.6 to 31.6 in the sodium fluoride group and from 20.5 to 25.76 in the amine fluoride group. No difference was found in the sediment-supernatant saliva ratio between the sodium fluoride and the amine fluoride groups 360 min after tooth brushing. For the assessment of fluoride kinetics in whole saliva it is necessary to pay attention to at least four factors: fluoride formulation, time after fluoride application, fluoride concentration in supernatant saliva and fluoride concentration in salivary sediment. This study was approved by the Ethical Committee of the University of Witten/Herdecke permission 21/2008.Archives of oral biology 02/2012; 57(7):870-6. · 1.65 Impact Factor
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ABSTRACT: INTRODUCTION: Detailed information about the size of the oral mucosa is scarce in the literature, and those studies that do exist do not take into account the size of the tongue or the enlargement of the surface by the papillae. Because of the various functions of the oral mucosa in the maintenance of oral health, knowledge of its true size may provide a better understanding of the physiology of the oral cavity and some oral diseases and direct future therapeutic strategies. The aim of this study was to determine the total size of the oral mucosa. METHODS: Five human adult cadaver heads were cut in the median sagittal plane, and the total area of the oral surface was determined using silicon casts. The surface of the tongue was measured with quantitative profilometry. Photographs of oral blood vessels were taken in different areas of the oral mucosa of adult test subjects using intravital microscopy, and the pictures were compared with vessel casts of the oral mucosal capillaries of a maccaca fasciculrais monkey, which was studied using a scanning electron microscope. RESULTS: The results showed that the dorsal side of the tongue comprises a large proportion of the total oral mucosal surface. The surface area of the epithelium increases moving from anterior to posterior on the tongue, and the number of underlying blood vessels increases proportionally. CONCLUSIONS: It can be concluded that the back of the tongue plays an important role in the oral resorption of drugs. Clinical relevance: The results may be of relevance for the delivery and development of oral drug application.Head & Face Medicine 03/2013; 9(1):8. · 0.87 Impact Factor
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ABSTRACT: Stress-related variations of fluoride concentration in supernatant saliva and salivary sediment, salivary cortisol, total protein and pH after acute mental stress were assessed. The hypothesis was that stress reactions have no influence on these parameters. Thirty-four male students were distributed into two groups: first received the stress exposure followed by the same protocol two weeks later but without stress exposure, second underwent the protocol without stress exposure followed by the stress exposure two weeks later. The stressor was a public speech followed by tooth brushing. Saliva was collected before, immediately after stress induction and immediately, at 10, 30 and 120 min. after tooth brushing. Cortisol concentrations, total protein, intraoral pH, and fluoride content in saliva were measured. The data were analyzed statistically. Salivary sediment was ca 4.33% by weight of whole unstimulated saliva. Fluoride bioavailability was higher in salivary sediment than in supernatant saliva. The weight and fluoride concentration was not altered during 2 hours after stress exposure. After a public speech, the salivary cortisol concentration significantly increased after 20 minutes compared to the baseline. The salivary protein concentration and pH also increased. Public speaking influences protein concentration and salivary pH but does not alter the fluoride concentration of saliva.Scientific Reports 05/2014; 4:4884. · 5.08 Impact Factor
RESEARCH ARTICLEOpen Access
Fluoride bioavailability in saliva and plaque
Ella A Naumova†, Phillip Kuehnl†, Philipp Hertenstein†, Ljubisa Markovic†, Rainer A Jordan†, Peter Gaengler†and
Wolfgang H Arnold*†
Background: Different fluoride formulations may have different effects on caries prevention. It was the aim of this
clinical study to assess the fluoride content, provided by NaF compared to amine fluoride, in saliva and plaque.
Methods: Eight trained volunteers brushed their teeth in the morning for 3 minutes with either NaF or amine
fluoride, and saliva and 3-day-plaque-regrowth was collected at 5 time intervals during 6 hours after tooth
brushing. The amount of collected saliva and plaque was measured, and the fluoride content was analysed using a
fluoride sensitive electrode. All subjects repeated all study cycles 5 times, and 3 cycles per subject underwent
statistical analysis using the Wilcoxon-Mann-Whitney test.
Results: Immediately after brushing the fluoride concentration in saliva increased rapidly and dropped to the
baseline level after 360 minutes. No difference was found between NaF and amine fluoride. All plaque fluoride
levels were elevated after 30 minutes until 120 minutes after tooth brushing, and decreasing after 360 minutes to
baseline. According to the highly individual profile of fluoride in saliva and plaque, both levels of bioavailability
correlated for the first 30 minutes, and the fluoride content of saliva and plaque was back to baseline after 6 hours.
Conclusions: Fluoride levels in saliva and plaque are interindividually highly variable. However, no significant
difference in bioavailability between NaF and amine fluoride, in saliva, or in plaque was found.
Already two decades ago it has been postulated that
site-specific aspects of salivary fluoride clearance may
have important implications for the site-specificity of
oral diseases . It is now well known that at least three
factors are influencing this site-specificity of oral patho-
biology: the different local composition and pathogeni-
city of oral biofilms (local microbiome), the site-specific
host response towards bacterial phylotypes as commen-
sals or pathogens (local immunity), and finally, the indi-
vidual variability of salivary and plaque clearance of
Whereas the first two factors are exclusively in the
focus of basic research, the kinetics of fluoride in oral
fluids are rather well documented. This is the reason why
clinical recommendations for the treatment of incipient
caries lesions or for the stagnation of lesions can be con-
cluded. Therefore, the bioavailability of fluoride in saliva,
and consequently in plaque fluid plays a crucial role in
preventing a net mineral deficit in enamel, cementum
and dentin due to caries challenge.
Bioavailability of fluoride is dependent upon various fac-
tors such as fluoride administration [2-7], fluoride formu-
lation, and salivary secretion rate [8-11]. Fluoride
bioavailability in plaque may also be influenced by the
compounds of the administered fluoride source. Recently
it has been demonstrated, that e. g. sodium lauryl sulphate
changes the structure of plaque biofilms which may have
an effect on fluoride uptake or release . Several studies
have demonstrated, that salivary fluoride concentration
increases dramatically after fluoride administration either
after tooth brushing or mouth rinsing with fluoridated
products, but is back to the baseline level two hours after
fluoride administration [4,9,13,14].
Fluoride concentration in saliva is the source for the
fluoride delivery to dental plaque. Recently it has been
demonstrated that elevated fluoride products like denti-
frices with 5000 ppm NaF or amine fluoride [10,15] or
oral hygiene tablets directly dissolved in saliva with 4350
ppm NaF enhance remineralization of advanced enamel
lesions  and result in increased bioavailability of
fluoride in saliva . Application of high concentrations
* Correspondence: Wolfgang.firstname.lastname@example.org
† Contributed equally
Faculty of Health, Department of Dentistry, University of Witten/Herdecke,
Alfred Herrhausenstrasse 50, 58448 Witten, Germany
Naumova et al. BMC Oral Health 2012, 12:3
© 2012 Naumova et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
of fluoride leads to the formation of a CaF2layer on the
enamel surface. It has been reported that this CaF2layer
dissolves rapidly and releases bioavailable fluoride .
No calcium-fluoride-like deposits were detected in
plaque shortly after a NaF mouth rinse , and the
authors concluded that the inability to form more per-
sistent CaF2deposits may account for the rapid loss of
fluoride in plaque after the use of topical fluoride
agents. Concerning the plaque clearance of fluoride
representing consequently the F-bioavailability over the
day and night time rather controversial results have
been reported: An experimental ex-vivo study demon-
strated a rapid and very substantial uptake of fluoride by
plaque after exposure to 1000 ppm NaF immersion ,
whereas from an in-vivo study it was concluded that ele-
vated salivary fluoride concentrations were not reflected
in dental plaque, measured 6 h after brushing (1400
ppm fluoride) and rinsing (250 ppm fluoride) . Other
data demonstrated after one hour post brushing (1074
ppm fluoride) a rapid fluoride uptake and 12 hours later
a clearance back to the placebo levels . More
detailed fluoride kinetics data in dental plaque are
As fluoride binding to the plaque reservoirs and the
release from the reservoir is rather complex, the source
of the fluoride may play an important role. It is well
known that different fluoride formulations lead to differ-
ent salivary fluoride concentrations after tooth brushing
. NaF is instantly dissociating in saliva. Sodium
monofluorphsphate (NaMFP) requires hydrolysis to
release free fluoride ions , and amine fluoride may
bind to organic constituents in saliva and plaque and
releases fluoride slower than the other two. Higher
fluoride concentrations may result in the formation of a
CaF2layer on the enamel surface which also may serve
as fluoride reservoir . The different dissolution prop-
erties may lead to different fluoride concentrations in
plaque, consequently affecting the caries protective
effect of plaque fluoride content.
It was, therefore, the aim of the present investigation
to follow up the fluoride bioavailability in whole saliva
and in individual plaque samples from baseline immedi-
ately after tooth brushing and up to 360 minutes, to
compare a NaF formulation dissolved directly in saliva
with an amine fluoride dentifrice formulation. The null
hypothesis that there is no difference in the fluoride
bioavailability after NaF or amine fluoride application
A power analysis with a power of 0.8 at a significance level
of p = 0.05 prior to the investigation resulted in a mini-
mum of 6 individuals and three samples per individual to
gain reliable data. The data for the power analysis relayed
on data obtained in a previous study . Eight healthy
subjects participated in this crossover study (7 male and 1
female subject, 24 - 65 years of age). They consented after
verbal and written information on the aim and perfor-
mance of the investigation and also received written
instructions and a schedule. Participants were further
asked to avoid fluoride-rich food products such as tea, fish
and specified mineral water during the period but had no
restriction concerning drinking water. All test subjects
were residents in the area with ≈ 0.2 ppm fluoride in the
drinking water and normally used fluoride containing den-
tifrices twice daily. The participants had good oral health.
Prior to the inclusion into the study salivary flow rate was
determined and only normal secretors (0.25 - 1.0 ml/min)
were included. The study protocol was approved by the
Ethical Committee of the University of Witten/Herdecke,
Germany (permission 21/2008).
NaF was administered as oral hygiene tablets DENT-
TABS®(Innovative Zahnpflegegesellschaft mbH, Berlin,
Germany) containing 1450 ppm fluoride per 1.0 g tablet.
The tablet had to be chewed before tooth brushing, and
the teeth were brushed with a wet tooth brush. Amine
fluoride was administered as dentifrice ELMEX®(Gaba,
Lörrach, Germany) containing 1400 ppm fluoride from
Before the experiments all participants received profes-
sional dental cleaning. Then they abstained from any oral
hygiene, and plaque was grown for three days prior to the
experiments. All participants brushed their teeth in the
lower jaw in the morning for 3 minutes with either NaF or
amine fluoride formulations. Whole saliva and plaque was
collected at 5 time intervals for 6 hours. Immediately
before brushing (T0) and 3 (T1), 30 (T2), 120 (T3) and
360 (T4) minutes after tooth brushing saliva was collected
by spitting into plastic tubes for 3 minutes. Plaque was
collected from the upper jaw teeth at the same time inter-
vals with a sterile curette from the approximal sites of
molars and premolars strictly from one given site. There-
fore, the plaque samples were not pooled. These 5 plaque
samples represented the 3-day plaque regrowth on 3 inter-
proximal buccal sites (teeth 14 - 17) and 2 interproximal
palatinal sites (teeth 14 - 16).
All subjects repeated every cycle 4 times with both for-
mulations (cross over). Individual cycles with plaque
amount less than 1 mg per sample were excluded and
consequently repeated. The plaque weight used was
between 1 mg and 5.4 mg. The washout period between
each cycle was one week. During the washout period the
Naumova et al. BMC Oral Health 2012, 12:3
Page 2 of 6
subjects were allowed to perform their individual oral
After removal of plaque the weight was determined with a
precision balance and the plaque samples were diluted in
500 μl TISAB III (Thermo Electron, Beverly, MA, USA).
To compare plaque and saliva values whole saliva samples
were weighted, and then centrifuged (B Centrifuge, Beck-
man Instruments Inc., Germany) for 10 min at 6000 rpm
in micro- centrifuge tubes. An aliquot of 1 ml was taken
and mixed with 1 ml of a TISAB II buffer solution
(Thermo Electron, Beverly, MA, USA). For fluoride ion
distribution during the measurement a magnetic stick stir-
rer (size 2 × 5 mm) was used. The salivary fluoride content
was analyzed using a fluoride-sensitive electrode (96-09
Orion, Thermo Electron, Beverly, MA, USA). All measure-
ments were repeated three times and the mean of the
measurements was calculated and used for further statisti-
For the measurement of the fluoride content the follow-
ing analytical techniques were used: direct calibration and
incremental techniques (the method of known addition
for low ionic strength samples with a fluoride concentra-
tion of less then 0.38 ppm) . Direct calibration was
performed in a series of prepared standards of 0.4, 4.0, 40
and 400 ppm fluoride.
The obtained data were processed with the Statistical
Package for Social Sciences (SPSS 15.0, Chicago, III.,
USA). The post-brushing values at the different time inter-
vals were compared with baseline levels using the non-
parametric singn test for related variables. As four tests
have been applied on the data for the time intervals, the
Bonferroni correction was applied and resulted in a
p-value of p < 0.0125 for those tests. For comparison of
the total amount of fluoride in saliva and plaque after NaF
or Amine fluoride administration, curves were plotted for
every test person for all time intervals and the area under
curve was calculated. These data were then compared
with the non parametric Wilcoxon-Mann-Whitney-Test
for independent variables. The level of significance for the
comparison between NaF and Amine fluoride was 0.05.
The baseline fluoride content of saliva ranged in the 8
cycles per subject, in a total of 64 measurements, from
0.02 ppm to 1.93 ppm. The mean fluoride content was
0.41 ppm ± 0.38 ppm for both study arms, and the
baseline levels for NaF and amine fluoride were statisti-
cally not different (Table 1). The salivary fluoride con-
centration for the NaF study arm immediately after
brushing was higher compared to amine fluoride (p =
0.017). The range of the fluoride content was 100.0 to
264.0 ppm for Na F and 70.0 to 183.0 ppm for amine
fluoride. Thirty minutes after brushing the fluoride con-
centration was still elevated about 10 fold compared to
the baseline values, but not significantly (p = 0.73). The
range for the NaF arm was 0.4 to 9.3 ppm for NaF and
0.3 to 8.1 ppm for amine fluoride Two hours after
brushing the fluoride content in saliva was back to base-
line and the rather high interindividual and intraindivi-
dual range was demonstrated also for the 6 hours
measurements after tooth brushing in both study arms
(Figure 1). Comparison of the total salivary fluoride con-
tent from baseline until 6 hours after tooth brushing
demonstrated a significantly higher fluoride in saliva for
NaF (p < 0.001) (Figure 2).
The baseline individual plaque fluoride content ranged
from 3.9 to 676 ppm, and the mean for all 8 subjects was
147.5 ± 171.1 ppm. All baseline levels of interproximal
plaque samples were statistically not different in both
study arms (Table 2). Immediately after brushing the
fluoride content in plaque did not increase, and the indi-
vidual range from subject to subject and from cycle to
cycle was as high as the baseline data.
The fluoride content in plaque increased 30 minutes
after tooth brushing, but the increase was not significant
(p = 0.152). The range of the NaF arm was 3 to 1063 ppm,
and for the amine fluoride arm was 24.3 to 1201 ppm
fluoride. Between 30 minutes and 2 hours the fluoride
content within plaque decreased slightly and after 6 hours
the fluoride content in plaque was close to the baseline
level with no significant differences (Figure 3). Six hours
after tooth brushing the mean fluoride content in plaque
and the high intraindividual and interindividual range
represented the baseline levels with no statistical differ-
ences (Figure 4)
Table 1 Salivary fluoride content (in ppm) after tooth brushing with NaF or amine fluoride
Baseline 3 minutes30 minutes120 minutes360 minutes
Mean STDp MeanSTDpMeanSTDpMeanSTDpMeanSTDp
Amine fluoride0.390.35 p =
120.725.81p = 0.0172.31.9p = 0.730.310.22p = 0.510.240.19p = 0.03
Naumova et al. BMC Oral Health 2012, 12:3
Page 3 of 6
Figure 1 Fluoride concentration in saliva. Fluoride concentration
in saliva at baseline, 30, 120 and 360 minutes after tooth brushing.
The boxplots demonstrate the high interindividual variability
demonstrated by the extend of the whiskers and the extremes of
the salivary fluoride content.
Figure 2 Total amount of fluoride in saliva. The boxplot graphic
of the total amount of fluoride over the whole measured time
period demonstrates a significantly higher salivary fluoride content
after NaF administration which can be seen by the higher value of
the median in the NaF group.
Table 2 Fluoride content (in ppm) in plaque after tooth brushing with NaF of amine fluoride
Baseline3 minutes30 minutes120 minutes 360 minutes
MeanSTDp MeanSTDp MeanSTDp MeanSTDp MeanSTDp
Amine fluoride147.5171.1 p =
128.5139.3 p = 0.5 192.6209.6 p = 0.56177.7 154.8p = 0.74 130.498.08 p = 0.39
NaF161217.9170.4 161.2220.2 217.9216.1 438.5164.3 186.04
Figure 3 Fluoride content in plaque. Fluoride content in plaque
after brushing with NaF or amine fluoride. The fluoride content in
plaque is increasing 30 and 120 minutes after using NaF or amine
fluoride and dropping to baseline level after 360 minutes. The
differences are not significant.
Figure 4 Variability of fluoride content in plaque. The boxplot
graphic demonstrated the high interinidividual variability of plaque
fluoride content after administration of NaF or amine fluoride
demonstrated by the extend of the whiskers and the extremes.
There is not significant difference between the different time
intervals and between NaF and amine fluoride.
Naumova et al. BMC Oral Health 2012, 12:3
Page 4 of 6
Several studies have shown that after fluoride administra-
tion, either with dentifrice or mouth rinse, salivary fluoride
concentration increases shortly after administration and
drops back to the baseline level after 3 to 6 hours
[8-11,23]. It is well known that fluoride penetrates into
plaque by diffusion , and thus becomes a fluoride
reservoir which stores fluoride for some time and releases
fluoride [18,24]. Plaque fluoride content depends mainly
upon the time of exposure to fluoride [20,25,26] and the
fluoride formulation [11,21,27,28].
The results of the present study demonstrated a peak
increase of salivary fluoride concentration immediately
after brushing and lasting for at least 30 min. This is about
the time for fluoride diffusion into the plaque biofilm ,
and consequently the plaque fluoride concentration is ele-
vated 30 min after brushing. Both formulations, NaF and
amine fluoride, demonstrated the same trend, whereas the
fluoride concentration after NaF administration was
slightly higher. These results are confirmed by the assess-
ment of penetration of fluoride into natural biofilms. From
the literature it is known that the fluoride uptake into pla-
que is restricted after short term exposure up to 120 sec,
whereas exposure for 30 min. demonstrated significantly
higher concentrations even in deep plaque layers towards
the enamel surface . The plaque fluoride content is of
great importance, since dental plaque bacteria are respon-
sible for causing caries. Fluoride is enhancing reminarali-
zation of the enamel surface. The fluoride content in
dental plaque may also be dependent upon the fluoride
All 8 subjects exhibited a rather normal distribution of
fluoride concentration at baseline and still 2.5 to 10 fold
increase after 30 min. throughout the two study arms
there are subjects with a rather constant fluoride bioa-
vailability in plaque (from baseline to increase after 30
min. and back to baseline after 360 min.) on the one
hand, and, on the other hand subjects with very variable
fluoride concentrations in plaque (as well as at baseline,
increase after 30 min. as after 360 min.). This clinical
study clearly demonstrated high intraindividual and
interindividual as well as site-specific differences in the
salivary and plaque fluoride bioavailability. According to
the research protocol and considering the limitations of
this study, no differences between NaF and amine fluor-
ide were observed for fluoride concentrations in saliva
and plaque. Thus the null hypothesis has been con-
firmed. The high interindividual variability has been
demonstrated before [9,29] and may be a reason for the
non significant differences in the fluoride content
between NaF and amine fluoride.
There are obviously many factors contributing to the
oral fluoride kinetics in an open organ system like the
oral cavity. Saliva secretion and content plays a major
role for bioavailability of fluoride [9,29]. But there are
other factors such as oral hygiene behaviour, brushing
time and frequency, fluoride formulation and bioavail-
ability of acting ions, dietary tradition and alimentary
fluoride sources and seemingly unknown factors which
contribute to the efficiency of bio available fluoride. To
enlighten the complex interplay between saliva, plaque
and fluoride bioavailability further in vivo and in vitro
studies experimental have to be carried out.
The authors would like to thank Mrs. Susanne Haussmann for her technical
EAN did the planning of the project and supervision; PK was responsible for
the plaque collection and fluoride measurement in the plaque samples; PH
was responsible for saliva collection and fluoride measurement in saliva: LM
provided the patients for plaque and saliva collection; RAJ also provided
patients for plaque and saliva collection; PG was advisor of the project; WHA
wrote manuscript and did the statistical calculation. All authors read and
approved the final manuscript.
The authors declare that they have no competing interests.
Received: 22 November 2010 Accepted: 9 January 2012
Published: 9 January 2012
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The pre-publication history for this paper can be accessed here:
Cite this article as: Naumova et al.: Fluoride bioavailability in saliva and
plaque. BMC Oral Health 2012 12:3.
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