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E
ffect
of
fluoride
on
salivary
immunoglobulins
and
sialic
acid
R
ev
A
ssoc
M
ed
B
ras
2017; 63(4):320-323
321
that there is a negative correlation between salivary sIgA
and dental caries.
9,10
Several studies have reported that car-
ies were particularly correlated with sIgA and sIgG.
11-13
In
addition, many studies demonstrated that high salivary
sIgA levels result in lower incidence of dental caries.
14-16
With the increasing use of fluoride products, dental
fluorosis has become more common. Dental fluorosis is
characterized by enamel hypomineralisation with increased
surface and subsurface porosity, which causes opacity,
pitting, or staining of the enamel. Information about
changes in sIgA, sIgG, and sialic acid levels in the presence
of dental fluorosis is limited.
The aim of our study was to evaluate the effects of
fluoride on salivary sIgA, sIgG, and sialic acid levels in
children with dental fluorosis and those with healthy teeth.
In addition, the effects of high levels of fluoride in drink-
ing water on the salivary content were evaluated.
M
ethod
The ethical committee of Süleyman Demirel University’s
Faculty of Medicine approved the study. All participants
and their parents/guardian were informed about the study
and a written consent was obtained for all children.
Isparta Province, Turkey, is a region of endemic fluo-
rosis with high fluoride levels (2.7–2.8 ppm) in drinking
water. Children in six primary schools were chosen to
participate. Children and their parents filled out a total
of 1,026 questionnaires with items regarding the duration
of residence in Isparta, place of birth, tooth-brushing and
daily dietary habits, systemic diseases, and socio-econom-
ic status. Children were examined for oral hygiene, dental
plaque index and caries status. The forms were reviewed
and 473 children were identified with similar features.
Children were examined and oral hygiene instructions
were given again at our clinic. Before starting the collection
of saliva, the criteria for inclusion were that: (a) the patients
should be healthy and free of systemic disease; (b) the
patients should not have consumed any medications for
at least 15 days before saliva collection; (c) the patients’
first permanent molars should have erupted fully; (d) there
should be equal portions of male and females participants;
(e) and they should have the same fluorosis index.
According to these criteria, 51 healthy 6 to 12 year-old
(9,15
±
1,17) children were randomly selected from the
primary schools enrolled in a dental-care programmain-
tained by the Department of Pediatric Dentistry. The
children were divided into two groups: group I comprised
26 children with dental fluorosis [Thylstrup–Fejerskov
Dental Fluorosis Index (TFI) = 4]
17
who lived in Isparta
(2.7–2.8 ppm), and group II consisted of 25 children with-
out dental fluorosis who were born in low-fluoride areas
and had lived in Isparta for only the previous two years.
The groups are summarized in Figure 1.
Saliva sample collection
A special diet was assigned to the participants for break-
fast and the teachers were instructed not to give any food/
beverage except water to the children after breakfast on
the day of saliva collection. The saliva was collected in the
morning (10-10:30 am) to minimize circadian rhythm
effects. The samples were collected in a quiet, isolated,
ventilated and lighted room, at room temperature.
After an initial swallow of saliva, the unstimulated
saliva was collected into sterilized polyethylene cups. For
saliva stimulation, participants were asked to chew on one
piece (1 g) of unsweetened, unflavored chewing gum for 2
minutes. The saliva produced in the first 30 seconds was
expectorated. The subjects spat 4 mL of saliva into the
second sterilized cup. The unstimulated saliva was used
for saliva immunoglobulin analyses and the stimulated
saliva was used for salivary fluoride and sialic acid analyses.
Analysis of the saliva samples
The salivary immunoglobulin levels were evaluated using
ELISAmethod. The salivary sialic acid levels were determined
using Sigma Sialic Acid Quantitation Kit (Sigma-Aldrich
Inc., Missouri, USA). The salivary levels of fluoride were
analyzed by fluoride electrode.
Statistical analysis
Results were analyzed statistically by Mann-Whitney-U
test and Kruskall-Wallis. All analyses were performed
using the SPSS statistical package for Windows version
13.0 (SPSS, Chicago, IL, USA). All levels of significance
were set at p<0.05.
Group I
Children with dental fluorosis and
living in high flouride area
n=26 (9.2308 ± 0.7646)
Group II
Children without dental fluorosis,
born in low fluoride areas and
living in Isparta for the last 2 years
n=25 (9.0800 ± 1.4977)
Study groups
n=51
FIGURE 1
The study and control groups.