66

Karama M T Al-Nuaimy

Department of Dental Basic Science
BDS, MSc (Asst Prof) College of Dentistry, University of Mosul

Isra'a H Al-Hamdani Department of Dental Basic Science
(BSc, MSc) College of Dentistry, University of Mosul

Nahla OM Tawfik Department of Dental Basic Science
(MBChB, MSc, PhD) College of Dentistry, University of Mosul

(

ABSTRACT
Aims: To assess the effect of acute psychological stress on some salivary glands functions. Materials
and Methods: Twenty-three undergraduate dental students participated in this study. They were asked
to provide two samples of unstimulated whole saliva for 5 minutes, the first sample before amid- year
oral academic examination and subsequently the second, one month later in a non stressful situation
(after holiday break) as control group. Salivary flow in one minute was determined, in addition, total
protein, albumin, uric acid and calcium levels in saliva were assessed using determination kits method
from Biolabo (France). Results: The results showed a significant stress mediated decrease in the sali-
vary calcium concentration (0.230.21 mmol/L) by -78.50% in comparison with non stressful condi-
tion. Salivary flow level (0.62 0.28 ml /min) and albumin concentration (16.3913.69 mg/100ml)
were decreased by (- 4.62% and -37.30% respectively), while uric acid (1.671.13 mmol/L) and total
protein concentrations (1.030.69 g/100ml) were elevated by 9.56% and 10.60% respectively. Not all
these changes were statically significant. Conclusions: These results suggest that the acute psychologi-
cal stress exerts its influence on salivary composition and this will increase the value of saliva as dy-
namic biological fluid in controlling the oral health.
Key Words: Saliva, stress, total protein, albumin, uric acid, calcium.

Nuaimy KM, Al-Hamdani IH, Tawfik NO. Effect of Stress on the Composition and Flow Rate of
Saliva. AlRafidain Dent J. 2012; 12(1): 66-70.
Received: 15/7/2010 Sent to Referees: 15/7/2010 Accepted for Publication: 26/10/2010

INTRODUCTION
Academic examination have been con-
sidered as one of the most acute stresses
experienced by students because passing
or failing usually has consequences for
one's career development.
(1-3)
Changes in
salivation often accompany the stress re-
sponse, therefore it is important to estab-
lish whether these changes is truly mirror
the physiological response to stress, or
merely confound altered salivary flow.
(3,4)

Salivary gland secretion is mainly under
autonomic nervous control, but various
hormones may also modulate salivary
composition.
(5)
Saliva consists of two
components that are secreted by inde-

Effect of Stress on the Composition and
Flow Rate of Saliva
I SSN: 18121217
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67
pendent mechanisms; first component in-
cludes ions, which is produced mainly by
parasympathetic stimulation and second
protein component, which is released
mainly in response to the sympathetic sti-
mulation.
(6)
Saliva plays an important role
in oral health monitoring, regulating and
maintaining the integrity of the oral hard
tissues and some soft tissues; many studies
have demonstrated the importance of sali-
vary calcium with regard to both dental
and gingival health.
(7)
The association be-
tween stress and the occurrence of oral
disease is based on clinical observations,
epidemiological research, and experiments
with animals.
(8-10)
Saliva possesses a mul-
tiplicity of immunological and nonimmu-
nological defense systems against toxins,
fungi, viruses and bacteria.
(7, 8)
The impor-
tance of saliva to the oral health becomes
evident in individuals with a reduced sali-
vary flow, particularly in the dry mouth
syndrome or xerostomia.
(6,11)
One of the
symptoms of xerostomia is a dramatic in-
crease of dental caries and other oral infec-
tions, accompanied by an increase of pa-
thogenic bacteria.
(12)
Human saliva lubri-
cates the oral cavity structures and protects
teeth and oral mucosa against potentially
injurious factors.
(6, 7)
Protection of the oral
tissues is, among other things, achieved by
the physical movement of saliva that ef-
fectively washes away many potentially
harmful microorganisms.
(6,8,10)
This study
of dental undergraduate students will as-
sess the effect of examination stress on
salivary flow and on concentration of
some salivary constituents and to assess
the differences between these parameters
before the examination, and after the holi-
day.

MATERIALS AND METHODS
This cross-sectional study was conducted
at the Faculty of Basic Science of Denti-
stry, College of Dentistry, University of
Mosul. During a mid year term test in phy-
siology. Twenty three undergraduate den-
tal students of either sex participated in
this study, age range from 21-23 years
with a mean age of 21 1.12years. None
of the participants was under medication
and all reported to be in good health, most
of the students were non-smoker (74%)
Students were requested in advance not to
eat or drink except for water one hour be-
fore saliva collection to minimize possible
food debris and stimulation of salivation.
The data were collected on two occasions;
the first occasion was obtained on the first
day of the exam week and the second
sample was collected after 6 weeks of the
first measurement, during a period of rest
(after mid-trimester holiday) as a control
Five minutes unstimulated whole saliva
was collected.
(13)
Deionized water was
used to rinse the mouth. The volume of the
saliva was measured and divided by five
minutes to obtain the salivary flow rate
expressed in ml/min After the collection,
the samples were centrifuged (4000 rpm,
for 10 minutes). The supernatant was di-
vided over several aliquots and frozen at (-
20C) until analysis Whole saliva was
assessed colorimetrically by a spectropho-
tometer. Calcium, uric acid, total protein
and albumin concentration were deter-
mined by (O-Cresolphtalein complexone,
Uricase, Biuret and Bromocresol methods
respectively) using a commercial kits (Bi-
olabo reagents, Biolabo SA France).
(14)

Data analysis was performed using
SPSS version 13.0 for Window. Data were
expressed as the mean standard devia-
tion (MSD), and percentage (%)
throughout the paper. Means were com-
pared using an paired sample Student's t-
test. The relation between the salivary
flow and biochemical parameters was ex-
plored by means of Pearson correlation
coefficient(r). P<0.05 was considered a
statistically significant difference.

RESULTS
In this study, the saliva of 23 dental stu-
dents was assessed on two occasions. Dur-
ing examination stress, the results showed
non significant increase of uric acid and
total protein concentrations (1.671.13
and1.030.69) by (10.60% and 9.57% re-
spectively) compared with the non stress-
ful sample. While salivary flow rate (0.62
0.28) and albumin levels (16.3913.69)
were decreased by (- 4.62%, - 37.30% re-
spectively), all these results were statisti-
cally non significant (Table 1).
Al Rafidain Dent J
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Nuaimy KM, Al-Hamdani I H, Tawfik NO

68
Table (1): Comparison between the effect of exam stress and non-stress (holiday samples)
situation on salivary flow rate and biochemical parameters.
Parameter
Exam Stress
Samples
Mean SD
Non Stress
Samples
Mean SD

% of
changes

P value
N=23 N=23
Salivary flow rate (ml /min) 0.62 0.28 0.650.30 -4.62% .216
Ca ( mmol/L) 0.23 0.21 1.070.99** -78.50% .001
Total protein (g/100ml) 1.030.69 0.940.59 9.57% .644
Uric acid (mmol/L ) 1.671.13 1.511.23 10.60% .546
Albumin ( mg/100ml) 16.3913.69 21.9210.76 -37.30% .216
Significant at ** p < 0.01

The concentration of calcium was sig-
nificantly decreased (-78.50%) at level of
p<0.01 in comparison with the non-
stressful condition (control group) as
shown in figure (1):

0
0.2
0.4
0.6
0.8
1
1.2
Exam Stress Non Stress
C
a

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

(
m
m
o
l
/
L
)

Figure (1): Comparison between mean Calcium concentration in exam stress sample and Non-
stress sample


Correlation analysis between these pa-
rameters in the first sample (stress in-
duced) showed that there were interrela-
tion between the parameters but its not
statistically significant (Table 2), while
correlation analysis between the same pa-
rameters in the second sample (holiday)
(Table 3) showed that salivary flow were
negatively correlated with uric acid con-
centration (r = - 0.46, p< 0.05) (Figure 2).

Table (2): Correlations between salivary flow rate and biochemical parameters in exam stress
situation
Parameter
Salivary
flow rate
(ml /min)
Ca
(mMol/L)
Total protein
(g/100ml)
Uric acid
(mMol/L)
Albumin
(mg/100ml)
Salivary flow rate
(ml/min)
-
Ca ( mmol/L) -.187 -
Total protein (g/100ml) -.261 .124 -
Uric acid (mmol/L) -.240 -.321 -.040
Albumin (mg/100ml) -.059 .186 -.340 -.091 -
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Effect of stress on salivary gland functions

69
Table (3): Correlations between salivary flow rate and biochemical parameters in Non Stress situ-
ation
Parameter Salivary flow
rate (ml /min)
Ca
(mMol/L)
Total protein
(g/100ml)
Uric acid
(mMol/L)
Albumin
(mg/100ml)
Salivary flow rate(ml/min) -
Ca (mmol/L) -.052 -
Total protein (g/100ml) -.198 .037 -
Uric acid (mmol/L) -.460* -.108 .321
Albumin (mg/100ml) -.070 .094 .261 .137 -



Figure (2): Correlation between salivary flow rate and uric acid concentration in Non-stress
Sample

DISCUSSION
The protective functions of saliva and the
association between oral disease and psy-
chological factors is considered. It is sur-
prising how little psychological stress can
alter the defense systems in saliva. In this
study, total protein concentration was ele-
vated, while salivary flow rate decreased
and this is in agreement with other stu-
dies.
(2-4)
Turner and Sugiya (2002) sug-
gested that Parasympathetic stimulation
produces copious saliva of low protein
concentration while sympathetic stimula-
tion produces little saliva but of high pro-
tein concentration and may thus give a
sensation of dryness.
(5,15)
Subjective oral
dryness and reduced unstimulated salivary
flow were significantly associated with
depression, trait anxiety, perceived stress
and state anxiety.
(10,11)
This elevation
could be caused by an increased sympa-
thetic tone and catecholamine output,
(4)
or
could be due to activation of the hypotha-
lamic-pituitary- adrenal axis and subse-
quent release of cortisol in saliva with
subsequent increase in total protein con-
tent, and secretory immunoglobulin A as
one defense mechanism(2). Calcium is one
of important inorganic content of salivary
fluid.
(16)
Secretion of calcium in saliva de-
pend upon salivary flow rate, so there is a
negative correlation between calcium con-
centration and salivary flow rate
(17)
and
this is in agreements with the results of the
study. Sewon et al. (1998) showed that a
positive correlation between high salivary
calcium content and periodontitis and be-
tween high salivary calcium level and the
number of intact teeth in selected groups
of subjects.
(17)
Change in the concentration
of some of the saliva constituent is co-
responsible for the dynamics of the
processes, which contribute to the devel-
opment of new equilibrium between teeth
remineralization and demineralization.
(7)

Uric acid is considered as one of the con-
stituents in the salivary fluid. The concen-
tration of uric in mixed saliva has been
reported as a range from 0.5 to 20.6
mg/100ml
(18)
and this is in agreement with
our results. In recent years it has been em-
phasized that the concentration of many of
the constituents of saliva varies with flow
rate and that composition of the saliva
produced by individual salivary gland dif-
fers from each other's (19, 20). Uric acid is
Al Rafidain Dent J
Vol. 12, No1, 2012

Nuaimy KM, Al-Hamdani I H, Tawfik NO

70
one of the antioxidant defenses and that
the elevation in the levels could be due to
this cause, uric acid content in saliva cor-
relate with plasma uric acid.
(20)


CONCLUSION
This study demonstrates that the effect
of stress (mid-year examination) on some
important constituent of saliva. The results
suggest that acute psychological stress
exerts its effect on salivary composition
and this will increase the value of saliva as
dynamic biological fluid in controlling the
oral health.

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Effect of stress on salivary gland functions