Free Radical Biology & Medicine, Vol. 34, No. 3, pp.

377384, 2003
Copyright 2003 Elsevier Science Inc.
Printed in the USA. All rights reserved
0891-5849/03/$see front matter

doi:10.1016/S0891-5849(02)01297-2

Original Contribution
INHIBITION OF ORAL PEROXIDASE ACTIVITY BY CIGARETTE SMOKE:
IN VIVO AND IN VITRO STUDIES
ABRAHAM Z. REZNICK,* IFAT KLEIN,* JASON P. EISERICH, CARROLL E. CROSS,

and

RAFAEL M. NAGLER

*Department of Anatomy and Cell Biology, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology,
Haifa, Israel; Division of Pulmonary-Critical Care Medicine, University of California at Davis, Davis, CA, USA; and Oral
Biochemistry Laboratory and Salivary Clinic, Department of Oral and Maxillofacial Surgery, Rambam Medical Center,
Haifa, Israel
(Received 15 January 2002; Revised 16 September 2002; Accepted 21 October 2002)

AbstractOral peroxidase (OPO), the pivotal enzyme in the salivary antioxidant system, seems to be of paramount
importance in the oral defense mechanism, especially against the attack of free radicals related to cigarette smoke (CS)
and the evolution of oral cancer. The major inducer of oral cancer is exposure to tobacco, which is responsible for
50 90% of cases worldwide. The purpose of our study was to elucidate the outcome of interaction between CS and OPO
in smokers and nonsmokers. After smoking a single cigarette, a sharp drop of OPO activity was observed in both groups:
42.5% in smokers and 58.5% in nonsmokers (p .05). After 30 min, the level of activity returned to 90 100% of the
presmoking level, presumably due to the secretion of new saliva into the oral cavity. The difference between the two
groups was also observed after exposure of saliva to one cigarette in smoking flasks (in vitro studies); however, as
expected, no recovery of activity was observed in either group. Similarly, the OPO activity loss was accompanied by
increased carbonylation of the salivary proteins, an indicator of the oxidative damage to proteins. These results may be
of great clinical importance, as heavy smokers smoke 20 cigarettes or more on a daily basis. Accordingly, most of the
time the oral epithelium of heavy smokers is essentially unprotected by OPO against the deleterious effects of
thiocyanate ions and hydroxyl radicals produced by unremoved hydrogen peroxide in the presence of the salivary
redox-active metal ions. This may pave the way for the CS-induced and saliva-mediated initiation and progression of
oral cancer. 2003 Elsevier Science Inc.
KeywordsAldehydes, Carbonyls, Cigarette smoke, Oxidative stress, Peroxidase, Saliva, Free radicals

INTRODUCTION

in these studies measured the activity of both enzymes.

In structural and antigenic characteristics, SPO resembles the lactoperoxidase found in bovine milk [4,5].
Despite the importance of peroxidase in saliva, it accounts for only 0.01% of the total salivary proteins and
exists in equilibrium between a monomeric state and
multimolecular aggregates [4,6]. OPO plays a dual role:
(i) it reduces the level of hydrogen peroxide (H2O2)
excreted into the oral cavity from the salivary glands by
bacteria and by leukocytes, and (ii) it increases specific
antibacterial activity by inhibiting the metabolism and
proliferation of various bacteria in the oral cavity [1].
In the reaction of the thiocyanate ion (SCN) H2O2
3 OSCN H2O, which is catalyzed by the peroxidase, H2O2 oxidizes SCN, a detoxification product of
cyanide secreted mainly from the parotid gland. In this
reaction, SCN acts as the electron-donating component,
similar to glutathione (GSH) in other biological systems

The peroxidase found in the oral cavity is a very important salivary antioxidant enzyme [1]. This oral peroxidase (OPO) is composed of two peroxidase enzymes,
salivary peroxidase (SPO) and myeloperoxidase (MPO).
The SPO secreted from the major salivary glands, mainly
the parotid gland [2], contributes 80% of OPO activity,
while MPO, produced by leukocytes in inflammatory
regions of the oral cavity [3], contributes the remaining
20% of OPO activity. The term OPO is used here to
denote the total activity of both isoforms, since the
2-nitrobenzoic acid-thiocyanate (NBS-SCN) assay used
Address correspondence to: Dr. Abraham Z. Reznick, TechnionIsrael Institute of Technology, The Bruce Rappaport Faculty of Medicine, Department of Anatomy and Cell Biology, P.O. Box 9649, 31096
Haifa, Israel; Tel: 972 (4) 829-5388; Fax: 972 (4) 829-5403;
E-Mail: reznick@tx.technion.ac.il.
377

378

A. Z. REZNICK et al.

[3,7,8]. Two potent antibacterial oxidizing products

evolve from this reaction: hypothiocyanous acid
(HOSCN) and its conjugated anion, hypothiocyanite
(OSCN). The cytotoxic properties of these salivary
oxidants depend on the extracellular pH, and HOSCN
can oxidize oxyhemoglobin into methemoglobin in
erythrocytes, while both HOSCN and OSCN can oxidize intracellular reduced GSH [9]. The cytotoxic antibacterial activity of HOSCN and OSCN stems from
their ability to react with sulfhydryl groups of bacterial
enzyme that are vital for glycolysis, such as hexokinase,
aldolase, and pyruvate kinase [3,7,9 11]. Under various
oral stimuli, the peroxidase may act as a scavenger of
H2O2 to produce molecular oxygen, but without producing OSCN plus HOSCN [12]. In any case, the accumulated antibacterial activity of the combination of peroxidase, H2O2, and SCN is much more potent than that
of H2O2 alone [13].
However, in contrast to the oral antibacterial characteristics of OPO, which have been studied thoroughly,
the possible anticarcinogenic role of OPO against the
most prevalent and lethal cancer of the oral cavity, squamous cell carcinoma (SCC), has scarcely been mentioned and never investigated. Oral SCC is the most
common malignancy of the head and neck, with a worldwide incidence of over 300,000 new cases annually [14].
The disease is characterized by a high rate of morbidity
and mortality (about 50%) and in this respect is similar to
malignant melanoma [14 17]. The major inducer of oral
SCC is exposure to tobacco, which is considered to be
responsible for 50 90% of cases worldwide [18,19]. The
incidence of oral SCC in cigarette smokers is four to
seven times higher than in nonsmokers [20,21]. Moreover, the higher CS-related risk for oral SCC is manifested by a reduction in the mean age of development of
the disease by 15 years, as compared with nonsmokers
[22].
The field cancerization concept is the currently accepted explanation for the carcinogenic effect of CS on
oral mucosa [17]. According to this theory, there is a
constant and direct attack of various CS reagents on the
oral epithelial cells, which gradually accumulate and
cause a step-wise malignant transformation. Free radicals, reactive oxygen species (ROS), and reactive nitrogen species in the inhaled CS have been suggested to
induce this gradually evolving process, initially expressed by dysplastic lesions of the mucosa, then transformed into in situ carcinoma lesions, eventually resulting in full-blown infiltrating and metastasizing oral SCC.
This circumstantial evidence emphasizes the possible
significance of the CS effect on salivary antioxidants in
the pathogenesis of oral cancer. Surprisingly, the effect
of CS on OPO activity has never been examined. This is

a major omission in light of two phenomena demonstrated recently:

1. Removal of H2O2 from the oral cavity, which is
carried out by the OPO, may have special significance, as redox-active metal ions are uniquely and
constantly secreted into the oral cavity via the parotid
saliva [23,24]; such metals, in the presence of H2O2,
may play a vicious role in the oral cavity by enhancing the production of the ultimately aggressive hydroxyl free radicals, which may be very injurious to
the neighboring oral epithelial cells.
2. CS may attack antioxidant enzymes rather than molecules; this was recently shown to severely reduce the
activity of various salivary antioxidant enzymes, but
not antioxidant molecules such as uric acid [2529].
The purpose of the current study was to analyze the in
vivo and in vitro effects of CS on OPO activity, concomitant with examining the effect of CS on protein
modification. The latter was measured by assessing levels of salivary protein carbonylation, a well-known indicator [30 32] of protein damage induced by exposure to
oxidative stress and/or CS [30]. Preliminary results on
the effect of CS on inactivation of OPO activity were
reported earlier [33].
MATERIALS AND METHODS

Experimental design, saliva collection, and smoking

Whole saliva was collected from healthy smoking
(over 20 cigarettes daily for at least 10 years) and nonsmoking volunteers under nonstimulatory conditions, as
described previously [33]. In the in vivo study, the OPO
activity of volunteers (smoking and nonsmoking) was
measured prior to and after smoking one cigarette. The
smoking volunteers were asked not to smoke for 1 h
prior to the experiment. The in vitro exposure to the CS
of one cigarette was performed as described previously
[25,27,30,31]. Following exposure to CS, 2 ml of saliva
were collected and immediately centrifuged at 800 g
for 10 min at 4C to remove squamous cells and cell
debris. The resulting supernatant was then used for determining peroxidase activity and for analysis of carbonyls in the salivary proteins.
Cigarettes
The cigarettes used in this study were popular commercial cigarettes containing 14 mg of tar and 0.9 mg of
nicotine (Time cigarettes, Dubek Ltd., Tel Aviv, Israel).
Exposure of saliva to CS: in vitro study
The in vitro study was carried out using a cigarette
combined with a vacuum system, as described previously

Cigarette smoke affects oral peroxidase

379

Table 1. Oral Peroxidase Activity (Time 0) in In Vivo and In Vitro Studies

In vivo studies (mU/ml)

In vitro studies (mU/ml)

Measurement

Smokers

Nonsmokers

Smokers

Nonsmokers

n
Range
Mean
SD
SEM

17
1801728
695.6
417.4
101.3

16
1931019
572.7
251.6
62.9

7
3971094
595.2
237.4
89.9

11
2361166
516.9
298.9
90.6

[25,27]. Saliva (4 to 5 ml) was placed in 50 ml flasks

with a sidearm to which the cigarettes were hooked. A
reproducible 50 mmHg vacuum was created in the flask
and, upon opening the vacuum, the CS of a whole lighted
cigarette was drawn into the flask in four or five puffs.
Salivary samples for OPO analysis were drawn before
smoking, after completion of one cigarette (about 10 min
after the first analysis), and at 30 and 60 min later while
flasks were incubated in a metabolic shaker at 37C.
Peroxidase activity
SPO activity was measured according to the NBSSCN assay [3]. Briefly, in this assay DTNB was reduced
to NBS by the addition of -mercaptoethanol. The disappearance of NBS while reacting with OSCN, the
product of OPO, was monitored at 412 nm at pH 5.6 [3].
One unit of enzyme activity was defined as the level of
enzyme activity needed to cleave 1 mol of NBS/min at
22C, using a molar extinction coefficient of 12,800 [3].
Western blot analysis of protein carbonyls
Salivary proteins were run on 10% SDS-PAGE as
described previously [25]. OxyBlot Kit (Intergen Co.,
Purchase, NY, USA) with antidinitrophenylhydrazine
(DNPH) antibodies was used to detect protein carbonyls,
since DNPH reacts with protein carbonyls on a one-toone basis. Thus, using antirabbit second antibody bound
to horseradish peroxidase and enhanced chemiluminescence detection, the level of DNPH was assessed [25].

Student t-test for differences in means was used to compare the means, and statistical significance was set at p
.05.
RESULTS

Peroxidase activity
Table 1 shows OPO activity at time 0 in both the
smoking and nonsmoking groups in the in vivo and in
vitro studies. The ranges and means were similar in both
groups, but a slight, nonsignificant decrease to 82 and
85% of activity was observed in the nonsmokers as
compared with the smokers in vivo and in vitro, respectively. The effect of smoking one cigarette on OPO
activity in the saliva of the smoking group as compared
with the nonsmoking group is shown in Fig. 1. Shortly
after smoking one cigarette and collecting 2 ml of saliva
from each volunteer, a sharp drop in OPO activity was
observed in both groups: a decrease of 42.5% in smokers
and of 58.5% in nonsmokers (p .05). The volunteers
were not allowed to smoke, and their OPO activity was
measured at 30 and 60 min after the onset of the experiment. After 30 min, the level of OPO activity returned to
90 100% of the initial presmoking level.
Another group of volunteers was chosen for a second
in vitro study. Two milliliters of saliva from each vol-

Thiocyanate determination
The level of SCN in smokers and nonsmokers was
determined using the method of Densen et al. [26].
Saliva was deproteinized with 10% trichloroacetic acid,
the supernatant was reacted with ferric nitrate and nitric
acid, and the color reaction was measured at 460 nm
using a standard solution of potassium thiocyanate.
Statistical analysis
The results for statistical analysis were compared
between the smoking and nonsmoking groups in both the
in vivo and in vitro studies. Data on the activities of each
group were observed and calculated. The two-sample

Fig. 1. In vivo effect of CS on OPO activity. The OPO activity of 17

smokers and 16 nonsmokers was measured prior to and after smoking
one cigarette.

380

A. Z. REZNICK et al.

Fig. 2. In vitro effect of smoking one cigarette on OPO activity in saliva

from seven smokers and 11 nonsmokers.

Fig. 3. OPO residual activity after in vitro exposure of the saliva of

three nonsmokers to one cigarette in control (no SCN) and increasing
concentrations of KSCN (0.55.0 mM). Each value represents an
average of three different volunteers SD.

unteer were placed in a smoking flask, and the saliva was

smoked with a single cigarette. OPO activity was measured prior to smoking (time 0) and at 10, 30, and 60 min
after smoking a single cigarette. Figure 2 shows the
results of smoking salivas of smokers and nonsmokers in
the flasks. Again, the loss of OPO activity was more
pronounced in the nonsmokers than in the smokers (p
.01). However, as would be expected, where no replacement of saliva was possible, there was no recovery of
OPO activity in either group.
To ascertain the reason that OPO activity was slightly
higher and more resistant to CS in smokers as compared
with nonsmokers, the effect of the exogenous addition of
SCN in the form of KSCN was measured in nonsmokers. The literature reports that the saliva of nonsmokers
contains between 0.3 and 1.5 mM of SCN, while the
level of SCN in the saliva of heavy smokers is 1.4 4.0
mM [34]. Using the ferric nitrate-nitric acid method, we
determined the ranges and means of SCN in our studies, which were found to be similar to the literature: (i) in
the range of 0.3151.23 mM in nonsmokers (0.725
2.44 mM, mean SD, n 10), and (ii) 1.272.38 mM
in smokers (1.82 0.50 mM, mean SD, n 9). Thus,
it was postulated that higher quantities of SCN in heavy
smokers might provide some protection against CS damage of OPO. Figure 3 shows the effect of adding KSCN
(range, 0.55.0 mM) to the saliva of three nonsmoking
volunteers, after overnight dialysis to remove endogenous SCN, in the in vitro system, and measuring OPO
activity before and after smoking one cigarette. It can be
seen that the addition of SCN increased OPO activity
considerably after smoking one cigarette and, indeed,
provided some protection of OPO activity as compared
with the controls.

4). Lane 1 of Fig. 4A indicates the intensity of carbonyl

detection of saliva in a representative nonsmoker before
and after smoking a single cigarette. Lanes 2, 3, and 4
show the intensity of carbonyl detection at 10, 30, and 60
min, respectively. The highest level of protein carbonyls
was observed shortly after the subject smoked a single
cigarette and provided 2 ml of saliva (lane 2); this
declined to presmoking levels after 30 and 60 min (lanes
3 and 4). Figure 4B depicts the results of Coomassie blue
staining of the same saliva. As can be seen, the protein
contents of all saliva samples were the same. However,
after exposure of saliva in vitro to one cigarette, the
intensity of carbonyl detection remained the same at 10,
30, and 60 min after smoking, as no recovery was observed (data not shown).
Finally, as noted above, OPO is composed of two
peroxidase enzymes: SPO and MPO. The question arose
whether the two components of OPO are affected similarly by CS. A compound called dapsone (4,4'-diaminodiphenylsulfone) has been shown to specifically inhibit
lactoperoxidase and not MPO at acidic pH [35]. The
OPO activity of five volunteers was examined in the
presence of increasing concentrations of dapsone (50
150 M). Figure 5 shows that dapsone reduced OPO
activity between 60 and 85%, while under the same
conditions plasma peroxidase activity (MPO) was not
affected at all. Figure 6 shows that CS caused a loss of
peroxidase activity at the same rate, whether or not it
contained dapsone (150 M), demonstrating that CS
seemed to be decreasing salivary MPO activity to the
same degree that it decreased salivary total peroxidase
activity.

Level of protein carbonyls

The levels of protein carbonyls were assessed by
Western blot analysis with anti-DNPH antibodies (Fig.

DISCUSSION

The salivary antioxidant system, in which the peroxidase is the pivotal enzyme, has been drawing increased

Cigarette smoke affects oral peroxidase

381

Fig. 4. Levels of protein carbonyls (A) and Coomassie blue staining for proteins (B) of saliva from a representative nonsmoker before
and after smoking one cigarette. Lane 1, before smoking (time 0); lane 2, 10 min after smoking one cigarette; lane 3, 30 min after
smoking one cigarette; and, lane 4, 60 min after smoking one cigarette. AMY amylase; PRPs protein-rich proteins; and LYS
lysozyme.

attention in recent years [2,28,3537]. This enzyme

seems to be of paramount importance in the oral defense
mechanism, especially against the attack of free radicals
related to CS and the evolution of oral cancer. In a study
published in 1997, saliva was shown to significantly
inhibit the initiation and progression of oral cancer in an
animal model induced by the carcinogen 4-nitroquinoline 1-oxide (4NQO) [38]. Further credence to this salivary anticarcinogenic capacity against oral cancer was
provided by Nishioka et al. [39], who, using the Ames
test, found that saliva inhibited the mutagenicity of the
well-known oral cancer inducers CS, 4NQO, and benzopyrene. Moreover, it was recently reported that patients
with oral lichen planus (a premalignant lesion) have a

Fig. 5. Effect of incubation of various concentrations of dapsone on

OPO activity in saliva of five subjects. For comparison, the effect of
dapsone on plasma is also shown. } subject 1; subject 2;
subject 3; subject 4; subject 5; and E plasma.

lower salivary antioxidant capability [40]. In addition,

Nair et al. [41] demonstrated that saliva inhibited the
production of ROS, such as the superoxide free radical
and H2O2 from betel quid tobacco, the most potent
inducer of oral cancer.
The special significance of SPO in cigarette smokers
relates to the observation that CS contains hydrogen
cyanide, which is metabolized by the liver to the SCN.
The SCN is specifically sequestered from the plasma by
the parotid gland and is secreted by this gland into the
oral cavity. Its concentration in the saliva of nonsmokers

Fig. 6. Effect of exposure to CS on saliva in the absence or presence of

dapsone (150 M). Saliva in which dapsone caused 40% loss of
peroxidase activity was exposed to CS. In the figure, the initial activities of the saliva containing dapsone (MPO active component) and the
saliva without dapsone (OPO) were adjusted to be equal. } OPO
exposed to CS; OPO exposed to CS in the presence of 150 M
dapsone; F control saliva exposed to air; and saliva in air in the
presence of 150 M dapsone.

382

A. Z. REZNICK et al.

Fig. 7. Pathway of cyanate metabolism (half-life 9.5 h).

ranges from 0.3 to 1.5 mM, while the respective range in

smokers is approximately 1.4 4.0 mM, depending on
the number of cigarettes smoked per day, with a prolonged half-life of 9.5 h [34]. Following SCN secretion
via the saliva, it reacts with the H2O2 in the oral cavity,
leading to elimination of the H2O2. This reaction is
catalyzed by the peroxidase. However, if the peroxidase
activity is significantly reduced, as occurs upon exposure
to CS, the H2O2 in the oral cavity is not eliminated and
remains available for further reaction with the redoxactive metal ions that are secreted via the parotid saliva,
as was discovered in the late 1990s [23,24] (Fig. 7).
Redox-active metals such as iron and copper in the
presence of H2O2 and other low reactive free radicals
found in CS, such as superoxide radicals, participate in
the deleterious Haber-Weiss and Fenton reactions, in
which hydroxyl free radicals are produced. These radicals are highly reactive and immediately react with
neighboring cellular macromolecules including DNA.
This may result in malignant transformation and may

explain the CS-induced role of free radicals in oral SCC

alongside other direct effects of carcinogens contained in
CS. Accordingly, the importance of the current demonstration of a CS-induced 60% reduction in OPO activity
(in the in vitro study) stems from the fact that, in this
condition, H2O2 is not removed from the oral cavity and,
hence, is available to participate in the DNA-injurious
reactions previously mentioned. The exact mechanism
by which CS leads to peroxidase loss of activity is now
being actively pursued and will be reported on in a
follow-up study.
In the in vivo studies, the CS-induced inactivation of
peroxidase activity was reversible, which may be explained by the continuous secretion of saliva into the oral
cavity (0.11.0 ml/min secretion rate) containing undamaged peroxidase, concomitantly with continuous removal
of the damaged enzyme by swallowing the affected saliva. The in vitro studies revealed that the loss of activity
was irreversible, as no new saliva could replace the
damaged enzyme. In any event, this peroxidase activity
recovery process is time-consuming, as demonstrated in
Fig. 1, taking approximately 30 min to fully regain
baseline values. Heavy smokers who smoke over 20
cigarettes per day have significantly reduced SPO activity values for long periods of time, depriving them from
being protected by SPO against CS and other free radicals that penetrate their oral cavity via food, drink, and/or
inhaled ingredients.
Further evidence for the mechanism suggested is presented in two reports by Shklar and Schwartz [42] and
Schwartz et al. [43]. In those studies, it was shown that
ascorbic acid acted as a cancer promoter rather than
suppressor in an animal model of oral cancer that was
induced by free radicals. This is in accordance with the
known pro-oxidant rather than antioxidant role of ascorbic acid in the presence of redox metals. In that model,
-carotene, a known antioxidant that inhibits peroxidation, inhibited cancer promotion and progression. Furthermore, we recently showed, in a system in which
saliva was exposed to CS, that ascorbic acid acts as a
pro-oxidant, enhancing the induced enzymatic activity
loss of amylase and lactate dehydrogenase [25,27].
Another interesting observation in both the in vivo
and in vitro studies was the relative stability of the
peroxidase in smokers vs. nonsmokers after exposure to
one cigarette, which may have resulted from partial
protection rendered by the higher concentrations of SCN
found in the saliva of smokers. Such a protective role
was demonstrated in the experiment in which we added
exogenous SCN to the saliva of nonsmokers, rendering
its peroxidase less vulnerable to CS. However, the continuous protective capacity of SCN, which may be
explained by the extended biological half-life of the
molecule (9.5 h) [44], was only partial and could not

Cigarette smoke affects oral peroxidase

prevent the immediate overwhelmingly injurious effect

of CS on peroxidase that did not recover during the first
30 min postsmoking.
Figure 4 depicts the levels of modified salivary proteins as measured by Western blot for carbonyls. It has
been shown that protein carbonyls in plasma exposed to
CS are mainly products of exposure to aldehydes in CS
[30] and that one cigarette can cause a huge increase in
protein carbonyls. In the current study, it was shown that
carbonylation was reversible in the in vivo but not in the
in vitro studies, again due to the secretion of new saliva
in the former but not in the latter. It is interesting to note
that the major salivary proteins amylase, acidic prolinerich proteins, and lysozyme were the ones most carbonylated by CS (Fig. 4).
In summary, the results of our study open new avenues for research aimed at elucidating the underlying
mechanisms of the immediate and overwhelming CSinduced inactivation of OPO or of the partial protection
rendered by SCN against it. From a clinical point of
view, our findings are important because the specific
peroxidase sensitivity to CS is unique as compared with
other salivary antioxidants, such as uric acid or GSH
molecules, which were not affected by smoking a single
cigarette [29,45]. In other words, it is not the general
reduction in salivary antioxidant capacity that may account for the much higher prevalence of oral pathologies
associated with the ROS attack in smokers (e.g., lethal
SCC, periodontitis), but, most likely, the unique injury to
the peroxidase as a crucial phenomenon. This specific
peroxidase activity loss leaves the oral epithelial cells
almost completely unprotected against the deleterious
effects of both SCN ions and hydroxyl free radicals
produced by unremoved H2O2 in the presence of salivary
redox-active metal ions. It has been shown that SCN ions
and free radicals may adversely react with DNA
[23,24,46,47], thus paving the way for CS induction and
saliva-mediated initiation and progression of oral cancer
[36].
Acknowledgements This work was supported by grants from the
Technion Vice President for Research and the Jan M. and Eugenia Krol
Foundation, Clifton, NJ, USA. The authors thank Ruth Toffler for her
excellent technical assistance and Ruth Singer for preparation of the
manuscript.

[4]
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[11]

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[13]

[14]
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[16]
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[18]

[19]
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[21]

[22]
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ABBREVIATIONS

DNPH dinitrophenylhydrazine
4NQO 4-nitroquinoline 1-oxide
GSH glutathione
HOSCN hypothiocyanous acid
H2O2 hydrogen peroxide
MPOmyeloperoxidase
NBS2-nitrobenzoic acid
OPO oral peroxidase
OSCN hypothiocyanite ion
ROSreactive oxygen species
SCCsquamous cell carcinoma
SCNthiocyanate
SCNthiocyanate ion
SPOsalivary peroxidase