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Tikrit University College of Dentistry

Salivary Sialic Acid Level and Oral Health
Statues in Sample in Tikrit City
A project study
Submitted to the College of dentistry Tikrit University in Partial Fulfillment of
the Requirements for the Degree of Bachelor in dentistry
By
Rahma Ghalib
Aisha Ahmed
Supervised By Dr. Raghad Tahseen Thanoon
Dr. Shatha Nasil Tawfeeq
هـ 1445 2024 م
بسم الله الرحمن الرحيم
ï´¿

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قَالُُوا سُبْحَانَكَ لاَ عِلْمَ

لَنَآ اِلاّ مََا عََّلمْتَنَآَّ
اِنَّك َانْتَ الْعَلِيمُ
الْحَكِيمُ ﴾
صَدَقَ اللهُ العَظيم
}البقرة/اية32{
Dedication
I dedicate my graduation to those who wished me success:_ My dear brothers
1 2
and sisters, to everyone who supported me and to everyone who wished me
3 4
success, to my dear doctors and the respected dean of the college of dentistry
who worked hard for us to reach this point of success and progress. And to my
family, friends, and colleagues, and a great thanks to the two greatest people,
5 6
my father, my mother, they raised us with their eyes that never sleep.

7
Acknowledgement
Praise be to God, Lord of the worlds, and prayers and peace be upon the most
honorable of the prophets and messengers, our Master Muhammad, his family,
his companions, and those who followed them with charity until the Day of
Judgment.
I thank God Almighty for his bounty for allowing me to accomplish this work
thanks to Him. Praise be to Him first and foremost
Then I thank those good guys who extended a helping hand to me during this
period, in the forefront of which is my professor overseeing the letter Dr.

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8
Raghad and Dr. Shatha , who spared no effort in helping me, as is his habit with
all students of knowledge.
Contents
Summary ................................................................................................. 6
CHAPTER 1 3
Introduction 3
CHAPTER 2 6
Literature review 6
2.1 pathogenesis 6
2.2 Oral cavity 7
CHAPTER 3 10
material and methods 10
CHAPTER 4 11
9
Results and discussion 11
Discussion 12
Conclusion 14
Reference 15

Summary
Background
Gingivitis and periodontitis are the most frequent chronic diseases worldwide.
Plaque-induced gingivitis affects 50% of individuals. Periodontitis may cause
10
gum recession, loss of gingival tissue, alveolar bone, and tooth, lowering
masticatory function and nutritional status if left untreated.
11
When some bacteria and their products colonise the gum, they release
proteolytic enzymes and ROS that increase host tissue damage biomarkers,

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causing periodontal disease. Smoking, poor nutrition, and low antioxidant (AO)
capacity may damage periodontal tissue from free radicals.
Smoking is a single, modifiable environmental risk factor that increases
periodontal disease prevalence and changes periodontal features.
The aim of the study
To identify diagnostic sialic acid fraction and its scavenger effect for Oral
Health Statues
Conclusion Statistical study indicates a significant correlation between oral
cavity characteristics and salivary sialic acid levels and oral health.
12
Statistical Analysis: The data were analysed using SPSS version 19.0. The
current research used descriptive analysis, analysis of variance, student T-test,
13
linear correlation, and multiple linear regression model.
The analysis is predicted to have a significance level of p < 0.05.
14
Key words: salivary, oral, significant, sialic acid.

3
3

3
CHAPTER 1
Introduction

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Periodontal diseases (gingivitis and periodontitis) are the most prevalent

15
chronic diseases affecting population worldwide. Gingivitis is inflammation of
the gum due to the accumulation of plaque, and affects 50% of the adult
population Periodontitis affects the supporting structures of the teeth and if
not promptly recognized and correctly managed can ultimately lead to gum
recession, loss of gingival tissue, underlying alveolar bone and tooth, resulting
in reduced masticatory function and subsequent alterations in dietary intake
and nutritional status [1-3]
Periodontal disease is initiated by the colonization of the gum by specific
bacteria and their products which causes abnormal host response, involving
the release of excess proteolytic enzymes and reactive oxygen species (ROS),
that cause increased levels of biomarkers for host tissue damage Tissue injury
from free radical production in periodontitis is related to low antioxidant (AO)
16
capacity and may be caused by a number of factors including smoking and poor
nutritional status [4,5]
Smoking is a single, modifiable environmental risk factor responsible for
excess prevalence of periodontal disease in the population and has a direct
influence on periodontal variables. Smoking effects include chronic reduction
of blood flow, altered neutrophil function, cytokine and growth factor
production, inhibition of fibroblast growth and attachment, and decreased
collagen production and vascularity It was demonstrated that smoking
increases the levels of free radicals and lipid peroxidation in periodontal
17
tissues. In addition to decreased antioxidant levels in blood, gingival tissue,
saliva, and gingival crevicular fluid (GCF) of periodontitis and gingivitis smokers
18
reported that socioeconomic disadvantages, poor oral hygiene habit, and bad
19
eating behavior associated with smoking and smoking related diseases [6-8].

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20
Laboratory tests of samples from plaque, saliva or gingival crevicular fluid are
more accurate than clinical measurements and are developed to measure
biomarkers (derived from bacterial structure or the host inflammatory system)
21
of periodontal diseases to detect of 'high-risk' individuals and an increased
probability of disease
Saliva is the first defense fluid and an important salivary biomarker is sialic
22 23
acids, they are family of nine carbon acidic monosaccharide, systemic
24 25
inflammatory marker, and component of salivary glycolipids, glycoproteins
including IgA and other immunological and acute phase proteins). Sialic acid
26 27 28
levels increased in periodontitis, because it is protective constituent of human
29 30
salivary mucin, and lipid bound sialic acid fraction can be used as diagnostic
31
parameter for periodontitis concluded that sialic acids of mucin acts as
scavengers for hydroxyl (OH) free radical and react directly with it. Therefore
32
this study was conducted to identify sialic acid fractions levels among smokers
33
as biomarkers for periodontal diseases and its prognoses [9,10].
CHAPTER 2
Literature review
2.1 pathogenesis
Many promising salivary biomarkers associated with PD have been reported
[11]. The pathogenesis of periodontitis is related to enzymatic alterations such
as malondialdehyde (MDA), sialic acid (SA), lactate dehydrogenase (LDH),
cortisol, β-glucuronidase (BetaG), interleukin 1β (IL-1β), antioxidants,
oxidative stress, superoxide dismutase (SOD), 8- hydroxydeoxyguanosine,
glutathione peroxidase (GPx), and 4hydroxynonenal [5–8]. SOD is an
antioxidant enzyme that is localized within human periodontal ligaments, and
it provides an important defense within gingival fibroblasts against superoxide.
However, plasma glutathione peroxidase, a selenium-containing peroxidase,

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comprises a major group of enzymes that remove the hydrogen peroxide

created by SOD in the cell [12]. IL-1β stimulates the expression of matrix
metalloproteinases (MMPs), which contribute to bone resorption and tissue
destruction. To date, 24 different MMPs have been cloned, and three of them
have been found in humans. Based on the substrate to be degraded, they are
divided into six types: collagenase, gelatinases (type collagenase),
stromelysins, matrilysins, membrane-type metalloproteinases, and others [13].
Among the MMPs, MMP-8 and MMP9 are in the spotlight as biomarkers for
periodontal disease. A kit that can test for MMP-8 in 5 min in an office has been
developed [13,14]. PD progression can be influenced by various risk factors
such as periodontal pathogens, host factors, anatomical factors, and iatrogenic
34
factors [15]. Among the associated risk factors, smoking is the secondlargest
risk factor for PD after dental plaque. Reports indicate that the prevalence of
periodontitis is 3–6 times higher in smokers than in nonsmokers, and the
increased risk is proportional to the duration of smoking and smoking rate.
Smokers exhibit more pronounced PD clinical findings than non-smokers, such
as deeper pockets, more extensive and severe loss of attachment, higher levels
of bone destruction, and higher rates of tooth loss [16]. In addition, smoking
negatively affects successful implant placement and non-surgical and surgical
treatment [17].
2.2 Oral cavity
Soft tissue (mucosal) surfaces such as those in the oral cavity, gastrointestinal
35
surfaces and other regions play critical roles in routinely protecting the
underlying regions and tissue i.e., blood vessels and structural components
from the environment [17]. A range of factors influence the mucosal surfaces of
the human mouth with its unique anatomical features that include the teeth
and tongue. Environmental factors such as those found in the diet along with

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localized influences due to the distinctive niches and regions that include the
unique structural features within the mouth comprise those routinely
impacting the oral mucosa [18].
In addition to the above influences, an important constituent of the human
mouth is its indigenous microbial populations that impact the oral mucosa. The
mucosal surface of the oral cavity is colonized by large densities of both gram-
positive and gram-negative bacteria with fungi and other constituents
representing additional residents. These microflora are found as biofilms in the
supragingival plaque on the exposed surfaces of teeth, as subgingival plaque
36
below the gumline and readily found within the other distinct niches of the oral
cavity such as the tongue and cheek surfaces. The salivary microbial
37
populations can be considered planktonic constituents that are able to
transport organisms between the oral surfaces.
Further to the above, the routine intake of diet and their nutritional features
facilitate microbial proliferation leading to a range of byproducts such as acids,
38
toxins, microbial cell wall constituents and including those with immunogenic
and other pathogenic characteristics. Taken together microbial factors
represent an important component of the stress and inflammatory burden of
the mouth. Identified widely in the literature with information drawn from
surveys and clinical studies are the relationships between the microbial load
within the human mouth and disease. Contemporary practices in clinical
dentistry are based on maintaining routine optimal oral hygiene to preserve oral
39
health. Selfcare measures based on toothbrushing with toothpaste are widely
accepted to cleanse the mouth and improve oral aesthetics [19]. Despite their
availability and educational measures to reduce the burden of oral diseases,
most populations report the significant impact of these diseases. Some of the
most common oral diseases reported are caries and periodontal disease. In the

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40
absence of adequate treatments these conditions can lead to tooth loss and
41
changes in aesthetics with longlasting impacts on the quality of life. Surveys
show that despite widespread access to excellent dental care, only about 10%
of UK adults register good oral health. Reversible conditions such as gingivitis
representing inflammation of the gums and structures that support the tooth
are other commonly reported oral conditions that are reported in 90% of
certain populations. The role of microbial influences on the initiation and
progression of these conditions represents an area of extensive laboratory and
clinical investigations [20].
CHAPTER 3
material and methods
42
Unstimulated salivary samples were collected from 100 healthy child, aged 5-
43 44
15 years for 5-minutes, between 9:00 - 11:00 A.M. This study was a analyzing
PD-related salivary biomarker factors associated with age. We conducted this
study per the standard method of the Preferred Reporting Items for
45,46
spectrophotometer and ELISA analzed the index tests (salivary biomarkers).
Plain text words (including synonyms or plural forms) and controlled vocabulary
of concept (e.g., Medical Subject Headings terms) were combined and used for
47
searches in the title and abstract fields for each other . The salivary factors
evaluated in this study may prove to be useful measures for gingival
inflammation in children and allow pediatric dentists to target preventive
measures appropriately.
Statistical analysis was conducted using SPSS version 19.0. The present study
used descriptive analysis, analysis of variance, student Ttest, linear
correlation, and multiple linear regression model. The analysis is expected to
have a significance level below 0.05.
CHAPTER 4

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Results and discussion

Table(1)shows the correlation between sialic acid and study parameters. A
48
positive strong correlation recorded between flow rate in children and dental
49 50 51 52
age (r = 0.400). On other hand .there was a statistically a significant correlation
53 54
between the salivary S.A and GI(p=0.04). It is obviously noted from table (2)
that children suffered from mild gingivitis recorded with lower salivary S.A
mean 61.5 u\l compared to those with moderate gingivitis 61.9 u\l and the
55
differences between the numerical value are non-significant when tested
statistically
56
Table 1 rate of parameters
Parameters
Sialic acid level (saliva)
Salivary flow rate
R = - 0.104 p= 0.3 [NS]
DMFs
R = 0.053 p = 0.59 [NS]
CI
R = 0 p< 0.001
FS
R =- 0.197 p = 0.042
Ds
R = 0.057 p = 0.56 S]
Table 2 salivary biomarker
Salivary Biomarker
mean ± standard
deviation
Significance

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Sialic acid (nmol/µL)

0.14 ± 0.02
p < 0.001
Activity of LDH
(nmol/min/mg)
896.56 ± 264.14
N/A
IL-1β (pg/mL)
251.35 ± 81.19
p < 0.0001
Cortisol (pg/mL)
417.16 ± 99.67
p < 0.0001
SOD (U/mL)
50.41 ± 4.25
p < 0.001
Urea(mg/dl)
5.6±1.2
N/A
Creatinine (mg/dl)
0.9±0.06
N/A
Discussion
57
In current study Reduced levels of antioxidant enzymes and elevated levels of
lipid peroxidation product could be used as diagnostic markers to measure
oxidative stress in PD associated with risk factors such as smoking a
58
significantly higher salivary cortisol and IL-1β; thus, they may have an

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59
increased risk of PD and PD severity [23] significantly altered enzymeactivity;
however, LDH and BetaG were reliable salivary biomarkers
Many promising salivary biomarkers associated with PD have been reported
[24]. The pathogenesis of periodontitis is related to enzymatic alterations such
as malondialdehyde (MDA), sialic acid (SA), lactate dehydrogenase (LDH),
cortisol, β-glucuronidase (BetaG), interleukin 1β (IL-1β), antioxidants,
oxidative stress, superoxide dismutase (SOD), 8- hydroxydeoxyguanosine,
glutathione peroxidase (GPx), and 4hydroxynonenal [5–8]. SOD is an
antioxidant enzyme that is localized within human periodontal ligaments, and
it provides an important defense within gingival fibroblasts against superoxide
[9]. However, plasma glutathione peroxidase, a selenium-containing
peroxidase, comprises a major group of enzymes that remove the hydrogen
peroxide created by SOD in the cell [10]. IL-1β stimulates the expression of
matrix metalloproteinases (MMPs), which contribute to bone resorption and
tissue destruction [11]. To date, 24 different MMPs have been cloned, and three
of them have been found in humans. Based on the substrate to be degraded,
they are divided into six types: collagenase, gelatinases (type collagenase),
stromelysins, matrilysins, membrane-type metalloproteinases, and others [25].
Among the MMPs, MMP-8 and MMP9 are in the spotlight as biomarkers for
periodontal disease. A kit that can test for MMP-8 in 5 min in an office has been
developed [13,14].
Meanwhile, saliva contains a unique and complex variety of enzymes and
proteins with important oral functions. The use of these enzymes for
diagnosing PD has unfortunately been hindered because the relevance of
60
protein and enzymes in saliva and disease etiology remain limited.
Furthermore, enzymatic alterations can be caused by various factors such as
temperature, pH, enzyme substrates, and the effect of inhibitors and activators

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[26]. In particular, tobacco compounds the damage activities of salivary

enzymes at the molecular level. However, saliva samples are non-invasive,
readily available, and inexpensive; therefore, saliva can be a valid alternative to
blood as a biomarker. Saliva is a favorable oral fluid to determine the health
state of the oral cavity, including the presence of PD [26]. Therefore, an
effective and reproducible salivary biomarker would be preferred over other
61
biomarkers. The aim of this study was to evaluate the evidence, using a
systematic review, and to highlight the future directions regarding the
diagnostic potential of salivary biomarkers associated with PD based on
smoking status.
Conclusion
62
There is a significant relationship between parameters of oral cavity and
63
salivary scalic acid with oral health according to the statistical analysis and the
renal function test there is no significant relationship with oral health in the
current study.

Reference
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peroxidation in young cigarette smokers compared to nonsmokers: impact of

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dietary intake. Nutr. J. 2007;6:39.

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Cavas L., Arpinar P., Yurdakoc K. Possible interactions between antioxidant
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Int. J. Sports Med. 2005;26(10):832–835. [
Chapple Iain L.C., Milward Mike R., Dietrich Thomas. The prevalence of
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Eguchi H., Ikeda Y., Ookawara T., Koyota S., Fujiwara N., Honke K., Wang P.G.,
Taniguchi N., Suzuki K. Modification of oligosaccharides by reactive oxygen
species decreases sialyl lewis x-mediated cell adhesion. Glycobiology. 2005;15
Beltrán-Aguilar Eugenio D., Eke Paul I., Thornton-Evans Gina, Petersen Poul E.
Recording and surveillance systems for periodontal diseases. Periodontol
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Jawzaly, J., 2010. A PhD thesis, College of Dentistry, Hawler Medicinal
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Jette Alan M., Feldman Henry A., Tennstedt Sharon L. Tobacco use: a
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Health. 1993;83(9)

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11.Jiang, Y.; Zhou, X.; Cheng, L.; Li, M. The impact of smoking on subgingival
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diseases. Int. J. Oral Implantol. 2021, 14, 385–399.
Srivastava, N.; Nayak, P.A.; Rana, S. Point of care: A novel approach to
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Ali, S.A.; Telgi, R.L.; Tirth, A.; Tantry, I.Q.; Aleem, A. Lactate dehydrogenase and
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19.Varki, A.; Gagneux, P. Multifarious roles of sialic acids in immunity. Ann. N. Y.
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Acknowledgment

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smoking-related

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monosaccharides

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compound/complex sentences

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etc.)

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