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

‫ م‬2024 1445 ‫هـ‬

 ‫بسم هللا الرحمن الرحيم‬

‫﴿ قَالُوا سُبْحَانَكَ الَ عِلْمَ لَنَآ اِالّ مَا عَلَّمْتَنَآ اِنَّكَ َانْتَ الْعَلِيمُ‬

‫الْحَكِيمُ ﴾‬

‫صَدَقَ اهللُ العَظيم‬

‫{البقرة‪/‬اية‪}32‬‬
 Dedication

I dedicate my graduation to those who wished me success:_

My dear brothers and sisters, to everyone who supported
me and to everyone who wished me 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, my father, my
mother, they raised us with their eyes that never sleep.
 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. 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
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 gum recession, loss of gingival tissue, alveolar
bone, and tooth, lowering masticatory function and nutritional status if
left untreated.
When some bacteria and their products colonise the gum, they release
proteolytic enzymes and ROS that increase host tissue damage
biomarkers, 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.

Statistical Analysis: The data were analysed using SPSS version 19.0.
The current research used descriptive analysis, analysis of variance,
student T-test, linear correlation, and multiple linear regression model.
The analysis is predicted to have a significance level of p < 0.05.
Key words: salivary, oral, significant, sialic acid.
CHAPTER 1

Introduction

Periodontal diseases (gingivitis and periodontitis) are the most prevalent

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) capacity
and may be caused by a number of factors including smoking and poor
nutritional status [4,5]

3
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 tissues. In addition to decreased antioxidant
levels in blood, gingival tissue, saliva, and gingival crevicular fluid
(GCF) of periodontitis and gingivitis smokers reported that
socioeconomic disadvantages, poor oral hygiene habit, and bad eating
behavior associated with smoking and smoking related diseases [6-8].

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) 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 acids, they are family of nine carbon acidic monosaccharide,
systemic inflammatory marker, and component of salivary glycolipids,
glycoproteins including IgA and other immunological and acute phase
proteins). Sialic acid levels increased in periodontitis, because it is
4
protective constituent of human salivary mucin, and lipid bound sialic
acid fraction can be used as diagnostic parameter for
periodontitis concluded that sialic acids of mucin acts as scavengers for
hydroxyl (OH) free radical and react directly with it. Therefore this
study was conducted to identify sialic acid fractions levels among
smokers as biomarkers for periodontal diseases and its prognoses [9,10].

5
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 4-
hydroxynonenal [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, 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

6
[13,14]. PD progression can be influenced by various risk factors such as
periodontal pathogens, host factors, anatomical factors, and iatrogenic
factors [15]. Among the associated risk factors, smoking is the second-
largest 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 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 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].

7
 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 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 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 by-
products such as acids, 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 health. Self-
care measures based on toothbrushing with toothpaste are widely
accepted to cleanse the mouth and improve oral aesthetics [19]. Despite

8
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 absence of adequate treatments these
conditions can lead to tooth loss and changes in aesthetics with long-
lasting 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].

9
CHAPTER 3

material and methods

Unstimulated salivary samples were collected from 100 healthy child,

aged 5-15 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 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 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 T-
test, linear correlation, and multiple linear regression model. The
analysis is expected to have a significance level below 0.05.

10
CHAPTER 4

Results and discussion

Table(1)shows the correlation between sialic acid and study parameters.

A positive strong correlation recorded between flow rate in children and
dental age (r = 0.400). On other hand .there was a statistically a
significant correlation 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 differences between the
numerical value are non-significant when tested statistically

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]

11
Table 2 salivary biomarker

Salivary Biomarker mean ± standard Significance

deviation
Sialic acid (nmol/µL) 0.14 ± 0.02 p < 0.001
Activity of LDH 896.56 ± 264.14 N/A
(nmol/min/mg)
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
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 significantly higher salivary cortisol and IL-1β; thus, they
may have an 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

12
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 4-
hydroxynonenal [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
protein and enzymes in saliva and disease etiology remain limited.

13
Furthermore, enzymatic alterations can be caused by various factors
such as temperature, pH, enzyme substrates, and the effect of inhibitors
and activators [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
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

There is a significant relationship between parameters of oral cavity and

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.

14
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