DEPARTMENT OF ORAL PATHOLOGY AND MICROBIOLOGY

2015-2016

SALIVA AS A
DIAGNOSTIC TOOL

PREPARED BY,
SHEPHINA MARY PHILIP
ROLL NO : 130020086
III YEAR
CIIDS $ RC
CONTENTS
 INTRODUCTION
 WHY USE SALIVA
 ADVANTAGE AS DIAGNOSTIC FLUID
 FUNCTIONAL AND CLINICAL UTILITY OF SALIVA
 SALIVA DIAGNOSTICS IN VARIOUS SYSTEMIC DISEASES
 METHODS OF SALIVA COLLECTION
 SALIVA BIOMARKERS IN DENTAL CARIES
SALIVARY IONS , PH, BUFFER CAPACITY, FLOW RATE, PROETINS , BACTERIA
 CARIES ACTIVITY TEST
LACTOBACILLUS COLONY TEST
SWAB TEST
S.MUTANS LEVEL TEST
S.MUTANS DIP SLIP METHOD
BUFFER CAPACITY TEST
SALIVARY REDUCTASE TEST
 SALIVARY TUMOUR MARKERS IN ORAL CANCER
PROTEIN MARKERS
GENOMIC MARKERS
SALIVARY MICROBES
 METHODS TO TEST SALIVA
SIALOMETRY
SIALOCHEMISTRY
SALIVARY HORMONE PROFILING
 ADVANCED STUDIES
OFNASET
INTRODUCTION

Early detection of disease plays a crucial role for

treatment planning and prognosis.
Saliva has great potential as a diagnostic fluid and
offers advantage over serum and other biological
fluids by an economic and non invasive collection.

The plethora of components in this fluid can act as

biomarkers for diagnosis of various and systemic
diseases.
..
..
WHY USE SALIVA ?????

HORMONES
PROTE
S
ENZYME
S
ANTIBOD ANTIMICRO
L
IES CONSTITUE

CYTOKIN
MUCOSAL TRANSUDATIONS

GINGIVAL CREVICULAR FLUID

SERUM $ BLOOD DERIVATIVES FRM ORAL WOUNDS

DESQUAMATED EPITHELIAL CELLS

BRONCHIAL $ NASAL SECRETIONS

BACTERIA AND ITS PRODUCTS

VIRUSES AND FUNGI

CELLULAR COMPONENTS

FOOD DEBRIS
ADVANTAGE AS DIAGNOSTIC
FLUID
Non invasive diagnosis of disease

Patient suffers no discomfort and little anxiety

East to collect

Relatively cheap technology as compared to other tests

Convenient for multisampling

Cost effective applicability for screening large population

Easy to store and ship

Unlike blood, saliva doesn’t clot and cant be manipulated easily.

Safer for health professionals than blood tests

FUNCTIONS AND CLINICAL
UTILITY OF SALIVA
SALIVA DIAGNOSTIC IN VARIOUS SYSTEMIC DISEASES
METHODS OF SALIVA
COLLECTION
 Whole Saliva
 Catheterization
 Parotid Saliva
 Submandibular and sub maxillary saliva
 .

SALIVARY
BIOMARKERS IN
DENTAL CARIES
 SALIVA
RY IONS
SALIVARY
SALIVARY
CARIOGENI
C BACTERIA pH

SALIVARY SALIVARY
FLOW BUFFER
RATE CAPACITY
SALIVARY
PROTEINS
SALIVARY IONS
CALCIUM AND PHOSPHATE IONS
 The inorganic phase of enamel consist of crystalline hydroxyapatite in
the form of calcium and phosphate complexes.
 These complexes usually dissociate as pH drops and result in free active
concentration of ions.
 This is an example of dynamic equilibrium such that the rate of
dissolution and precipitation are equal to one another.
 At equilibrium saliva as a solution is saturated and the ion activity
product (IAP) is same as solubility product (Ksp).
 If IAP= Ksp, saturation index (SI) is zero, which means mineral is in
equilibrium with solution.
 If IAP <Ksp, SI is negative, saliva is unsaturated and the teeth would
solubilise.
 Under normal circumstances, saliva is supersaturated with respect to
enamel apatite which tends to precipitate apatite in the surface enamel
of carious lesions.
AMMONIA
 The saliva of caries immune persons exhibited greater ammonia
content than saliva from persons with caries.
 The ammonia of saliva from caries susceptible individuals was about
0-8 mg/100 ml, where as the same in caries immune individuals was
about 4.0-10 mg/100ml.

pH OF SALIVA
 There is an apparent relation of acidic pH of saliva to dental caries.
 Studies have shown larger quantities and faster rates of acid
production in caries active individuals than in caries free individuals.
 The pH at which any particular saliva ceases to be saturated with
calcium and phosphate is referred to as the ‘ critical pH’, below this
value, inorganic material of the tooth may dissolve.
BUFFER CAPACITY
 The quantitative assessment of resistance to ph changes is referred
to as buffer capacity.
 Low buffering capacity is associated with caries development
because of its impaired neutralization of plaque acids and reduced
remineralization of early enamel lesions.
 individuals with high buffering capacity are often caries resistant.

FLOW RATE
 Low salivary flow rate is a risk for caries incidence.
 The most common alterations in salivary flow rate involve reduced
secretion, which may be influenced by medications, pathological
changes in the salivary glands, and age etc.
 It is considered a potential risk factor when the unstimulated salivary
flow rate is lower than 0.30ml/min and the stimulated salivary flow is
lower than 0.7ml/min.
SALIVARY PROTEINS
 Caries rampant group exhibited a significant reduction in the salivary
level of basic proteins and a significant increase in amylase
compared to caries free group.
 Diminished levels of MUC7 , one of the predominant mucins in
saliva, is found to be significantly associated with elevated S.mutans
titers.
 Low salivary levels of alpha –defensins HNPI -3 may present a
biological factor that contributes to caries susceptibility in children.
 Lower salivary levels of statherin and cystatin S may be potential
risk indicators for caries development.
SALIVARY CARIOGENIC BACTERIA
 Saliva could act as an oral circulating fluid for bacterial transmission
and act as reservoir for bacterial colonization.
 There are about 700 oral microbial species living in saliva and some
could serve as biomarkers of the health and disease status of the oral
cavity.
 Level of certain bacterial species in saliva can reflect their presence
in plaque.
 Studies have proved that increased proportions of mutants
streptococci and lactobacilli in saliva are correlated with increased
caries initiation and progression as well as the presence of root
caries.
CARIES ACTIVITY TEST
Lactobacillus colony test
It measures the number of aciduric bacteria in a patients saliva for
assessing the caries activity. If number is > 10,000 caries activity is
marked.

Swab test
This test is developed to find the rate of acid production. The
aciduric and acidogenic elements of the oral flora are measured. A
ph of 4.1 and below denotes marked caries activity.

Salivary s.mutans level test

Levels of s.mutans greater than 100000 are indicative of an
acceptable cariogenic challenge because colonization does not
occur until the level of s.mutans reaches 4.5*10000/ml.
s.mutans Dip- slide method
This test classifies salivary samples according to estimates of
s.mutans colonies growing on modified mitis salivarius agar [MSA]
The colony density is compared with a model chart and classified as
zero[negligeble], 1[less than 1 lakh], 2[1-10 lakh]

Buffer capacity test

This test evaluates the quantity of acid required to lower the pH of
saliva through an arbitrary pH interval. This test relates the buffering
capacity of saliva and caries activity.

Salivary reductase test

The activity of the reductase enzyme present in salivary bacteria is
measured. The caries free adults exhibits low or negative scores.
SALIVARY
TUMOUR
MARKERS IN
ORAL CANCER
PROTEIN GENOMIC
MARKERS MARKERS

SALIVARY
MICROBES
PROTEIN MARKERS
 Over expression of proteins in OSCC :
PROTEIN FUNCTIONS
M2BP Tumor antigen
MRP14 Calcium binding protein
CD59 Antibody mediated killing
Prifilin 1 Regulator of
microfilament system
Catalase Protects against oxidative
stress

 Salivary concentration of salivary epithelial markers which included

CA125, CA19-9, TPA, CEA was increased in cancer patients than in
normal controls.
 Salivary soluble CD44 levels which are adhesion molecules in t
lymphocytes had increased value in head and neck carcinoma than
in controls
 P53 gene located on chromosome 17p was observed in DNA
extracted from saliva of OSCC patients suggesting a potential use as
a biomarker for oral cancer detection.
 Hyaluronic acid responsible for regulation of cell adhesion,
migration and proliferation was seen increased in OSCC patients.

GENOMIC MARKERS
 Tumor specific genomic markers consisting of DNA and RNA
markers can be identified for detection of oral cancer considering
that the initiation and progression of malignant tumors is driven by
the accumulation of specific genetic alterations.
 DNA shows tumor specific characteristics such as tumor somatic
mutation s in tumor suppressor genes and p53, microsatellite
alteration, mitochondrial DNA mutations and presence of tumor
related viral RNA.
 Salivary mRNA showing significant changes in oral cancer :
SALIVARY MICROBES
 Oral cavity plays host to a wide array of microorganisms inclusive of
variety of bacteria, viruses, and fungi.
 Role of bacteria in oral cancer is currently being investigated to
determine if the role is a casual or it is a co –incidental.
 Salivary counts of 40 common oral bacteria were determined in
OSCC and normal counts using DNA hybridization technique. It was
found that there is an elevated level of P.gingivalis, P. melaninogenic
and S.mitis in saliva of OSCC patients.
 Candida species are normal commensals of the oral cavity and
evidence suggest their involvement in oral cancer development
through nitrosation of nitrosobenzene compounds.
METHODS TO TEST SALIVA
SIALOMETRY
Sialometry is a measure of saliva flow.
 The following tests can measure saliva production:
A. Whole unstimulated saliva production.
B. Stimulated whole saliva production.
C. Stimulated parotid saliva production.
SIALOCHEMISTRY
 Sialochemistry is the chemical analysis of saliva.
 Diseases of the salivary glands can be determined.
 Systemic diseases where salivary glands are involved like sjogrens
syndrome can be accurately determined.

SALIVARY HORMONE PROFILING

 Saliva hormone testing is an accurate and reliable way of measuring
hormones.
 Saliva hormone testing can be used to identify existing hormone excesses
or deficiencies.
 The saliva test for cortisol, the adrenal hormone for stress responses are
highly reliable.
 Saliva tests can also be accurately determine levels of progesterone and
testosterone.
ADVANCED STUDIES
OFNASET
 The Oral Fluid Nanosensor Test.
 The intended use of the OFNASET is for the point of care multiplex
detection of salivary biomarkers for oral cancer.
 It combines cutting edge technologies, such as self assembled layers
monolayers (SAM), bionanotechnology, cyclic enzymatic
amplification and micro fluidics.
LAB ON CHIP
 It refers to technologies which allow operations which normally
require a laboratory synthesis and analysis of chemicals on a very
miniaturized scale within a portable or handheld device.
 It integrates many different areas of technology such as micro
fluidics and nanofluidics.
 Micro fluidics is defined as the manipulation of the flow of very small
quantities of fluid within channels in the micrometer range.
 Nanofluidics deals with the movement s of individual
macromolecules in the solution.
REFERENCES

 SHAFER, TEXT BOOK OF ORAL PATHOLOGY,

7TH EDITION, ELSEVIER PUBLICATION
 IOSR JOURNAL OF DENTAL AND MEDICAL
SCIENCES, VOLUME 11, ISSUE 6 (NOV-DEC
2013 ), PP 96-99