SALIVA

Dr. Nitika Jain

 contents
• Introduction
• Salivary gland anatomy
• Secretion of saliva
• Composition of saliva
– Organic components
– Inorganic components
• Functions of saliva
8/12/2012 SALIVA 2
 What is saliva
• Saliva serves multiple and important
functions. Three major, paired salivary
glands produce the majority of saliva: the
parotid, the submandibular, and the
sublingual glands. In addition, 600-1,000
minor salivary glands line the oral cavity
and oropharynx, contributing a small
portion of total salivary production.
8/12/2012 SALIVA 3
 Major salivary glands
• Parotid
• Sub – mandibular
• Sub - lingual

8/12/2012 SALIVA 4
 Salivary gland - secretion
• Serous: very thin and watery
o parotid gland
o lingual glands of von Ebner
(serous glands of von Ebner)

Serous acini

8/12/2012 SALIVA 5
• Mucous: very thick and viscous
o palatine glands
o posterior lingual glands
o labial buccal glands

Mucous acini

8/12/2012 SALIVA 6
• Mixed secretions: mix of the two
o Sublingual glands
– Mostly mucous with some serous
o Submandibular glands
– Mostly serous with some mucous
o Anterior lingual glands
– Mixed secretion

Mixed: mostly serous acini

(dark), partially mucous acini
8/12/2012 SALIVA (light cells) 7
 Parotid gland

8/12/2012 SALIVA 8
 Parotid gland
• Largest of all the salivary glands
• Purely serous gland which produces thin,
watery, amylase rich saliva
• Superficial portion lies in front of the
external ear and deeper portion lies behind
the ramus of the mandible
• Stensons duct
– Open out adjacent to maxillary second molar
8/12/2012 SALIVA 9
• It is 5.8 cm in the craniocaudal dimension,
and 3.4 cm in the ventral-dorsal
dimension.
• Weight is 14.28 g. It is
irregular, wedge
shaped, and unilobular.

8/12/2012 SALIVA 10
• Superior border – Zygoma
• Posterior border – External Auditory
Canal
• Inferior border – Styloid Process, Styloid
Process musculature, Internal Carotid
Artery, Jugular Veins
• Anterior border – a diagonal line drawn
from the Zygomatic to external auditory
canal.
8/12/2012 SALIVA 11
• Stensen’s duct arises from the anterior
border of the Parotid and parallels the
Zygomatic arch, 1.5 cm inferior to the
inferior margin of the arch. Stensen’s duct
runs superficial to the masseter muscle,
then turns medially 90 degrees to pierce
the Buccinator muscle at the level of the
second maxillary molar where it opens
onto the oral cavity.
8/12/2012 SALIVA 12
 Sub – mandibular gland

8/12/2012 SALIVA 13
 Sub – mandibular gland
• Second largest salivary gland
• Produces 65-70% of total saliva output
• The duct is called Wharton’s duct
• Wharton’s duct exits on the floor of the
mouth opposing the lingual surface of the
tongue

8/12/2012 SALIVA 14
• Located in a depression on the lingual side
of the mandibular body
• Innervated by parasympathetic nerve
endings and possesses NO sympathetic
receptors
• The parasympathetic fibers arrive through
the facial and glossopharyngeal nerves
• Mixed secretion – mostly serous
8/12/2012 SALIVA 15
 Sub – lingual gland

8/12/2012 SALIVA 16
 Sub – lingual gland
– Smallest of the major glands
– Produce less than 5% of total saliva output
• Saliva delivered via the ducts of Bartholin
• The Bartholin ducts exit on the base of the lingual
surface of the tongue
– Innervated by parasympathetic fibers
– Little or no sympathetic influence
– Mixed secretion – mostly mucous

8/12/2012 SALIVA 17
 Minor salivary glands
• Minor salivary glands are found
throughout the mouth:
–Lips
–Buccal mucosa (cheeks)
–Alveolar mucosa (palate)
–Tongue dorsum and ventrum
–Floor of the mouth
• Together, they play a large role in salivary
production.
8/12/2012 SALIVA 18
 Histology of salivary gland
• The acini are the primary secretory organs
but the saliva is modified as it passes
through the intercalated, striated, and
excretory ducts before being discharged
into the mouth and oropharynx.

8/12/2012 SALIVA 19
• The serous cells contain numerous
proteinaceous secretory (zymogen)
granules. These granules contain high
levels of amylase. In addition, the
secretory cells produce
kallikrein,lactoferrin, and lysozyme.

8/12/2012 SALIVA 20
8/12/2012 SALIVA 21
8/12/2012 SALIVA 22
 Saliva
• General characterstics:
Consistency - slightly cloudy due to
presence of mucins and cells
Reaction - usually slightly acidic (pH 6.02-
7.05).On standing or boiling, it loses Co2
and becomes alkaline.
Specific gravity - 1.002-1.012
Freezing point - 0.07-0.34° Celsius
8/12/2012 SALIVA 23
• Percentage contribution of different
salivary glands during unstimulated saliva:

8/12/2012 SALIVA 24
 Unstimulated flow
• resting salivary flow―no external stimulus
oTypically 0.2 mL – 0.3 mL per minute
oLess than 0.1 mL per minute means the person
has hyposalivation
Hyposalivation – not producing enough saliva

8/12/2012 SALIVA 25
 Stimulated Flow
• response to a stimulus, usually taste,
chewing, or medication eg, at mealtime
oTypically 1.5 mL – 2 mL per minute
oLess than 0.7 mL per minute is considered
hyposalivation

8/12/2012 SALIVA 26
The average person produces approximately
0.5 L – 1.5 L per day
•Salivary flow peaks in the afternoon
•Salivary flow decreases at night.
•There is a difference in the quality between
stimulated and unstimulated saliva

8/12/2012 SALIVA 27
 Ions and salivary flow
As saliva passes
through the salivary
ducts, cations
(sodium and chloride)
are reabsorbed into
the adjacent
blood vessels.
8/12/2012 SALIVA 28
As saliva passes through the salivary ducts, cations
(sodium and chloride) are reabsorbed into the
adjacent blood vessels. In exchange, bicarbonates
and potassium are transferred from the blood 29
Stimulated Salivary Flow
• Saliva passes through the salivary duct very rapidly
o It impedes the exchange of sodium and chloride for
potassium and bicarbonate

Unstimulated Salivary Flow

• Has a high content of potassium and bicarbonate
o The quality of unstimulated saliva will change when
flow increases because of a stimulus (chewing gum,
thinking about lemons, looking at a food you crave)

8/12/2012 SALIVA 30
 composition
• Water content - 99.5%
• Solids - 0.5%
 Inorganic content - 0.2%
 Organic content - 0.3%
 Gases - 1ml oxygen/100ml
- 2.5ml nitrogen/100ml
- 50ml carbondioxide/100ml
 Cellular elements

8/12/2012 SALIVA 31
Organic Components of Saliva
Enzymes:
• Amylase – converting starch into glucose
and fructose
• Lysozymes – prevents bacterial infections
in the mouth
• Histatins – prevents fungal infections
• Secretory IgA – immunity mediator

8/12/2012 SALIVA 32
• Lactoperoxidases – stimulation of minor
salivary glands
• RNase and Dnase – cellular maintenance
• Lipase – initiates digestion of fat
• Kallikrein – vasoreactive substances

8/12/2012 SALIVA 33
 Cellular Composition

• The cellular composition consists of:

• Epithelial cells
• Neutrophils
• Lymphocytes
• Bacterial flora

8/12/2012 SALIVA 34
8/12/2012 SALIVA 35
 -Amylase

• Present in parotid saliva at conc. of 60-120 mg/ 100ml in

submandibular saliva at approx. 25 mg/100 ml.

• Very little amylase activity in the sublingual and minor

glandular secretions.

• 6 isoenzyme forms exist; alpha-amylase (ptyalin) is Ca++

dependent and readily inactivated by a pH of 4 or less.

• The enzyme hydrolyses the alpha 1:4 glycosidic bond between

glucose units in the polysaccharide chain of starch.

8/12/2012 SALIVA 36
 Lysozyme
• An antibacterial enzyme.

• The mean concentration in whole saliva

– resting: 22 mg/100ml.
– Stimulated: 11 mg/100 ml.
• Lysozyme acts on the B (1-4) bond between N-acetyl-muramic
acid and N-acetyl glucosamine in the Gram positive bacterial cell
wall component.

• Lysozyme may also be bactericidal,

• Inhibits mucosal colonization by microbial aggregation.

8/12/2012 SALIVA 37
 Kallikrein
•Splits serum beta-globulin into bradykinin
•Functional vasodilatation to supply an actively secreting gland.
Dextranases
Increased whole saliva dextranase levels may be associated with
impaired oral hygiene and over consumption of sucrose and related
fermentable carbohydrates which support the growth of organisms
producing dextranases.

Invertases
•High invertase activity is based on the involvement of several
enzymes chiefly derived from dental plaque S.Mitis and S.Salivarius.
High invertase activity ---- consume high sucrose and it usually
•parallels with high lactobacillus and streptococcus counts of plaque.

8/12/2012 SALIVA 38
 Glycoprotein (Mucins)

• Lubricant

• Types—MG1 & MG2

• Polypeptide chain that stick together

• Low solubility, High viscosity, Strong adhesiveness

• Aids in mastication, speech, swallowing by

lubrication

8/12/2012 SALIVA 39
 Glycoprotein (Mucins)
• Preserve mucosal integrity

• Protective barrier against excessive wear

• Antibacterial action by selective adhesion of

microbes to oral tissue surface

• Barrier against acid penetration

8/12/2012 SALIVA 40
 MG1

• High molecular-wt

• Adsorbs tightly to tooth surface--enamel

pellicle formation--Protection from acid
challenges

• High in caries susceptible patients

8/12/2012 SALIVA 41
 MG2
• Low molecular wt

• Binds to enamel but get displaces easily

• Promotes the aggregation and clearance of oral

bacteria (S.mutans)

• High in caries resistant cases

8/12/2012 SALIVA 42
 Secretory leucocyte
proteinase inhibitor (SLPI)
Proteinase inhibitory property

Antimicrobial and antiviral

Imp. Role in wound healing

8/12/2012 SALIVA 43
 Tissue inhibitors of
metalloproteinase
Remodeling of extracellular matrix in inflammation

Growth promoting activity

Stimulation of osteoclastic bone resorption

8/12/2012 SALIVA 44
 Immunoglobulins

• Secretory IgA is the predominant immunoglobulin - 20mg/100ml

• 90% of the total parotid lgA

• 85% of whole saliva lgA

• 30-35% of which is derived from minor glands, IgG (1.5 mg/100ml) &
IgM (0.2 mg/100ml)

• Secretory IgA is synthesized by plasma cells within the glands in addition to

the mucosal epithelial cells.

• Secretory IgA --- non-lymphoid-derived glycoprotein designated as the

secretory component.

8/12/2012 SALIVA 45
 Immunoglobulins

• This IgA exhibits 3 possible functions:

-Inhibition of bacterial colonization, probably by

agglutination.

- Binding to specific bacterial antigens involved with

adherence.

-Affecting
8/12/2012 specific enzymesSALIVA
essential for bacterial metabolism.46
 Structural features of salivary
proteins
• Proline – rich proteins

• Statherins

• Cystatins

• Histatins

8/12/2012 SALIVA 47
 Proline-rich protein (Glycoprotein)

 70% of total secretory proteins

 Acidic (Large), Basic (Small)

 Present in enamel pellicle

 Larger PRP promote bacterial attachment

 Smaller reduces the initial bacterial attachment

 A.Viscosus, S.mutans, S.Gordoni

8/12/2012 SALIVA 48
 Statherin (Phosphoprotein)

• Is a small phosphoprotien (12000 daltons) relatively

rich in tyrosine and proline which has the property of

inhibiting Hydroxyapatite crystal growth.

• Potential precursor of enamel pellicle

• Inhibit spontaneous precipitation of cal. phosphate

in saturated solution

8/12/2012 SALIVA 49
 Cystatins

• Several cystatins are phosphorylated and

bind to HA

• Inhibit crystal growth of Cal. Phosphate

salts
8/12/2012 SALIVA 50
 Histatin
• Parotid and Submandibular saliva
• Bind to HA, precursor of acquired pellicle
• kills C.Albican in yeast form and mycelia form
• Bacteriostatic
• Inhibit hem agglutination and thereby
colonization

8/12/2012 SALIVA 51
 Other organic compounds

 Free Amino acids- (Below 0.1 mg/100 ml)

o Too low to provide nutrient source for
bacterial growth
 Urea (12-20 mg /100 ml)
o Hydrolyzed by bacteria with the release of
Ammonia---Rise in pH
 Glucose (0.5- 1 mg/ 100ml)
o Too low for bacterial growth
o Increase in DM
8/12/2012 SALIVA 52
  Lipids

 Cholesterol, fatty acid

glycerides, phospholipids

 Corticosteroids

 Cortisol and cortisine

 1-2 mg/100ml

 Vitamins

 Water soluble vitamins

8/12/2012 SALIVA 53
 Function of saliva
Protective properties:
– Lubrication
– Maintenance of mucous membrane integrity
– Soft tissue repair
– Dilution and clearance
– Aggregation
– Action of lactoferrin
– Salivary peroxidase system
– Antifungal activitySALIVA
8/12/2012 54
 Function of saliva
– Antiviral activity

– Buffering capacity of saliva

– Maintenance of tooth integrity

• Digestion

• Taste

•8/12/2012
Saturation SALIVA 55
 Lubrication
• Coat the food, the oral soft and hard tissues.
• Allows food to travel through the digestive system
surfaces with minimal friction.

• Without appropriate lubrication, food is retained and

impacted around the teeth,

• Both mg1 and mg2 can provide fluid layers with high-
film strength

8/12/2012 SALIVA 56
 Maintenance of mucous
membrane integrity
 Salivary mucins possess rheological properties that include low
solubility, high viscosity, elasticity, and adhesiveness.

 Provide an effective barrier against desiccation and

environmental factors

 Protect the underlying cells from sudden changes in osmotic

pressure.

 Second line of defense against protease activity-cysteine

containing phosphoprotien

8/12/2012 SALIVA 57
 Soft tissue repair
• EGF involved in oral wound healing

• Hormone like properties in stimulating epithelial cells

• Parotid gland is the major source

• VEGF also known as vasculotropin---- an angiogenic

cytokine

• PAF ---- a potent phospholipid mediator

8/12/2012 SALIVA 58
 Dilution and clearance
 Saliva dilutes and eliminates dietary sugars and acids

 This process is dependent on flow rate and swallowing

frequency

 Oral sugar clearance extensively prolonged when

unstimulated whole saliva flow rate is below .2ml/min

8/12/2012 SALIVA 59
 Aggregation
 Inhibit bacterial attachment

 Inhibits the adherence of these cariogenic organisms to

teeth and protection against caries.

Expectorated
or swallowed
Hinder
effective
adherence
Clumping of
bacteria
Histidine-rich peptide has growth-inhibitory and
bactericidal effects on oral bacteria.
8/12/2012 SALIVA 60
 Action of lactoferrin

• Lactoferrin, the exocrine gland secretion

• The bacteriostatic properties are attributed to the ability of the

unsaturated protein to bind two iron atoms per molecule

• Lactoferrin is capable of both a bacteriostatic and a bactericidal

effect on S mutans that is distinct from simple iron deprivation.

8/12/2012 SALIVA 61
 Action of salivary peroxidase

• The antimicrobial effect of salivary peroxidase against S mutans

is significantly enhanced by interaction with secretory lgA.

• Interaction with each other and with high molecular weight

mucin. This mucin serve to concentrate a defense force on the
mucosa against the external environment, entrapping and
incapacitating microorganisms.

8/12/2012 SALIVA 62
 Antifungal activity
• Parotid fluid has an antifungal capacity, reflecting properties of
both the neutral and the basic histidine rich peptides.

• Pollock and others showed that the basic peptides could cause
99% loss of viability of Candida albicans at levels of 25 mg/ml,

• Oppenheim and others found that the neutral histidine rich

peptide was a potent inhibitor of C. albicans germination at
levels as low as 2 mg/ml.

8/12/2012 SALIVA 63
 Antiviral activity
• Antibodies (secretory lgA) can directly neutralize viruses.

• Mucins are also effective antiviral molecules.

• A major function of saliva is to prevent the establishment of

unwanted species in the first place

8/12/2012 SALIVA 64
 Buffering capacity
Resistance to pH changes at an arbitrary point.

 3 buffer systems :
pH

o Carbonic acid / bicarbonate

o Phosphate Buffer
capacity
Secretion
rate
o Protein
CA CA
o CO2 + H2O H2CO3 H+ + HCO3-

o Concentration of bicarbonates is highest in parotid saliva

8/12/2012 SALIVA 65
 Maintenance of tooth integrity
• Physical flow of saliva (the hydrokinetic property) coupled with
muscular activity,

• Small decrease in the resting salivary flow rate can greatly

prolong sugar clearance time.

• Interaction with saliva provides a post eruptive maturation

through the diffusion of ions

8/12/2012 SALIVA 66
• This enrichment of the crystal structure increases hardness,
decreases permeability, increases resistance to caries.

• The original pellicle is replaced by a constantly replenished

salivary film selectively absorbed proteins with a high

affinity for hydroxyapatite provides a protective barrier

8/12/2012 SALIVA 67
8/12/2012 SALIVA 68
 Digestion
SALIVARY AMYLASE

STARCH DIGESTION

STARCH CLEARANCE

LIBERATING MALTOSE
8/12/2012 SALIVA 69
Mechanism of saliva formation
Saliva is formed in 2
stages :

• A primary secretion
occurs in the acini

• Then modified as it
passes through the ducts

8/12/2012 SALIVA 70
8/12/2012 SALIVA 71
 METHOD OF COLLECTING
SALIVA
Passive drool

Oral swab

Infant swab

Spitting method

Suction method

8/12/2012 SALIVA 72
• • Avoid alcohol for 12 hours before
sample collection.
• • Avoid eating major meal within 60
minutes of sample collection.
• • Avoid dairy products for 20 minutes
before sample collection.

8/12/2012 SALIVA 73
• Avoid foods with high sugar or acidity, or
high caffeine content, immediately before
sample collection, since they may
compromise the assay by lowering saliva
pH and increasing bacterial growth.
• • Rinse mouth with water to remove food
residue before sample collection. Wait at
least 10 minutes after rinsing before
collecting saliva toSALIVA
8/12/2012
avoid sample dilution. 74
 Prior to saliva collection
• Participants should rinse with water 10
minutes prior to collection.
• Cut plastic drinking straws into 2-inch (5
cm) pieces.

8/12/2012 SALIVA 75
 PASSIVE DROOL
• Passive drool is highly recommended
because it is cost effective and approved
for use with almost all analytes. To avoid
problems with analyte retention or the
introduction of contaminants, use only
high quality polypropylene vials for
collection, such as our 2 ml cryovials. The
vials used must seal tightly and be able to
withstand temperatures as low as -80ºC.
8/12/2012 SALIVA 76
8/12/2012 SALIVA 77
 Instructions to the patient
• Allow saliva to pool in the mouth.
• With head tilted forward, participants
should drool down the straw and collect
saliva in the cryovial. (It is normal for
saliva to foam, advise using a vial with
twice the capacity of the desired sample
volume.)

8/12/2012 SALIVA 78
8/12/2012 SALIVA 79
• Repeat as often as necessary until
sufficient sample is collected. One mL
(excluding foam) is adequate for most
tests. Collection of samples to be analyzed
for multiple analytes may require larger
vials.

8/12/2012 SALIVA 80
 Salimetrics Oral Swab (SOS)
• Participants who are not willing or able to
drool saliva into a vial.
If the saliva samples are to be analyzed for
cortisol, testosterone, α-amylase,
chromogranin A, cotinine, C-reactive
protein, or SIgA, the Oral Swab is an
excellent alternative to passive drool
because of its ease of use. The SOS also
helps filter mucus from the sample, help
improve immunoassay
8/12/2012 SALIVA results. 81
8/12/2012 SALIVA 82
8/12/2012 SALIVA 83
8/12/2012 SALIVA 84
 Children and infant swab

8/12/2012 SALIVA 85
8/12/2012 SALIVA 86
8/12/2012 SALIVA 87
 To be continued…
• Saliva as a diagnostic marker
• Diseases
• Xerostomia
• Conclusion

8/12/2012 SALIVA 88