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Saliva lacks the drama of blood , the emotion of

tears and toil of sweat but still remains one of the

most imp. fluids in the human body (Mandel,

1990)

Its status in the oral cavity is at par with that of

blood i.e. to remove waste,supply nutrients and

protect the cells

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Saliva is composed of more than

99% water and less than 1%solids,mostly electrolytes and

proteins,the latter giving saliva its

characteristic viscosity.

The term saliva refers to the mixed

fluid in the mouth in contact with

the teeth and oral mucosa,which is

often called whole saliva

Normally the daily production of

whole saliva ranges from 0.5 to 1.0

litres

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90% of the whole saliva is

produced by three paired

major salivary glands:

Parotid Gland

Submandibular gland

Sublingual gland

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Secretions from many minor salivary glands in the

oral mucosa (labial, lingual, palatal,buccal,glossopalatine and retromolar glands) also

contribute (less than 10%) to the saliva secretion

In addition,whole saliva contains contributions fromnon-glandular sources such as gingival crevicular

fluid in an amount that depends on the periodontal

status of the patient

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Whole saliva,in contrast to glandular saliva,alsocontains vast amounts of epithelial cells from

the oral mucosa and millions of bacteria.

These components give whole saliva its cloudyappearance,which is different from glandular

saliva, which is transparent like water.

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Multifunctionality

SalivarySalivary

FamiliesFamilies

AntiAnti--

BacterialBacterialBufferingBuffering

DigestionDigestion

MineralMineral--

izationization

LubricatLubricat--

ion &Viscoion &Visco--

elasticityelasticity

TissueTissue

CoatingCoating

AntiAnti--

FungalFungal

AntiAnti--

ViralViral

Carbonic anhydrases,Carbonic anhydrases,

HistatinsHistatins

Amylases,Amylases,

Mucins, LipaseMucins, Lipase

Cystatins,Cystatins,

Histatins,P

rolineHistatins,P

roline--rich proteins,rich proteins,

StatherinsStatherins

Mucins, StatherinsMucins, Statherins

Amylases,Amylases,

Cystatins, Mucins,Cystatins, Mucins,

ProlineProline--rich proteins, Statherinsrich proteins, Statherins

HistatinsHistatins

Cystatins,Cystatins,

MucinsMucins

Amylases, Cystatins,Amylases, Cystatins,

Histatins, Mucins,Histatins, Mucins,PeroxidasesPeroxidases

adapted from M.J. Levine, 1993adapted from M.J. Levine, 1993

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MAJOR FUNCTIONS OF

SALIVA

Solvent

Buffering

Lubrication Remineralization

Digestion

Anti-bacterial

Anti-fungal

Temperature regulation Production of growth factors and other

regulatory peptides

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Fluid or lubricant: Saliva coats the mucosa & helps

to protect against mechanical wear ,chemical

erosion and thermal irritation.It also assists smooth

airflow,speech & swallowing.

Buffering: Saliva helps to neutralise plaque pH after

eating thus reducing time for demineralizationcaused by bacterial acids produced during sugar

metabolism

remineralization :Saliva is supersaturated withions,which facilitate remineralization of teeth

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Digestion :breakdown of starch-amylase

Fat-lingual lipaseMoistening and lubricative properties of saliva:

allow the formation & swallowing of food bolus

Antimicrobial action: Lysozyme, lactoferrin,sialoperoxidase help against pathogenic

microorganisms specifically Immunoglobulins and

secretory IgA also act against microorganisms.

Cleansing: Clears food and aids swallowing.

.

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Agglutination: immunoglobulins and secretory IgA

cause agglutination of specific microorganisms,

preventing their adherence to oral tissues.

Mucins as well as specific agglutinins also

aggregate microorganisms

Pellicle formation: Derived from salivary proteins,it

forms a protective diffusion barrier to acids fromplaque.

taste: Saliva has a low threshold concentration of

sodiumchloride , sugar, urea etc allowingperception of taste to occur. It acts as a solvent

allowing mixing and interaction of food with

taste buds

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Water balance: Osmoreceptors act as per state of

hydration of the body to transmit information to

the hypothalamus

Tissue repair: A variety of growth factors & other

biologically active peptides and proteins are

present in small quantities in saliva.

under experimental conditions,many of these

promote tissue growth & differentiation,wound

healing and other beneficial effects

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Calcium & Phosphate :

Help to prevent dissolution of dental enamel

Calcium

1.4 mmol/lt.(1.7 mmol/lt.in stimulated saliva)

50% in ionic form

sublingual > submandibular > parotid

Phosphate

6 mmol/lt.(4 mmol/lt.in stimulated saliva)

90% in ionic form

pH around 6 -hydroxyapatite is unlikely to dissolve

Increase of pH -precipitation of calcium salts => dental

calculus

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H2 CO3

Buffer

Low in unstimulated saliva, increases with flow

rate

Pushes pH of stimulated saliva up to 8 pH 5.6 critical for dissolution of enamel

Defence against acids produced by cariogenic

bacteria

Derived actively from CO2 by carbonicanhydrase

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Other ions Fluoride

Low concentration, similar to plasma

Thiocyanate

Antibacterial (oxidated to hypothiocyanite

OSCN- by active oxygen produced frombacterial peroxides by lactoperoxidase)

Higher conc. => lower incidence of caries

Smokers - increased conc.

Sodium, potassium, chloride, SO4

Lead, cadmium, copper, Mg May reflect systemic concentrations -

diagnostics

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Mucins

Proline-rich proteins

Amylase

Lipase

Peroxidase

Lysozyme

Lactoferrin

Secretory IgA

Histatins

Statherin

Blood group substances, kallikrein, sugars, steroid hormones, amino

acids, ammonia, urea, uric acid, clotting factors & lipids

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Mucins Lubrication

Hydrophillic, entraining water (resistsdehydration)

Unique rheological properties (e.g.,

viscoelasticity, adhesiveness, lowresistnce to proteolytic degradation andlow solubility)

Two majormucins (MG1 and MG2)

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Amylases Calciummetalloenzyme

Hydrolyzes E(1-4) bonds of starches suchas amylose and amylopectin

Several salivary isoenzymes

Maltose is the major end-product (20% is

glucose)

Appears to have digestive function -inactivated in stomach, provides

disaccharides for acid-producing bacteria

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Lingual Lipase Secreted by lingual glands and parotis

Involved in first phase offat digestion

Hydrolyzes medium- to long-chaintriglycerides

Important in digestion ofmilk fat innew-born

Unlike othermammalian lipases, it ishighly hydrophobic and readily entersfat globules

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Statherins

Calcium phosphate salts of dental enamel

are soluble under typical conditions of pH

and ionic strength

Supersaturation of calcium phosphates

maintain enamel integrity

Statherins prevent precipitation or

crystallization of supersaturated calcium

phosphate in ductal saliva and oral fluid

Produced by acinar cells in salivary glands

Also an effective lubricant

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Proline-rich Proteins (PRPs)

40% of AAs is proline

Inhibitors of calcium phosphate crystalgrowth

Part of pellicula dentis

Subdivided into three groups

Acidic 45%

Basic 30%

Glycosylated 25%

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Lactoferrin Iron-binding protein

Nutritional immunity (iron starvation)

Some microorganisms (e.g., E. coli) haveadapted to this mechanismby producing

enterochelins. bind iron more effectively than

lactoferrin

iron-rich enterochelins are thenreabsorbed by bacteria

Lactoferrin, with or without iron, can bedegraded by some bacterial proteases.

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Lysozyme Present in numerous organs and most

body fluids

Also called muramidase

hydrolysis ofF(1-4) bond between N-acetylmuramic acid and N-acetylglucosamine in thepeptidoglycan layer ofbacteria.

Gram negative bacteria generally

more resistant than gram positivebecause of outer LPS layer

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Histatins

secretory Leukocyte

protease inhibitors

A group of small histidine-rich

proteins

Potent inhibitors ofCandidaalbicans growth

Have antiviral properties

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Cystatins Are inhibitors of cysteine-proteases

Are ubiquitous in many body fluids

Considered to be protective against

unwanted proteolysis bacterial proteases

lysed leukocytes

May play inhibit proteases inperiodontal tissues

Also have an effect on calcium

phosphate precipitation

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Salivary

peroxidasesystems

Sialoperoxidase (SP, salivary

peroxidase)

Produced in acinar cells of parotid

glands

Also present in submandibular saliva

Readily adsorbed to various surfaces of

mouth

enamel, salivary sediment, bacteria, dental

plaque

Myeloperoxidase (MP)

From leukocytes entering via gingivalcrevice

15-20% of total peroxidase in whole

saliva

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Functions Salivary Components Involved

(1) Protective functions

Lubrication Mucins, proline-rich glycoproteins, water

Antimicrobial

Amylase, complement, defensins,

lysozyme, lactoferrin, lactoperoxidase,

mucins, cystatins, histatins, proline-rich

glycoproteins, secretory IgA, secretory

leukocyte protease inhibitor, statherin,

thrombospondin

Growth factors

Epidermal growth factor (EGF),

transforming growth factor-alpha (TGF-

), transforming growth factor-beta

(TGF-), fibroblast growth factor (FGF),

insulin-like growth factor (IGF-I & IGF-II),

nerve growth factor (NGF)

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Mucosal integrity Mucins, electrolytes, water

Lavage/cleansing Water

Buffering

Bicarbonate, phosphate

ions, proteins

Remineralization

Calcium, phosphate,

statherin, anionic proline-

rich proteins

(2) Food- and speech-related functions

Food preparation Water,mucins

Digestion

Amylases, lipase,

ribonuclease, proteases,

water, mucins

Taste Water, gustin

Speech Water,mucins

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Each SG consists of a large no of acini

Each acinus is lined by a single layer of

epithelial cells Acinar/ end peice

cells From the acinus anintercalated duct arises opens

into the striated duct finally

several open into the excretory duct

oral cavity.

Myoepithelial cells surround the acini

& intercalated ducts ( contain actin &

myosin)

Contractions of these lead to expulsion

of secretions within acinus & duct.

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Salivon

Secretory unit of saliva

Acinus + Intercalated duct +

striated duct

The Intercalated & striated

ducts are more than passive

condiutsbu

t actively involvedin formation of final saliva.

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Production ofSaliva

The production of saliva is an activeprocess occurring in 2 phases:

1) Primary secretion occurs in the acinar

cells. This results in a product similar in

composition and osmolality to plasma.

2) Ductal secretion results in a hypotonic

salivary fluid. It also results in decreased

sodium and increased potassium in the endproduct

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THE SECRETORY UNIT

The basic building block of all salivary glands

ACINI - waterandionsderived fromplasma

Saliva formedinaciniflowsdownDUCTStoemptyintotheoralcavity.

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Primary saliva

S

ecretedby acinar cells and si

milar incomposition to plasma (isotonic)

Fluid & electrolyte secretion:

This process is drivenby transepithelial Cl-

movt.

The acinar tight jns provide a cation-selective

pathway for Na+ flux down its electrical

gradient into the acinar lumen

The resultant osmotic gradient for NaCl causes

watermovt., via water channels & across

tight jns, to produce an isotonic, plasma lke

primary fluid.

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Fliud secretion in major salivary glnds is

largely initiated in response to

stimulation ofmuscarinic receptors on

the cell surface

ACh associated with its receptors G

protein activation & consequently, an

elevation of intracellular [Ca+] through a

PLC/IP3- dependent pathway

Inturn, this increase in intracellular [Ca+]

triggers the opening of apical Ci-

channels.

Na & then water follow Cl- into the

acinar lumen

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Water/electrolyte

secretion

Water secretion is driven by osmotic

changes

Mediated by ionic fluxes

Frombasolateral surfaces to the

apex (lumen)

Involves ion pumps and channels

Basolateral

Na+-K+-ATPase

Ca2+ activated K+ channel

Na+-K+-2Cl--cotransporter (NKCCl)

Na+-H+ exchanger

Cl-- HCO3- exchanger, plus

Carbonic anhydrase

Lumenal Ca2+ activated Cl- channel

HCO3- channel (Ca2+ activated?) ,

plus Carbonic anhydrase

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Alternative mechanisms

Na+-K+-ATPase

Ca2+ activated K+

channel

Na+-K+-2Cl--

cotransporter

Ca2+ activated Cl-

channel

Na+-H+ exchanger

Cl-- HCO3- exchanger

Carbonic anhydrase

Na+-H+ exchanger

HCO3- channel

Carbonic anhydrase

Adapted from Turner and Sugiya, Oral Dis. 2:3-11, 2002

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Ductal modification

S

aliva entering the lum

en is isotonic Saliva entering the mouth is hypotonic

Reabsorption of Na+ and Cl- by striated duct cells

Similar to distal tubules of kidneys

Ion pumps and channels

Lum

enal Na+-H+ exchanger

Cl-- HCO3- exchanger

HCO3- channel

Na+-K+ exchanger

Na

+

-Cl

-

-cotransporter Basolateral

Na+-K+-ATPase

Cl- channel

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Striated duct cell

Cl-

3 Na+

2 K+ATP

Cl-

Na+

Na+

Na+

K+

H+

Cl-

HCO3-

HCO3-

Lumen Interstitium

Nucleus

Mitochondria

Basolateral

membrane folds

Carbonic

anhydrase

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The final electrolyte composition of saliva

varies depending on the salivary flow rate

At high flow rates,saliva is in contact with

the ductal epithelium for shorter time &

Na & Cl conc increase & K conc

decrease

At low flow rates,the electrolyte conc.

change in the opposite direction

The HCO conc. increases with increased

flow rates,reflecting the increased

secretion of HCO by the acinar cells to

drive fluid secretion

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Protein secretion

A parallel process to water/ion secretion

Both occur side by side in the same secretory

cell

Multiple methods of secretion coexist in the

same acinar cells

There is complex cross-talk between

pathways

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Classic exocytosis pathway(stored granule

exocytosis)

Endoplasmic reticulum - translation,glycosylation

Golgi - more extensive glycosylation

Condensing vacuole - packaging, condensation

Immature granule - sorting, major branchingpoint

Secretory granule - protein storage

F-adrenergic stimulation

D

ocking, membrane fusion, exocytosis Time taken from synthesis to exocytosis is

about 3-5 hrs

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Classic exocytosis

Immediate response to NA:

Docking and fusion of preformed

granulesRelease of contents

Long-term response to NA:Transcription

Translation

Glycosylation

New granules

http://www.liv.ac.uk/~petesmif/teaching/1bds_mb/p4/14.gif

(Noradrenaline)

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Secretory granules

Complex internal structure

Multiple types of proteins, compacted and

folded

Membrane proteins that mediate docking and

fusion

V(esicle)-SNARES on granule membranes

T(arget)-SNARES on inner side of cell

apical membrane

A Ca2+ -dependent process

Example of cross talk between pathways

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The other protein pathways Constitutive-like pathway

Branches off from immature granules/golgicomplex

Proteins carried in vesicles to apex - fuseand open

Always active - no stimu

lation required Minor regulated pathway

Branches off from immature granules

Proteins carried in vesicles to apex - fuseand open

Triggered by low levels of M3 cholinergicagonists

Vesicle membranes contain t-SNARES forgranules

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Both are sources of proteins in basal and restingsecretions

Vesicle contents are different from granulecontents

Explains different protein composition afterstimulation

In ths mechanism some proteins travel inopposite direction; to the interstitium

In addition; transcytosis is also seen whichindicates passage of substances through acinarcells; like IgA; from interstitial tissue throughthe cell from BL to the apical cell membrane

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The flow of saliva is regulated predominantly

by the ANS

Although both sympathetic & prasympathetic

stimulation produces saliva, the

parasympathetic is dominant.

Parasympathetic: Ach & VIP

Sympathetic: NA

Postganglionic fibers ofboth the divisions

innervate the secretory cells

Myoepthelial, arteriolar smooth muscle cells,

intercalated & striated duct cells also recieve

direct innervation

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The receptors for these reside directly on

cell membrane and the NT is non synaptic

The axons can be hypolemmal or

epilemmal

The release of NT from the nerve

terminals adjacent to the parenchymal

cells stimulates them to discharge their

secretory granules, secretes water &

electrolytes & contraction ofmyoepthelial

cells.

The molecular events that occur during

this process is called Stimulus-secretion

coupling

Functions of ANS

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Functions of ANS

Parasympathetic

Fluid formation Glandularmetabolism & growth

Transport activity in acinar & ductal cells

Vasodilatation

Sympathetic

Exocytosis & protein metabolismmodulation

Constriction ofblood vessels

Dual

Stimulation of salivary flow

Constriction ofmyoepithelial cells

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Muscarinic messages


The Phospholipase C - IP3pathway sends the

message

Intracellular (and extracellular)

Ca2+ flux is a major effector

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Adrenergic messages


The adenylate cyclase - cAMP

pathway sends the message

Effectors are activated by a

phophorylation cascade

(Noradrenaline)

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There are 2 types of salivary secretions:

Spontaneous : occurs all the time w/o any

known stimulus and keeps ourmouth

moist all the time

Stimulated: can be :

Conditioned

unconditioned

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Unconditioned

Inborn

Eg: place lemon juice on the tongue of a

new born baby - there is salivation

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Conditioned

Requires previous training

Pavlovs experiment

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Salivary gland secretions:

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Salivary gland secretions:

Parotid gland:

Proteinaceous , watery serous secretion

2/3rd of salivary flow during gustatory &

olfactory stimuli

Organic (proteins inc. Enzymes; amylase) &

inorganic materials are higher

Submandibular gland:

High mucin content, viscous/ serous

secretion

High basal flow rate

Ca is higher

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Sublingual

Highermucin content

5% of salivary flow

Minor salivary gland secretion

Purely mucous glands

5% of salivary flow

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Secretion of salva is minimum at birth&

does not contain salivary amylase

The volume of saliva increases by 2-3

months & salvary amylase appears when

the infant is given complex CHO in diet

In old age the secretory reserves become

decreased though the constituents appear

to be stable

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Salivary Flow

The average volume of saliva secreted in

a 24 hour period is 1-1.5 liters (approx 1

cc/minute), most of which is secreted

during meals.

The basal salivary flow rate=0.001-0.2ml/minute/gland.

With stimulation, salivary flow rate=0.18-

1.7ml/min/gland.

Salivary flow rate from the minor salivary

glands is independent of stimulation,

constitu

ting7-8% of total salivary o

utput.

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Under normal conditions, the ph of

unstimulated saliva is about neutral(mean

value ph 6.8)

Upon stimulation the conc. Of HCO3

increases, resulting in higher ph (mean

value 7..4)

The specific gravity of saliva : 1.01-1.02

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In the UNSTIMULATED state the relative

contribution of the major salivary glands is

as follows:

1) Submandibular gland=69%

2) Parotid gland=26%

3) Sublingual gland=5%

In the STIMULATED state the relative

contribution of the major salivary glands is

as follows:

1) Parotid gland=69%

2) Submandibular gland=26%

3) Sublingual gland=5%

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Though the Sublingual glands and minor

salivary glands contribute only about 10%

of all saliva, together they produce the

majority ofmucous and are critical in

maintaining the mucin layer over the oralmucosa.

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Saliva Collectors

Whole saliva :

1. Draining method

2. Spitting

3. Suction4. Absorbent

Parotid :

1. Cannulation2. Lashley/Carlson-

Crittenden cup

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Submandibular/sublingual

:

1. Cannulation

2. S

egregator( individu

alprosthesis)

3. Suction

4. Wolff apparatus

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Saliva as a Mirror of the Body

Tissue fluid levels of natural substances, as well asmolecules introduced for therapeutic, dependencyor recreational purposes

Emotional status

Hor monal status

Immunological status

Neurological status

Nutritional and metabolic influences

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Clinical Situations Affecting

Salivary Secretions

Digitalis toxicity

Drug monitoring

Environmental pollutants

Ovulation Immunodeficiency

Pharmacological agents Dry mouth side effects, drugs with parasympathetic,

sympathetic and ganglionic blocking effects Direct effects- hypersensitivity or idiosyncratic reaction

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Applications ofSialochemistry

Diseases of the salivary glands

Systemic diseases where salivary glands are

involved

Clinical situations in which salivary flow and

chemistry are helpful in diagnosis ormonitoring

patient progress

Diagnostic Aids in Clinical Situations

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Diagnostic Aids in Clinical Situations

Digitalis toxicity (calcium and potassium)

Affective disorders (prostaglandin)

Immunodeficiency (sIgA)

Stomatitis in chemotherapy (albumin)

Cigarette usage (cotinine)

Gastric cancer (nitrates and nitrites)

Forensic medicine (blood group substance)

Coeliac disease (anti-IgA gliadin)

Liver f unction (caffeine clearance)

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Malignancy

P53 Tumor suppressor antigen inactivation in certain cancers leads to accumulation.

Oral squamous cell carcinoma leads to anti-p53antibodies in saliva

Salivary Defensin-1 levels elevated in oral SCC (made

by PMNs). C-er bB-2 (erb) Tumormarker associated with breast

carcinoma.

CA 125 (ovarian cancermarker) associated withelevated salivary levels

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Drug and Hormone Monitoring

Psychiatrists studying methadone: advantages usingsaliva

humanitarian- less discomfort

clinical- patient acceptance of repeated testing

children and patients with limiting coping abilities economic (do it yourself tests)

HIV therapy

Epilepsy

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Drugs

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Salivary Anti

bodies and Antigens

Advantages in large scale studies

Viral Screening

Antigen Detection

H. pylori(PCR of saliva)

Antibody Screening

rubella

hepatitis A and B Shigella

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Disorders of salivary secretion

Disorders of salivary secretion &

composition can be generally termed:

Dyschylia

Hypo/hypersecretion

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Hyperfunction/ptyalism/sialorrhoe

a/hypersialia

Drugs: bethanicol, clozapine, lithium,

physostigmine, pilocarpine, risperidone

Oral conditions: teething, ill-fitting

prosthesis, mucosal ulcerations

Other conditions: CVA, GERD, heavy

metal poisoning, hyperhydration, nausea,

obstructive esophagitis, parkinsons

disease, secretory phase ofmenstrutiation

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Xerostomia/hyposecretion

Drugs: antcholinergics, antihistamines,

antihypertensives, oncological

chemotherapy, sedatives & chemotherapy

Radiation & radioisotopes

Oral conditions: sg benign& malignant

tumors, infections

Other conditions: amyloidosis, bells palsy,

cystic fibrosis, diabetes, graft Vs Host

disease, granulomatous diseases, HV, latestage liver disease, malnutrition, sjorens

syndrome, psychologcal factors, Sjogrens

syndrome, thyroid disease

References:

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References:

1. Orbans Oral Histology and Embryology

2. Salivary Diagnostics : By David T. Wong

3. Concise medical Physiology: Chaudhary

4. Otorhinolaryngology, Head and Neck Surgery :By Matti Anniko, Manuel Bernal-Sprekelsen, PATRICK BRADLEY

5. Salivary gland diseases: surgical and medical management :By Robert Lee Witt

6. ANATOMY AND PHYSIOLOGY OF THE SALIVARY GLANDSSOURCE: GrandRounds Presentation, UTMB, Dept. ofOtolaryngology DATE: January 24, 2001

Resident Physician: Frederick S. Rosen, MD Faculty Physician: Byron J. Bailey,

MD

7. Physiology of saliva: DENT 5302;Topics in Dental Biochemistry;Dr. Joel Rudney

8. Saliva as a Diagnostic Fluid: Dennis E. Lopatin, Ph.D.;University of Michigan

9. www.google.com

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