Saliva

• 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
• 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
90% of the whole saliva is
produced by three paired
major salivary glands:

Parotid Gland

Submandibular gland

Sublingual gland
• 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 from non-glandular sources
such as gingival crevicular fluid in an
amount that depends on the periodontal
status of the patient
• Whole saliva,in contrast to glandular
saliva,also contains vast amounts of
epithelial cells from the oral mucosa
and millions of bacteria.

• These components give whole saliva its

cloudy appearance,which is different
from glandular saliva, which is
transparent like water.
Functions
Functions
Amylases, Cystatins,
Multifunctionality
Carbonic anhydrases,
Histatins, Mucins, Histatins
Peroxidases Anti-
Buffering
Bacterial

Cystatins, Amylases,
Mucins Mucins, Lipase
Anti-
Viral Digestion
Salivary
Families
Anti- Mineral-
Fungal ization
Cystatins,
Histatins
Histatins, Proline-
Lubricat- rich proteins,
Tissue ion &Visco- Statherins
Coating elasticity
Amylases,
Cystatins, Mucins,
Proline-rich proteins, Statherins
Mucins, Statherins
adapted from M.J. Levine, 1993
MAJOR FUNCTIONS OF
SALIVA

• Solvent
• Buffering
• Lubrication
• Remineralization
• Digestion
• Anti-bacterial
• Anti-fungal
• Temperature regulation
• Production of growth factors and other
regulatory peptides
• 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 demineralization caused by bacterial
acids produced during sugar metabolism

• remineralization :Saliva is supersaturated

with ions,which facilitate remineralization
of teeth
Digestion :breakdown of starch-amylase
•Fat-lingual lipase
•Moistening 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.

.
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 from plaque.

taste: Saliva has a low threshold

concentration of sodiumchloride , sugar,
urea etc allowing perception of taste to
occur. It acts as a solvent allowing
mixing and interaction of food with
taste buds
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
Sialochemistry
Inorganic Components
• 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
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 carbonic
anhydrase
Other ions • Fluoride
• Low concentration, similar to plasma
• Thiocyanate
• Antibacterial (oxidated to
hypothiocyanite OSCN- by active
oxygen produced from bacterial
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
Organic components
• 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
Mucins • Lubrication

• Hydrophillic, entraining water (resists

dehydration)

• Unique rheological properties (e.g.,

viscoelasticity, adhesiveness, low
resistnce to proteolytic degradation and
low solubility)

• Two major mucins (MG1 and MG2)

 • Calcium metalloenzyme
Amylases
• Hydrolyzes (1-4) bonds of starches such
as 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
Lingual Lipase • Secreted by lingual glands and parotis

• Involved in first phase of fat digestion

• Hydrolyzes medium- to long-chain

triglycerides

• Important in digestion of milk fat in

new-born

• Unlike other mammalian lipases, it is

highly hydrophobic and readily enters
fat globules
 • Calcium phosphate salts of dental enamel
are soluble under typical conditions of pH
Statherins
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

Proline-rich Proteins (PRPs)
• 40% of AAs is proline

• Inhibitors of calcium phosphate crystal

growth

• Part of pellicula dentis

• Subdivided into three groups

• Acidic 45%
• Basic 30%
• Glycosylated 25%
Lactoferrin • Iron-binding protein

• „Nutritional“ immunity (iron starvation)

• Some microorganisms (e.g., E. coli) have

adapted to this mechanism by producing
enterochelins.
• bind iron more effectively than
lactoferrin
• iron-rich enterochelins are then
reabsorbed by bacteria

• Lactoferrin, with or without iron, can be

degraded by some bacterial proteases.
Lysozyme • Present in numerous organs and most
body fluids

• Also called muramidase

• hydrolysis of (1-4) bond between N-

acetylmuramic acid and N-
acetylglucosamine in the
peptidoglycan layer of bacteria.
• Gram negative bacteria generally
more resistant than gram positive
because of outer LPS layer
Histatins • A group of small histidine-rich
proteins
• Potent inhibitors of Candida
albicans growth

secretory Leukocyte • Have antiviral properties

protease inhibitors
 • Are inhibitors of cysteine-proteases
Cystatins
• Are ubiquitous in many body fluids

• Considered to be protective against

unwanted proteolysis
• bacterial proteases
• lysed leukocytes

• May play inhibit proteases in

periodontal tissues

• Also have an effect on calcium

phosphate precipitation
 • Sialoperoxidase (SP, salivary
Salivary
peroxidase)
peroxidase • Produced in acinar cells of parotid
systems 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 gingival
crevice
• 15-20% of total peroxidase in whole
saliva
Functions Salivary Components Involved
(1) Protective functions

Lubrication Mucins, proline-rich glycoproteins, water

Amylase, complement, defensins,

lysozyme, lactoferrin, lactoperoxidase,
mucins, cystatins, histatins, proline-rich
glycoproteins, secretory IgA, secretory
leukocyte protease inhibitor, statherin,
Antimicrobial thrombospondin

Epidermal growth factor (EGF),

transforming growth factor-alpha (TGF-
α), transforming growth factor-beta
(TGF-β), fibroblast growth factor (FGF),
insulin-like growth factor (IGF-I & IGF-II),
Growth factors nerve growth factor (NGF)
Mucosal integrity Mucins, electrolytes, water
Lavage/cleansing Water
Bicarbonate, phosphate
Buffering ions, proteins
Calcium, phosphate,
statherin, anionic proline-
Remineralization rich proteins

(2) Food- and speech-related functions

Food preparation Water, mucins
Amylases, lipase,
ribonuclease, proteases,
Digestion water, mucins
Taste Water, gustin
Speech Water, mucins
PHYSIOLOGY OF
SALIVA
SECREATION
• 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 an intercalated 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.
Salivon

• Secretory unit of saliva

• Acinus + Intercalated duct +

striated duct

• The Intercalated & striated

ducts are more than passive
condiuts but actively involved
in formation of final saliva.
 Production of Saliva
• The production of saliva is an active
process 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 end
product
 THE SECRETORY UNIT
The basic building block of all salivary glands

 ACINI - water and Saliva formed in acini

ions derived from flows down DUCTS to
plasma empty into the oral
cavity.
 Primary saliva
• Secreted by acinar cells and similar in
composition to plasma (isotonic)

• Fluid & electrolyte secretion:

This process is driven by 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
water movt., via water channels & across
tight jns, to produce an isotonic, plasma lke
primary fluid.
• Fliud secretion in major salivary glnds is
largely initiated in response to
stimulation of muscarinic 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
 • Water secretion is driven by osmotic
Water/electrolyte changes
secretion • Mediated by ionic fluxes
• From basolateral 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
Alternative mechanisms

Na+-H+ exchanger
Cl-- HCO3- exchanger
Carbonic anhydrase

Na+-H+ exchanger
HCO3- channel
Carbonic anhydrase
Na+-K+-ATPase
Ca2+ activated K+
channel
Na+-K+-2Cl--
cotransporter
Ca2+ activated Cl-
channel Adapted from Turner and Sugiya, Oral Dis. 2:3-11, 2002
Ductal modification
• Saliva entering the lumen 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
• Lumenal
• Na+-H+ exchanger
• Cl-- HCO3- exchanger
• HCO3- channel
• Na+-K+ exchanger
• Na+-Cl--cotransporter
• Basolateral
• Na+-K+-ATPase
• Cl- channel
 Striated duct cell
Lumen Interstitium

Nucleus
Na+
H+
Cl- 3 Na+
HCO3- ATP
Carbonic 2K +

HCO3- anhydrase

Na+
Cl-
K+
Na+
Cl- Basolateral
Mitochondria membrane folds
• 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 HCO₃⁻ conc. increases with

increased flow rates,reflecting the
increased secretion of HCO₃⁻ by the
acinar cells to drive fluid secretion
 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
• Classic exocytosis pathway(stored
granule exocytosis)
• Endoplasmic reticulum - translation,
glycosylation
• Golgi - more extensive glycosylation
• Condensing vacuole - packaging,
condensation
• Immature granule - sorting, major
branching point
• Secretory granule - protein storage
• -adrenergic stimulation
• Docking, membrane fusion,
exocytosis
• Time taken from synthesis to exocytosis
is about 3-5 hrs
 Classic exocytosis
(Noradrenaline)

Immediate response to NA:

Docking and fusion of preformed
granules
Release of contents

Long-term response to NA:

Transcription
Translation
Glycosylation
New granules

http://www.liv.ac.uk/~petesmif/teaching/1bds_mb/p4/14.gif
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
The other protein
pathways • Constitutive-like pathway
• Branches off from immature granules/golgi
complex
• Proteins carried in vesicles to apex - fuse
and open
• Always active - no stimulation required
• Minor regulated pathway
• Branches off from immature granules
• Proteins carried in vesicles to apex - fuse
and open
• Triggered by low levels of M3 cholinergic
agonists
• Vesicle membranes contain t-SNARES for
granules
• Both are sources of proteins in basal
and resting secretions

• Vesicle contents are different from

granule contents

• Explains different protein

composition after stimulation

• In ths mechanism some proteins travel

in opposite direction; to the interstitium

• In addition; transcytosis is also seen

which indicates passage of substances
through acinar cells; like IgA; from
interstitial tissue through the cell from
BL to the apical cell membrane
Regulation of secretion
• 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 of both the divisions

innervate the secretory cells

• Myoepthelial, arteriolar smooth muscle cells,

intercalated & striated duct cells also recieve
direct innervation
• 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 of myoepthelial
cells.

• The molecular events that occur during

this process is called Stimulus-secretion
coupling
 Functions of ANS
• Parasympathetic
 Fluid formation
 Glandular metabolism & growth
 Transport activity in acinar & ductal cells
 Vasodilatation

• Sympathetic
 Exocytosis & protein metabolism modulation
 Constriction of blood vessels

• Dual
 Stimulation of salivary flow
 Constriction of myoepithelial cells
 Muscarinic messages

The Phospholipase C - IP3

pathway sends the
message

Intracellular (and extracellular)

Ca2+ flux is a major effector

http://www.liv.ac.uk/~petesmif/teaching/1bds_mb/p4/15.gif
 Adrenergic messages
(Noradrenaline)

The adenylate cyclase - cAMP

pathway sends the message

Effectors are activated by a

phophorylation cascade

http://www.liv.ac.uk/~petesmif/teaching/1bds_mb/p4/16.gif
There are 2 types of salivary secretions:

• Spontaneous : occurs all the time w/o any

known stimulus and keeps our mouth
moist all the time

• Stimulated : can be :
 Conditioned
 unconditioned
Unconditioned

• Inborn

• Eg: place lemon juice on the tongue of a

new born baby - there is salivation
 Conditioned

• Requires previous training

• Pavlov’s experiment
sialometry
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
• Sublingual
 Higher mucin content
 5% of salivary flow

• Minor salivary gland secretion

 Purely mucous glands
 5% of salivary flow
• 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
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=0.001-0.2

ml/minute/gland.

• With stimulation, salivary flow rate=0.18-

1.7 ml/min/gland.

• Salivary flow rate from the minor salivary

glands is independent of stimulation,
constituting 7-8% of total salivary output.
• 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

• 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%
• Though the Sublingual glands and minor
salivary glands contribute only about 10%
of all saliva, together they produce the
majority of mucous and are critical in
maintaining the mucin layer over the oral
mucosa.
Saliva Collectors
• Whole saliva :
1. Draining method
2. Spitting
3. Suction
4. Absorbent

• Parotid :
1. Cannulation
2. Lashley/Carlson-
Crittenden cup
• Submandibular/sublingual
:
1. Cannulation
2. Segregator( individual
prosthesis)
3. Suction
4. Wolff apparatus
Clinical implications
 Saliva as a Mirror of the Body
• Tissue fluid levels of natural substances, as well as
molecules introduced for therapeutic, dependency
or recreational purposes
• Emotional status
• Hormonal status
• Immunological status
• Neurological status
• Nutritional and metabolic influences
 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
Applications of Sialochemistry

• Diseases of the salivary glands

• Systemic diseases where salivary glands are
involved
• Clinical situations in which salivary flow and
chemistry are helpful in diagnosis or monitoring
patient progress
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 function (caffeine clearance)
 Malignancy
• P53 Tumor suppressor antigen
• inactivation in certain cancers leads to accumulation.
Oral squamous cell carcinoma leads to anti-p53
antibodies in saliva
• Salivary Defensin-1 levels elevated in oral SCC (made
by PMN’s).
• C-erbB-2 (erb) Tumor marker associated with breast
carcinoma.
• CA 125 (ovarian cancer marker) associated with
elevated salivary levels
 Drug and Hormone Monitoring
• Psychiatrists studying methadone: advantages using
saliva
• humanitarian- less discomfort
• clinical- patient acceptance of repeated testing
• children and patients with limiting coping abilities
• economic (do it yourself tests)
• HIV therapy
• Epilepsy
Drugs
Salivary Antibodies and Antigens
• Advantages in large scale studies
• Viral Screening
• Antigen Detection
• H. pylori (PCR of saliva)
• Antibody Screening
• rubella
• hepatitis A and B
• Shigella
Disorders of salivary secretion

• Disorders of salivary secretion &

composition can be generally termed:
Dyschylia

• Hypo/hypersecretion
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, parkinson’s
disease, secretory phase of menstrutiation
Xerostomia/hyposecretion
• Drugs: antcholinergics, antihistamines,
antihypertensives, oncological
chemotherapy, sedatives & chemotherapy
• Radiation & radioisotopes
• Oral conditions: sg benign& malignant
tumors, infections
• Other conditions: amyloidosis, bell’s palsy,
cystic fibrosis, diabetes, graft Vs Host
disease, granulomatous diseases, HV, late
stage liver disease, malnutrition, sjorens
syndrome, psychologcal factors, Sjogrens
syndrome, thyroid disease
References:
1. Orban’s 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 GLANDS SOURCE: Grand

Rounds Presentation, UTMB, Dept. of Otolaryngology 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