Craniofacial Biology Lecture 3 & 4

Saliva and Salivation I/II

CLINICAL CASE: 28 year old male, swallowing of the right submandibular area, started gradually
5 days prior, no trauma history, tender on palpation but no redness or warmth of the skin,
changes its size, usually 2-3 times a day.
Translation:
1. It can be a lymph node, an abscess, a salivary gland stone, tumor, cyst
2. It is acute (5 days), there cannot be a tumor
3. It is not an inflammation or abscess because there is no warmth or redness
4. It cannot be a lymph node or cyst because it changes in size daily
5. It has to be a salivary gland associated.
Background Concepts:
• The anatomy of the gland: under and behind the mylohyoid
• Biochemistry/physiology: most likely gland to have stone: submandibular because saliva
viscosity, duct shape (long and bent)
• Pathology: salivary glands can swell due to mucus plug, stone, trauma
• If it is a stone, should be able to palpate and show on X-ray
• What is the diagnosis? Salivary stone
• What concepts would I need to know to diagnose it?
o Location of the gland and unique aspect of its duct
o Nature of submandibular salivary secretion- viscous
o Epidemiology of salivary stone- most frequent site is the submandibular gland

CLINICAL CASE: 39 year old female, received radiation and chemotherapy 6 months before for
cancer of the nasopharynx, concurrently developed very sensitive mucosa, rampant caries,
complains of painful tongue, “nasty appearance” of teeth, cracked tongue, intraoral
examination: a dry mucosa of the tongue, cracked lips, plaque deposit on teeth.
Translation:
1. Radiation and chemotherapy in the region of salivary glands causes dry mouth within a
week
2. Mucosa sensitivity and rampant caries are a result of the absence of saliva to protect it.
3. Tongue complaints- due to higher friction of tongue to hard palate
4. Confirmed dry mouth due to lack of saliva to protect it.
• What is the diagnosis? Radiation/chemotherapy induced xerostomia
• What concepts would I need to know to diagnose it?
o Role of saliva as a protective, buffering lubricating, antimicrobial, antiviral,
antifungal, re-mineralizing fluid
Concept 1: Saliva as a diagnostic fluid
• Normal salivary flow rate: 0.5 mL/min
• Chairside Diagnostic Test Kits in Periodontics
o Microbiological
o Genetic- gene polymorphisms are
considered to be risk factors for the
initiation or progress of periodontal
disease
o Biochemical

Concept 2: The Challenge to Protect the Oral Cavity

• Oral cavity- shared by two systems
• Protection against physical, chemical, and biological agents
• Exposed mineralized tissue

Concept 3: Key Functions of Saliva

• Protective
o Antibacterial- the most common protective function
§ Agglutination, bactericidal, bacteriostatic
• Antifungal
• Antiviral
• Against physical agents (lubricant, pellicle)
• Against chemical agents (acid, base)
• Repair of oral tissues (EGF, NGF)
• Mastication, swallowing
• Buffering agent- bicarbonate, critical pH
• Ion reservoir, inhibitor of crystal growth
§ High calcium phosphate concentration
• As a diagnostic tool
• Excretion
• Chemical communication (pheromonal, HLA)
• Paracrine- von Ebner’s saliva

Concept 4: Composition of Saliva

• Water (99%)
• Ions (Ca2+, HPO43-, HCO3-, Na+, K+, SCN-)
• Organic molecules
o Proteins (0.3%)- about 900 proteins, 600 shared P/SM
o Lipids
o Carbohydrates
o Small molecules: (urea, ammonia, steroids, volatile compounds)
Concept 5: General Principles of Salivary Proteins
• Redundancy
• Multifunctionality
• Overexpression
o So we can survive even at low flow rate

Salivary Proteins- Size and Functions

Salivary Proteins with Antibacterial Function

• Lysozyme
o Fleming sneezed into a petri dish, 1922s, saw
wherever the saliva went, there was death of
bacteria.
o First enzyme that was sequenced
o Found in several exocrine secretions
o N-acetyl muramic acid (NAM)- N-acetyl
glucosamine (NAG)
o Repeating NAM-NAG makeup bacterial cell
walls
o Has antibacterial and anti-HIV function
• Lactoferrin
o Iron binding- transferrin (ferric form) @ pH > 4.0
o Mr 76,000, binds two HCO3-
o Has bactericidal effect
o Anti-HIV
• Salivary peroxidase
o Present in saliva, milk, tears
o Mechanism: inhibition of bacterial -SH to S-S (disulfide) or sulfonyl states
§ SCN- + H2O2 = HSCN- + OHSCN
§ Thiocyanate + hydrogen peroxide = hypothiocyanite +
hypothyocyanous acid (which are toxic to bacteria)
 o SCN- from blood
o H2O2 from leukocytes and from bacteria, rate limiting factor
o antibacterial
• sIgA (salivary immunoglobulins)
o Secretory Component = Polymeric Immunoglobulin Receptor
o J chain + Secretory Component (SC) (S-S bridge)
o Secretory Component protects against proteolytic cleavage,
babies are protected by their mother’s immunoglobulin, but
then salivary glands start to secrete it
o Antibacterial, antiviral

Salivary Antiviral & Antifungal Proteins

• Antiviral
o Gp340, Lysozyme, Cystatins, Mucins, sIgA, Defensins
• Antifungal
o Histidine-Rich Proteins (histatins)

Salivary Proteins with Digestive Function

• Salivary Amylase (4-5 isoforms) hydrolyzes internal alpha-1,4-glucoside bonds of starch
to the disaccharide maltose which is broken down into glucose by maltase
• The main amylase in digestion is the pancreatic amylase which is secreted into the
duodenum
• Also functions in tissue-coating and antibacterial

Concept 6: Calcium and Phosphate in Saliva

• How is it possible? Place candies in a jar and they stick to one another like calcium
phosphate in a solution. Once you reach the max capacity, no more candies can be
placed without sticking to each other.
o Solution: space them
• How can you increase the concentration of the supersaturated calcium phosphate in
saliva without precipitating out?
o “Wrapping” calcium phosphate molecules individually with salivary proteins-
provides “space”
o Imagine each candy is the calcium phosphate molecule
o Imagine each wrap as the statherin and proline-rich protein cover that protects
against demineralization
• Calcium-Phosphate Salts in Saliva and Teeth (from lowest to highest pH)
o Dicalcium phosphate (acidic, amorphous) that will precipitate out at pH < 6.2 (ex:
calculus)
o Tricalcium phosphate
o Octacalcium phosphate
o Decacalcium phosphate (hydroxyapatite) (Basic, crystalline)- found in enamel
>6.2
• Calcium-phosphate salts dissolution- critical pH
 o pH < 5.5 Hydroxyapatite (enamel starts dissolving)
o pH < 4.5 Fluorapatite (aka stabilized hydroxyapatite by fluoride)
• Primary Crystal Growth- spontaneous crystal growth
o Staterin (Tyrosine-Rich Protein)- the only one, inhibits crystal growth
• Secondary Crystal Growth- seeded crystal growth
o Staterin- main
o Acidic proline-rich proteins (histatins)- less active
o Histidine-rich proteins (histatins)- less active
o Cysteine-rich protein (Cystatins)- less active

Concept 7: Acquired Mucosal and Enamel Pellicle (acquired salivary pellicle)

• Protection against mechanical insults
• Lubrication
• Mucins
o High and low molecular weight mucins (MG1,
MG2)
o Can bind water bc the protein has
carbohydrates outside
o Incompressible → makes a physical boundary
• Present throughout the entire GI system
• Heavily glycosylated to retain water
• Facilitates subsurface remineralization of the enamel
• Contains a number of salivary proteins
• Forms within a few minutes after tooth cleaning
• Attached to the surface of enamel.

1. Saliva forms a protective protein

pellicle on all oral surfaces-
mucosal or dental pellicle
2. Bacteria can also bind to the
pellicle
3. Oral mucosa sheds, removing
bacteria
4. Enamel does not shed. Dental
pellicle becomes a bacterial
binding surface.
Acquired Enamel Pellicle
• Subsurface lesion, intact enamel→
remineralization
• Supersaturated CaP (Calcium phosphate)
• Open surface lesion = no remain
• Allows for the initial colonization of bacteria
onto the enamel surface

Sialolith- Salivary Stone

• Most frequent at submandibular gland.

Major points of the lecture

• Salivary gland and secretory pathology are real and affect:
o Oral health, quality of life, success of your treatment
• Salivary composition reflects the adaptation to a wide variety of functions, primarily
protective, including anti: bacterial, viral, fungal, and crystal growth
• Three principles in the design of salivary proteins:
o Redundancy (overlap)
o Overexpression
o Multifunctionality
• Crystal growth inhibition- pellicle formation