Microbiology and dentistry

Wenyuan Shi
Professor
School of Dentistry
Microbiology, Immunology and Molecular Genetics

Tel: 310-825-8356
Office: CHS 20-114
E-mail: wenyuan@ucla.edu
 Microbial pathogenesis

Bacteria Parasites Viruses Fungi

The Importance of Studying Bacterial Pathogenesis
• A human body has 1 X 1013 eukaryotic
cells and 1 X 1014 bacterial cells
• Microbial infections are the most
epidemic diseases and the leading cause
of death
– Diarrhea and enteric bacteria
– Tuberculosis and Mycobacterium
– Ulcer and Helicobacter infection
– Urinary tract infection
– STD

How microbiology is related to dentistry?

The dirty mouth!

100,000,000,000,000 bacteria/per mouth

>700 bacterial species
The first microbes observed

• Anton Van Leeuwenhoek

(1632-1723) developed
the microscope and was
the first to discover oral
bacterial flora: “I didn’t
clean my teeth for three
days and then took the
material that had lodged
in small amounts on the
gums above my front
teeth…. I found a few
living animalcules..”
Microbiology and dentistry

G+
Supragingival plaque

G-
Subgingival plaque

•79% of the population infected

•Annual dental spending $85 billion

 Dental Caries

Dental Caries is the localized destruction of the hard tissues of the tooth
 What causes dental caries?

• Pre-microbiology era
– Dental caries is the death (decay) of a tissue

• Microbiology period era

– Dental caries is a microbe related disease
W.D. Miller and his "chemico-parasitic" theory

SUSCEPTIBLE
SUSCEPTIBLE
HOST
HOST

PLAQUE
PLAQUE FERMENTABLE
FERMENTABLE
CARBOHYDRATE
CARBOHYDRATE

DEMINERALIZATION
DEMINERALIZATION
ACID
ACID
PRODUCTION
PRODUCTION
Plaque is a complex microbial community
How to identify bacteria within dental plaque?

• Culture methods
– take saliva or plaque, dilute and plate on
appropriate plates, grow to single colonies,
identify by microscopic and biochemical methods

• 16S DNA/RNA based detection

– use 2 oligo-nucleotide primers universal to ALL
bacteria 16S rDNA, PCR amplification of the total
saliva or plaque DNA pool, clone the PCR product
and sequence, phylogenetic analysis using computer
database
The supragingival plaque
The supragingival plaque
Supra-gingival plaque species as analyzed by 16s DNA sequencing
Current knowledge about bacteria in plaque
• Both culture and DNA/RNA-based techniques
are used for identification and quantification of
oral microorganisms
• Overall, there are ~700 species exist in the oral
cavity
• ~20% of these 700 species have been cultivated
• Both Gram-positive and Gram-negative exist
• Some archaea are found
• Most anaerobic or facultative anaerobic

Who are the bad guys?

The first isolation of cariogenic bacteria

Clark, 1924
***************

Isolation of cariogenic bacteria from caries lesions

Discovery of Mutans streptococci

 Keyes and Fitzgerald, 1962’s
Re-isolation of “Mutans streptococci”:
• Streptococcus mutans (human) (same
species Clark isolated in England in 1924)
• Streptococcus sobrinus (human)
• Streptococcus rattus (rats)
• Streptococcus cricetus
• Streptococcus ferus
• Streptococcus macacae
• Streptococcus downeii
Quantification of bacteria from healthy and carious sites by DNA analysis
 The “cariogenic bacteria” – bacteria
associated with dental caries

Mutans streptococci (S. mutans) – caries initiation

Actinomyces – early colonizers and root caries

• A. odontolyticus
• A. naeslundii genospecies 2
• A. isrealii
• A. gerensceriae

Lactobacilli (L. casei) – caries progression

 The virulence factors of
cariogenic bacteria

1. Acid production (acidogenicity)

• Lower the pH to below 5.5, the critical pH. Drives
the dissolution of calcium phosphate
(hydroxyapatite) of the tooth enamel
• Inhibit the growth of beneficial bacteria, promote the
growth of aciduric bacteria.
• Further lower the pH, promote progression of the
carious lesion
 The virulence factors of
cariogenic bacteria

2. Acid tolerance (aciduricity)

• Allows the cariogenic bacteria to thrive under acidic

conditions while other beneficial bacteria are
inhibited. This results in dominance of the plaque
by cariogenic bacteria
 The virulence factors of
cariogenic bacteria
3. Glucan formation
• Allows the cariogenic bacteria to stick onto the teeth
and form a biofilm

• Glucan mediated biofilms are more resistant to

mechanical removal

• Bacteria in these biofilms are more resistant to

antimicrobial treatments
 Dental Caries
Carbohydrates (Sucrose)

Cariogenic bacteria such as S. mutans

Glucans/Levans Acids
Plaque formation Demineralization
Dental caries is a bacterial infectious disease
 Transmission

• Mother – Child (vertical transmission) -

true for most oral bacteria
• Persons in close contact to the
baby
• Horizontal transfer (between
spouses) is rare, only observed in
some periodontal pathogens (i.e. P.
gingivalis)
• The most common vehicle is
saliva
New problem: everybody has S. mutans!

0.27
Fraction

0
1 10 20 50

Number of S. mutans in Saliva(104)

Why not every body who has S. mutans develop
dental caries?

• S. mutans is not present in high portions

• Acid produced is neutralized urea or
ammonia produced by other bacteria in
the plaque
• S. mutans is away from the tooth surface
so acid produced is diffused
The ecologic plaque hypothesis
Both pathogenic and commensal (non-
harmful) bacteria exist in a natural plaque. At
sound site, the pathogenic bacteria may exist
in low numbers to cause any clinical effect, or
they may exist in higher numbers, but the acid
produced is neutralized by the action of other
bacteria. Disease is a result of a shift in the
balance of the residence microflora driven by
a change in the local environment (frequent
sugar intake etc).
 The ecological development of Caries
dental caries
Demineralization
More sugar
Inhibition of beneficial
Acid producing bacteria
bacteria Lower pH Overgrowth of
Acids cariogenic bacteria

Sugar Neutral pH
Base producing
Bacteria: S. sanguis,
S. oralis
Remineralization

Health
 Human genome project

Oral bacteria genome projects

Metagenomics of oral cavity

 Complex genetic make-up

Human genome only has 200,000 genes.

Each oral bacterium has 2000-6000 genes,

with over 1 million bacterial genes in oral cavity
The dental plaque is a complex
multispecies biofilm
 Dental plaque formation
an ordered, complex process
Transmission Attachment & Growth of
Colonization Pioneer species
Mother to Child Pioneer colonizers:
S. oralis
S. mitis
S. salivarius Environmental
S. sanguis Modification
S. anginosus
pH, Eh
S. gordonii
Expose new receptors
Generate new nutrients

Microbial
Climax Increased species succession
community diversity G+, G-
(periodontal pathogens)
Oral bacteria have the ability to count the population size
Extensive chemical warfare between bacterial species

Competition/coexistence between S. mutans & S. sanguinis

Sm Ss Sm Ss Sm Ss
 Xylitol

Sugars
Cariogenic Tooth
Bacteria Decay
Antimicrobial agents Fluoride
Future dentistry (minimal invasive)

Diagnosis Treatment/prevention