DENTAL PLAQUE

CONTENTS
PART - I PART - II
1. Introduction 8. Dental plaque as a biofilm
2. Classification of soft deposits 9. Physiological properties
3. Definitions of dental plaque 10. Microbial specificity of
4. History of dental plaque periodontal disease
5. Classification of dental
plaque 11. Detection of dental plaque

6. Composition & Structure of 12. Conclusion

dental plaque 13. References
7. Formation of dental plaque

Page  2
 INTRODUCTION
 Human fetus is sterile.
 Colonization starts at birth.
 Within hours – facultative & aerobic bacteria.
 2nd day – anaerobic bacteria.
 Within 2 weeks – mature microbiota estd in gut.
 After weaning - 1014 microorganisms with 400 different type
of bacteria.
 There are 10 times more bacteria than human
cells.

Page  3
  Establishing microbiota - harmony with the host.
 Constant renewal - prevents the accumulation of
microorganisms.
 Teeth provide hard, non-shedding surfaces - accumulation &
metabolism of bacteria on hard oral surfaces is considered the
primary cause of dental caries, gingivitis, periodontitis and peri-
implant infections.
 In the oral cavity, the bacterial deposits have been termed dental
plaque or bacterial plaque.
 In 1 mm3 of dental plaque weighing approximately 1mg, approx
1011 bacteria are present. [Socransky et al ,1953]

Page  4
Classification of soft deposits
ACQUIRED
PELLICLE
A non-cellular thin film

An organized transparent deposit which is

DENTAL
primarily composed of bacteria and their
PLAQUE
products

Soft, whitish deposit with no specific

MATERIAL
architecture, which can be removed by
ALBA
water spray.

Retained food which is usually removed by

FOOD DEBRIS
saliva and oral muscular action.
Page  5
Schwartz et al 1969
 DEFINITIONS
Davies et al (1963) defined plaque as a soft concentrated mass containing
mainly of large variety of bacteria together with certain amount of cellular
debris which develops within a short time after tooth brushing.

According to Schwartz & Massler (1969) – Plaque is a dense microbial

Layer consisting of coherent mass of filamentous, rod like and coccoidal
microorganisms embedded in an inter microbial matrix which accumulates
on tooth surface.

Bowen W.H. (1976) defined dental plaque clinically as a structured,

resilient,yellow-grayish substance that adheres tenaciously to the
intraoral hard surfaces, including removable and fixed restorations.

Page  6
 According to WHO (1978) Plaque is a specific but highly variable
structural entity resulting from colonization and growth of
microorganisms on surfaces of teeth and consisting of numerous
microbial species and stains embedded in a extracellular matrix.

According to the GPT, 4th Edition An organized mass, consisting

mainly of microorganisms, that adheres to teeth, prostheses, and oral
surfaces and is found in the gingival crevice and periodontal pockets.
Other components include an organic, polysaccharide-protein matrix
consisting of bacterial by-products such as enzymes, food debris,
desquamated cells, and inorganic components such as calcium and
phosphate

According to Carranza, 11th Edition Dental plaque is defined

clinically as a structured, resilient yellow-grayish substance that
adheres tenaciously to the intraoral hard surfaces, including
Page  r7emovable and fixed restorations.
 HISTORY
 J Leon Williams (1897) – Described dental plaque

 GV Black (1899) – Coined term “gelatinous dental plaque”

 W D Miller (1902) - Bacterial plaque

 Wild (1941) - Shortened Black’s terminology to the term ‘Plaque’

 Waerhaug (1950) Described the importance of bacterial plaque in the

etiology of periodontal disease

 Loe et al (1965), Landmark study on plaque , saying that plaque is main

etiological agent in periodontal disease.
Page  8
 Schei (1959), Russel (1967) - Epidemiological studies-
Positive correlation between the amount of bacterial plaque and the severity
of gingivitis

 W.Loesche (1976) - Modern theories of specificity – “Specific plaque

hypothesis”

 Socransky 1979 - Modern Version of Specific Plaque Hypothesis

 Thelaide 1986 - Unified Theory

 PD Marsh & Martin (1999) - Ecological plaque hypothesis

 Costerton (1999) - Evolved Biofilm

 Hajishengallis et al 2012 - Keystone Pathogenic Hpothesis

Page  9
 CLASSIFICATION
I. GRANTS CLASSIFICATION- ACCORDING TO LOCATION

A. Coronal plaque- Coronal to the gingival margin

B. Gingival plaque- forms on the external surface of the oral
epithelium and attached gingiva

C. Sub gingival plaque- located between the periodontal

attachment and the gingival margin, within the sulcus or
pocket.

D. Fissure plaque- develops in pits and fissures

E. Peri-implant plaque.
Page  10
 II. GLICKMAN’S CLASSIFICATION- ACCORDING TO
LOCATION

Page  11 11
 SUPRAGINGIVAL PLAQUE SUBGINGIVAL PLAQUE

Page  12
 TOOTH UNATTACHED TISSUE
ATTACHED ATTACHED

Gram +ve, Gram negative Gram negative

Few Gram –ve rods, filaments, rods, filaments,
rods and cocci, spirochetes spirochetes

Does not Extend to JE Extend to JE

extend to JE

Calculus Gingivitis Gingivitis,

formation, root periodontitis
caries
- May penetrate
May penetrate epithelium and
cementum connective
Page  13 tissue 13
  Pavel Godoroja and Olga Dulghieru 2004
The dental plaque is differentiated into two categories by
Supra-gingival
plaque Sub-gingival plaque

Gingival third of the Tooth adherent

crown of the tooth zone

Inter-proximal areas Epithelial adherent

zone

Pits and fissures and also on

other such surface with
Page  14 irregularities. Non adherent zone
 Composition of dental plaque
PLAQUE

SOLIDS 20-30% WATER 70-80%

Micro-
• Bacterial
organisms • Non-bacterial
70%

Intercellular
• Organic material
Matrix • Inorganic materials
20%-30%
MICROORGANISM

INTERCELLULAR MATRIX
15
Page  15
 BACTERIAL PORTION
 70 to 80 % of total solid plaque volume.
 1 gm of plaque contains approximately 2 X 1011 bacteria.
(Socransky SS,1953), (Schroeder, De Boever-1970)
Bacteria Facultative Anaerobic

Gram +ve  Strep.mutans

 Strep.sanguis

 A.viscosus

Gram -ve  A.actinomycetemcomitans  P.Gingivalis

 Capnocytophypa sp.  F.nucleatum

 Ekinella corrodens  P.intermedia

 B.forsythus

 C.rectus

Spirochetes  T.denticola
Page  16
 NON BACTERIAL PORTION

YEAST
VIRUSES

NON
BACTERIAL

PROTOZOA MYCOPLASMA

Page  17
 INTERCELLULAR MATRIX

 Accounts for 20% to 30% of the plaque mass

 Organic and inorganic material..
 Derived from – Saliva , Gingival crevicular fluid and Bacterial products.

ORGANIC
CONTENT

CARBOHYDRATES PROTEINS LIPIDS

30% 30% 15%

Page  18
 CALCIUM

INORGANIC PHOSPHORUS
CONTENT

SODIUM
OTHER
POTASSIUM
MINERALS
FLOURIDE

Page  19
 MICROSCOPIC STRUCTURE
SUPRAGINGIVAL PLAQUE
 Typically demonstrates a stratified organization of the bacterial
morphotypes.
 Gram-positive cocci and short rods predominate at the tooth
surface
 Gram-negative rods and filaments ,spirochetes predominate in
the outer surface of the mature plaque mass.
 Supra gingival plaque can have a structured architecture
polymer containing channel or pores have been observed that
link the plaque/oral environment interface to the tooth surface (
Wood et al 2000,Auschillet al 2001,Zaura Arite et al 2001)

Page  20
 Thin section of supragingival plaque

GRAM POSITIVE BACTERIA IN

PALISADING ARRANGEMENT

Page  21
 SUBGINGIVAL PLAQUE
 Between sub gingival plaque and the tooth an electron dense
organic material is interposed , termed as cuticle.
 Gingival crevicular fluid, -contains many substances that the
bacteria may use as nutrients
 Host inflammatory cells and mediators have influence on the
establishment and growth of bacteria in this region.

DENTA PLAQUE UNDER

Page  22
X 400 MAGNIFICATION
 Thin section of plaque in a deep pocket

RODS

COCCI

FILAMENTS

Page  23
 DEVELOPMENT OF DENTAL
PLAQUE

The formation of the

pellicle on the tooth
surface
Initial adhesion and
attachment of
bacteria

Colonization and
plaque maturation

Page  24 12/2 /2011

 I. Formation of the pellicle
 Vigorous tooth brushing – nanoseconds – acquired pellicle .
 Acquired pellicle - a homogenous, membranous, acellular film that
covers the tooth surface and frequently form the interface between
the tooth ,the dental plaque and calculus. (Schluger)
 `A fully established pellicle - 30 min, within 24 hr- 0.8 µm in
diameter.
 Derived from components of saliva and crevicular fluid as well as
bacterial and host tissue cell products and food debris.

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Transmission electron micrograph (TEM) of the acquired pellicle on an enamel surface
 ULTRA STRUCTURE OF DENTALPELLICLE
 Salivary pellicle can be detected on clean enamel surfaces
within 1 minute.
 By 2 hours, the pellicle is essentially in equilibrium.
 Thickness - 30 - 100 nm
 2 hr pellicle: Granular structures which form globules, that
connect to the Hydroxyapatite surface via stalk like structures.
 24 hrs Later: Globular structures get covered up by fibrillar
particles : 500 - 900 nm thick
 36 hrs Later: The pellicle becomes smooth, globular
(Panacea for Periodontology: Basic Tissue, Etiology and Pathogenesis
By Dr. Priyam Mishra)

Page  26 12/2 /2011

  Studies shows ( 2 hours) enamel pellicle, its amino acids
composition differs from that of saliva, indicating that the
pellicle forms by selective adsorption of the environmental
macromolecules. (Scannapieo FA et al , “ saliva and dental pellicles’”
contemporary periodontics, 1990)

 Mechanism involved are:

 Electrostatic forces
 Van der waals
 Hydrophobic forces

Page  27 12/2 /2011

 CHEMICAL COMPOSITION OF ACQUIRED PELLICLE
(Mayhell & Butller 1976, Sonju 1975)
Amino acids - Pellicle contains more hydrophobic and less neutral
4.6% amino acids than whole saliva (ie more leucine,
alamine, tyrosine and sereine than saliva)

Hexosamines - Glucosamine - 18%, Galactosamine -18%

2.7%
Carbohydrates - Glucose - 20%, Galactose - 27%
14% Mannose - 9% Fructose - 18%

Salivary Molecules Mucins

Proline rich proteins - statherins
Cystatins, Amylases
Ductal & stromal products
Page  28 Lactoferrin & Lysozyme
 ATTACHEMENT
SIGNIFICANCE
CALCULUS: A OF PELLICLE
of calculus
attachment.

NIDUS FOR PROTECTIVE

BACTERIA: provide barrier
Plaque formation against acids thus
by adherence of may reduce dental
microorganisms. caries attack.

LUBRICATIONke
p surface moist
prevent drying.
Page  29
 II. Initial Adhesion and Attachment of
Bacteria

COLONIZATION OF
SURFACE & BIOFILM
FORMATION

ATTACHMENT

INITIALADHESION

TRANSPORT TO
SURFACE

Page  30
 PHASE I. Transport to the surface

Active bacterial RANDOM CONTACTS

movement OCCUR THROUGH:
(chemotactic
activity)

Brownian
Liquid flow motion ( 40
µm/hour)

Sedimentation of
organisms
Page  31 12/27/2011
PHASE II. INITIAL ADHESION
Colonization
of the
Attachment surface and
biofilm
formation
Initial
 Reversible adhesion of the adhesion

bacterium and the surface. Transport

to surface

 Initiated by interactions b/w bacterium and surface through long

range and short range forces, including
 Van der Waals attractive forces
 Electrostatic repulsive forces

Physical phase
Page  32
 DLVO theory

 Derjaguin, Landau, Verwey & Overbeek (DLVO) have postulated

that above a separation distance of 1nm, the summation of previous
two forces describes total range interaction also called as Total
Gibbs Energy (GTOT).
 The result of summation (GTOT= GA+GE) is function of a
separation distance between negatively charged particle and a
negatively charged surface in a medium ionic strength suspension
medium.

Page  33
  Three stages

1. Secondary minimum (reversible attraction)

2. Positive maximum (energy barrier)
3. Primary minimum (irreversible attraction)

Page  34
 PHASE III. Attachment

 A firm anchorage between bacterium and surface will be

established by specific interactions ( ionic, covalent, or hydrogen
bonding)
 This follows direct contact or bridging true extra cellular
filamentous appendages (with length up to 10nm).

Page  35 12/27/2011
  On a rough surface, bacteria are better protected against shear
forces so that a change from reversible to irreversible bonding
occurs more easily and more frequently.
 The bonding between bacteria and pellicle is mediated by
specific extracellular proteinaceous components (adhesions) of
the organism and complementary receptors (proteins,
glycoproteins, polysaccharides) on the surface (pellicle) and is
species specific.

Page  36
  Streptococci (mainly S. sanguis) – Primary colonizer - binds to
acidic proline-rich-proteins
 α-amylase Receptors in pellicle
 sialic acid.
 Actinomyces - Primary colonizers, eg A. viscosus possesses
fimbrae - adhesins - specifically bind to proline-rich proteins of
dental pellicle.
 A. viscosus - reognises cryptic segments [cryptiotopes] of proline
rich proteins, which are only available in adsorbed molecules.
( with lock &key mech.)
( Mergenhagen et al 1987)

Page  37
Selected Bacterial Adhesins & Target Substrates
ATTACHMENT SUBSTRATE BACTERIAL ADHESIN SUBSTRATE
SURFACE SPECIES RECEPTOR
Tooth Saliva coated A.Viscsus Proline rich
surfaces S.Mitis Fimbriae proteins.
F.Nucleatum Saliva treated
hydroxyapatite
Tissue Epithelial cells P.Gingivalis Fimbriae Galactosyl
A.Viscosus Fimbriae residues
Fibroblasts T.Denticola Surface protein Galactosyl /
PMN`S A.Viscosus Fimbriae Mannose
residues
A.Naeslundii
Fibrinogen/
Connective tissue P.Gingivalis Membrane protein fibronectin
P.intermedia
Pre existing S.Sanguis C.Ochracea Heat sensitive Rhamnose/
plaque mass A.Naeslundii protein fucose/ N-acetyl
A.Israelii neura acid
S.Sanguis P.Loescheii Fimbrial protein Galactosyl
residues
A.Israelii
P.Gingivalis F.Nucleatum Outer membrane
Page  38
protein
 III. Colonization and plaque
maturation
Colonization
of the
Attachment surface and
biofilm
formation
Initial
adhesion

Transport
to surface

Mainly by 2 COAGREGGATION. COADHESION

mechanisms

Page  39 12/2 /2011

 PRIMARY COLONIZERS

 They provide new binding sites for adhesion by other oral bacteria.
 The early colonizers (e.g., streptococci and Actinomyces species) use
oxygen and lower the reduction-oxidation potential of the
environment, which then favors the growth of anaerobic species.

SECONDARY COLONIZERS

 They do not initially colonize the clean tooth surface but adhere to
bacteria already in the plaque mass.
 Including Prevotella intermedia, Prevotella loescheii,
Capnocytophaga spp., Fusobacterium nucleatum, and
Porphyromonas gingivalis.

Page  40
12/27/2011
  Co-aggregation is the interaction
between planktonic micro-organisms
of a different strain or species

 Co-adhesion is the interaction

between a sessile, already adhering
organism and planktonic micro-
oganisms of a different strain or
species

Page  41
 Co- Aggregation

 It was described by Gibbsons & Nygaard

 Cell to cell recognition of a genetically distinct partner cell type.
 Occurs primarily through

1. Highly specific interaction of

protiens and carbohydrate
molecules located on the bacterial
cell surfaces.

2. Less specific interaction resulting

from hydrophobic electrostatic &
van der waals forces.

Page  42
  Well characterized interaction include the coaggregation of:

• Fusobacterium nucleatum S. sanguis,

• Prevotella loescheii A. viscosus
• Capnocytophaga ochraceus A. viscosus

• Streptococci show intrageneric co-aggregation  bind to

the nascent monolayer of already bound streptococci.

 Later stages – coaggregation between different Gram negative

species seen – F. nucleatum & P. gingivalis or T. denticola.

Page  43 43
 Coaggregation Bridges:
 Formed when the common partner bears two or more types of coaggregation
mediators.
 Mediators can be various types of receptor polysaccharides, or various types
of adhesins, or a mixture of the two.
 Bridging is usually considered to be a cooperative event that brings three or
more cell types into close proximity and fosters symbiotic relationships.
 Bridging can also be an antagonistic event which brings together organisms
that compete with each other for nutrient or other needs.

Page  44
  Thus most coaggregation among strains of different genera are
mediated by lectin-like adhesin & can be inhibited by lactose &
other glycosides.

Page  45
 • F.nucleatum is central to the mechanism - since this
organism can co aggregate with numerous other species.
• Examples
F.nucleatum:
 S.sanguis
 P. loescheii
 A.viscous
 Capnocytophaga
 P.gingivalis
 B.forsythus
 T.denticola
Page  46 12/2 /2011
Page  47
 COAGGREGATION COMPETITION:
Competition occurs when multiple cell types recognize the same
coaggregation mediator on the common coaggregation partner.

Model depicting competition for binding sites

on Streptococcus oralis .

Page  48
  Corncob formation:
 Feature of plaque present on teeth
associated with gingivitis .
 Rod-shaped bacterial cells eg.
Bacterionema matruchotii or
Actinomyces sp. that forms inner core
of the structure and coccal cells eg.
Streptococci or P. gingivalis that attach
Described by Gibbsons and Nygaard
along the surface of the rod shaped
cells.

 Test tube brush:

 Composed of a central axis of a filamentous
bacterium with perpendicularly associated
short filaments.
 Commonly seen in the subgingival plaque of
teeth associated with periodontitis
 Detected between filaments of bacteria to
Page  49 which gram –ve rods adhere.
E V.parvula
CLOSELY ASSOCIATED
A A.odontolyticus COMPLEXES IN THE ORAL
R S.mitis CAVITY
L
S.oralis
Y
S.sanguis C.rectus
C P.intermedia
O Streptococcus sps P.nigrescens P.gingivalis
P.micros E.nodatum
L S.gorondi,
S.intermedius F.nucleatum T.forsythus
O T.denticola
N C.showae
I
Z
E E.corrodens
Capnocytophaga sps
R A.actinomycetocomitans LATE COLONIZERS
S

Page  50 12/2 /2011

Socransky et al (1998)
 Distribution of different complexes in subgingival plaque
sample
Kigure et al (1995)
Page  51
 CO-ADHESION

• Some bacteria are unable to bind directly to the conditioning

film, but are able to interact with molecules on bacteria that
are already attached (co-adhesion), also by adhesin-receptor
interactions.
• One bacterium, Fusobacterium nucleatum, can co-adhere with
almost all other bacteria found in dental plaque, and is
considered to be a key bridging organism between early and
later colonisers.

Page  52
 Development of dental plaque on a clean enamel surface. Coccal bacteria attach to the enamel pellicle as pioneer species
(A) and multiply to form microcolonies (B), eventually resulting in confluent growth (biofilm formation) embedded in a
matrix of extracellular polymers of bacterial and salivary origin (C). With time, the diversity of the microflora increases, and
rod and filament-shaped bacteria colonize (D and E). In the climax community, many unusual associations between
different bacterial populations can be seen, including ‘corn-cob’ formations (F). (Magnification approx. ×1150)
Page  53
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