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Guided By : Presented By :
Dr. Hiral Parikh Dr. Priya Patel
Dr. Charu Agrawal
Dr. Shilpa Duseja
Dr. Mishal Shah
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 CONTENTS
Introduction
Classification of soft deposits
Definitions of dental plaque
History of dental plaque
Classification of dental plaque
Composition & Structure of dental plaque
Formation of dental plaque
Dental plaque as a biofilm
Physiological properties
Microbial specificity of periodontal disease
Detection of dental plaque
Conclusion
Page 3 References
 INTRODUCTION
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]

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

An organized transparent deposit which is

DENTAL
primarily composed of bacteria and their
PLAQUE
products

Soft, whitish deposit with no specific

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

Retained food which is usually removed by

FOOD DEBRIS
saliva and oral muscular action.
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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.

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 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 removable
7 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 Blacks terminology to the term Plaque

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

etiology of periodontal disease

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 Loe et al (1965), Landmark study on plaque , saying that plaque is main
etiological agent in periodontal disease.

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 Hypothesis

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 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.
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 II. GLICKMANS CLASSIFICATION- ACCORDING TO
LOCATION

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 SUPRAGINGIVAL PLAQUE SUBGINGIVAL PLAQUE

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 TOOTH ATTACHED UNATTACHED TISSUE ATTACHED

Gram +ve, Gram negative rods, Gram negative rods,

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

Does not extend to JE Extend to JE Extend to JE

Calculus formation, root Gingivitis Gingivitis, periodontitis

caries

- May penetrate
May penetrate cementum epithelium and
connective tissue

Page 13 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
Page 15
 PLAQUE

SOLIDS 20-30% WATER 70-80%

Micro-
Bacterial
organisms Non-bacterial
70%

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

INTERCELLULAR MATRIX
16
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 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
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 NON BACTERIAL PORTION

YEAST
VIRUSES

NON
BACTERIAL

PROTOZOA MYCOPLASMA

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 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%

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 CALCIUM

INORGANIC PHOSPHORUS
CONTENT

SODIUM
OTHER POTASSIUM
MINERALS FLOURIDE

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 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)

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 Thin section of supragingival plaque

GRAM POSITIVE BACTERIA IN

PALISADING ARRANGEMENT

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 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

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X 400 MAGNIFICATION
 Thin section of plaque in a deep pocket

RODS

COCCI

FILAMENTS

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 DEVELOPMENT OF DENTAL
PLAQUE

The formation of the

pellicle on the tooth
surface
Initial adhesion and
attachment of
bacteria

Colonization and
plaque maturation

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 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 m 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 DENTAL PELLICLE
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)

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 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

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 CHEMICAL COMPOSITION OF ACQUIRED PELLICLE
(Mayhell & Butller 1976, Sonju 1975)
Amino acids - Pellicle contains more hydrophobic and
4.6% less neutral 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 Mucins
Molecules
Proline rich proteins - statherins
Cystatins, Amylases
Ductal & stromal products
Page 29 Lactoferrin & Lysozyme
 ATTACHEMENT
SIGNIFICANCE
OF CALCULUS: OF PELLICLE
A mode of calculus
attachment.

NIDUS FOR PROTECTIVE

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

LUBRICATION
keep surface moist
prevent drying.
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 II. Initial Adhesion and
Attachment of Bacteria

COLONIZATION OF
SURFACE & BIOFILM
FORMATION

ATTACHMENT

INITIAL ADHESION

TRANSPORT TO
SURFACE

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 PHASE I. Transport to the surface

RANDOM
Active bacterial CONTACTS OCCUR
movement THROUGH:
(chemotactic
activity)

Brownian
Liquid flow motion ( 40
m/hour)

Sedimentation of
organisms
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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
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 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.

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 Three stages

1. Secondary minimum (reversible attraction)

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

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 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).

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 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.

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 Streptococci (mainly S. sanguis) Primary colonizer - binds to
acidic proline-rich-proteins
-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 - recognises cryptic segments [cryptiotopes] of
proline rich proteins, which are only available in adsorbed
molecules. ( with lock &key mech.)
( Mergenhagen et al 1987)

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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 39
protein
 III. Colonization and plaque
maturation
Colonization
of the
Attachment surface and
biofilm
formation
Initial
adhesion

Transport
to surface

Mainly by 2
COAGGREGATION COADHESION
mechanisms

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 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.

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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 42
 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 43
 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 44 44
 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.

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

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Page 47
 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
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 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 49
 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 50 which gram ve rods adhere.
E Actinomyces sps V.parvula
CLOSELY ASSOCIATED
A A.odontolyticus COMPLEXES IN THE ORAL
R CAVITY
L
Y
S.mitis C.rectus
C S.oralis P.intermedia
O S.sanguis P.nigrescens P.gingivalis
P.micros E.nodatum
L F.nucleatum T.forsythus
O Streptococcus sps
S.gorondi, T.denticola
N S.intermedius C.showae
I
Z
E E.corrodens
R Capnocytophaga sps LATE COLONIZERS
S A.actinomycetocomitans

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Socransky et al (1998)
 Distribution of different complexes in subgingival plaque
sample
Kigure et al (1995)
Page 52
 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.

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 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 54
 Matrix embedded microbial populations, adherent to each other
and/or to surfaces or interfaces (Costerton et al. 1999)
Biofilms consist of one or more communities of microorganisms,
embedded in a glycocalyx, that are attached to a solid surface.
(Sigmund S. Socransky & Anne D. Haffajee. 2001)

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 A microbial biofilm is considered a community that meets the
following four basic criteria:

AUTOPOIESIS HOMEOSTASIS

SYNERGY COMMUNALITY

Page 56
 BENEFITS OF MICROBIAL COMMUNIY LIFESTYLE
Schematic representation of the types of interaction that
occur in a microbial community, such as dental plaque

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 PROPERTIES OF BIOFILM
ATTACHMENT
OF BACTERIA

MICROBIAL PHYSIOLOGICAL
INTERACTIONS HETEROGENICITY

QUORUM
STRUCTURE OF SENSING
BIOFILM

GENE TRANSFER ANTIBIOTIC

RESISTANCE
Page 58
 1. STRUCTURE OF A BIOFILM
Biofilms are composed of microcolonies of bacterial cells (1520%
by volume) that are non-randomly distributed in a matrix or
glycocalyx (7580% volume).

The bacterial vitality varies

throughout the biofilm,
with the most viable
bacteria present in the
central part of plaque , and
lining the voids and
channels (Auschill et al
2001)

Page 59
 Structure of the Biofilm depends on environmental parameters
under which they are formed. These include:
Surface and interface properties
Nutrient availability
Composition of the microbial community
Hydrodynamics

The biofilm matrix is

penetrated by fluid
channels that conduct the
flow of nutrients, waste
products, enzymes,
metabolites, and oxygen.

Page 60
 The bacteria in a biofilm use a communication system termed
quorum sensing that involves sending out chemical signals .
These chemical signals trigger the bacteria to produce potentially
harmful proteins and enzymes, virulence factors that help the
intraoral biofilm bypass host defense systems

Page 61
EXOPOLYSACCHARIDES the backbone of the biofilm
The bulk of the biofilm consists of the matrix or glycocalyx and
Maintaining the
is composed predominantly of integrity
water andof the biofilm
aqueous solutes
The dry material is a mixture of exopolysaccharides, proteins,
salts, andPreventing
cell material.
desiccation and attack by harmful agents.
Exopolysaccharides (EPS), which are produced by the bacteria in
the biofilm, are the major components of the biofilm making up
Binds essential nutrients to create a local nutritionally rich
5095% of the dry weight. environment.

Acts as a buffer

Assists in retention of extracellular enzymes enhancing substrate

utilization by bacterial cells
Page 62
 2. ATTACHMENT OF BACTERIA

The key characteristic of a biofilm - the microcolonies within the

biofilm attach to a solid surface.
Many bacterial species possess surface structures such as fimbriae
and fibrils that aid in their attachment to different surfaces.
Fimbriae have been detected on a number of oral species including
P. gingivalis, A. actinomycetemcomitans and some strains of
streptococci.
Oral species that possess fibrils include S. salivarius, the S. mitis
group, Pr. intermedia, Pr. nigrescens, and Streptococcus mutans.
Sigmund S. Socransky & Anne D. Haffajee. Dental Biofilms: Difficult Therapeutic
Targets Periodontology 2000 2001;28:1255.

Page 63
 VARIABLES IMPORTANT IN CELL ATTACHMENT
AND BIOFILM FORMATION

PROPERTIES OF PROPERTIES OF PROPERTIES OF

THE THE BULK THE CELL
SUBSTRATUM FLUID
Texture or Flow velocity Cell surface
roughness pH hydrophobicity
Hydrophobicity Temperature Fimbriae
Conditioning Cations Presence Flagella
Extracellular
polymeric
substances of
antimicrobial
agents
Page 64
 3. PHYSIOLOGICAL HETEROGENEITY
Cells of the same microbial species can exhibit extremely
different physiologic states in a biofilm even though separated
by as little as 10 m.

Studies to date indicate that

sessile cells growing in
mixed biofilms can exist in
an almost infinite range of
chemical and physical
microhabitats within
microbial communities.

Page 65
Clinical Periodontology and Implant Dentistry by Jan Lindhe, 5th Edition.
 4. MICROBIAL INTERACTIONS

The residents in the microbial community display extensive

interactions while forming biofilm structures, carrying out
physiological functions, and inducing microbial pathogenesis.
These interactions, include
1. Competition between bacteria for nutrients
2. Synergistic interactions which may stimulate the growth or survival of
one or more residents
3. Production of an antagonist by one resident which inhibits the growth of
another
4. Neutralization of a virulence factor produced by one organism by
another resident
5. Interference in the growth-dependent signaling mechanisms of one
organism by another
Page 66
 NUTRIENTS AS THE BASIS FOR BACTERIAL
INTERSPECIES INTERACTION WITHIN BIOFILM

Page 67
 GENERAL METABOLIC PRODUCTS WHICH
INFLUENCE BIOFILM RESIDENT INTERACTIONS
Antagonistic effect - S. sanguinis group are producers of H 2O2 , a
nonspecific antimicrobial agent - an antagonistic effect on other
coresidents, such as S. mutans.
Synergistic effect - lactic acid produced by S. mutans can be readily
metabolized by members of the Veillonella family.
Co-operative metabolic interactions -
F. nucleatum & P. intermedia Using glutamic
Generate
grow at pH range of 5.0 to and aspartic
Ammonia +
7.0. P. gingivalis susceptible acids (GCF &
organic acids.
to - pH levels < 6.5. saliva)

prevents in pH even in Acid-sensitive species - P.

This contributes
presence of lactic acid gingivalis are protected
to a more
bacteria & fermentable against acid attack
neutral pH
Page 68 carbohydrates
 BACTERIOCINS
Proteinaceous bactericidal substances produced by bacteria to
inhibit the growth of closely related bacterial species or strains
(Hojo et al 2009)
Regulated by genetic and environmental factors
Enable bacteria to select their neighbours, promote the
establishment of a community with specific bacterial species.
Inhibition of growth of P.gingivalis, T.forsythia, S.salivarius,
S.sanguinis by bacteriocin produced by L.paracaesi
Nigrescin, produced by P.nigrescens display bactericidal effect
against P.gingivalis, P.intermedia, T.forsythia, Actinomyces spp.
Bacteriocin production also reported by P.intermedia, A.a,
C.ochracea, F.nucleatum, E.corrodens, H.influenzae
Page 69
 Bacterial co-aggregation influences localization within
biofilms

Gibbons and coworkers(1970), demonstrated that strains of P.

gingivalis aggregated with several oral streptococci, this might
be an important mechanism - P. gingivalis could become
incorporated into dental plaque that is composed primarily of
the initial streptococcal colonizers.
T. denticola does not appear to form biofilms on most inert
surfaces, in contrast to P. gingivalis, which does so readily.
However, in the presence of P. gingivalis , T. denticola is
incorporated into the biofilm.
T. forsythia is a weak colonizer of inert surfaces but will
become incorporated into biofilms in the presence of F.
nucleatum.
Page 70
 5. QUORUM SENSING
It is defined as the cell density dependent regulation of gene
expression in response to soluble signals called autoinducers (Bassler
1999)
It has been defined by Miller (2001) as the regulation of gene
expression in response to fluctuations in cell population density.
Quorum sensing can occur within a single bacterial species as well
as b/w diverse species.

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 Quorum sensing has been described in both G+ve & G-ve bacteria.
Cell-cell communication may occur b/w and within bacterial
species (Miller, 2001)
Quorum sensing-controlled behaviors are those that only occur
when bacteria are at high cell population densities.

Page 72
 QUORUM SENSING MOLECULES

Three types of molecules :

1. Acyl-homoserine lactones (AHLs) - signaling molecules
used by many G-ve bacteria, it synthesized by Lux-I family
protiens.
2. Autoinducer peptides (AIPs) - signaling molecules used by
G +ve bacteria
3. Autoinducer-2 (AI-2) - used by both G-ve & G+ve
bacteria, chemically it is furanosyl borate diester. Synthsized
by protein Lux-S.

Schauder, S. and B. L. Bassler (2001). "The languages of

bacteria."
Page 73
Page 74
 This communication controls various functions reflecting the
needs of a specific bacterial species to inhabit a particular niche
such as the production of virulence factors, or by the transmission
and acquisition of the generic information needed to produce
virulence factors from other species in the biofilm development
(Passador et al., 1993; Reading et al., 2006).

Several strains of P. intermedia, T. forsythia, F. nucleatum and P.

gingivalis were found to produce quorum sensing signal
molecules (Frias et al., 2001; Sharma et al., 2005).

Page 75
Page 76
 Strategies for quorum sensing inhibition
3 strategies can be applied

Anti- activator
Targeting signal
Interference
Targeting AHL signal
Bacterial
Targeting the signal
proteins
generation of signal
dissemination components
receptor
AHL receptor Transgenic
Signal precursor Signal precursor Signal precursor
degradation
X Furanones plants
enzymes L-canavanine Synthetic
RNASignal
dependant Signal
Human analogues
Signal
regulation X
hormones
X
Signal receptor Signal receptor Signal receptor
77

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 6. ANTIBIOTIC RESISTANCE

Bacteria growing in a biofilm are

highly resistant to antibiotics, up
to 1,000-1,500 times more
resistant than the same bacteria
not growing in a biofilm.

MIC of chlorhexidine and amine fluoride was 300 and 75

times greater respectively, when S.sobrinus was grown in
biofilm compared to planktonic cells
Biofilms of P.gingivalis tolerated 160 times the MIC of
metronidazole than planktonic cells
Page 78
Page 79
 Bacteria replicate only slowly in an established biofilm
The
and,biofilm matrix mayare
as a consequence, restrict the penetration
inherently of a
less susceptible
charged antimicrobial .
agent (diffusion-reaction theory)
than faster dividing cells.

In addition,
Agent samples
may also bind of
to, gingival crevicular
and inhibit, fluid at
the organisms
(GCF)
the can of
surface contain sufficient
the biofilm, lactamase
leaving cells in to
the depths
inactivate the concentrations of
of the biofilm relatively unaffected. antibiotic delivered
to the site

The novel phenotype

A susceptible pathogenexpressed in a biofilm
can be rendered may result
resistant if
in the drug target
neighbouring, being modified
non-pathogenic or not
cells expressed,
produce a or the
organism may
neutralising oruse alternative metabolic
drug-degrading enzyme. strategies

Page 80
 7. EXCHANGE OF GENETIC
INFORMATION

Conjugation,
transformation and
transduction have all
been shown to occur in
naturally occurring mixed
species biofilms. Clinical
Periodontology and Implant
Dentistry by Jan Lindhe, 5th
Edition.

Page 81
 Cells also communicate and interact with one another in biofilms
via horizontal gene transfer.
Gene transfer between Treponema denticola and S. gordonii has
also been demonstrated in the laboratory. Wang BY, Chi B,
Kuramitsu HK. Genetic exchange between Treponema denticola
and Streptococcus gordonii in biofilms. Oral Microbiol Immunol
2002: 17: 108 112.
The presence of pathogenicity islands in periodontal pathogens
such as P. gingivalis is also indirect evidence for horizontal gene
transfer having occurred in plaque biofilms at some distant time in
the past, and may explain the evolution of more virulent strains.
Chen T, Hosogi Y, Nishikawa K, Abbey K, Fleischmann RD,
Walling J, Duncan MJ. Comparative whole-genome analysis of
virulent and avirulent strains of Porphyromonas gingivalis. J
Bacteriol 2004: 186: 54735479.
Page 82
Detachment
Can be Movement of Individual cells or Biofilm en masse
Brading et al have emphasized the importance of physical
forces in detachment, stating that the three main processes for
detachment are (JADA 1996)
erosion or shearing (continuous removal of small portions of
the biofilm)
sloughing (rapid and massive
removal), and
abrasion (detachment due to
collision of particles from the
bulk fluid with the biofilm)
Page 83
 Individual Cell Transfer En masse transfer

Erosion - detachment of single cells in a This possibility has been

continuous predictable fashion demonstrated in vitro studies of
mixed biofilm that showed
Sloughing - sporadic detachment of movement of intact biofilm
large groups of cells or structures across solid surfaces
Intermediate process whereby large while remaining attached to them.
pieces of biofilm are shed from the Advantage - formation of the
biofilm in a predictable manner, resulting biofilm is not reliant on planktonic
in detached clusters consisting of about cells, which are known to be more
104 cells. susceptible to antimicrobial agents

Stoodley 1991

Page 84
 Factors affecting biofilm
development and behavior
1. ROLE OF SALIVA
Saliva contains mixture of glycoproteins mucin.
Bacteria enzymes (glycosidases) split off carb. utilized
as nutrients.
Remaining protein contributes to plaque matrix
Neuraminidase separates sialic acid from salivary
glycoprotein.
Loss of sialic acid - salivary viscosity
- Formation of precipitate factor in
plaque formation
Page 85
 2. ROLE OF INGESTED NUTRIENTS
Most readily utilized nutrients diffuse easily into plaque sucrose,
glucose, fructose, maltose & less amt. of lactose.
Dextran greater quantity, adhesive properties , relative insolubility &
resistance to destruction by bactera.
Levan Used as carbohydrate nutrient by plaque bacteria in absence of
exogenous sources.
3. DIET AND PLAQUE FORMATION
Consistency affects the rate of plaque formation : Forms rapidly on soft
diets, hard chewy food retard it.
Dietary supplements of sucrose plaque formation and affect its bacterial
composition. i.e ECM
Plaque formation occurs on high protein fat diets and carbohydrate - free
diets but in smaller amounts.

Page 86
 IMPORTANCE OF FOOD CHAINS
Stimulation of growth of other bacteria (eg) stimulation of
growth of T.denticola by butyric acid produced by P.gingivalis
Increasing the virulence of organisms (eg) more virulent strains
of P.gingivalis in the presence of S.gordonii.
Removal of toxic metabolites (eg) protection from hydrogen
peroxide by A.neaslundii
Utilization of metabolic products for maintaining structural
integrity (eg) succinic acid produced by T.denticola integrated
onto the cell wall of P.gingivalis

Page 87
 MICROBIAL SPECIFICITY

Modern
Non Ecological Keystone
Specific Version of
Specific Unified Plaque Pathogenic
Plaque Specific
Plaque Theory Hypothesis Hpothesis
Hypothesis Plaque
Hypothesis Thelaide PD Marsh Hajishenga
Loesche Hypothesis
Early 1986 & Martin llis et al
1976 Socransky
1930s 1999 2012
1979

Page 88
 NON SPECIFIC PLAQUE HYPOTHESIS
Non specific plaque hypothesis was proposed by WALTER LOESCHE(1976).
The nonspecific plaque hypothesis maintains that periodontal disease results
from the elaboration of noxious products by the entire plaque flora.
Thus it lead to concept that control of periodontal disease depends on
control of the amount of plaque accumulation.

SPECIFIC PLAQUE HYPOTHESIS

Specific plaque hypothesis states that only certain plaque is pathogenic, and
its pathogenicity depends on the presence of or increase in specific
microorganisms. NEWMAN MG , SOCRANSKY SS (1977),ADI et al
,(1978)
Plaque harboring specific bacterial pathogens results in periodontal disease.
A. actinomycetemcomitans as a pathogen in localized aggressive periodontitis.

Page 89
 ECOLOGICAL PLAQUE HYPOTHESIS
A change in a key environmental factor (or factors) will trigger a shift in
the balance of the resident plaque microflora, and this might predispose a
site to disease. ( PD Marsh 1994)

This hypothesis is based on the theory that the unique local

microenvironment influences the composition of the oral microflora.

This hypothesis postulated dynamic relationship between environmental

cause & ecological shifts within the biofilm.

It also introduced the concept that the disease can be prevented not only
by inhibiting the putative pathogens, but also interfering with the
environmental factors driving the selection & enrichment of these
bacteria.

Page 90
 Low GCF Gram +ve Gingival health
Gingival health flow bacteria
Reduced Reduced
plaque inflammation
Environmental Ecological
Stress change shift

Increased High GCF, Gram -ve

Increased
plaque bleeding, anaerobes
inflammation
raised pH &
temperature,
Gingivitis low Eh Periodontitis

A schematic representation of the ecological plaque hypothesis in relation to

periodontal disease
Plaque biofilm accumulation can produce an inflammatory host response; this causes changes in the local
environmental conditions and introduces novel host proteins and glycoproteins that favour the growth of
proteolytic and anaerobic G ve bacteria. In order to prevent or control disease, the underlying factors
responsible for driving the selection of the putative pathogens must be addressed, otherwise disease will
Page 91
recur.
 THEORY DRAWBACK
Non-Specific Plaque Individuals with longstanding plaque and gingivitis do not
Hypothesis(NSPH), develop periodontitis, while others, with minimal plaque, had
lower resistance to disease.

The Specific Plaque Many of the organisms observed in periodontal health were also
Hypothesis(SPH), observed at diseased sites (Slots, 1977)

The Ecological Plaque Does not address the role of genetic factors of the host that
hypothesis (EPH) contribute to the composition of dental plaque and to
susceptibility to disease

The Keystone Pathogen P.Gingivalis is one of the easily culturable micro organisms in
Hypothesis(KPH). plaque. Over 700 bacterial speices are found in dental plaque.
So it can be that any one of the uncultured micro-organisms can
also create conditions ideal to the growth of periodontal
pathogens.

Page 92
 MICROBIAL
SHIFT/DYSBIOSIS

This model proposes that

Concept that some
diseases are due to a
decrease in the number of
beneficial symbionts and different microbial members
or an increase in the
number of pathogens.
periodontitis is initiated by
a dysbiotic microbial
community (rather than by
select periodontal
pathogens) within which
or specific gene
combinations have distinct
roles that synergize to
shape a microbiota that
causes disease.

Page 93
 MICROBIAL SHIFT LEADING TO PERIODONTITIS

GRAM +VE AEROBES GRAM -VE ANAEROBES

Gradually changes the symbiotic hostmicrobe relationship to a

pathogenic one.

Prevotella intermedia P. Gingivalis

Fusobacterium Tannerella forsythia

nucleatum
Treponema denticola

Page 94
 Biofilm and host in conflict
Oral microflora has a harmonious and +vely beneficial
relationship with the host - microbial homeostasis.
Introduction of complex
Inflammatory
Plaque host molecules
response by GCF flow
Accumulation (transferrin, Hb) into
Host
GCF

Used as a nutrient This proteolytic local ph

These get source by the metabolism
proteolytic G ve in the redox
catabolized leads to potential,
Bac

Promotes Favours growth at the Re-arrangement in

upregulation of expense of beneficial community structure &
virulence factors species(i.e. the selective in the
(e.g. Gingipain If sustained proportions of the
competitiveness of
activity by P. the potential anaerobic & proteolytic
Gingivalis) pathogens) components

Page 95
 POSSIBLE STRATEGIES TO
CONTROL ORAL BIOFILM
1. Inhibiting Adherence with Antagonists
2. Passive Immunization
3. Replacement Therapy
4. Regulating the Levels of Nonpathogenic Bacteria to Influence
Virulence
5. Probiotic Approaches
6. Interference with Signaling Mechanisms
7. Targeted Antimicrobial Therapy via a Novel STAMP
Technology

Page 96
 CONTROL Addition of base generating nutrients (arginine)
OF Reduction of GCF flow through anti-
NUTRIENTS inflammatory agents
Inhibition of key microbial enzymes

CONTROL Sugar substitutes

OF BIOFILM Antimicrobial agents
pH Fluorides
Stimulate base production

CONTROL Redox agents

OF REDOX eg: methylene blue
POTENTIAL Oxygenating agents

Page 97
 METHODS OF DETECTION
OF DENTAL PLAQUE

VISUAL

DISCLOSING PERIODONTAL
AGENTS PROBE OR
EXPLORER

Page 98
 1. DIRECT VISION : -
Thin plaque may be translucent & therefore not visible
Stained plaque may be acquired e.g- tobacco stained
Thick plaque tooth may appear dull & dirty

2. USE OF EXPLORER : -
Tactile Examination when calcification has started it appears slightly
rough, otherwise it may feel slippery due to coating of soft , slimy
plaque
Removal Of Plaque when no plaque is visible , an explorer can be
passed over the tooth surface & when plaque is present it will adhere to
explorer tip.this technique is used when evaluating plaque index.
This can be done by running the explorer or probe along the gingival 3
rd of the tooth

Page 99
 3. Disclosing Agents:
1) Two tone
2) Erythrosine
3) Bismark
4) Benders
5) Basic Fuschin
6) Disclosing Tablets Dental Mart, oral B

Page 100
 CONCLUSION
Dental plaque biofilm cannot be eliminated permanently.
However, the pathogenic nature of the dental plaque biofilm
can be reduced by reducing the bioburden (total microbial load
and different pathogenic isolates within that dental plaque
biofilm) and maintaining a normal flora with appropriate oral
hygiene methods that include daily brushing,flossing and
rinsing with antimicrobial mouthrinses.
This can result in the prevention or management of the
associated sequelae, including the development of periodontal
diseases and possibly the impact of periodontal diseases on
specific systemic disorders.

Page 101
 REFERENCES
Newman, Takei, Klokkevold, Carranza; Clinical Periodontology;
10th Edition: Elseveir
Jan Lindhe, Niklaus P. Lang, Thorkildkarring; Clinical
Periodontology And Implant Dentistry: 5th Edition: Blackwell
Socransky SS, Haffajee AD. Dental biofilms: difficult therapeutic
targets. Peridntol 2000 2002 : 28; 12-55.
Marsh PD, Moter A, Devine DA. Dental plaque biofilms:
commuinties, conflict and control. Periodontol 2000 2011; 55: 16-
35
Max A. Listgarten , The structure of dental plaque, Periodontology
2000, Vol. 5, 1994, 52-65

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 Every Ending is really just
A NEW BEGINNING..

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