PRESENTED BY : DATE: 8/7/11

G.SRINIVAS 2ND YEAR POST GRADUATE

CONTENTS
y Introduction y Definition

-Dental plaque: -Biofilm:

y Types and Composition of Dental Plaque

y

Formation of Dental Plaque

y Coaggregation y Microscopic Structure and Physiologic Properties

of Dental Plaque

y Significance of the plaque Environment y Microbial specificity of periodontal diseases y What makes plaque pathogenic? y Microorganisms Associated with Specific

Periodontal Diseases: -Periodontal health -Periodontal diseases y Transmission of periodontal plaque bacteria y References

INTRODUCTION
y Dental plaque is a general term for the diverse microbial

community (predominantly bacteria)

y Plaque develops naturally on teeth, and forms part of the

defence systems of the host by helping to prevent colonisation of enamel by exogenous (and often pathogenic) microorganisms (colonisation resistance).
y Plaque is an example of a biofilm y Plaque is found preferentially at protected and stagnant

surfaces, and these are at the greatest risk of disease.

DEFINITION

Dental plaque:
y Is defined clinically as a structured, resilient, yellow-grayish

substance that adheres tenaciously to the intraoral hard surfaces including removable and fixed restorations.*

*Bowen WH: Nature of plaque. Oral sci Rev 1976;9:3

y Because of their unique structure, biofilms are

notoriously resistant to -surfactants -antibiotics -opsonization and -Complement mediated phagocytosis and killing

TYPES & COMPOSITION OF DENTAL PLAQUE

y Types of dental plaque: y Based on its relationship to the gingival margin y Dental plaque is broadly classified as y Supragingival plaque -coronal plaque

-marginal plaque
y Subgingival plaque= 1.Attached plaque

a. tooth associated b. tissue associated 2.Unattached plaque

DEFERENCE BETWEEN SUPRA & SUBGINGIVAL PLAQUE

Supragingival
y Found at or above the

subgingival
y Found below the gingival

gingival margin. y Contains 50% of matrix. y Bacteria are mostly Gram positive. y Motile bacteria are few. y Aerobic environment. y Predominantly carbohydrate metabolism.

margin. y Contains little or no matrix. y Bacteria are mostly Gram negative. y Motile bacteria are common. y Anaerobic environment. y Predominantly protein metabolism.

y SUPRA GINGIVAL PLAQUE:

Tooth surface:Gram positive cocci Short rods. Outer surface:Gram negative rods &filaments Spirochetes.

y SUBGINGIVAL PLAQUE: y Tooth associated:Gram +ve rods &cocci

S.mitis,S.sanguis,A.viscosus A.naeslundii& Eubacterium sps

y Tissue assosiated:Gram ve rods& cocci, filaments

flagellated rods, spirochetes S.oralis,S.intermedius, Peptostreptococcus,Porphyromonas Prevotella,Fusobacterium.

y Diagram depicting the plaque-bacteria association with

tooth surface and periodontal tissues.

y Morphologic studies indicate a differentiation of tooth

associated and tissue-associated regions of subgingival plaque Listgarten MA

y in certain cases bacteria are found within the host

tissues. -Saglie FR, Carranza FA

COMPOSITION
y Dental plaque is composed primarily of

microorganisms---70-80% bacteria
y One gram of plaque (wet weight) contains

approximately 2 x 10 11 bacteria--------Socransky SS
y The intercellular matrix, estimated to account for 20%

to 30% of the plaque mass

y consists of organic and inorganic materials derived

from saliva, gingival crevicular fluid, and bacterial products.

y Organic constituents = polysaccharides

proteins glycoproteins, lipid material y Glycoproteins from saliva are an important component of the pellicle that initially coats a clean tooth surface.

y Inorganic components = calcium and phosphorus

sodium, potassium, and fluoride

FORMATION OF DENTAL PLAQUE

y The location and rate of plaque formation vary among

individuals

y determining factors include = oral hygiene as well as

host factors such as diet or salivary composition and flow rate . --Manganiello AD, Socransky SS

y Three major phases:

1) Formation of the pellicle 2) Initial adhesion& attachment of bacteria 3) Colonization & plaque maturation

FORMATION OF THE PELLICLE

y Within nanoseconds after a vigorously polishing the teeth a

thin saliva derived layer called the acquired pellicle covers the tooth surface.
y Pellicle consists of glycoproteins

proline rich proteins phospho proteins histidine rich proteins enzymes y Formation of pellicle involves electrostatic, vanderwaals and hydrophobic forces.

INITIAL ADHESION &ATTACHMENT OF BACTERIA

This includes 4 phases
y Phase 1:Transport to surface

This stage involves the initial transport of bacterium to the tooth surface. This occurs through - brownian motion, - sedimentation of microorganisms , -liquid flow or - active bacterial movement.

y Phase 2:Initial adhesion:

This stage results in initial reversible adhesion of the bacterium to the tooth surface. This occurs through vander waals attractive forces & electrostative repulsive forces.
y Phase 3:Attachment:

A firm anchorage between bacterium and surface will be established by specific interactions(covalent,ionic or hydrogen bonding )
y Phase 4:Colonization of the surface and biofilm

formation:

3.COLONIZATION & PLAQUE MATURATION

Firmly attached microorganisms start growing and newly formed bacterial clusters remain attached resulting in a biofilm

COAGGREGATION

y Cell-to-cell recognition of genetically distinct partner

cell types

Kolenbrander PE

y Interactions of secondary colonizers with primary

colonizers Primary colonizers: Steptococci,Actinomycetes

y Cumulative evidence suggests that such adhesion

influences the development of complex multi-species biofilms.

Secondary colonizers: P. intermedia, Capnocytophaga, F. nucleatum, Porphyromonas

y Examples of coaggregation:

Corncob formation: Steptococci adhere to filaments of Bacterionema matruchotii or Actinomyces

y Testube brush: Gram ve rods adhere to filamentus bacteria

y Experimental gingivitis model (Loe et al. 1965) (a) Human

volunteer with clean teeth and clinically healthy gingival tissues at the start of the period of experimental plaque accumulation. (b) Same human volunteer after 21 days of abolished oral hygiene practices leading to plaque deposits covering almost all tooth surfaces and consequently developing a generalized marginal gingival inflammation.

MICROSCOPIC STRUCTURE & PHYSIOLOGIC PROPERTIES OF DENTAL PLAQUE

y Supragingival plaque : y Gram-positive cocci and short rods predominate at the tooth

surface,
y Whereas gram-negative rods and filaments as well as

spirochetes predominate in the outer surface of the mature plaque mass.

y Highly specific cell-to-cell interactions are also evident from

the "corncob" structures" often observed. y Corncob formations have been observed between rod-shaped bacterial cells (e.g., Bacterionema matruchotii or F. nucleatum)

y Subgingival plaque: y The tooth-associated (attached) plaque is characterized by

gram-positive rods and cocci, including Streptococcus mitis, S. sanguis, A.viscosus, Actinomyces naeslundii, Eubacterium spp.
y The apical border o f the plaque mass is separated from the

junctional epithelium by a layer o f host leukocytes.

y Left Diagrammatic representation of the histologic

structure of subgingival plaque. Right, Histologic section of subgingival plaque. Arrow with box, Sulcular epithelium. White arrow, Predominantly gram-negative unattached zone. Black arrow, Tooth surface. Asterisk, Predominantly gram-positive attached zone.(Listgarten

y The bacteria o f this apical tooth-associated region show an

increased concentration of gram-negative rods.

y Tissue associated plaque is more loosely organized than the

very dense tooth-associated region.

y It contains primarily gram-negative rods and cocci, as well as

large numbers of filaments, flagellated rods, and spirochetes.

y Tooth associated plaque is similar to supragingival plaque. y Tissue associated plaque is covered with flagellated bacteria

without a well defined extracellular matrix .

y Long-standing supragingival plaque near the gingival

margin demonstrates "corncob" arrangement. A central gram negative filamentous core supports the outer coccal cells, which are firmly attached by interbacterial adherence or coaggregation

MICROBIAL COMPLEXES
y Recent analyses of more than 13,000 plaque samples of 40

subgingival microorganisms using a DNA-hybridization methodology was used to define "complexes" of periodontal microorganisms.
y The composition of the different complexes is based on the

frequency with which microorganisms are recovered together.*

y *Socransky SS, Haffajee AD, Cugini MA, et al: Microbial

complexes in subgingival plaque. J Clin Periodontol 1998; 25:134.

y the early colonizers are either independent of defined

complexes (A. naeslundii, A. viscosus) or members of the yellow (Streptococcus spp.) or purple complexes (A. odontolyticus).
y secondary colonizers fell into the green, orange or red

complexes.
y The green and orange complexes include species recognized

as pathogens in periodontal and nonperiodontal infections.

y The red complex is of particular interest because it is

associated with bleeding on probing, which is an important clinical parameter of destructive periodontal diseases.*
y The existence of complexes of species in plaque is another

reflection of bacterial interdependency in the biofilm environment. *Socransky SS, Haffajee AD, Cugini MA, et al: Microbial complexes in subgingival plaque. J Clin Periodontol 1998; 25:134.

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MICROBIAL SPECIFICITY OF PERIODONTAL DISEASES

y Walter Loesche proposed two hypothesis in 1976. y 1.Non specific hypothesis y 2.Specific hypothesis

Nonspecific Plaque Hypothesis (NPH):

y The NPH maintains that periodontal disease results from the

"elaboration of noxious products by the entire plaque flora*. *Loesche WJ: Importance of nutrition in gingival crevice microbial ecology. Periodontics 1968; 6:245.

y According to this thinking, when only small amounts of

plaque are present, the noxious products are neutralized by the host.
y NPH is the concept that control of periodontal disease

depends on control of the amount of plaque accumulation.

y Specific Plaque Hypothesis:(SP) y The SP states that only certain plaque is pathogenic, and its

pathogenicity depends on the presence of or increase in specific microorganisms *. y This concept predicts that plaque harboring specific bacterial pathogens results in periodontal disease. y *Loesche WJ: Importance of nutrition in gingival crevice microbial ecology. Periodontics 1968; 6:245.

y The resistance of bacteria to antimicrobial agents is

significantly increased in the biofilm environment. ----Allison DG, Costerton JW, Helmerhorst EJ
y The resistance to antimicrobials inherent to biofilm bacteria

may relate to
y 1.limited diffusion of substances into the biofilm matrix,

y 2.the slow rate of cell growth in the biofilm environment,

possibly to altered properties of bacteria in response to growth on a surface ---Costerton JW

WHAT MAKES PLAQUE PATHOGENIC ?

y The following are the possible pathogenic mechanisms

by which the plaque microorganisms can cause periodontal disease. a. physical nature of plaque b. invasion of tissues by bacteria c. release of toxic and inflammatory substances d. role of bacterial specificity

MICROORGANISMS ASSOCIATED WITH SPECIPHIC PERIODONTAL DISEASES

y The microbiota associated with periodontal health and

disease has been studied with a wide variety of techniques for sampling, cultivation, and/or detection of bacteria by DNA hybridization--- Colombo AP, Haffajee AD, Dzink JL, Socransky SS, Loesche WJ.

PERIODONTAL HEALTH

y Actinomyces (viscosus and naeslundii) y Streptococcus (S. mitis and S. sangius) y Veillonella parvula y Small amounts of Gram-negative species are also

found.

MICROBIAL SHIFT DURING DISEASE

y Comparing the microbiota in health, gingivitis , and

periodontitis , the following microbial shifts can be identified: -From G+ve to G-ve -From cocci to rods (and at a later stage to spirochetes). -From non motile to motile organisms. -From facultative anaerobes to obligate anaerobes. -From fermentating to proteolytic species.

CHRONIC GINGIVITIS
y Gram-positive (56%), Gram-negative (44%) organisms

are found. Predominant gram-positive species include, S.sangius, S.mitis, S.oralis, A.viscosus, A.naeslundii, Peptostreptococcus micros.
y Gram-negative organisms are:

Fusobacterium nucleatum Prevotella intermedia Veillonella parvula as well as Hemophilus, Capnocytophaga and Campylobacter species

Pregnancy-associated gingivitis -Prevotella intermedia Acute necrotizing ulcerative gingivitis -Spirochetes -Prevotella intermedia

ADULT PERIODONTITIS
y Porphyromonas gingivalis y Bacteroides forsythus y Prevotella intermedia y Campylobacter rectus y Eikenella corrodens y Fusobacterium nucleatum y Actinobacillus actinomycetemcomitans y Peptostreptococcus micros y Treponema, and y Eubacterium species.

Viruses such as: y EBV-1 (Ebstein-Barr virus) y HCMV (Human cytomegalovirus) Localized juvenile periodontitis y Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis y Eikenella corrodens y Campylobacter rectus y Fusobacterium nucleatum y Bacteroides y Eubacterium y Capnocytophaga y Herpes virus.

Generalized juvenile periodontitis y Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis y Prevotella intermedia y Capnocytophaga y Eikenella corrodens y Neisseria Refractory periodontitis y Actinobacillus actinomycetemcomitans y Bacteroides Forsythus y Porphyromonas gingivalis y Prevotella intermedia y Wolinella recta

Abscesses of the periodontium

y Fusobacterium nucleatum y Prevotella intermedia y Peptostreptococcus micros y Bacteroides forsythus y Porphyromonas gingivalis

TRANSMITION OF PERIODONTAL PLAQUE BACTERIA

y Several transmission studies have demonstrated that the

same clonal types of P gingivalis, Aa, P. intermedia, Porphyrornonas endodontalis and Streptococcus mutans are present in family members ---Petit MD, van Steenbergen TJ, Scholte LM
y But different clonal types are found in unrelated individuals y This suggests that transmission most likely occurs through

intimate contact
y It would be interesting to determine what clonal type was

present in spouses before they were married.

y Assessment of plaque can be done by using 1.plaque index

2. PHP index

y Plaque control can be done by

1.mechanical methods 2.chemical methods

BIOFILM
Introduction:
y Commonly encountered as a layer on slime clogging

drainpipes y biofilms appear in our everyday life in more than one form. y Bacteria suspended in water or aqueous cultures are "Planktonic . y In such aqueous environments when bacteria adhere to surfaces and become "Sessile", secreting a slimy-glue like substance for anchorage, they form a biofilm* * Costerton JW, Lewandowski Z, Caldwell DE, Korber DR, Lappin -Scott HM. Microbial biofilms. Ann Rev Microbiol 1995; 49:711-45.

Definition:
y Matrix ---enclosed bacterial populations adherent to each

other and or/to surface or interfaces

Costerton.

y The term biofilm describes the relatively undefinable

microbial community associated with a tooth surface or any other hard, non-shedding material (Wilderer & Charaklis 1989)

y A single bacterial species can form a biofilm, y But in natural environment often biofilms are formed from

various species of bacteria, fungi, algae, protazoa, debris along with corrosion products Adhesion to surfaces.

y provides considerable advantage for the biofilm forming

bacteria, such as protection from anti-microbial agents, exchange of nutrients, metabolites or genetic material from close proximity to other micro organisms.

FORMATION OF BIOFILM
y Pioneering studies by Cholodny, Henrici and Zo Bell

commenced more than 50-60 years ago.

y Usually the methodology involved was unmersion of glass

slides into natural environments and observing the biofilms developed under microscope.
y Biofilm formation usually commences with the colonization

of a surface by bacteria.

y The adhesion and attraction of the bacteria to the surface

may be brought about by different mechanisms. -surface charge, -gravity, -Brownian motion and - chemo attraction, y After attraction, attachment of the bacteria to the surface occurs by a two-step process comprised of reversible binding* y 1. The reversible binding is usually brought about by weak Vander Waal forces y 2. stronger attachment can form by a combination of both physical or chemical forces. y Jenkinson HF, Lappin-Scott HM. Biofilms adhere to stay. Trends Microbiol. 2001; 9(1):9-10.

y Factors influencing bacterial attachment to surfaces :

-Nutrient availability at the surface, -nutrient concentration, -pH available, - temperature, - electrolyte concentration, -the flux of materials and surface types such as : a) High surface energy materials: These are negatively charged hydrophilic materials such as glass, metal or minerals. b) Low surface energy materials : These are either low positively or low negatively charged hydrophobic materials such as plastics made up of organic polymers.

y Limiting of a biofilm is brought about by the removal of

sessile cells or sections of the biofilm, as a result of erosion, sloughing or abrasion processes. l. Erosion leads to a continuous loss of cells from biofilm. Thickness of biofilm, fluid shear stress and fluid velocity all affect the rate of erosion. 2. Sloughing is observed in cases of bulky biofilms and involves a large and rapid loss of material from the biofilm. 3. Abrasion occurs when some object repeatedly collides with the biofilm.

BIOFILMS IN MEDICAL SYSTEMS

y Orthopedic implants (Screws, pins, plates, and other

prosthetic devices) : y Ocular lenses, heart values, vascular grafts, intra uterine devices, y temporary indwelling catheters, intra-venous catheters ports/caths and reverse osmosis membrane filters

REFERENCE
y Carranza ; clinical periodontology-9th & 10th edition y Jan Lindhe ; clinical periodontology& Implantology y Max A ,Listgarten; The structure of dental plaque.

Periodontal 2000;5:1994
y RICHARD P . DARVEAAUN, NET ANNER& ROYC . PAGE; the microbial challenge in periodontitis Periodontology 2000, Vol.

14, 1997, 12-32 y Sigmund S, Socransky, Anne D. Haffajee. Dental biofilms: difficult therapeutic targets; periodontal 2000;28:2002

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