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Plaque and Biofilm

Plaque and Biofilm

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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 .

Unattached plaque . tissue associated 2.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.

y Anaerobic environment. y Predominantly protein metabolism. y Bacteria are mostly Gram positive. . y Contains little or no matrix. y Aerobic environment. y Motile bacteria are common. margin. y Contains 50% of matrix. y Motile bacteria are few. y Bacteria are mostly Gram negative.DEFERENCE BETWEEN SUPRA & SUBGINGIVAL PLAQUE Supragingival y Found at or above the subgingival y Found below the gingival gingival margin. y Predominantly carbohydrate metabolism.

y SUPRA GINGIVAL PLAQUE: Tooth surface:Gram positive cocci Short rods. Outer surface:Gram negative rods &filaments Spirochetes. .

.naeslundii& Eubacterium sps y Tissue assosiated:Gram ve rods& cocci.intermedius.oralis. Peptostreptococcus.mitis.Porphyromonas Prevotella.S.S.sanguis.viscosus A. spirochetes S.A.Fusobacterium. filaments flagellated rods.y SUBGINGIVAL PLAQUE: y Tooth associated:Gram +ve rods &cocci S.

.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.

and fluoride . lipid material y Glycoproteins from saliva are an important component of the pellicle that initially coats a clean tooth surface.y Organic constituents = polysaccharides proteins glycoproteins. y Inorganic components = calcium and phosphorus sodium. potassium.

--Manganiello AD. Socransky SS .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 .

y Three major phases: 1) Formation of the pellicle 2) Initial adhesion& attachment of bacteria 3) Colonization & plaque maturation .

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

sedimentation of microorganisms .brownian motion. . This occurs through . -liquid flow or .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. .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. .

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

. (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.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.

nucleatum) .MICROSCOPIC STRUCTURE & PHYSIOLOGIC PROPERTIES OF DENTAL PLAQUE y Supragingival plaque : y Gram-positive cocci and short rods predominate at the tooth surface. y Corncob formations have been observed between rod-shaped bacterial cells (e. y Highly specific cell-to-cell interactions are also evident from the "corncob" structures" often observed. Bacterionema matruchotii or F. y Whereas gram-negative rods and filaments as well as spirochetes predominate in the outer surface of the mature plaque mass..g.

A.viscosus. y The apical border o f the plaque mass is separated from the junctional epithelium by a layer o f host leukocytes. sanguis. Actinomyces naeslundii. Eubacterium spp. .y Subgingival plaque: y The tooth-associated (attached) plaque is characterized by gram-positive rods and cocci. including Streptococcus mitis. S.

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

y Tooth associated plaque is similar to supragingival plaque.y The bacteria o f this apical tooth-associated region show an increased concentration of gram-negative rods. flagellated rods. . and spirochetes. y Tissue associated plaque is more loosely organized than the very dense tooth-associated region. y It contains primarily gram-negative rods and cocci. y Tissue associated plaque is covered with flagellated bacteria without a well defined extracellular matrix . as well as large numbers of filaments.

A central gram negative filamentous core supports the outer coccal cells. which are firmly attached by interbacterial adherence or coaggregation .y Long-standing supragingival plaque near the gingival margin demonstrates "corncob" arrangement.

Haffajee AD. et al: Microbial complexes in subgingival plaque. y The composition of the different complexes is based on the frequency with which microorganisms are recovered together. .* y *Socransky SS. Cugini MA. 25:134.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. J Clin Periodontol 1998.

A. viscosus) or members of the yellow (Streptococcus spp.y the early colonizers are either independent of defined complexes (A. naeslundii. y The green and orange complexes include species recognized as pathogens in periodontal and nonperiodontal infections.) or purple complexes (A. . orange or red complexes. odontolyticus). y secondary colonizers fell into the green.

J Clin Periodontol 1998. 25:134. Haffajee AD. which is an important clinical parameter of destructive periodontal diseases.y The red complex is of particular interest because it is associated with bleeding on probing. *Socransky SS. Cugini MA. et al: Microbial complexes in subgingival plaque.* y The existence of complexes of species in plaque is another reflection of bacterial interdependency in the biofilm environment. .

7/11/2011 Template copyright www.com 2005 32 .brainybetty.

. y 1.MICROBIAL SPECIFICITY OF PERIODONTAL DISEASES y Walter Loesche proposed two hypothesis in 1976. *Loesche WJ: Importance of nutrition in gingival crevice microbial ecology. 6:245.Non specific hypothesis y 2. Periodontics 1968.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*.

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. . Periodontics 1968.y According to this thinking. 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. 6:245. the noxious products are neutralized by the host. when only small amounts of plaque are present. y *Loesche WJ: Importance of nutrition in gingival crevice microbial ecology.

y The resistance of bacteria to antimicrobial agents is significantly increased in the biofilm environment. Costerton JW.limited diffusion of substances into the biofilm matrix. possibly to altered properties of bacteria in response to growth on a surface ---Costerton JW . y 2. ----Allison DG. Helmerhorst EJ y The resistance to antimicrobials inherent to biofilm bacteria may relate to y 1.the slow rate of cell growth in the biofilm environment.

invasion of tissues by bacteria c. a. role of bacterial specificity . release of toxic and inflammatory substances d.WHAT MAKES PLAQUE PATHOGENIC ? y The following are the possible pathogenic mechanisms by which the plaque microorganisms can cause periodontal disease. physical nature of plaque b.

cultivation. Dzink JL. Socransky SS.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. and/or detection of bacteria by DNA hybridization--. Loesche WJ.Colombo AP. Haffajee AD. .

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

-From non motile to motile organisms.MICROBIAL SHIFT DURING DISEASE y Comparing the microbiota in health. gingivitis . and periodontitis . -From fermentating to proteolytic species. 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 facultative anaerobes to obligate anaerobes. .

mitis.sangius. S. Gram-negative (44%) organisms are found. Predominant gram-positive species include. Capnocytophaga and Campylobacter species .CHRONIC GINGIVITIS y Gram-positive (56%). A.oralis.naeslundii. S. y Gram-negative organisms are: Fusobacterium nucleatum Prevotella intermedia Veillonella parvula as well as Hemophilus. S.viscosus. Peptostreptococcus micros. A.

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 .

.

. intermedia. 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. Porphyrornonas endodontalis and Streptococcus mutans are present in family members ---Petit MD. P. van Steenbergen TJ.TRANSMITION OF PERIODONTAL PLAQUE BACTERIA y Several transmission studies have demonstrated that the same clonal types of P gingivalis. Aa.

.

y Assessment of plaque can be done by using 1. PHP index y Plaque control can be done by 1.mechanical methods 2.plaque index 2.chemical methods .

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

non-shedding material (Wilderer & Charaklis 1989) .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.

algae. debris along with corrosion products Adhesion to surfaces. metabolites or genetic material from close proximity to other micro organisms. such as protection from anti-microbial agents. exchange of nutrients. fungi.y A single bacterial species can form a biofilm. y provides considerable advantage for the biofilm forming bacteria. . protazoa. y But in natural environment often biofilms are formed from various species of bacteria.

y Biofilm formation usually commences with the colonization of a surface by bacteria. .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.

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

b) Low surface energy materials : These are either low positively or low negatively charged hydrophobic materials such as plastics made up of organic polymers. . -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. . -pH available. .electrolyte concentration.y Factors influencing bacterial attachment to surfaces : -Nutrient availability at the surface.temperature. -nutrient concentration.

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

y temporary indwelling catheters.BIOFILMS IN MEDICAL SYSTEMS y Orthopedic implants (Screws. intra uterine devices. intra-venous catheters ports/caths and reverse osmosis membrane filters . and other prosthetic devices) : y Ocular lenses. pins. vascular grafts. heart values. plates.

Vol. clinical periodontology-9th & 10th edition y Jan Lindhe .REFERENCE y Carranza . clinical periodontology& Implantology y Max A . Socransky. the microbial challenge in periodontitis Periodontology 2000. Periodontal 2000. NET ANNER& ROYC . Anne D. The structure of dental plaque. 12-32 y Sigmund S. Dental biofilms: difficult therapeutic targets.28:2002 . Haffajee.5:1994 y RICHARD P . periodontal 2000. DARVEAAUN.Listgarten. 14. 1997. PAGE.

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