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ENDODONTIC BIOFILM
G.SPARSHA REDDY CONTENTS INTRODUCTION
HISTORY
DEFINITION
MICROBIOLOGY OF ROOT CANAL
ROUTES OF MICROBIAL ENTRY
MODES OF COLONIZATION
CHARACTERISTICS OF BIOFILM ECOLOGICAL
DETERMINANTS CLASSIFICATION OF BIOFILM
METHODS FOR MICROBIAL IDENTIFICATION
CURRENT THERAPEUTIC OPTIONS FOR ENDODONTIC BIOFILM
HISTORY
ANTONY VAN LEEUWENHOEK 1683
D.Miller- Father of Oral Microbiology described the clinical effects of gangrenous tooth pulps as centers of infection
In 1911 theory of focal
infection emerged by Dr.Hunter DEFINITION
Biofilm can be defined as a sessile multicellular microbial community
characterized by cells that are firmly attached to a surface and enmeshed
in a self- produced matrix of extracellular polymeric substance (EPS),
usually a polysaccharide-( Textbook of Endodontology Wiley
Blackwell Second Edition)
BASIC CRITERIA FOR BIOFILM
Caldwell et al. highlighted four characteristics of biofilm as follows:
Autopoiesis – Must possess the ability to self-organize
Homeostasis – Should resist environmental perturbations
Synergy – Must be more effective in association than in isolation
Communality – Should respond to environmental changes as a unit rather than as single
individuals . MICROBIOLOGY
• A fully developed biofilm is described as a heterogeneous
arrangement of microbial cells on a solid surface.
• It is composed of matrix material consisting of proteins,
polysaccharides, nucleic acids, and salt, which makes up 85% by volume, while 15% is made up of cells TEXTBOOK OF ENDODONTOLOGY-WILEY BLACKWELL COHENS PATHWAY OF PULP-11TH EDITION ROUTES
TEXTBOOK OF ENDODONTOLOGY-WILEY BLACKWELL
MODES OF COLONIZATION CHARACTERISTICS OF BIOFILM
Protection of biofilm bacteria from environmental
threats Enhanced tolerance to antimicrobials Quorum Sensing ECOLOGICAL DETERMINANTS CLASSIFICATION
2. Adhesion and colonization of planktonic microorganisms
3. Bacterial growth and biofilm expansion
4. Detachment of biofilm microorganisms into their surroundings.
Microbial Ecology and the Root Canal Ecosystem METHODS FOR MICROBIAL IDENTIFICATION SODIUM HYPOCHLORITE
1. Complete dissolution of cells with absence of visual evidence
2. Bacterial cells are disrupted and separated from the biofilm and are
nonviable
3. Bacterial cells remain adherent within the biofilm but are nonviable
4. Bacterial cells are disrupted and separated from the biofilm but are
viable
5. Bacterial cells remain adherent within the biofilm and are still viable.
ROOT CANAL IRRIGANTS-D.KANDASWAMY 2010—JOURNAL OF CONSERVATIVE DENTISTRY
ULTRASONICS TEMPERATURE ALTERING THE PH CHX • Spratt et al. have evaluated the effectiveness of 2.25% NaOCl, 0.2% CHX, 10% povidone iodine
against monoculture biofilms of P intermedia, P miros, S intermedius, F nucleatum, and E faecalis.
• They reported that NaOCl was the most effective antimicrobial agent, followed by the iodine solution.
Clegg et al. evaluated the ex vivo effectiveness against apical dentine biofilms of three concentrations
of NaOCl (6%, 3%, and 1%), 2% CHX, and Mixture of Tetracycline acid and detergents (MTAD).
• They reported that the 6% NaOCl and 3% NaOCl were capable of disrupting and removing the
biofilm, the 1% NaOCl and the MTAD were capable of disrupting the biofilm but did not eliminate
the bacteria, and the 2% CHX was not capable of disrupting the biofilm. TETRACLEAN • Tetraclean is a mixture of doxycycline hyclate (at a lower concentration than in MTAD), an acid, and a detergent.
• It is able to eliminate microorganisms and smear layer in dentinal tubules of infected root canals with a final 5-min rinse. Comparison of antimicrobial efficacy of 5.25% NaOCl,
MTAD, and Tetraclean® against E faecalis biofilm showed that only 5.25% NaOCl could
consistently disgregate and remove the biofilm at every time interval.
• However, treatment with Tetraclean®caused a high degree of biofilm disgregation in every
considered time interval (5, 30, and 60 min at 20°C) as compared with MTAD. Silver diamine fluoride
• A 3.8% w/v silver diamine fluoride (Ag[NH3]2F) solution has been developed for intracanal irrigation. This represents a 1:10 dilution of the original 38% Ag(NH 3)2 F solution used for
root canal infection.
• The study on the antibacterial effect of 3.8% Ag(NH3)2F against a E faecalis biofilm model concluded that Ag(NH3)2F has potential for use as an antimicrobial root canal irrigant or
interappointment medicament to reduce bacterial loads.
• E faecalis was completely killed by Ag(NH3)2F after exposure to these agents for 60 min. The silver deposits were found to occlude tubular orifices after removal of the smear layer. MTAD • MTAD is composed of three constituents that are expected to act synergistically against bacteria. The bactericidal effect of MTAD was inferior to 1%-6% NaOCl against E
faecalis biofilms.
• The antibacterial activity of MTAD might also be inhibited by the buffering effect of dentin and the serum albumin present in the root canal.
• .In the MTAD preparation, the citric acid may serve to remove the smear layer, allowing doxycycline to enter the dentinal tubules and exert an antibacterial effect.The recently
revised protocol for clinical use of MTAD advises an initial irrigation for 20 min with
1.3% NaOCl, followed by a 5-min final rinse with MTAD.
BIOFILMS IN ENDODONTICS 2017—INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES ADVANCED AGENTS
NANOPARTICLES BIOACTIVE GLASS OZONE LASER BIOACTIVE GLASS • Bioactive glass (BAG) consists of SiO2, Na2O, CaO2, and P2O5 at different concentrations. Stoor
et al. attributed the antibacterial mechanism of BAG to its high pH, osmotic effects and Ca/P
precipitation
• Zehnder et al. demonstrated that compared CH, BAG showed significantly less antibacterial
effects as an intracanal medicament. In addition, Gubler et al. showed that BAG did not
effectively prevent recontamination of instrumented root canals
• Incorporation of nanometric BAG fillers into polyisoprene (PI) and polycaprolactone (PCL) root
filling materials, rendered the resulting composite material bioactive and permitted improved
mineralization OZONE • Ozone (O3) is an energized, unstable gaseous form of oxygen that readily dissociates back into oxygen
(O2), liberating a reactive form of oxygen, aka the singlet oxygen (O 1). The singlet oxygen is capable of
oxidizing cells.
• Aqueous ozone (up to 20 mg/mL) showed essentially no toxicity to oral cells in vitro . Hems et al. showed
that ozone had an antibacterial effect on planktonic E. faecalis cells and those suspended in fluid, but little
effect on cells embedded in a biofilm structure .
• the antibacterial efficacy of ozone was not comparable with that of NaOCl. Huth et al. assessed the
antimicrobial efficacy of aqueous (1.25–20 mg/mL) and gaseous ozone (1–53 g/m 3) as an alternative
antiseptic against endodontic pathogens in suspension and in a biofilm model
NANOPARTICLES
• Kishen et al. indicated that treatment of root dentin with ZnO nanoparticles, ZnO–CS mixed nanoparticles, CS-layer- ZnO nanoparticles, or CS nanoparticles, produced an 80–95% reduction in the adherence of E. faecalis to dentin .
• root dentin treated with CHX and then with nanoparticles, shows the maximum reduction (97%) in bacterial adherence.
• Chitosan (CS) is a natural non-toxic biopolymer derived from the deacetylation of chitin. It binds to negatively charged surfaces and has excellent antimicrobial and antifungal activities. The exact mechanisms of the antibacterial
action of CS and its derivatives have still not been elucidated.
• Kishen et al. examined the antimicrobial properties of ZnO and resin-based root canal sealers loaded with CS and ZnO nanoparticles—improves the direct and diffusible antibacterial effects in root canal sealers based on a direct
antibacterial assay and a membrane- restricted antibacterial assay, respectively.
PAD PAD is a unique combination of a photosensitizer solution and low-power laser light.
The photosensitizer, which is mostly colored, adheres to or gets absorbed by
microbial cells. The low-power laser will destruct the target area and inactivate the microbial invaders. The photosensitizer then binds to microbial cell walls or even enters the cells. Further, the Laser light activates the photosensitizer and creates a cascade of energy transfer and variable chemical reactions in which singlet oxygen and free radicals play an important role. MICROBUBBLE EMULSION Halford et al., were the first to employ a microbubble emulsion to enhance the effect of sonic and ultrasonic agitation of sodium hypochlorite.
the technique employs unstable gas-filled microbubbles that expand when exposed to ultrasonic waves. The dynamics thereby induced in the fluid would help in detaching surface adherent bacteria or biofilm destruction.
In addition, it may also generate reactive oxygen species to exhibit an antibacterial
effect. Microbubble emulsion in combination with ultrasonic agitation was shown to be superior than with sonic agitation. ENZYMATIC IRRIGATION Enzymatic irrigation was introduced by Niazi and coworkers, who evaluated the effectiveness of 1% trypsin and 1% proteinase K, with or without ultrasonic activation, on a multi-species biofilm.
Trypsin with ultrasonic activation was able to effectively kill both aerobic and anaerobic bacteria and has the capability of disrupting the biofilm.
Agents that interfere with the cell wall, such as D-amino acids, specifically D-leucine has been demonstrated to bring about efficient dispersal of Enterococcus fecalis biofilms. It has been suggested that the dispersal of biofilms by sub-toxic concentrations of this agent reduces the success of resistant organisms. CONCLUSION
The most common endodontic infection is caused by the surface-associated growth of
microorganisms.
It is important to apply the biofilm concept to endodontic microbiology to understand the
pathogenic potential of the root canal microbiota as well as to form the basis for new
approaches for disinfection. It is foremost to understand that how the biofilm formed by root
