Documente Academic
Documente Profesional
Documente Cultură
antimicrobial
photodynamic therapy
Dr Saji George
• Antibiofilm concepts
Biofilm?
Fluid Phase
Protective Matrix
Micro Organisms
Micro
Colonies
Fluid Channels
Solid Substratum
Rivers
Pipelines
Food processing appliances
Ship hull
Biofilm- definition, structure, occurrence, formation and
characteristics
Stages of Biofilm Formation
Phenotypic EPS
variation. production,
Altered Gene
Coaggregation and
expressions.
Coadhesion of planktonic
cells
Planktonic cells
Disperse Modification
bacterial by Mineral
cells to the accumulation
bulk fluid
Mature biofilm with new bacterial cells emerging
Biofilm- definition, structure, occurrence, formation and
characteristics
(2)sequestration to a
nutrient-rich area
(colonization)
(3) utilization of
cooperative benefits
(community)
Share many
common features
Community of interacting
organism
Elevated tower like
structures
Voids acting as water
channel-primitive circulatory
system
Transport of materials
Signalling
Biofilm- throughoccurrence, formation and
definition, structure,
Biofilm- definition, structure, occurrence, formation and
characteristics
Microbial Interactions
Coaggregatio
n and
Nutrient
Coadhession
exchange and
Facilitate
Biofilm
sharing Physical
Metaboli gene
Communicatio
c transfer
n
Determine
Spatial
relationship
Tolerance to
extremes of pH,
salinity, Nutrients Genetic
and Antibiotics
Increased
rate of Specific gene
Gene transfer
activation
transformatio
across MOs
n
Biofilm- definition, structure, occurrence, formation and
characteristics
Nutrient sharing
Eubacteria,
Peptostreptococci,
F.nucleatum
Fo
e s Hy rm
tid dr ate
p og o
Pe en r
Campylobacter rectus
P.intermedia, & P. gingivalis Hemin
Biofilm- definition, structure, occurrence, formation and
characteristics
Heterogeneity in biofilm
The status of nutrients,
oxygen,and pH vary in
different parts of biofilm
The heterogeneity of
various physicochemical
factors facilitate the
coexistance of microbes
with varying physiological
Biofilm- definition, structure, occurrence, formation and
characteristics
Quorum sensing - cell-to-cell
signaling
Gram-Negative
signaling transfer
molecules - acyl
homoserine lactone
(AHL)
Biofilm- definition, structure, occurrence, formation and
characteristics
Biofilm matrix
Extracellular polymeric
substances (EPS)
Carbohydrates, Proteins,
Lipids, Nucleic acids
Functions
Trap nutrients- micro
nutrients
Protection from
antimicrobial agents
Prevent from being washed
away
Protection from water loss
Your Body Is a Planet: 90% of the cells
within us are not ours but microbes'.
(Josie Glausiusz)
Rober Koch –
First isolated pure culture of bacteria…and
corelated to disease
“Until the late seventies, no one even knew biofilms existed. Scientists
thought most of the bacterial world was made up of free-floating
bacteria. They developed antibiotics and vaccines using bacteria
floating in a test tube. In many cases, the medicines just didn't work —
prostatitis, middle ear infections in children and periodontal disease to
name a few. As it turns out, scientists were targeting the wrong kind of
bacteria”.
Heat
Quaternary ammonium compounds
Iodine, Chlorine etc
Resistance
Constituents of Matrix
Enzymes present in
Matrix
Fight against
biofilm- antibiofilm
strategies
Antibiofilm concepts
Anti-biofilm coatings
Use of Furanones- ( Bavega et al, Givskov et al., J Bacteriol.
1996; 178:6618-6622).
Surface modification
Modify the solid surfaces to prevent bacterial
adhesion (antibacterial nano-particles)
Replacement therapy
Replace potential pathogenic micro-organisms with
genetically modified organisms that are less virulent
Immunization
The aim is to inhibit adhesion or reduce the virulence
of putative microbial etiologic agents.
Antibiofilm concepts
cc c c
c c
PS Biomolecules
PS
O**
PS**
PS PS O2 Biomolecules
Secondary radicals
production from the LPS of Dahl et al. Journal Of Bacteriology, Apr. 1989, p.
2188-2194
Antimicrobial photodynamic therapy, definition
and principle
Type of bacteria and
Susceptibility to PDT
Polysaccharides
Cell membrane
MIX
Bacteria internalized by
THP1
100% elimination of
internalized bacteria at the
cost of 70% THP1 cells with
damaged cell membrane!!!
(unpublished data)
PDT for eradicating biofilm- model studies
and result
Antimicrobial Photodynamic
Therapy for root canal disinfection
Concept of APDT in Root Canal
Disinfection
1 wee k
10
Antimicrobial 4 wee ks
log numebr of bacteria
treatment in four 8
canal biofilm is 4
less effective than
2
1 week old biofilm
0
Modified
Control Conventional ANILAD RCT
PDT
Lim et al. Communicated. PDT
PDT for eradicating biofilm- model studies
and result
The ultrastructure of matured root canal
biofilm –limited accessibility to the biofilm
core
Confocal microscopy
Biofilm
Dentine
Fluid phase
Photosensitiz
er
Bacteria
mediated
dentine
dissolution
Dentine
surface
Scanning Microscopy of
biofilm subjected to PDT.
C D
PDT using a
photosensitizing
formulation containing
a matrix disrupting E F
Discussion
PDT can break down the matrix components of biofilm
(Wainwright et al., 2002).
Summary
Summary
2. Eco-friendly treatment of
toxic effluents Biofilm on clay
based substrate
for waste water
treatment
3. Biofilm technology in
solid state fermentation-
production of therapeutic
Acknowledgement
Faculty of Dentistry
Funding from the National University of Singapore
ARF, Grant No. FY 04 R-224-000-021-101
Dr Anil Kishen
n k
A/P Neo Chiew Lian, Jennifer
a
Dr Song Keang Peng (Monasch University
h
T ou
Malaysia)
Dr Raj Kumar (University of Florida)
Mrs Shibi George
Dr Sum Chee Peng
Mr Zhang Xu
Y
Dr Annie Shrestha
Biofilm is formed on diverse materials
t i
s
Treatment of biofilm mediated human infections
e
is a huge clinical task and newer therapeutic
strategies are being screened for preventing
u s?
and eradicating biofilm bacteria
Q n
Photodynamic therapy involving the use of
o
photosensitizer and light has been tried as an
effective antimicrobial therapeutic strategy