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Microbiology



Dental  Caries
                 Dr. Ali Yaldrum
                 SEGi UC, KD
Learning  Outcomes
At  the  end  of  this  session  students  should  be  

!   Describe  rela4onship  between  plaque  and  oral  diseases
!   Describe  role  of  plaque  in  development  of  caries
!   Define  Dental  Caries
!   Describe  the  ae4ology  and  the  role  different  factors  
    play  in  ini4a4on  and  progression  of  the  disease
!   Describe  the  role  played  by  different  microorganisms
Relationship    of

Plaque bacteria   to   Caries
Plaque & Caries




Historically  for  any  microbe  to  be  considered  responsible  
  for  a  given  condi4on,  Koch s postulate was  applied
Koch s Postulate


1.  The  microbes  should  be  found  in  all  cases  of  the  disease  with  a  
    distribu4on  corresponding  to  the  observed  lesions
2.  The  microbes  should  be  grown  on  laboratory  media  for  several  
    subcultures
3.  A  pure  subculture  should  produce  the  disease  in  the  suscep4ble  
    animal
4.  A  high  an4body  4ter  to  the  microbe  should  be  detected  during  
    infec4on;  this  may  provide  protec4on  on  subsequent  reinfec4on
Koch s Postulate


1.  A  microbe  should  be  present  in  sufficient  numbers  to  ini4ate  
    disease
2.  The  microbe  should  generate  high  levels  of  specific  an4bodies
3.  The  microbe  should  produce  relevant  virulence  factors
4.  The  microbes  should  cause  disease  in  an  appropriate  animal  
    model
5.  Elimina4on  of    the  microbe  should  result  in  clinical  improvement
Role  of  Plaque




!   The   Specific  Plaque  Hypothesis (fig.1)
!   The   Non-­‐Specific  Plaque  Hypothesis   (fig.2)
!   The   Ecological  Plaque  Hypothesis (fig.3)
Specific Plaque Hypothesis

               Plaque



                     {   few species out of diverse resident
                         flora are actively involved in disease



                 X   problem
                               (   disease can still occur in absence of these
                                                   organisms
                                                                                 )
              Disease
fig.1
Non-Specific Plaque Hypothesis

                 Plaque



                     {  out come of the overall activity of
                        the total plaque microflora




                              (                                                    )
                                  plaque mediated diseases are polymicrobial but one
                    problem

                                  specific organism predominate

                Disease
fig.2
Ecological Plaque Hypothesis

                Plaque



                    {
                        organisms associated with disease
                        are already present on teeth but very
                        low to detect




fig.3
                Disease      (   disease only initiates as a result of shift in
                                 the balance of resident microf lora              )
Dental Caries
Dental Caries




!   It  is  bacterial  disease  of  calcified  4ssue  of  the  teeth  characterized  
    by   demineraliza4on   of   the   inorganic   and   destruc4on   of   the  
    organic  substance  of  the  tooth.

                 Dental Caries is known as disease of civilization
Dental Caries

The Early theories

!  The legend of Worms
!  Endogenous Theory
!  Chemical Theory
!  Parasitic Theory
!  Chemoparasitic Theory
Dental Caries




!  Chemoparasitic theory
Miller  in  1890  suggested  that  oral  bacteria  converted  dietary  
carbohydrates  into  acid  which  solubilized  the  calcium  phosphate  of  
enamel  to  produce  a  caries  lesion
Aetiology of Dental Caries
Saliva
                                                Caries
                    Susceptible Surface

                                     P laque




                                                     Saliva
        Saliva




                 Time                Bacteria
                         Sugar


                            Saliva

fig.4            aetiology of caries
Role of Saliva
Role of Saliva


Saliva   plays   a   vital   role   in   the   maintenance   of   oral   health  
and  the  integrity  of  oral  4ssues
Saliva  contains:
      • exfoliated  epithelial  cells
      • oral  microorganisms
      • food  residues  (carbohydrates)
      • an4microbial  factors
      • minerals
Role of Saliva


!   Mechanical   washing   ac4on   of   saliva   is   very   effec4ve   for  
    cleaning  of  teeth
!   high  buffering  capacity  neutralizes  acid
!   supersaturated   with   Calcium   and   phosphorus   helps   in  
    remineraliza4on

      Salivary  flow  rate:  0.8  mm  /min  on  upper  labial  surface
                                                            5.0  -­‐  8.0  mm/min  on  lower  lingual  surface
Susceptible Surface
Susceptible Surface


Some   areas   of   the   tooth   are   more   suscep4ble   to   developing   caries  
than  others,  known  as     susceptible areas 

Suscep4bility  is  related  to  several  factors
    !   Mineral  &  Fluoride  Content
    !   Structure  of  that  par4cular  area
Susceptible Surface


Susceptible tooth surfaces includes


  !   Pits  &  Fissures  (fig.5)
  !   Approximal  Enamel  cervical  to  contact  point  (fig.6)
  !   Exposed  root  surface  (fig.7 & 8)
  !   Margins  of  Restora4on  (fig.6  &  9)
  !   Cervical  margins  of  tooth  (fig.10)
pits & fissures caries




fig.5
Caries under restoration   Approximal Caries




fig.6
Root surface Caries




fig.7
Root Surface Caries




fig.8
Caries under restoration   Amalgam restoration




fig.9
white spot lesions




fig.9
From white spot to cavity formation




white spot lesion                                                   Cavitation


    earliest clinical evidence (reversible)       Later clinical stage (irreversible)
Time   &

Sugar Substrate
Sugar Substrate



!   Direct   rela4onship   between   intake   of   Carbohydrates  
    (sugar)  &  Caries
!   Glucose,  Fructose  &  Sucrose  are  cariogenic  
! Sucrose  is  the  most  cariogenic  sugar
     •   fermentable
     • easily  penetrates  plaque
Time & Sugar Substrate



There  is  a  direct  rela4onship  between  caries  and:
      !  Frequency  of  sucrose  consump4on
      !  s4ckiness  of  the  sucrose

Both   affects   the   amount   of   4me   sugar   stays   into   contact  
with  the  teeth  surface
Plaque Bacteria
Plaque Bacteria




Dental  Caries  does  not  occur   in vivo   if  microorganisms  in  
the  form  of  dental  plaque  are  absent;  making  it  clear  that  
dental  caries  is  a  plaque  mediated  disease
Plaque Bacteria




!   Streptococcus  mutans
!   Lactobacillus  species
!   Ac4nomyces  Species
!   Veillonella  species
Streptococcus species



The  term  mutans  streptococci  refers  to  a  group  of  7  
different  species,  but  two  species  are  most  commonly  
found  in  humans
!   S.  mutans  (serotypes  c/e/f)
!   S.  sobrinus  (serotypes  d/g)
Streptococcus species


Factors related to Cariogenicity of S. mutans
! S.mutans  counts  in  saliva  and  at  site  of  lesion
!   Isolated  from  site  before  the  development  of  caries
!   Efficient  sugar  metabolism  &  transport  capability
! Acidogenic  &  acidouric
!   Liele  effect  of  pH  on  growth  and  metabolism
!   Produc4on  of  EPS  &  IPS
Streptococcus species



S.  mutans  been  implicated  in  
!   Pits  &  fissure  caries  (strong  evidence)
!   Root  surface  caries
!   All  forms  of  caries
!   Some  strains  are  more  pathogenic  than  others
Streptococcus species



!   S.  Sobrinus  role  is  uncertain
!   Frequently  isolated  from  plaque  but  in  lesser  numbers
!   Shares  the  same  cariogenic  proper4es  as  S.mutans
Lactobacillus species



Lactobacilli  can  be  divided  into  two  groups
    !   Homofermenta4ve  species
    !   Heterofermenta4ve  species
Lactobacillus species

Homofermentative species
!   L.  acidophilus  &  L.  casei
!   Mainly  produce  lac4c  acid  from  glucose  fermenta4on

Heterofermentative species
!   L.  fermentum
!   Produce  lac4c  acid  and  significant  amounts  of  acetate
Lactobacillus species



Most  commonly  isolated  species  from  oral  cavity  are
!   L.  casei
!   L.  fermentum
!   L.  acidophilus
Lactobacillus species


Factors related to Cariogenicity of Lactobacillus
species
!   Increased  number  in  carious  cavi4es  affec4ng  enamel  &  
    root  surface
!   Number  in  saliva  reflect  caries  ac4vity
! Acidogenic  and  acidouric
!   Few  strains  can  produce  EPS
Lactobacillus species


!   Have  a  low  affinity  for  tooth  surface
!   Low  in  numbers  at  healthy  site
!   Implicated  in  progression  of  caries  deeper  into  den4ne  
    rather  than  ini4a4on
!   Implicated  in  ae4ology  of  root  surface  caries
Actinomyces species



Ac8nomyces  spp.  form  a  major  and  complex  part  of  the
oral  microflora,  par4cularly  at  the  approximal  site  and
gingival  crevice
Actinomyces species


! Ac8nomyces  odontoly8cus  been  associated  with    very  
  early  stages  of  tooth  demineraliza4on

! Ac8nomyces  naeslundii  been  associated  with  
  development  of  root  surface  caries
CariesVaccination
Caries Vaccination



In  rats  ,  experimental  dental  caries  have  been  reduced  by  
immunizing   with   cell-­‐wall   associated   an4gens   or  
glucosyltransferase  from  S.mutans
Caries Vaccination


The  following  mechanism  have  been  suggested

! Inhib4on  by  secretory  IgA  of  bacterial  coloniza4on  of  
    teeth
!   Interference  with  bacterial  metabolism
!   Enhancement  of  phagocy4c  ac4vity  in  gingival  crevices
Summary
!   Dental  Caries  is  a  mul4factorial  disease,  strongly  
    associated  with  the  dental  plaque
!   Ecological  Plaque  Hypothesis  caries  forms  as  a  result  of  
    disturbance  in  the  normal  balance  of  Oral  Biota
!   The  main  factors  involved  are  suscep4ble  tooth  surfaces,  
    4me,  sugar  substrate  and  plaque  bacteria
!   Pits  &  fissure  are  the  most  suscep4ble  areas
!   Frequency  of  sugar  intake  is  more  decisive  than  total  
    consump4on
Summary
!   Sucrose  is  the  most  cariogenic  sugar
! Mutans  streptococci  are  implicated  in  ini4a4on  of  pits  &  
    fissure  and  root  caries
!   Lactobacillus  spp  are  associated  with  progression  of  
    caries
References

!   Philip   D.   Marsh,   Michael   V   Mar4n,   Plaque   mediated  
    diseases-­‐  Dental  Caries  and  Periodontal  diseases   in  Oral  
    Microbiology,  5th  Edi4on,  Churchil  Livingstone,  2009,  pp  
    104-­‐117


!   J.   Bagg,   T.   W.   Macfarlane,   I.   R.   Poxton   and   A.   J.   Smith,  
     Dental   Caries    in   Essen4als   of   Microbiology   for   Dental  
    Students,   2nd   Edi4on,   Oxford   University   Press,   2006  
    pp238-­‐248

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Microbiology of Dental caries

  • 1. Microbiology Dental  Caries Dr. Ali Yaldrum SEGi UC, KD
  • 2.
  • 3. Learning  Outcomes At  the  end  of  this  session  students  should  be   !  Describe  rela4onship  between  plaque  and  oral  diseases !  Describe  role  of  plaque  in  development  of  caries !  Define  Dental  Caries !  Describe  the  ae4ology  and  the  role  different  factors   play  in  ini4a4on  and  progression  of  the  disease !   Describe  the  role  played  by  different  microorganisms
  • 4. Relationship of Plaque bacteria to Caries
  • 5. Plaque & Caries Historically  for  any  microbe  to  be  considered  responsible   for  a  given  condi4on,  Koch s postulate was  applied
  • 6. Koch s Postulate 1.  The  microbes  should  be  found  in  all  cases  of  the  disease  with  a   distribu4on  corresponding  to  the  observed  lesions 2.  The  microbes  should  be  grown  on  laboratory  media  for  several   subcultures 3.  A  pure  subculture  should  produce  the  disease  in  the  suscep4ble   animal 4.  A  high  an4body  4ter  to  the  microbe  should  be  detected  during   infec4on;  this  may  provide  protec4on  on  subsequent  reinfec4on
  • 7. Koch s Postulate 1.  A  microbe  should  be  present  in  sufficient  numbers  to  ini4ate   disease 2.  The  microbe  should  generate  high  levels  of  specific  an4bodies 3.  The  microbe  should  produce  relevant  virulence  factors 4.  The  microbes  should  cause  disease  in  an  appropriate  animal   model 5.  Elimina4on  of    the  microbe  should  result  in  clinical  improvement
  • 8. Role  of  Plaque !   The   Specific  Plaque  Hypothesis (fig.1) !   The   Non-­‐Specific  Plaque  Hypothesis  (fig.2) !   The   Ecological  Plaque  Hypothesis (fig.3)
  • 9. Specific Plaque Hypothesis Plaque { few species out of diverse resident flora are actively involved in disease X problem ( disease can still occur in absence of these organisms ) Disease fig.1
  • 10. Non-Specific Plaque Hypothesis Plaque { out come of the overall activity of the total plaque microflora ( ) plaque mediated diseases are polymicrobial but one problem specific organism predominate Disease fig.2
  • 11. Ecological Plaque Hypothesis Plaque { organisms associated with disease are already present on teeth but very low to detect fig.3 Disease ( disease only initiates as a result of shift in the balance of resident microf lora )
  • 13. Dental Caries !   It  is  bacterial  disease  of  calcified  4ssue  of  the  teeth  characterized   by   demineraliza4on   of   the   inorganic   and   destruc4on   of   the   organic  substance  of  the  tooth. Dental Caries is known as disease of civilization
  • 14. Dental Caries The Early theories !  The legend of Worms !  Endogenous Theory !  Chemical Theory !  Parasitic Theory !  Chemoparasitic Theory
  • 15. Dental Caries !  Chemoparasitic theory Miller  in  1890  suggested  that  oral  bacteria  converted  dietary   carbohydrates  into  acid  which  solubilized  the  calcium  phosphate  of   enamel  to  produce  a  caries  lesion
  • 17. Saliva Caries Susceptible Surface P laque Saliva Saliva Time Bacteria Sugar Saliva fig.4 aetiology of caries
  • 19. Role of Saliva Saliva   plays   a   vital   role   in   the   maintenance   of   oral   health   and  the  integrity  of  oral  4ssues Saliva  contains: • exfoliated  epithelial  cells • oral  microorganisms • food  residues  (carbohydrates) • an4microbial  factors • minerals
  • 20. Role of Saliva !   Mechanical   washing   ac4on   of   saliva   is   very   effec4ve   for   cleaning  of  teeth !   high  buffering  capacity  neutralizes  acid !   supersaturated   with   Calcium   and   phosphorus   helps   in   remineraliza4on Salivary  flow  rate:  0.8  mm  /min  on  upper  labial  surface                                              5.0  -­‐  8.0  mm/min  on  lower  lingual  surface
  • 22. Susceptible Surface Some   areas   of   the   tooth   are   more   suscep4ble   to   developing   caries   than  others,  known  as   susceptible areas Suscep4bility  is  related  to  several  factors !   Mineral  &  Fluoride  Content !   Structure  of  that  par4cular  area
  • 23. Susceptible Surface Susceptible tooth surfaces includes !   Pits  &  Fissures  (fig.5) !   Approximal  Enamel  cervical  to  contact  point  (fig.6) !   Exposed  root  surface  (fig.7 & 8) !   Margins  of  Restora4on  (fig.6  &  9) !   Cervical  margins  of  tooth  (fig.10)
  • 24. pits & fissures caries fig.5
  • 25. Caries under restoration Approximal Caries fig.6
  • 28. Caries under restoration Amalgam restoration fig.9
  • 30. From white spot to cavity formation white spot lesion Cavitation earliest clinical evidence (reversible) Later clinical stage (irreversible)
  • 31. Time & Sugar Substrate
  • 32. Sugar Substrate !   Direct   rela4onship   between   intake   of   Carbohydrates   (sugar)  &  Caries !   Glucose,  Fructose  &  Sucrose  are  cariogenic   ! Sucrose  is  the  most  cariogenic  sugar •   fermentable • easily  penetrates  plaque
  • 33. Time & Sugar Substrate There  is  a  direct  rela4onship  between  caries  and: !  Frequency  of  sucrose  consump4on !  s4ckiness  of  the  sucrose Both   affects   the   amount   of   4me   sugar   stays   into   contact   with  the  teeth  surface
  • 35. Plaque Bacteria Dental  Caries  does  not  occur   in vivo   if  microorganisms  in   the  form  of  dental  plaque  are  absent;  making  it  clear  that   dental  caries  is  a  plaque  mediated  disease
  • 36. Plaque Bacteria !   Streptococcus  mutans !   Lactobacillus  species !   Ac4nomyces  Species !   Veillonella  species
  • 37. Streptococcus species The  term  mutans  streptococci  refers  to  a  group  of  7   different  species,  but  two  species  are  most  commonly   found  in  humans !   S.  mutans  (serotypes  c/e/f) !   S.  sobrinus  (serotypes  d/g)
  • 38. Streptococcus species Factors related to Cariogenicity of S. mutans ! S.mutans  counts  in  saliva  and  at  site  of  lesion !   Isolated  from  site  before  the  development  of  caries !   Efficient  sugar  metabolism  &  transport  capability ! Acidogenic  &  acidouric !   Liele  effect  of  pH  on  growth  and  metabolism !   Produc4on  of  EPS  &  IPS
  • 39. Streptococcus species S.  mutans  been  implicated  in   !   Pits  &  fissure  caries  (strong  evidence) !   Root  surface  caries !   All  forms  of  caries !   Some  strains  are  more  pathogenic  than  others
  • 40. Streptococcus species !   S.  Sobrinus  role  is  uncertain !   Frequently  isolated  from  plaque  but  in  lesser  numbers !   Shares  the  same  cariogenic  proper4es  as  S.mutans
  • 41. Lactobacillus species Lactobacilli  can  be  divided  into  two  groups !   Homofermenta4ve  species !   Heterofermenta4ve  species
  • 42. Lactobacillus species Homofermentative species !   L.  acidophilus  &  L.  casei !   Mainly  produce  lac4c  acid  from  glucose  fermenta4on Heterofermentative species !   L.  fermentum !   Produce  lac4c  acid  and  significant  amounts  of  acetate
  • 43. Lactobacillus species Most  commonly  isolated  species  from  oral  cavity  are !   L.  casei !   L.  fermentum !   L.  acidophilus
  • 44. Lactobacillus species Factors related to Cariogenicity of Lactobacillus species !   Increased  number  in  carious  cavi4es  affec4ng  enamel  &   root  surface !   Number  in  saliva  reflect  caries  ac4vity ! Acidogenic  and  acidouric !   Few  strains  can  produce  EPS
  • 45. Lactobacillus species !   Have  a  low  affinity  for  tooth  surface !   Low  in  numbers  at  healthy  site !   Implicated  in  progression  of  caries  deeper  into  den4ne   rather  than  ini4a4on !   Implicated  in  ae4ology  of  root  surface  caries
  • 46. Actinomyces species Ac8nomyces  spp.  form  a  major  and  complex  part  of  the oral  microflora,  par4cularly  at  the  approximal  site  and gingival  crevice
  • 47. Actinomyces species ! Ac8nomyces  odontoly8cus  been  associated  with    very   early  stages  of  tooth  demineraliza4on ! Ac8nomyces  naeslundii  been  associated  with   development  of  root  surface  caries
  • 49. Caries Vaccination In  rats  ,  experimental  dental  caries  have  been  reduced  by   immunizing   with   cell-­‐wall   associated   an4gens   or   glucosyltransferase  from  S.mutans
  • 50. Caries Vaccination The  following  mechanism  have  been  suggested ! Inhib4on  by  secretory  IgA  of  bacterial  coloniza4on  of   teeth !   Interference  with  bacterial  metabolism !   Enhancement  of  phagocy4c  ac4vity  in  gingival  crevices
  • 51. Summary !   Dental  Caries  is  a  mul4factorial  disease,  strongly   associated  with  the  dental  plaque !   Ecological  Plaque  Hypothesis  caries  forms  as  a  result  of   disturbance  in  the  normal  balance  of  Oral  Biota !   The  main  factors  involved  are  suscep4ble  tooth  surfaces,   4me,  sugar  substrate  and  plaque  bacteria !   Pits  &  fissure  are  the  most  suscep4ble  areas !   Frequency  of  sugar  intake  is  more  decisive  than  total   consump4on
  • 52. Summary !   Sucrose  is  the  most  cariogenic  sugar ! Mutans  streptococci  are  implicated  in  ini4a4on  of  pits  &   fissure  and  root  caries !   Lactobacillus  spp  are  associated  with  progression  of   caries
  • 53. References !   Philip   D.   Marsh,   Michael   V   Mar4n,   Plaque   mediated   diseases-­‐  Dental  Caries  and  Periodontal  diseases  in  Oral   Microbiology,  5th  Edi4on,  Churchil  Livingstone,  2009,  pp   104-­‐117 !   J.   Bagg,   T.   W.   Macfarlane,   I.   R.   Poxton   and   A.   J.   Smith,   Dental   Caries   in   Essen4als   of   Microbiology   for   Dental   Students,   2nd   Edi4on,   Oxford   University   Press,   2006   pp238-­‐248