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1. Introduction
 1) Saliva is supersaturated with calcium and phosphate
    - remineralization at normal pH

 2) Action of Fluoride in oral cavity
    - Reduce the enamel solubility by substitution of hydroxyl
       group to form (Ca10(PO4)6F2) or (Ca10(PO4)6(OH)2-xFx)
    - Incorporation of fluoride into dental material
       i) inhibit the growth of relevant bacteria
                            -
       ii) recharged with F       significant cariostatic benefits
               -
    - High [F ] result in CaF2 like deposit : more acid labile
3) What is Chitosan
   - copolyme of glucosamine and N-acetyl glucosamine




                        Structure of Chitosan

   - exhibits moderate to good mucoadhesion
   - retained on the oral mucosa for several hours in vivo
   - antibacterial activity

4) Use of Chitosan microparticles
   - local delivery to oral cavity (ex, tetracycline, chorhexidine. etc)
   - by ionotropic gelation, emulsion polymerisatios forms
5) The aim of this work
   1) spray drying : method for manufacture and characterise
     chitosan microparticles containing NaF at low cost
   2) particle size distribution
   3) fluoride loading
   4) in vitro fluoride release
   5) particle morphology
   6) in vitro bioadhesion of discs prepared from compressed
     chitosan / fluoride microparticles

6) Null hypotheses using in vitro models
   1) Conc. of glutaraldehyde and chitosan – no affect on
      particle size, fluoride loading and fluoride release
   2) Use of glutaraldehyde – no affect on bioadhesive
      properties
2. Materials And Methods
 1) Microparticle manufacture
   a) Chitosan was dissolved in 1.0% acetic acid (40 ml), sealed,
      stirred overnight
   b) NaF sln. was prepared in DI water (10 ml)
   c) b) was added drop wise to a) over period of 60s
   d) spray drying the 3 sln. using a bench top mini spray drier
2) Microparticle characterisation
   2.1) Particle size distribution – particle sizer

  2.2) Enverionmental scanning electron microscypy (ESEM)

  2.3) Fluoride analysis – fluoride ion selective electrode

  2.4) Determination of total fluoride loading
                                        O
  2.5) in vitro fluoride release – at 32 C for 360 min

  2.6) texture probe analysis – chitosan disc on porcine oesophagi
3. Result
 3.1 Microparticle manufacture
    - fixed [chitosan]:[fluoride] = 2:1
    - two [chitosan]: 1.0 and 2.0 %
    - increasing chitosan conc. to 2.0 % reduced yield
    - 2.0 % chitosan only – 19 % yield
    - 2.0 % chitosan + 0.4 % NaF – 42 % yield
3.2 Particle size distribution
   - Increasing [chitosan] effected predictable increase in VMD
   - D[v, 90] shows greater polydispersity in 2.0 % chitosan
   - Addition of glutaraldehyde had no significant impact
      on size distribution (p>0.05)
3.3 Fluoride loading
   - Fluoride content and entrapment efficiency were enhanced
                                    -
     when conc. of [chitosan] and [F ] was increased
   - Addn. of glutaraldehyde had no significant effect on
     fluoride content and entraptment efficiency
3.4 Fluoride release profiles


                                  2




   - Sample 2 exhibited the greatest cumulative fluoride release
     with 77.4 % total fluoride released after 360 min.
- Microparticles without glutaraldehyde swelled relatively
  quickly (sample 1 and 4)

- Microparticles with high glutaraldehyde conc. showed
  minimal swelling due to the high cross-linking density
  (sample 3 and 6)

- For microparticles without or low conc. of glutaraldehyde
  (0.0010 %) (sample 1 and 4; 2 and 5, respectively),
  summed the cumulative fluoride release at 360 min was
  90 %
- Eeffect of pH on release of fluoride was not significant

                                                    pH 7.0
                                                    pH 4.0
                                                    pH 5.5
3.5 Microparticle morphology
  - Addition of NaF and glutaraldehyde had minimal effect on
    the morphology of isolated microparticles
3.6 Bioadhesion to porcine oesophageal mucosa
  - Both F max and WOA were dramatically decreased
4. Discussion
  1) Employed low cost spray drying technique to
     manufacture bioadhesive chitosan/fluoride matrix
     microparticles

  2) No fluoride incoporated chitosan microparticles
    using spray drying method was investigated

  3) Chitosan and fluoride conc. were primary
    determinant on size distribution

  4) Increasing the chitosan conc. resulted in
    significant increases in particle size
5) Micro particles could be formulated into an
   anhydrous dentifrice from which the release of
   fluoride would be initiated by the addn. of water

6) Alternative delivery formats could include
   aerosolised chitosan/fluoride microparticles

7) Based on current aerosol systems, 10 mg of
   chitosan/fluoride microparticles would introduce
   up to 630 ㎍ of fluoride
8) Compressed chitosan/fluoride discs method can
   be applied

9) Incoporation of NaF showed unexpected reduction
   in bioadhesive ability
9. Conclusion
  1) Bioadhesive chitosan/fluoride microparticles using
     a low cost spray drying technique was successful

  2) Recovered yield were inversely proportional to the
     viscosity of the original dispersion

  3) Isolated chitosan/fluoride microparticles have
     potential utility as vehicles to enhance fluoride
     retention and promote controlled fluoride delivery
     in the oral cavity

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Chitosan

  • 2. 1. Introduction 1) Saliva is supersaturated with calcium and phosphate - remineralization at normal pH 2) Action of Fluoride in oral cavity - Reduce the enamel solubility by substitution of hydroxyl group to form (Ca10(PO4)6F2) or (Ca10(PO4)6(OH)2-xFx) - Incorporation of fluoride into dental material i) inhibit the growth of relevant bacteria - ii) recharged with F significant cariostatic benefits - - High [F ] result in CaF2 like deposit : more acid labile
  • 3. 3) What is Chitosan - copolyme of glucosamine and N-acetyl glucosamine Structure of Chitosan - exhibits moderate to good mucoadhesion - retained on the oral mucosa for several hours in vivo - antibacterial activity 4) Use of Chitosan microparticles - local delivery to oral cavity (ex, tetracycline, chorhexidine. etc) - by ionotropic gelation, emulsion polymerisatios forms
  • 4. 5) The aim of this work 1) spray drying : method for manufacture and characterise chitosan microparticles containing NaF at low cost 2) particle size distribution 3) fluoride loading 4) in vitro fluoride release 5) particle morphology 6) in vitro bioadhesion of discs prepared from compressed chitosan / fluoride microparticles 6) Null hypotheses using in vitro models 1) Conc. of glutaraldehyde and chitosan – no affect on particle size, fluoride loading and fluoride release 2) Use of glutaraldehyde – no affect on bioadhesive properties
  • 5. 2. Materials And Methods 1) Microparticle manufacture a) Chitosan was dissolved in 1.0% acetic acid (40 ml), sealed, stirred overnight b) NaF sln. was prepared in DI water (10 ml) c) b) was added drop wise to a) over period of 60s d) spray drying the 3 sln. using a bench top mini spray drier
  • 6. 2) Microparticle characterisation 2.1) Particle size distribution – particle sizer 2.2) Enverionmental scanning electron microscypy (ESEM) 2.3) Fluoride analysis – fluoride ion selective electrode 2.4) Determination of total fluoride loading O 2.5) in vitro fluoride release – at 32 C for 360 min 2.6) texture probe analysis – chitosan disc on porcine oesophagi
  • 7. 3. Result 3.1 Microparticle manufacture - fixed [chitosan]:[fluoride] = 2:1 - two [chitosan]: 1.0 and 2.0 % - increasing chitosan conc. to 2.0 % reduced yield - 2.0 % chitosan only – 19 % yield - 2.0 % chitosan + 0.4 % NaF – 42 % yield
  • 8. 3.2 Particle size distribution - Increasing [chitosan] effected predictable increase in VMD - D[v, 90] shows greater polydispersity in 2.0 % chitosan - Addition of glutaraldehyde had no significant impact on size distribution (p>0.05)
  • 9. 3.3 Fluoride loading - Fluoride content and entrapment efficiency were enhanced - when conc. of [chitosan] and [F ] was increased - Addn. of glutaraldehyde had no significant effect on fluoride content and entraptment efficiency
  • 10. 3.4 Fluoride release profiles 2 - Sample 2 exhibited the greatest cumulative fluoride release with 77.4 % total fluoride released after 360 min.
  • 11. - Microparticles without glutaraldehyde swelled relatively quickly (sample 1 and 4) - Microparticles with high glutaraldehyde conc. showed minimal swelling due to the high cross-linking density (sample 3 and 6) - For microparticles without or low conc. of glutaraldehyde (0.0010 %) (sample 1 and 4; 2 and 5, respectively), summed the cumulative fluoride release at 360 min was 90 %
  • 12. - Eeffect of pH on release of fluoride was not significant pH 7.0 pH 4.0 pH 5.5
  • 13. 3.5 Microparticle morphology - Addition of NaF and glutaraldehyde had minimal effect on the morphology of isolated microparticles
  • 14. 3.6 Bioadhesion to porcine oesophageal mucosa - Both F max and WOA were dramatically decreased
  • 15. 4. Discussion 1) Employed low cost spray drying technique to manufacture bioadhesive chitosan/fluoride matrix microparticles 2) No fluoride incoporated chitosan microparticles using spray drying method was investigated 3) Chitosan and fluoride conc. were primary determinant on size distribution 4) Increasing the chitosan conc. resulted in significant increases in particle size
  • 16. 5) Micro particles could be formulated into an anhydrous dentifrice from which the release of fluoride would be initiated by the addn. of water 6) Alternative delivery formats could include aerosolised chitosan/fluoride microparticles 7) Based on current aerosol systems, 10 mg of chitosan/fluoride microparticles would introduce up to 630 ㎍ of fluoride
  • 17. 8) Compressed chitosan/fluoride discs method can be applied 9) Incoporation of NaF showed unexpected reduction in bioadhesive ability
  • 18. 9. Conclusion 1) Bioadhesive chitosan/fluoride microparticles using a low cost spray drying technique was successful 2) Recovered yield were inversely proportional to the viscosity of the original dispersion 3) Isolated chitosan/fluoride microparticles have potential utility as vehicles to enhance fluoride retention and promote controlled fluoride delivery in the oral cavity