chitosan nanoparticles synthesis and application in various fields i.e. biocompatible fruit preservatives, water treatment with non toxic substrate, cotton functionalization, etc.

2. Content :
• What is chitosan ?
• Chitosan Nanoparticle
• Properties of Chitosan Nanoparticle
• Synthesis of Chitosan based- Nanoparticles
• Application of chitosan nanoparticles
• Drug Delivery
• Waste Water Treatment
• Biomedical Applications
• Tissue engineering
• Wound dressing
• Chitosan nanoparticle based coating in fruits and Vegetables
• Green finish in Multifunctionalization of cotton fabric

3. What is chitosan ?
• It is safe nontoxic and is biodegradable polymer made from chitin.
• Chitosan is the 2nd most important and abundant biopolymer.
• Chitosan is positively charged, making it able to adhere to negative charged
surface.
• Chitosan is soluble in diverse acids to interact with polycations to form
complexes and Gels.
“Cellulose Fibres Functionalised by Chitosan: Characterization and Application” Strnad et al.

4. Sources of Chitin
“Cellulose Fibres Functionalised by Chitosan: Characterization and Application” Strnad et al.

5. Chitin to Chitosan
(1,4)-2-Amino-2-desoxy- beta-D-glucan
“Cellulose Fibres Functionalised by Chitosan: Characterization and Application” Strnad et al.
Synthesis of Chitosan
By Deacylation
(in aq NaOH
45% at 80OC)

6. Chitosan nanoparticles
• Chitosan is soluble in acidic conditions - in solution the free amino groups on its
polymeric chains can protonate, giving it a positive charge.
• Chitosan nanoparticles can be formed by incorporating a polyanion such as
tripolyphosphate (TPP) into a chitosan solution under constant stirring.
• Non toxic
• Non immunogenic
• Biodegradable
• Biocompatibility
• High charge density
• Antimicrobial property
• Absorb toxic metals
• Good adhesion
• Ability to coagulate
• Immunostimulating activity.
Properties Chitosan nanoparticles

7. Synthesis of
Chitosan
Nanoparticle
Ionotropic gelation
Microemulsion
Reverse miiceller
Coacervation
Ultrasonication
Microfluidic
Desolvation
method
Co-precipitation

8. Ionotropic gelation
“Chitosan-tripolyphosphate nanoparticles functionalized with apH-responsive amphiphile improved the in vitro antineoplastic effectsof doxorubicin “ Librelotto et
al. .Colloids and Surfaces B: Biointerfaces 2016, 147, 326–335 .

9. 1.Drug Loading
Drug Loading
During
Preparation of
Nps
After
preparation of
Nps
Chitosan nanoparticles for Drug Delivery

10. • Both water soluble and water – insoluble drug can be loaded
• Mix drug with CS
solutionStep 1
• Forms
homogeneous
mixture
Step 2
• Particles can
produced by any
method
Step 3
Water soluble drug

11. Water – insoluble drug
Resulting droplets can be hardened by
using a suitable cross-linking agent
Emulsified in CS solution to
form an oil-in-water (o/w) type
emulsion
Drug dissolved in
suitable solvent
Method 2
Multiple Emulsion Technique
Method 1
Drugs precipitate in
acidic pH solutions
Drug loaded by soaking
in CS particles

12. 2.Drug Release
In vitro release also depends upon
1. pH of media
2. Polarity of particle
3. Presence of enzymes in the dissolution media
Figure : Mechanism of drug release from
particulate systems.
‘Recent advances on chitosan-based micro- and nanoparticles in drug delivery’ S.A. Agnihotri et al. Journal of Controlled Release 2004,100, 5–28.

13. Multi-Stimuli Smart chitosan Nps for Targeted and Drug Delivery
Chitosan Nps (crosslinked with thioglycolic
acid) Functionalized with :
• Magnetic Nps ( e.g Fe3O4 Nps , CdTe
QDs) for guided motion under external
magnetic field.
• Fluorescent particle ( red dye RITC) for
imaging.
• Receptors –folate (FA) receptor
• Pharmaceutical drug (hydrophobic drug)
• Intracellular microenvironments of tumor
tissues are in a reductive state due to the
intracellular higher levels of glutathione
(GSH) than extracellular .
• It reduce disulphide bonds to free thiols.
• Used to controlled delivery and release
anticancer drugs
‘Multi-stimuli responsive smart chitosan-based nanocapsules for targeted drug delivery and triggered drug release’ X. Cui et al.Ultrasonics Sonochemistry
2017, 38, 145–153.
Scheme : The sonochemical synthesis schematic of multi-stimuli responsive
smart chitosan nanoparticle.

14. ‘Multi-stimuli responsive smart chitosan-based nanocapsules for targeted drug delivery and triggered drug release’ X. Cui et al.Ultrasonics Sonochemistry 2017, 38, 145–153.
Figure :Release profiles of drug: (a) without GSH, (b)with 10 µM GSH, and
(c) with 10 mM GSH.

15. Chitosan-based nanoparticles (CSNPs) for pH-sensitive
drug delivery systems
• Cationic property causes it to approach cell membranes more easily
and allows ionic cross-linkage with multivalent anions.
• Presence of amino groups in large quantities on chains of CS, its
ionization makes CS molecules pH-sensitive.
• Not modified with PEG and poloxamer.

16. Chitosan tripolyphosphate
nanoparticles (NPs)
 Loaded DOX (doxorubin, leads
to apoptosis of cell ) -
encapsulation by electrostatic
interactions.
 Anionic diacyl lysine-based
surfactant with sodium counterion
(77KS) as a pH-sensitive adjuvant
is also incorporated.
Figure : Schematic representation of a proposed mechanism for the
cellular uptake and drug release of pH-responsive CS-NPs.
“Chitosan-tripolyphosphate nanoparticles functionalized with apH-responsive amphiphile improved the in vitro antineoplastic effectsof doxorubicin “ Librelotto et
al. .Colloids and Surfaces B: Biointerfaces 2016, 147, 326–335 .

17. Chitosan coated magnetic nps :
Superparamagnetic iron oxide nanoparticles (SPIONPs, Fe3O4 NPs)
Responsive to both physical (ultrasound)and chemical (pH) stimuli.
Preparation of magnetic composite drug :
1. Preparation of chitosan-conjugated particles (chitosan–magnetite) using a co-precipitation method
because the functional groups at the surface of iron oxide particles easily react with those of chitosan or
its derivatives
2. Ciprofloxacin was loaded onto the nanocarrier via H-bonding interactions
The in vitro drug loading at pH 4.8 and release kinetics at pH 7.4 studies revealed that the drug delivery system can take
99% of ciprofloxacin load and quantitatively release the drug
Figure : Schematic illustration of preparation of the CS–Fe3O4 nanoparticles and drug loading
“Analytical characteristics and application of novel chitosan coatedmagnetic nanoparticles as an efficient drug delivery system forciprofloxacin. Enhanced drug
release kinetics by low-frequencyultrasounds” S. Kariminia et al.. Journal of Pharmaceutical and Biomedical Analysis 2016,129 , 450–457 .

18. Chitosan nanoparticles for Waste Water Treatment
 Modified chitosan nanomaterials used for
 Used for removal of Cr6+, Fe3+, Zn2+, Cu2+, congo red, methyl orange, methyl blue, etc.
 Chitosan nanoparticles have become potential materials of choice for water treatment because of :
a. Small size.
b. Large surface area.
c. Quantum size effects.
 Nanochitosan particles prepared by ionotropic gelation of chitosan and tripolyphosphate cross–linking
agent were tested for Pb(II) adsorption.
 Maximum adsorption capacity of 398.0 mg g−1 was reported from Langmuir isotherm.
 Chitosan nanorods explored for the adsorption of Cr(VI) and observed an uptake of 323.6 mg g−1 by the
adsorbent.
 The use of external magnetic field helps in easy recovery and reuse of the target contaminants from the
solution.

19. Removal of heavy metal ion using chitosan magnetic nanocomposite :
Figure: An illustration for the carboxymethylation and binding onto Fe3O4 nanoparticles of chitosan.
Carboxymethylated chitosan–
conjugated Fe3O4 nanoparticles
for the capture of Co(II) ions.
This nano–adsorbent comprised
Fe3O4 as cores and chitosan as ion
exchange groups.
“Magnetic chitosan nanoparticles: Studies on chitosan binding and adsorption of Co(II) ions” Chang et al.. Reactive & Functional Polymers 2006 ,66, 335–341.

20. Figure: Synthesis route of Fe3O4 nanoparticles of chitosan and their application for
removal of of Co(II) with the help of an external magnetic field
“Magnetic chitosan nanoparticles: Studies on chitosan binding and adsorption of Co(II) ions” Chang et al.. Reactive & Functional Polymers 2006 ,66, 335–341.

21. Figure: Effect of pH on the adsorption of Co(II) ions by the magnetic chitosan nano adsorbent at 25º C and an initial
concentration of 1500 mg/l.
• The magnetic chitosan nano-adsorbent was shown to be quite efficient for the removal of
Co(II) ions in the examined pH range of 3–7. The maximum adsorption capacity occurred at pH
5.5.
• Chitosan and its derivatives are capable of adsorbing a number of metal cations and can be
utilized as the adsorbent material for the removal of metal cations.
“Magnetic chitosan nanoparticles: Studies on chitosan binding and adsorption of Co(II) ions” Chang et al.. Reactive & Functional Polymers 2006 ,66, 335–341.

22. Dye removal using chitosan magnetic nanocomposite :
 Adsorption mechanism of anionic dyes with nano–sized chitosan as well as
modified nanochiosan materials was found to be because of the electrostatic
interaction between negatively charged dye ions and protonated amino groups of
chitosan.
 For Cationic dyes The chelating interaction between dye molecules and Fe(III)
center may play a leading role instead of electrostatic interactions , using chitosan-
Fe(III) hydrogel .
Dye
Anionic Cationic

23. Figure: Proposed interactions mode between cationic dyes
and chitosan–Fe(III) hydrogel.
The proposed mechanism of adsorption of
anionic dye follows the following steps:
1. The dissolution of dye in aqueous solution
and dissociation of its sulfonate groups
(D–SO3Na) and the conversion to anionic
dye ions as shown:
D − SO3Na → D − SO3
− + Na+
2. Protonation of amino groups of chitosan
takes place as represented by the equation:
R − NH2 + H+ ↔ R − NH+
3
3. Finally , the ionic interaction between the
oppositely charged moieties in solution
R − NH+
3 + D − SO3
− ↔ R − NH3. . .O3S − D
“Fast and highly efficient removal of dyes under alkaline conditions using magnetic chitosan-Fe(III) hydrogel” Shen et al 3 ..
Water Research 2011, 4 5 ,5200-5210.

24. Figure : Mechanism of CS–PMAA nanoparticle
formation .
Synthesis pf chitosan nanoparticles using
Methacrylic acid
• Chitosan nanoparticles were synthesized through
polymerization of CS with methacrylic acid (MAA)
using K2S2O8 as initiator.
• CS was dissolved in an aqueous MAA solution
(0.5%, wt/v) for 12 h under magnetic stirring. 0.2
mmol of K2S2O8 was added to the CS–MAA
solution under continuous stirring at 70 ◦C for 1 h,
leading to the formation of CS–PMAA
nanoparticles by ice cooling the mixture.
Figure : TEM micrograph
of CS/PMAA
nanoparticles prepared
with 0.2
wt%ofchitosan(110nm).
"Highly stable, edible cellulose films incorporating chitosan nanoparticles." de Moura, Márcia R., et al. Journal of food science 76.2 (2011).

25. Utilization of CsNPs as Green
finish in Multifunctionalization of
cotton fabric.
• When Chitosan nanoparticles were used as a
finish for cotton fabrics with a crosslinking
agent, there was improved dyeability, thermal
stability, antibacterial activity and UV
protection.
• Antibacterial and UV protection were further
increased by post treatment of fabric with
copper sulfate.
"Utilization of chitosan nanoparticles as a green finish in multifunctionalization of cotton textile." Hebeish, Ali et al, International journal of
biological macromolecules 60 (2013): 10-17.
Figure : Mechanism of Cs/GPTMS coated cotton fabric

26. Highly Stable, Edible Cellulose Films
Incorporating Chitosan Nanoparticles
• Film was prepared by mixing the 0.2%w/w
CsNPs with 2%w/w cmc solution in distilled
water under magnetic stirring for 12 h without
changing the pH of mixture.
• The flasks were allowed to rest for 6 h to degas
and prevent microbubble formation within the
films.
• The solutions were then poured on an acrylic
plate for film preparation by casting and stoved
at 120oC to get film.
Figure: CMC films formulated with (A) and
without (B) nanoparticles.
"Highly stable, edible cellulose films incorporating chitosan nanoparticles." de Moura, Márcia R., et al. Journal of food science 76.2 (2011).

27. Chitosan nanoparticle based coating in fruits and
Vegetables
• The effect of 1% chitosan nanoparticle film.
• After 32 day, the red index, decay incidence, weight loss, and
respiration rate of the coated jujubes were low as compared to
that of controlled.
• The lower PAL activity and higher activities of scavenger
antioxidant enzymes (i.e., SOD, POD, and CAT) of the coated
jujubes can be attributed to the compound coating.
• Increased MDA in the coated jujubes was restrained. Composite
coating had shown to be superior in preserving total flavonoid
than chitosan coating alone. But no differences were observed
in terms of vitamin C loss and total polyphenol content between
composite coating and control.

28. Nanofibrils Structure of Chitosan for Biomedical
Applications
“Nanofibrous Structure of Chitosan for Biomedical Applications” C. Mahoney
et al.. J Nanomedic Biotherapeu Discover 2012, 1,2.
Figure : Preparation of chitosan /PET solution for
electrospining
• Chitosan is electrospun in blended form with
different biological polymers to from
nanofibres.
• Electrospinning of chitosan is done by
blending chitosan with
• PEO Polyethlene oxide
• PVA polyvinyl alcohol
• PLA Polylactic acid
• PCL polycaprolactone

29. Tissue engineering
• Electrospun polyblend of chitosan/PCL
nanofibers are fabricated into tubular
constructs to suit as a nerve guide for
peripheral nerve regeneration in
scaffolds
• Severed nerve endings cannot be
repaired with sutures.
• Synthetic nerve guides can take the place
of autografts, to redirect nerve growth
across the critical gap.
• Nerve conduit materials prepared with
chitosan/PCL polyblend nanofibers have
demonstrated strong mechanical
properties, capable of being sutured to
the nerve ends and maintaining structural
stability in vivo.
“Application of Chitosan Based Coating in Fruit and Vegetable Preservation”Jianglian and Shaoying. J Food Process Technol 2013, 4,5.

30. Wound dressing
• In the area of wound healing, care, and management, antibacterial resistance of
microorganisms is a major concern.
• Chitosan is most suitable for this application because it meets several requirements
such as histocompatibility, biodegradability, lack of antigenicity and also
promotes wound healing.
• A polyblend nanofibrous membrane of chitosan/collagen was found to induce cell
migration and proliferation while assisting in wound heal. It is reported that
nanofibrous membrane have beneficial effects better than gauze and commercial
collagen sponge when conducting animal studies.
• Nanofibers have greater water-retention capacity because of very high-specific
surface area and are very soft, so that the dressing will not chafe the wound.
“Application of Chitosan Based Coating in Fruit and Vegetable Preservation”Jianglian and Shaoying. J Food Process Technol 2013, 4,5.

31. Chitosan nanofibre based drug release
“Application of Chitosan Based Coating in Fruit and Vegetable Preservation”Jianglian and Shaoying. J Food Process Technol 2013, 4,5.

32. REFERENCES
• “Recent advances on chitosan-based micro- and nanoparticles in drug delivery’”S.A. Agnihotri et al..
Journal of Controlled Release 2004,100, 5–28.
• “Potential applications of cellulose and chitosan nanoparticles/composites in wastewater treatment: A
review” S. Olivera et al. Carbohydrate Polymers 2016, 153, 600–618.
• “Mesoporous silica and chitosan based pH-sensitive smart nanoparticles for tumor targeted drug delivery’P.
Deveci et al.. J Incl Phenom Macrocycl Chem 2017, 89,15–27.
• “Multi-stimuli responsive smart chitosan-based microcapsules for targeted drug delivery and triggered drug
release” X. Cui et al. Ultrasonics Sonochemistry 2017, 38, 145–153.
• “Nanofibrous Structure of Chitosan for Biomedical Applications”C. Mahoney et al.. J Nanomedic
Biotherapeu Discover 2012, 1, 2.
• “Application of Chitosan Based Coating in Fruit and Vegetable Preservation” Jianglian and Shaoying. J
Food Process Technol 2013, 4,5.
• "Highly stable, edible cellulose films incorporating chitosan nanoparticles." de Moura, Márcia R., et al.
Journal of food science 76.2 (2011).

33. THANK YOU