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Synthesis of hydroxyapatite
nanoparticles
By
Ahmed Mostafa Hussein
Assistant lecturer, Dental Biomaterials Department
Faculty of Dentistry, Mansoura University
December 2015
1
Introduction
Hydroxyapatite (HAp) is the major mineral constituent of
vertebrate bones and teeth.
It has been well documented that HAp nanoparticles can
significantly increase the biocompatibility and bioactivity of
man-made biomaterials.
Bone is a composite consisting mainly of calcium phosphate
(69%), water (9%) & collagen (20%). Other organic substances,
such as proteins & polysaccharides are present in small
amounts.
2
The collagen, which gives the bone its elastic resistance, acts as
a matrix for the deposition and growth of minerals.
Among the CaP salts, hydroxyapatite (Ca10(PO4)6(OH)2 (HAp)
is the most similar to the mineral part of bone.
 The HAp crystals have the shape of needles 40–60 nm in
length, 20 nm in width, and 1.5–5 nm in thickness.
3
Applications of CaP in dentistry
1) Replacement for bony and periodontal defects & alveolar ridge
2) Tissue engineering systems
3) Bioactive coating on metallic osseous implants
4) Filler for reinforcing dental resins
5) Stimulate formation of reparative dentin better than Ca(OH)2
6) Repair of mechanical bifurcation perforation
7) Apical barrier formation
8) Pulp capping
4
5
A)Titanium implant
B)Titanium implant coated with HAp
Nanosized HAp: has a grain size less than 100 nm in at least one
direction similar to the mineral found in hard tissues.
6
Advantages of nanosized HAp
1. Nanosized HAp has higher surface area and surface roughness
resulting in superior surface functional properties of nanosized
HAp compared to its microphase counterpart.
2. Mimic the bone mineral in composition and structure.
3. Promote osteointegration and subsequent bone tissue formation.
4. The best material to use for bone replacement and regeneration.
7
5. Enhanced resorbability and much higher bioactivity than
micron-sized ceramics.
6. Capability of decreasing apoptotic cell death and hence
improving cell proliferation and cellular activity related to bone
growth.
7. Improved cell proliferation and differentiation.
8. Better cell adhesion and cell-matrix interactions.
8
Challenges during synthesis of nanosized HAp
1. Formation of phase impurities (other CaP salts) during synthesis
of HAp particles.
2. Difficulties in controlling size, size distribution, morphology,
crystallinity, stoichiometry and degree of particle agglomeration.
9
Notes
* Stoichiometry: definite proportions or quantitative relationship
or ratios between constituents.
* Hydroxyapatite: Ca10(PO4)6(OH)2
Ca/P = 10/6 = 1.67
10
11
Main calcium phosphate salts
The most common methods for synthesis of HAp
1. Chemical precipitation
2. Combination procedures
3. Hydrothermal method
4. Synthesis from biogenic sources
5. Sol-gel method
12
13
Methods for synthesis of hydroxyapatite nanoparticles
14
Methods for synthesis of hydroxyapatite nanoparticles
15
Methods for synthesis of hydroxyapatite nanoparticles
1. Dry methods
Dry methods do not use a solvent, unlike wet methods.
Advantages
1) Simple procedure
2) Low cost: relatively inexpensive raw materials
3) Produce highly crystalline HAp
4) Suitable for mass production of HAp powder
5) Not strongly influenced by the processing parameters
6) Mostly, do not require precisely controlled conditions
16
Disadvantages
1) Large size of particles (in case of solid-state synthesis)
2) Low phase purity of HAp (in case of mechano-chemical
process)
Types of dry methods
1.1. Solid-state
1.2. Mechanochemical
17
1.1. Solid-state synthesis
* Precursors are first milled and then calcined at a very high
temperature (e.g. 1000 °C).
* The precursors can be calcium- and phosphate-containing
chemicals of various types or a previously prepared CaP salt.
* The high temperature of calcination leads to formation of a
highly crystallized structure.
18
Advantages
1) Simple procedure.
2) Low cost
3) Suitable for mass production of HAp powder
4) The method of choice for commercial production
Disadvantages
1) Heterogeneity in phase composition owing to the small diffusion
of ions during the reaction.
2) Unattractive both scientifically & technologically
19
20
Solid-state synthesis of HAp
1.2. Mechanochemical method
* Sometimes known as mechanical alloying.
* Used for fabrication of nanocrystalline alloys and ceramics.
* The materials are ground on a planetary mill while the molar
ratio between the reagents is kept at the stoichiometric ratio.
* Increasing the milling time leads to a decrease in crystallite size.
21
* The method consists mainly of mixing Ca and P, maintaining
Ca/P ratio and pH.
* The powder mixture is placed in a ball mill and is subjected to
high energy collision from the balls, and thus mechanical force is
used to achieve chemical processing and transformation.
* Milling media such as Zirconia, alumina, stainless steel etc.
* Maintaining the ball mass ratio is critical.
22
Disadvantages
* Contaminations
* Long processing time
* No control on particle morphology
* Agglomerates
23
24
25
Mechanochemical method
2. Wet methods
Aqueous solutions of phosphate and calcium ions.
Advantages
1. Precise control over the morphology and size of particles
2. The most popular in scientific researches
3. The most promising methods for the synthesis of nanosized
HAp.
26
Disadvantages
1) Difficulties in controlling the crystallinity and phase purity of
nanoparticles
2) Time-consuming makes some wet procedures unsuitable for
production of large quantities of HAp
3) Low preparation temperature (compared to dry methods)
resulting in:
* Generation of CaP phases other than HAp
* Lowering of the crystallinity
* Various ions in the aqueous solution can be incorporated into
the crystal structure.
27
2.1. Conventional chemical precipitation
* The simplest route for synthesis of nanosized HAp.
* Note: At room temperature and pH 4.2, HAp is the least soluble
and usually the most stable CaP phase in an aqueous solution.
* The precipitation reaction is usually conducted at pH values
higher than 4.2 and temperatures ranging from room temperature
to temperatures close to the boiling point of water.
28
* Ca2+ source: calcium hydroxide or calcium nitrate
* PO43- source: orthophosphoric acid or diammonium hydrogen
phosphate
* A typical procedure involves the dropwise addition of one reagent
to another under continuous and gentle stirring, while the molar ratio
of elements (Ca/P) is kept at stoichiometry according to its ratio in
HAp (1.67).
* The resultant suspension may be aged under atmospheric pressure
or immediately washed, filtered, dried and crushed into a powder.
29
* It is the most common method.
Advantages
1) Simplicity
2) Ready availability
3) Relatively inexpensive raw materials.
4) Low reaction temperatures
5) Low operating costs
30
31
Conventional chemical precipitation method
32
Conventional chemical precipitation method
2.2. Hydrolysis method
* HAp nanoparticles can be prepared by the hydrolysis of other
CaP phases, including dicalcium phosphate anhydrous (DCPA),
dicalcium phosphate dihydrate (DCPD) and tricalcium phosphate
(TCP).
* Hydrolysis of octacalcium phosphate (OCP) has not been of
great interest for the preparation of HAp particles, probably
because of the slow rate of OCP hydrolysis and/or the ability of
OCP to incorporate impurities.
33
* Aqueous hydrolysis of CaP phases into HAp usually proceeds by
dissolution and precipitation processes.
* Among the CaP salts, acidic phases such as DCPA and DCPD are
thermodynamically less stable under pH values higher than 6–7
and undergo transformation into a more stable CaP e.g. HAp.
34
2.3. Sol-gel method
* The conventional sol-gel process involves the preparation of a
3D inorganic network by mixing alkoxides (or other suitable
precursors) in either an aqueous or an organic phase.
* This is followed by aging at room temperature, gelation, drying
on a hot plate.
* And finally removing of organic residues from the resulting dried
gel using post-heat treatment (calcination).
* The crystallite size and crystallinity of which can increase with
increasing the calcination temperature.
35
* In other words, Ca and P precursors are mixed & pH is fixed
(adjusted) with ammonia or ammonium hydroxide followed by
ageing, filtration, drying and calcinations.
* To avoid cracking in a target 3-D monolith structure it may be
necessary to age the gel before drying.
36
Advantages
1) Low temperature formation
2) Improving the chemical homogeneity of the resulting powder
3) A stoichiometric structure with a large surface area and a small
cluster size
4) The bioresorbability of the sol-gel HAp is higher than
conventional powder and is close to biological apatite.
37
Disadvantages
1) High cost of some of the starting materials, especially alkoxide-
based precursors.
2) Generation of secondary phase (usually calcium oxide, CaO).
3) Secondary CaO phase is harmful to the biocompatibility of
HAp.
Note: Attempts have been made to remove the coexisting CaO,
either through washing of the calcined powder using a dilute acid
solution (mainly HCl) or through increasing the aging time.
38
39
Sol-gel method
40
Sol-gel method
2.4. Hydrothermal method
* One of the most common methods for preparation of HAp.
* The third most popular method after the conventional
precipitation and combination methods.
* Reaction of chemicals in an aqueous solution at elevated
temperature and pressure.
* Hydrothermal synthesis can be considered a chemical
precipitation in which the aging step is conducted at a high
temperature (typically above the boiling point of water) inside
an autoclave or pressure vessel.
41
* Performed at lower temperatures than solid-state reactions.
* Other low temperature methods such as wet chemical
precipitation and sol-gel synthesis require post heat treatment to
crystallise the HA; whereas crystalline HA can be produced in one
step via hydrothermal synthesis.
* The initial stage is mixing the calcium and phosphorous
precursors followed maintaining the Ca/P ratio at a constant value
of 1.67. The solution is heated in a sealed vessel (autoclave).
42
* The mixing is then allowed to age, and subsequently washed and
filtered. Finally it is dried in an oven and calcined.
* The amount of HAp is limited to the size of the reaction vessel.
* The starting reagents and H2O should occupy 50–60% of the
autoclave volume.
43
Advantages
The formed HAp is relatively stoichiometric and highly
crystalline.
Disadvantages
1) Need expensive equipments
2) Poor capability to control the morphology and size of HAp
nanoparticles
44
45
Teflon-lined stainless steel autoclave
for hydrothermal synthesis of HAp
46
HAp prepared by hydrothermal method
47
Effect of pH, temperature, and
duration of hydrothermal treatment
on phase, morphology, and
particle size of the CaP powder
2.5. Emulsion method
* Two immiscible liquids (such as water and organic) stabilized by
the presence of surfactants.
* Note: Immiscible: incapable of mixing or attaining homogenity
* Surfactants can reduce the surface tension of the immiscible
liquids, resulting in a dispersed phase confined to nanometer scale.
* Surfactants can subsequently be removed easily by calcination.
48
Advantages
1) Reduces nanoparticles agglomeration.
2) Reduces the particle size.
3) Controls the morphology (spheres, rods, discs, etc.).
4) Simple procedure
5) Low synthesis temperature (around room temperature)
6) Occurs without any high-temperature requirement
49
2.6. Sonochemical method
* Chemical reactions activated by powerful ultrasound radiation.
* The first step is mixing Ca and P precursor, maintaining Ca/P
ratio and pH as a constant value followed by the passage of
ultrasonic waves, then drying and calcination.
Advantages
1) Accelerates the reactions & the growth of HAp crystals.
2) More uniform, smaller & purer crystals with minimal
agglomeration.
3) Single phase HAp could be synthesized after 15-60 min
sonication.
50
51
Sonochemical method
3. High-temerature processes
Used to burn or partially burn the precursors.
Advantages
1) Avoid undesirable CaP phases.
2) Produce HAp with high crystallinity & good chemical
homogeneity.
* High-temperature processes can be performed by 2 techniques:
1) Combustion: has received more attention.
2) Pyrolysis
52
3.1. Combustion
* A conventional process used to prepare various oxide ceramics.
* Combustion processing of HAp involves a very rapid exothermic
reaction.
* At first, the aqueous solutions of Ca(NO3)2 and (NH4)2HPO4 are
first mixed, followed by adding concentrated HNO3 (an oxidizing
agent) to dissolve the resulting white precipitate. A single or a
mixture of two or more fuels (e.g. citric acid, succinic acid, urea
and glycine) is subsequently incorporated into the resulting
solution.
53
* The reaction can be initiated by heating the mixture in a furnace
at a low temperature (e.g. 300 °C), then a sudden increase in
temperature to a maximum value.
* The final step is the fast cooling of mixture to induce maximum
nucleation and to prevent any further particle growth.
* Different fuels have proved to be capable of delivering different
flame temperatures ranging from 100 to 900 °C (e.g. citric acid:
150 °C; succinic acid: 425 °C; urea: 800 °C; glycine: 890 °C).
54
* A vigorous exothermic reaction occurs between the fuel and
oxidizer; the gaseous products of this reaction spontaneously
combust. This gives rise to a very high local temperature that
causes the formation of a solid calcium phosphate powder. This
process can be completed in less than 20 minutes.
* Depending on the fuel used, the product formed may be either
crystalline or amorphous. Both require a calcination step; to
remove organic residues and crystallize the phase formed,
respectively.
55
Advantages
1) Quickly produce powder with high purity in a single step
operation.
2) Inexpensive raw materials
3) Relatively simple
4) Good chemical homogeneity
56
57
Combustion method
3.2. Pyrolysis (spray pyrolysis)
* In general, during HAp synthesis, some post treatments and/or
long-term aging under elevated temperatures may be required to
achieve a high crystalline product.
* By contrast, Rapid pyrolysis produces stoichiometric,
homogeneous, and highly crystalline HAp.
* Pyrolysis method can also be classified under a broad category
known as aerosol methods (or gas-phase methods)
58
* Particles can be formed from reactants in a gas phase generated
by physical evaporation of liquid precursors.
* Spray pyrolysis involves spraying the precursor solutions into a
flame or a hot zone of an electric furnace using an ultrasonic
generator.
* This is then followed by reaction of the generated vapors and
gases at high temperatures to produce final powder, typically in an
aggregated and agglomerated form.
59
* In fact, the high temperature leads to complete evaporation of
precursor droplets followed by nucleation and growth of
nanoparticles in the gas phase.
Disadvantages
1) Small decomposition of HAp into α-TCP, because of high
temperature of flame (usually above 2000 °C).
2) Secondary aggregates are usually formed.
3) Poor control over the processing variables.
60
61
Spray pyrolysis
4. Synthesis from biogenic sources
* Biogenic sources: e.g. biowastes, eggshell and exoskeleton of
marine organisms.
* Economical and environmental benefits of waste recovery.
* Accepted better by the living organs, because of its
physicochemical similarity to the human bone apatite.
* Calcium carbonate originated from marine species usually
exhibits porosity and interconnectivity similar to that found in
human bone.
62
* The larger and higher macroporosity of the structure, similar to
the natural cancellous bone, has been reported to result in earlier
bone mineralization during the implantation; and hence corals-
generated HAps are claimed to be more beneficial to bone repair
applications.
* Expected to attract more attention in the near future.
* Can produce HAp blocks or particles.
63
64
Synthesis
of HAp
from
biogenic
sources
65
Cuttlebone, its porosity & interconnectivity
5. Combination procedures
* Two or more distinct procedures to synthesize HAp
nanoparticles.
* Combinations of hydrothermal-mechanochemical, hydrothermal-
hydrolysis and hydrothermal-emulsion have received more
attention.
66
* Combinations of the microwave irradiation with different
methods e.g. solid-state and sonochemical methods has been
reported.
* Combination procedures may improve the characteristics of HAp
nanoparticles.
67
Microwave-assisted irradiation method
* Increased reaction kinetics and rapid initial heating coupled with
reduced reaction times when compared to conventional heating
methods.
* The usual steps of mixing Ca & P, maintaining Ca/P ratio and pH
are all followed, and then passing microwave radiation followed
by heat treatment.
68
References
1. Sadat-Shojai M, Khorasani MT, Dinpanah-Khoshdargi E,
Jamshidi A. Synthesis methods for nanosized hydroxyapatite with
diverse structures. Acta Biomaterialia 2013; 9: 7591–621.
2. Anitha P, Pandya HM. Comprehensive review of preparation
methodologies of nano hydroxyapatite. Journal of Environmental
Nanotechnology 2014; 3: 101–21.
3. Cox S. Synthesis method of hydroxyapatite.
http://www.mandplabs.com/interface/uploads/files/whitepapers/hydroxyapatite-white-paper.pdf
69
70
Thank you

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Synthesis of hydroxyapatite nanoparticles

  • 1. Synthesis of hydroxyapatite nanoparticles By Ahmed Mostafa Hussein Assistant lecturer, Dental Biomaterials Department Faculty of Dentistry, Mansoura University December 2015 1
  • 2. Introduction Hydroxyapatite (HAp) is the major mineral constituent of vertebrate bones and teeth. It has been well documented that HAp nanoparticles can significantly increase the biocompatibility and bioactivity of man-made biomaterials. Bone is a composite consisting mainly of calcium phosphate (69%), water (9%) & collagen (20%). Other organic substances, such as proteins & polysaccharides are present in small amounts. 2
  • 3. The collagen, which gives the bone its elastic resistance, acts as a matrix for the deposition and growth of minerals. Among the CaP salts, hydroxyapatite (Ca10(PO4)6(OH)2 (HAp) is the most similar to the mineral part of bone.  The HAp crystals have the shape of needles 40–60 nm in length, 20 nm in width, and 1.5–5 nm in thickness. 3
  • 4. Applications of CaP in dentistry 1) Replacement for bony and periodontal defects & alveolar ridge 2) Tissue engineering systems 3) Bioactive coating on metallic osseous implants 4) Filler for reinforcing dental resins 5) Stimulate formation of reparative dentin better than Ca(OH)2 6) Repair of mechanical bifurcation perforation 7) Apical barrier formation 8) Pulp capping 4
  • 6. Nanosized HAp: has a grain size less than 100 nm in at least one direction similar to the mineral found in hard tissues. 6
  • 7. Advantages of nanosized HAp 1. Nanosized HAp has higher surface area and surface roughness resulting in superior surface functional properties of nanosized HAp compared to its microphase counterpart. 2. Mimic the bone mineral in composition and structure. 3. Promote osteointegration and subsequent bone tissue formation. 4. The best material to use for bone replacement and regeneration. 7
  • 8. 5. Enhanced resorbability and much higher bioactivity than micron-sized ceramics. 6. Capability of decreasing apoptotic cell death and hence improving cell proliferation and cellular activity related to bone growth. 7. Improved cell proliferation and differentiation. 8. Better cell adhesion and cell-matrix interactions. 8
  • 9. Challenges during synthesis of nanosized HAp 1. Formation of phase impurities (other CaP salts) during synthesis of HAp particles. 2. Difficulties in controlling size, size distribution, morphology, crystallinity, stoichiometry and degree of particle agglomeration. 9
  • 10. Notes * Stoichiometry: definite proportions or quantitative relationship or ratios between constituents. * Hydroxyapatite: Ca10(PO4)6(OH)2 Ca/P = 10/6 = 1.67 10
  • 12. The most common methods for synthesis of HAp 1. Chemical precipitation 2. Combination procedures 3. Hydrothermal method 4. Synthesis from biogenic sources 5. Sol-gel method 12
  • 13. 13 Methods for synthesis of hydroxyapatite nanoparticles
  • 14. 14 Methods for synthesis of hydroxyapatite nanoparticles
  • 15. 15 Methods for synthesis of hydroxyapatite nanoparticles
  • 16. 1. Dry methods Dry methods do not use a solvent, unlike wet methods. Advantages 1) Simple procedure 2) Low cost: relatively inexpensive raw materials 3) Produce highly crystalline HAp 4) Suitable for mass production of HAp powder 5) Not strongly influenced by the processing parameters 6) Mostly, do not require precisely controlled conditions 16
  • 17. Disadvantages 1) Large size of particles (in case of solid-state synthesis) 2) Low phase purity of HAp (in case of mechano-chemical process) Types of dry methods 1.1. Solid-state 1.2. Mechanochemical 17
  • 18. 1.1. Solid-state synthesis * Precursors are first milled and then calcined at a very high temperature (e.g. 1000 °C). * The precursors can be calcium- and phosphate-containing chemicals of various types or a previously prepared CaP salt. * The high temperature of calcination leads to formation of a highly crystallized structure. 18
  • 19. Advantages 1) Simple procedure. 2) Low cost 3) Suitable for mass production of HAp powder 4) The method of choice for commercial production Disadvantages 1) Heterogeneity in phase composition owing to the small diffusion of ions during the reaction. 2) Unattractive both scientifically & technologically 19
  • 21. 1.2. Mechanochemical method * Sometimes known as mechanical alloying. * Used for fabrication of nanocrystalline alloys and ceramics. * The materials are ground on a planetary mill while the molar ratio between the reagents is kept at the stoichiometric ratio. * Increasing the milling time leads to a decrease in crystallite size. 21
  • 22. * The method consists mainly of mixing Ca and P, maintaining Ca/P ratio and pH. * The powder mixture is placed in a ball mill and is subjected to high energy collision from the balls, and thus mechanical force is used to achieve chemical processing and transformation. * Milling media such as Zirconia, alumina, stainless steel etc. * Maintaining the ball mass ratio is critical. 22
  • 23. Disadvantages * Contaminations * Long processing time * No control on particle morphology * Agglomerates 23
  • 24. 24
  • 26. 2. Wet methods Aqueous solutions of phosphate and calcium ions. Advantages 1. Precise control over the morphology and size of particles 2. The most popular in scientific researches 3. The most promising methods for the synthesis of nanosized HAp. 26
  • 27. Disadvantages 1) Difficulties in controlling the crystallinity and phase purity of nanoparticles 2) Time-consuming makes some wet procedures unsuitable for production of large quantities of HAp 3) Low preparation temperature (compared to dry methods) resulting in: * Generation of CaP phases other than HAp * Lowering of the crystallinity * Various ions in the aqueous solution can be incorporated into the crystal structure. 27
  • 28. 2.1. Conventional chemical precipitation * The simplest route for synthesis of nanosized HAp. * Note: At room temperature and pH 4.2, HAp is the least soluble and usually the most stable CaP phase in an aqueous solution. * The precipitation reaction is usually conducted at pH values higher than 4.2 and temperatures ranging from room temperature to temperatures close to the boiling point of water. 28
  • 29. * Ca2+ source: calcium hydroxide or calcium nitrate * PO43- source: orthophosphoric acid or diammonium hydrogen phosphate * A typical procedure involves the dropwise addition of one reagent to another under continuous and gentle stirring, while the molar ratio of elements (Ca/P) is kept at stoichiometry according to its ratio in HAp (1.67). * The resultant suspension may be aged under atmospheric pressure or immediately washed, filtered, dried and crushed into a powder. 29
  • 30. * It is the most common method. Advantages 1) Simplicity 2) Ready availability 3) Relatively inexpensive raw materials. 4) Low reaction temperatures 5) Low operating costs 30
  • 33. 2.2. Hydrolysis method * HAp nanoparticles can be prepared by the hydrolysis of other CaP phases, including dicalcium phosphate anhydrous (DCPA), dicalcium phosphate dihydrate (DCPD) and tricalcium phosphate (TCP). * Hydrolysis of octacalcium phosphate (OCP) has not been of great interest for the preparation of HAp particles, probably because of the slow rate of OCP hydrolysis and/or the ability of OCP to incorporate impurities. 33
  • 34. * Aqueous hydrolysis of CaP phases into HAp usually proceeds by dissolution and precipitation processes. * Among the CaP salts, acidic phases such as DCPA and DCPD are thermodynamically less stable under pH values higher than 6–7 and undergo transformation into a more stable CaP e.g. HAp. 34
  • 35. 2.3. Sol-gel method * The conventional sol-gel process involves the preparation of a 3D inorganic network by mixing alkoxides (or other suitable precursors) in either an aqueous or an organic phase. * This is followed by aging at room temperature, gelation, drying on a hot plate. * And finally removing of organic residues from the resulting dried gel using post-heat treatment (calcination). * The crystallite size and crystallinity of which can increase with increasing the calcination temperature. 35
  • 36. * In other words, Ca and P precursors are mixed & pH is fixed (adjusted) with ammonia or ammonium hydroxide followed by ageing, filtration, drying and calcinations. * To avoid cracking in a target 3-D monolith structure it may be necessary to age the gel before drying. 36
  • 37. Advantages 1) Low temperature formation 2) Improving the chemical homogeneity of the resulting powder 3) A stoichiometric structure with a large surface area and a small cluster size 4) The bioresorbability of the sol-gel HAp is higher than conventional powder and is close to biological apatite. 37
  • 38. Disadvantages 1) High cost of some of the starting materials, especially alkoxide- based precursors. 2) Generation of secondary phase (usually calcium oxide, CaO). 3) Secondary CaO phase is harmful to the biocompatibility of HAp. Note: Attempts have been made to remove the coexisting CaO, either through washing of the calcined powder using a dilute acid solution (mainly HCl) or through increasing the aging time. 38
  • 41. 2.4. Hydrothermal method * One of the most common methods for preparation of HAp. * The third most popular method after the conventional precipitation and combination methods. * Reaction of chemicals in an aqueous solution at elevated temperature and pressure. * Hydrothermal synthesis can be considered a chemical precipitation in which the aging step is conducted at a high temperature (typically above the boiling point of water) inside an autoclave or pressure vessel. 41
  • 42. * Performed at lower temperatures than solid-state reactions. * Other low temperature methods such as wet chemical precipitation and sol-gel synthesis require post heat treatment to crystallise the HA; whereas crystalline HA can be produced in one step via hydrothermal synthesis. * The initial stage is mixing the calcium and phosphorous precursors followed maintaining the Ca/P ratio at a constant value of 1.67. The solution is heated in a sealed vessel (autoclave). 42
  • 43. * The mixing is then allowed to age, and subsequently washed and filtered. Finally it is dried in an oven and calcined. * The amount of HAp is limited to the size of the reaction vessel. * The starting reagents and H2O should occupy 50–60% of the autoclave volume. 43
  • 44. Advantages The formed HAp is relatively stoichiometric and highly crystalline. Disadvantages 1) Need expensive equipments 2) Poor capability to control the morphology and size of HAp nanoparticles 44
  • 45. 45 Teflon-lined stainless steel autoclave for hydrothermal synthesis of HAp
  • 46. 46 HAp prepared by hydrothermal method
  • 47. 47 Effect of pH, temperature, and duration of hydrothermal treatment on phase, morphology, and particle size of the CaP powder
  • 48. 2.5. Emulsion method * Two immiscible liquids (such as water and organic) stabilized by the presence of surfactants. * Note: Immiscible: incapable of mixing or attaining homogenity * Surfactants can reduce the surface tension of the immiscible liquids, resulting in a dispersed phase confined to nanometer scale. * Surfactants can subsequently be removed easily by calcination. 48
  • 49. Advantages 1) Reduces nanoparticles agglomeration. 2) Reduces the particle size. 3) Controls the morphology (spheres, rods, discs, etc.). 4) Simple procedure 5) Low synthesis temperature (around room temperature) 6) Occurs without any high-temperature requirement 49
  • 50. 2.6. Sonochemical method * Chemical reactions activated by powerful ultrasound radiation. * The first step is mixing Ca and P precursor, maintaining Ca/P ratio and pH as a constant value followed by the passage of ultrasonic waves, then drying and calcination. Advantages 1) Accelerates the reactions & the growth of HAp crystals. 2) More uniform, smaller & purer crystals with minimal agglomeration. 3) Single phase HAp could be synthesized after 15-60 min sonication. 50
  • 52. 3. High-temerature processes Used to burn or partially burn the precursors. Advantages 1) Avoid undesirable CaP phases. 2) Produce HAp with high crystallinity & good chemical homogeneity. * High-temperature processes can be performed by 2 techniques: 1) Combustion: has received more attention. 2) Pyrolysis 52
  • 53. 3.1. Combustion * A conventional process used to prepare various oxide ceramics. * Combustion processing of HAp involves a very rapid exothermic reaction. * At first, the aqueous solutions of Ca(NO3)2 and (NH4)2HPO4 are first mixed, followed by adding concentrated HNO3 (an oxidizing agent) to dissolve the resulting white precipitate. A single or a mixture of two or more fuels (e.g. citric acid, succinic acid, urea and glycine) is subsequently incorporated into the resulting solution. 53
  • 54. * The reaction can be initiated by heating the mixture in a furnace at a low temperature (e.g. 300 °C), then a sudden increase in temperature to a maximum value. * The final step is the fast cooling of mixture to induce maximum nucleation and to prevent any further particle growth. * Different fuels have proved to be capable of delivering different flame temperatures ranging from 100 to 900 °C (e.g. citric acid: 150 °C; succinic acid: 425 °C; urea: 800 °C; glycine: 890 °C). 54
  • 55. * A vigorous exothermic reaction occurs between the fuel and oxidizer; the gaseous products of this reaction spontaneously combust. This gives rise to a very high local temperature that causes the formation of a solid calcium phosphate powder. This process can be completed in less than 20 minutes. * Depending on the fuel used, the product formed may be either crystalline or amorphous. Both require a calcination step; to remove organic residues and crystallize the phase formed, respectively. 55
  • 56. Advantages 1) Quickly produce powder with high purity in a single step operation. 2) Inexpensive raw materials 3) Relatively simple 4) Good chemical homogeneity 56
  • 58. 3.2. Pyrolysis (spray pyrolysis) * In general, during HAp synthesis, some post treatments and/or long-term aging under elevated temperatures may be required to achieve a high crystalline product. * By contrast, Rapid pyrolysis produces stoichiometric, homogeneous, and highly crystalline HAp. * Pyrolysis method can also be classified under a broad category known as aerosol methods (or gas-phase methods) 58
  • 59. * Particles can be formed from reactants in a gas phase generated by physical evaporation of liquid precursors. * Spray pyrolysis involves spraying the precursor solutions into a flame or a hot zone of an electric furnace using an ultrasonic generator. * This is then followed by reaction of the generated vapors and gases at high temperatures to produce final powder, typically in an aggregated and agglomerated form. 59
  • 60. * In fact, the high temperature leads to complete evaporation of precursor droplets followed by nucleation and growth of nanoparticles in the gas phase. Disadvantages 1) Small decomposition of HAp into α-TCP, because of high temperature of flame (usually above 2000 °C). 2) Secondary aggregates are usually formed. 3) Poor control over the processing variables. 60
  • 62. 4. Synthesis from biogenic sources * Biogenic sources: e.g. biowastes, eggshell and exoskeleton of marine organisms. * Economical and environmental benefits of waste recovery. * Accepted better by the living organs, because of its physicochemical similarity to the human bone apatite. * Calcium carbonate originated from marine species usually exhibits porosity and interconnectivity similar to that found in human bone. 62
  • 63. * The larger and higher macroporosity of the structure, similar to the natural cancellous bone, has been reported to result in earlier bone mineralization during the implantation; and hence corals- generated HAps are claimed to be more beneficial to bone repair applications. * Expected to attract more attention in the near future. * Can produce HAp blocks or particles. 63
  • 65. 65 Cuttlebone, its porosity & interconnectivity
  • 66. 5. Combination procedures * Two or more distinct procedures to synthesize HAp nanoparticles. * Combinations of hydrothermal-mechanochemical, hydrothermal- hydrolysis and hydrothermal-emulsion have received more attention. 66
  • 67. * Combinations of the microwave irradiation with different methods e.g. solid-state and sonochemical methods has been reported. * Combination procedures may improve the characteristics of HAp nanoparticles. 67
  • 68. Microwave-assisted irradiation method * Increased reaction kinetics and rapid initial heating coupled with reduced reaction times when compared to conventional heating methods. * The usual steps of mixing Ca & P, maintaining Ca/P ratio and pH are all followed, and then passing microwave radiation followed by heat treatment. 68
  • 69. References 1. Sadat-Shojai M, Khorasani MT, Dinpanah-Khoshdargi E, Jamshidi A. Synthesis methods for nanosized hydroxyapatite with diverse structures. Acta Biomaterialia 2013; 9: 7591–621. 2. Anitha P, Pandya HM. Comprehensive review of preparation methodologies of nano hydroxyapatite. Journal of Environmental Nanotechnology 2014; 3: 101–21. 3. Cox S. Synthesis method of hydroxyapatite. http://www.mandplabs.com/interface/uploads/files/whitepapers/hydroxyapatite-white-paper.pdf 69