Review of Literature

CHAPTER II
REVIEW OF LITERATURE

An extensive research has been carried out on the magnetic particles during

the past few decades. Among the various magnetic particles the magnetite has been

intensively investigated for their potential applications as MRI contrast agent,

immunoassays, and targeted drug delivery vehicles. These applications demand

the magnetic materials to possess specific size, shapes, surface characteristics

and magnetic properties. The magnetite in nano sized form exhibits

superparamagnetic behavior at room temperature. In other words, they strongly

magnetize under an applied magnetic field but retain no permanent magnetism once

the field is removed [25, 26, 128].

2.1 SYNTHESIS OF MAGNETITE NANOPARTICLES

A number of synthetic methods have already been reported in literatures for

the preparation of magnetite nanoparticles. Among them the co-precipitation

technique is possibly the simplest and most competent chemical pathway to obtain

magnetite nanoparticles. The main advantage of the co-precipitation process is that it

can be easily scaled up for bulk preparation. However, the control of particle size

distribution is limited, because only kinetic factors control the growth of the crystal.

Size controlled magnetite nanoparticles of range 10-40 nm were prepared through

co-precipitation method [129]. The magnetite nanorods with anisotropic property

have been synthesized by reverse co precipitation technique with the support of

magnetic field [130]. The magnetic fluid has been synthesized from

magnetite nanoparticles and hydrophilic surfactant Tween 80 through

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co-precipitation for the applications in MRI and magnetic fluid hyperthermia [131].

In the co-precipitation process, two stages are involved (i) a short burst of nucleation

occurs when the concentration of the species reaches critical supersaturation and

(ii) slow growth of the nuclei by diffusion of the solute to the surface of the crystal.

To produce monodisperse iron oxide nanoparticles, these two stages should be

separate, i.e., nucleation should be avoided during the period of growth [128].

J.H. Wu et al., [25] has reported that the magnetite nano crystals of size 2-4

nm can be synthesized by a chemical co-precipitation method in which the particle

size was controlled by the reaction temperature. The mechanism of the first stage of

formation of magnetite nanoparticle synthesized by chemical precipitation technique

was suggested by Ali-Zade and he has also concluded that the rate of nanoparticle

formation is high in its initial period of time and then found decreasing due to the

decrease in the number of combining molecules in the solution. Thus the rate of

nanoparticle growth depends on its size because the mean size of nanoparticles

depends on the physical properties of the medium (viscosity, temperature, etc.) [26].

Another significant procedure for the synthesis of magnetite nanoparticle is

microemulsion method which involves the thermodynamic stable isotropic dispersion of

two immiscible liquids, where the microdomain of either or both liquids is stabilized by

an interfacial film of surfactant molecules [52]. Certain amount of water is added to a

large amount of non-polar solvent (oil) in the presence of appropriate amphiphilic

surfactant molecules, the homogeneously distributed droplets of water is stabilized by

the surfactant molecules and thus act as micro or nano-reactors for the nucleation as

well as controlled growth of iron oxide nanoparticles.

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Magnetic nanoparticles have been synthesized in a controlled manner using

the microemulsion method in which the particle size and shapes usually vary over a

wide range. Moreover, the working window for the synthesis in microemulsion is

usually quite narrow and the yield of nanoparticles is low compared to thermal

decomposition and co-precipitation methods.

In water-in-oil microemulsions, the aqueous phase is dispersed as

microdroplets (typically 1-50 nm in diameter) surrounded by a monolayer of

surfactant molecules in the continuous hydrocarbon phase. The size of the reverse

micelle is determined by the molar ratio of water to surfactant [27]. By mixing two

identical water-in-oil microemulsion containing the desired reactants,

the microdroplets will continuously collide, coalesce, and break again, finally a

precipitate forms in the micelles. By the addition of solvent, such as acetone or

ethanol to the microemulsion, the precipitate can be extracted by filtering or

centrifuging the mixture. Thus the microemulsion process helps in the formation of

nanoreactor for the nucleation and growth of magnetite nanoparticles [28].

But, large amounts of solvent are necessary to synthesize appreciable amounts of

material. Hence it is not a very efficient process for the formation of magnetite

nanoparticles [76].

Further the thermal decomposition of organometallic compounds for the

synthesis of magnetite nanoparticles is the quite interesting technique [132, 133].

Li et al., [132] has reported that the synthesis of single crystalline magnetite

nanocrystals with octahedron shapes enclosed by (111) planes via single step

thermal decomposition route. The average particle size of the range 8 to 430 nm can

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be finely controlled by modulating the precursor/surfactant ratio. This method offers

a simple way for obtaining monosized magnetite nano octahedra that span over both

superparamagnetic and ferrimagnetic regimes. The surfactant/precursor ratio and the

concentration of the surfactant are the two crucial parameters for synthesizing the

octahedron shaped nanoparticles with uniform sizes.

Phase transformation of maghemite nanoparticles to magnetite nanoparticles

is reported by H.T. Hai et al. [133]. The transformation is explained by the

reduction of maghemite nanoparticles in solution by exploiting the reservoir of

reduction gas (CO) produced from the incomplete combustion reaction of organic

substances in the reactor. Further this technique is interesting as the size, shape and

monodispersity of synthesized magnetite nanoparticles has been retained as such.

Synthesis of single-crystalline magnetite nanocrystals with octahedron shapes

enclosed by (111) planes via a one-step thermal decomposition route is reported.

A non-aqueous route has been reported for the preparation of nanocrystalline

magnetite. In a reaction process, iron (III) acetylacetonate was dissolved in benzyl

alcohol and treated in an autoclave between 175 and 200 C. This approach leads to

monocrystalline magnetite particles with size ranging from 12 to 25 nm [29].

Hydrothermal treatment of magnetite nanoparticle could alter the physical property

like magnetic properties due to the alteration of stoichiometric ratio of (Fe2.95O4)

from its original state (Fe3O4) by this treatment [30].

Vereda et al., [31] have reported the synthesis of single-domain

monocrystalline magnetite particles by the oxidative aging of Fe(OH)2 in presence

of potassium nitrate. High-temperature (210 C) solution-phase hydrolysis is

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reported for the synthesis of colloidal magnetite nanocrystals with the controlled

size of 16 nm and mono size distribution [134]. The size controlled monodisperse

Fe3O4 nanoparticles leading to 20 nm have been reported by high-temperature

(265 C) reaction of Fe(acac)3 in the presence of alcohol, oleic acid, and oleylamine

[41].

The electrochemical technique has control over the particle size in the

synthesis of magnetite nanoparticles by regulating the imposed electro-oxidation

current density (i) or potential (E) to the system. Furthermore, if the synthesis is

performed in the presence of a surfactant, it is possible to avoid the aggregation of

particles. The distances between the cathode and anode play a vital role in this

method. Electrodeposition of magnetite on gold single crystal has been reported by

Nikiforov et al., for the application of spin polarized devices [135]. Cabrera et al.,

[44] has reported the synthesis of magnetite nanoparticles by electro oxidation of

iron in the presence of an amine surfactant that resulted in quite uniform and

spherical particles with mean size between 30 and 20 nm. Further there is a

restriction that the pure Fe3O4 nanoparticles can be achieved only when the potential

is lower than 6 V. The increase in potential or current density leads to the possibility

of formation of higher size homogeneity of the nanoparticles and possibility of

formation of metallic iron as an impurity. In continuance of this work, a report [136]

has been found on the electrochemical synthesis of magnetite nanoparticles in

surfactant free medium with Fe2SO4 as the electrolyte. Both the methods failed to

achieve the monodispersed magnetite nanoparticle by electrochemical method.

Further stabilizing the colloidal solution of MNP by these procedures is difficult and

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also separation of MNP from this solution leads to the oxidation of magnetite to

maghemite and other oxides.

Electrosynthesis of magnetite nanoparticle have been developed by using

complexing agents like NaCl, Na2SO4, NaSCN, Na2S2O3 and sodium citrate in the

aqueous electrolyte [32]. The complexing agents enhance the charge transfer

kinetics and decrease the particle size to the nanometer level. Electro-precipitation

of magnetite nanoparticle in Fe(NO3)3.9H2O ethanol solution has been reported by

Marques et.al. [33]. Two carbon electrodes were employed as cathode and anode

respectively. The electrolysis was carried out at constant 62 V. The magnetite

particles were collected as platelets on the surface of cathode. This work seems to be

working on high voltage and further the synthesized MNP can agglomerate with

each other due to the absence of repulsing agent or surfactant or chelating agents in

electrolyte. Electrochemical synthesis of magnetic nanoparticles within porous silica

microspheres has been carried out by Wang et al. [137]. The mechanism of iron

nanoparticles formation within the mesopores appears to occur through the initial

binding and coating of the pore walls to form wormlike structures within the

mesoporous template. The magnetization behavior of the iron/silica composite

microspheres reveals the iron/silica structure formation in silica spheres and exhibit

essential characteristics of ferromagnetic behavior.

Magnetite particles with an average size of 39 nm and good monodispersity

have been synthesized by coprecipitation at 70 C from ferrous Fe2+ and ferric

Fe3+ ions by a tetra methyl ammonium hydroxide solution, followed by

hydrothermal treatment at 250 C. Further this report explains the conversion of

magnetite to other iron oxide phases at elevated temperature [30].

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Water soluble secondary structural magnetite nanoparticles have been

reported by Yang et,al., [138] through single step hydrothermal technique.

According to this report the single particles get agglomerated which results in the

formation of secondary structures of magnetite. The magnetite nanoparticle could

immobilize on the polyamide fiber by hydrothermal technique for potential

applications [139].

Recent development in the synthesis of magnetite nanoparticles is achieved

by sonochemical technique [140]. In this process, a high energy ultrasonication

creates acoustic cavitations that provide localized heat with a temperature of about

5000 K. At high temperature, the formation and growth of nuclei and the implosive

collapse of bubbles take place and hence monodisperse nanoparticles of a variety of

shapes can be prepared by this method. The ultrasonication helps in preventing

magnetic nanoparticles from aggregation because of the magnetic interactions.

However, at the same time, ultrasonication increases the system temperature as well

as the reaction rate, which would lead to aggregation of the particles.

Sonochemical synthesis of magnetite nanoparticles are investigated by many

researchers. This route yields homogenous solution, good crystallinity of magnetite

and rapid process of magnetite nanoparticle production. The assynthesized

magnetite nanoparticles are hydrophilic and dispersable in ethanol/water mixtures

[141]. Ultrasonic assisted chemical co-precipitation of magnetite nanoparticles

requires no protecting gas hence the MNP can be easily implemented [142].

Ultrasonic assisted in situ synthesis of magnetite nanoparticles is reported by

Feng et al. [143]. Magnetite nano whiskers have been prepared by ultrasonic aided

reduction in concentrated sodium hydroxide solution [144].

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Magnetite nanoparticles were synthesized and then self-assembled into

microparticles via a solvothermal method. In this method, the Fe3O4 microparticles

were assembled with the size range of 80 nm [145]. One-pot solvothermal synthesis

of magnetite nanostructures have been reported with the diverse nanoparticles in

which the hybrid micelle self-assembly mechanism has been developed and further

the assynthesized MNP demonstrates MRI capability of both in vitro and in vivo

[146]. Cauliflower-like Fe3O4 architectures were successfully prepared by a simple

solvothermal route and these architectures were in the range of 200-300 nm in size.

They show a low saturation magnetization of 65 emu/g and an enhanced coercive

force of 247 Oe. These Fe3O4 architectures may have potential applications in

catalysis and biological fields [147]. Further using the solvothermal technique

the various nanostructure of magnetite has been reported instantly and magnetite

hollow spheres with an average diameter of 240 nm have been prepared by this

method [148].

We have reported the cubic magnetite nanoparticles with the size range of

15 6 nm by electro-oxidation of iron in an aqueous medium of ferrous perchlorate.

This electrochemical technique is quite interesting since no micelles or stabilizing

agent or surfactant was used to stop cluster growth, which occurs commonly in

many conventional chemical routes. The MNP shows superparamagnetic property

and hence it could be used for synthesis of VUSPIO for cancer treatment [47].

2.2 SURFACE FUNCTIONALIZATION OF MNP

Although there have been many significant developments in the synthesis of

magnetic nanoparticles, sustaining the stability of these particles for a long time

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without agglomeration or precipitation or oxidation is an important issue. Stability is

a crucial requirement for almost any application of magnetic nanoparticles in

determining the physical properties. Thus, the surface functionalization of MNP

plays a key role in stabilizing magnetite nanoparticles against agglomeration.

The stabilization of the iron oxide particles is decisive to obtain magnetic

colloidal ferrofluids that are stable against aggregation in both biological medium

and magnetic field. The stability of a magnetic colloidal suspension results from the

equilibrium between attractive and repulsive forces. In magnetite, the surface iron

atoms act as Lewis acids and coordinate with molecules that donate lone-pair

electrons. Therefore, in aqueous solutions, the Fe atoms coordinate with water,

which dissociate readily to leave the iron oxide surface hydroxyl functionalized and

these hydroxyl groups are amphoteric and may react with acids or bases. Dependent

upon the pH of the solution, the surface of the magnetite will be positive or negative.

The isoelectric point is observed at pH 6.8. Stabilization of magnetic particles can be

achieved by playing on one or both of the two repulsive forces: electrostatic and

steric repulsion [149, 150].

2.2.1 Functionalization by Polymers

Several approaches have been developed to coat iron oxide nanoparticles,

including in situ coatings and post-synthesis coatings. In the first approach,

nanoparticles are coated during the synthesis. For example, Josephson et al., [151]

have developed a coprecipitation process in the presence of dextran.

The post-synthesis coating method consists of grafting the polymer on the

synthesized magnetic particles [152-154] (polymeric surfactants). The most

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commonly used polymer coating materials reported in the literature are dextran,

carboxymethylated dextran, carboxydextran, starch, arabinogalactan,

glycosaminoglycan, sulfonated styrene-divinylbenzene, polyethylene glycol (PEG),

polyvinyl alcohol (PVA), poloxamers, and polyoxamines [155, 156].

2.2.2 Functionalization by Silica

Silica has been exploited as a coating material for magnetic nanoparticles

[157-162]. Usually, an inert silica coating on the surface of magnetite nanoparticles

prevents their aggregation in liquid, improves their chemical stability, and provides

better protection against toxicity [163]. This coating stabilizes the magnetite

nanoparticles in two different ways [164]. One is by shielding the magnetic dipole

interaction with the silica shell. On the other hand, the silica nanoparticles are

negatively charged. Therefore, the silica coating enhances the coulomb repulsion of

the magnetic nanoparticles.

2.2.3 Functionalization by Rare Metal

Precious metals can be deposited on magnetic nanoparticles through

reactions in microemulsion, redox transmetalation, hydroxylamine seeding.

Gold seems to be an ideal coating owing to its low reactivity. However, it was found

that the direct coating of magnetite nanoparticles with gold is very difficult, because

of the dissimilar nature of the two surfaces. Hence the Au coated magnetic

nanoparticles could be prepared by reverse microemulsion technique where CTAB

was used as surfactant, 1-butanol as a co-surfactant, and octane as the continuous oil

phase [165-177].

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2.3 SYNTHESIS OF HYDROXYAPATITE

During the past few decades, considerable research efforts have been bound

towards the synthesis of various bioceramics for biomedical applications.

Among different classes of bioceramics, HAP is the most emerging bioceramic,

which is widely used in varied biomedical applications. Several methods have been

utilized for the synthesis of this bioceramic which include precipitation technique

[178-180], sol-gel approach [181], hydrothermal technique [182], microwave

irradiation [183-185], ultrasonic [186], ultrasonication assisted mixed template

method [187], freezing method [188], ultrasonic coupled sol-gel method [189].

In precipitation method [178-180], the chemical reactions take place between

calcium and phosphorus ions under a controlled pH and temperature of the solution.

There are many shortcomings in this method in which the precipitated powder is

calcined at high temperature in order to obtain a stoichiometric apatitic structure.

Slow titration and diluted solutions must be used to improve the chemical

homogeneity and stoichiometry within the system. Careful control of the solution

condition is always required in the wet precipitation methods. There are many

reports which states that the decrease of solution pH below about 9 could lead

to the formation of calcium deficient apatite structure. In some cases,

a well-crystallized HAP phase was developed only on approaching a calcination

temperature of 1200 oC [190].

The sol-gel approach provides significantly easier conditions for the

synthesis of HAP. Sol-gel synthesis of HAP ceramics has recently attracted much

attention. Sol-gel process refers to a low-temperature method using chemical

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precursors that can produce ceramics and glasses with better purity and homogeneity

[191, 192]. This process is becoming a common technique to produce ultra fine and

pure ceramic powders, fibers, coatings, thin films, and porous membranes.

It is clear from the reports that the sol-gel method of synthesizing

hydroxyapatite requires two critical factors like appropriate aging time and

temperature of the precursor solution in developing the apatitic phase. Appropriate

aging is required to complete the chemical reaction between calcium and

phosphorus precursors. During the chemical reaction between calcium and

phosphorus precursors, an intermediate compound is formed. This compound

determines the resulting apatite phase after calcination. A preparation time of

24 h or longer is generally reported in literature as required to form a desirable

intermediate compound.

There are many reports on the synthesis of HAP by hydrothermal method [193,

194]. This method yields HAP with high degree of crystallinity. Though HAP powders

can be synthesized with high crystalline nature, it involves long reaction times and thus

leads to agglomeration which hinders this method for the synthesis of HAP [195, 196].

But the researchers have attempted for the synthesis by the addition of various

complexing agent which is responsible for the morphological changes in the resultant

HAP [197]. Eventhough synthesis of HAP is well known by several chemical routes

such as hydrothermal [198, 199], solid-state reaction [200], solgel [201], emulsion and

microemulsion synthesis [202] and microwave irradiation [203, 204], wet, dry or

hydrothermal route, all these methods are tedious and time consuming.

The most reported method of synthesizing HAP is precipitation process.

This process is simple, low cost and suitable for industrial production but the

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product is generally obtained as large size crystalline particles or agglomerates of

nanoparticles. Further, when a calcium phosphate material is precipitated, the used

molar ratio of Ca/P, the synthesis temperature, the method of agitation, the pH of the

reaction mixture, the rate of adding the reagents, the ageing time, etc. are variables

that affect the final characteristics of the precipitated powder [205, 206]. But for

some specific biomedical applications it is necessary to produce HAP crystallites

with reduced particle size and high surface areas, or controlled morphology, or both.

On the other hand, recent investigations reveal that the reactivity of chemical species

in solution being involved in a synthesis process can be stimulated by ultrasonic

irradiation (UI) of the reaction mixture. It has been recognized that this type of

treatment causes cavitations in an aqueous medium inducing formation, growth and

collapse of micro bubbles. This intense agitation process leads to events of

dissolution and precipitation of solids through which a reduction of particle size and

surface activation of solid materials is produced [207-209].

2.3.1 Advantages of Ultrasonication in the Synthesis of HAP

Kim and his coworkers have reported that the hydroxyapatite particles were

synthesized with the aid of ultrasonic radiation. It was observed that the

hydroxyapatite particles synthesized by this method also possess high phase purity

and good thermal stability. They have also found that the morphology of the

particles can be controlled by the variation of the ultrasonic irradiation time [210].

Fang et al., [211] have compared the synthesis of HAP from a mixture of

Ca4(PO4)2O and CaHPO4 (H2O) (brushite) at 38 0.5 C. The reaction time required

to prepare hydroxyapatite without sonication was 3 h whereas the reaction time was

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reduced to 15 min with sonication. The morphology of the obtained product was

also different. From the reports it was observed that the nanohydroxyapatite particles

synthesized with the aid of ultrasonic radiation possess high phase purity and good

thermal stability.

Gopi et al., [189] have demonstrated the synthesis of hydroxyapatite by

ultrasonic coupled solgel synthesis. The powders synthesized by ultrasonic coupled

solgel synthesis showed remarkable reduction in the particle size when compared

with the conventional solgel method. Hence these powders can be used in the

biomedical applications. Recently, he has also reported the synthesis of

hydroxyapatite nanoparticles by a novel ultrasonic assisted mixed template direct

method [187]. The effect of ultrasonic irradiation time on the crystallinity and the

size of HAP nanoparticles in presence of glycineacrylic acid results in a hollow

sphere template were investigated. As a result of the ultrasonic treatment HAP

nanoparticles with decreased crystallinity and size was achieved. From the

inspection of the above results it is confirmed that the size of the HAP nanoparticles

decreases with increasing ultrasonic irradiation time. Hence it is well proved that the

desired size and morphology of HAP can be obtained for the biomedical

applications.

2.4 FUNCTIONALIZATION OF MNP WITH HAP

Functionalization of the MNPs, for practical application, is indispensable to

improve functions such as biomolecular recognition and stability in tissues and cells.

Many researchers have explored the coating materials on MNP by means of the

introduction of functional groups, ligandreceptor reactions and with the improved

structural stability and solubility in tissues, such as dextran [212], polyethylene

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glycol [213], silica [214], gold [215] and hydroxyapatite (HAP, Ca10(PO4)6(OH)2)

[56, 216]. HAP shows excellent biocompatibility and has been investigated as a drug

delivery carrier [217]. Functionalization performs a better role in improving the

chemical properties of individual HAP nanoparticles.

HAP is used in orthopedic and dental surgeries and odontology for wadding

[218] or surface [219] coverings, despite their weak mechanical properties

[220, 221]. Due to these disadvantages considerable interest has recently been

shown in the substitution of calcium in the HAP structure by metals [222, 223] or

ions, as for instance, silicon and magnesium [224], lead [225], titanium and

zirconium [226] and aluminum [227] as reinforcing agents to improve the

mechanical properties of the HAP. The presence of iron in the apatite structure

seems to be important because iron is a vital element in the circulatory system and

essential for the functioning of numerous proteins in cells [66, 67].

Bioceramic materials containing iron oxide receives considerable

technological applications in biomedicine. As an example, ferrimagnetic bioglass

ceramics (FBC) was introduced for hyperthermic treatment of bone cancer

[55, 68-70]. Recent report shows that the presence of iron oxide in hydroxyapatite

will enhance the radiopacity and the osteoblast proliferation activity than pure HAP

[57]. HAP with different iron concentrations obtained by solgel method at different

temperatures has also been used for hyperthermia treatment and bone tumor.

Furthermore the micro hardness (mechanical strength) of the apatite structure can be

achieved by heat treating with iron oxide [58]. The superparamagnetic

hydroxyapatite has the potential to serve as non-viral vector for the delivery of genes

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to select sites in the brain and also shows the elevated binding affinity with plasmid

DNA [57]. The Fe3O4 doped HAP exhibit a better solubility in physiological

solution rather than HAP [58]. There are many reports on the synthesis of pure HAP

and MNP [228-230] but only a few researchers have worked on the functionalization

of the magnetic nanoparticle with HAP [55, 57, 70]. Recently, the synthesis of

HAP-ferrite composite by ultrasonic spray pyrolysis at high temperature of 500 C

has been reported using a suspension composed of crushed ferrite particles and

calcium/phosphate precursors [231]. Wu et al., have reported the synthesis of

magnetic HAP through co-precipitation method [56] and the product has been aged

for 1020 h and before it was dried to get the resultant product. The obtained

product was in the aggregated form which limits its application in biomedicine. Poly

vinyl alcohol (PVA) stabilized ferrofluids hydroxyapatite nanocomposite has been

reported by Mir et al., through wet coprecipitation method [232] and the product has

been aged for seven days. The method is time consuming and this prolonged

duration results in aggregated product. The synthesis of m-HAP through wet

coprecipitation method need stabilizer, more time consuming and the product exists

in aggregated form. Therefore this method is not an efficient technique to synthesize

a monodispersed m- HAP.

Recently we reported the ultrasonic assisted synthesis of magnetic

hydroxyapatite nanocomposite with excellent surface morphology. The ultrasonic

irradiation technique is simple, rapid and efficient. The reaction could be performed

at ambient condition like room temperature. The sonochemical route does not need

any stabilizers or template hence the product is free from impurities and it can be

used for synthesis of m-HAP nanocomposite at large scale [186].

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