1.1. Introduction To Nanotechnology: Chapter-1

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1.1. Introduction to nanotechnology

The nanoscience and nanotechnology are a relatively recent developing
trend and showing widespread activities in scientific research. They have their
roots in the idea of some scientists of the last century. In 1959, Richard
Feynman, the American physicist assertively delivered legendary talk entitled
“There's Plenty of Room at the Bottom.” It makes others inspired to bring new
revelations in the field of the nanotechnology. The process which enables the
manipulation of individual atoms and molecules by using proper tools and
techniques resulting in another proportionally smaller set on the desired scale
had been described by Feynman [1-2]. The term “nanotechnology” was
originally coined by the Japanese scientist, Norio Taniguchi of the Tokyo
University of Science, in a 1974 [3]. However, the term was unused until 1981,
when Eric Drexler published his first paper on the nanotechnology. He was
unaware of the fact that the nanotechnology term was prior used by Taniguchi
[4-6]. The concept of nanotechnology had been developed and popularized by
K. Eric Drexler. He founded the field of molecular nanotechnology, which
includes engineered nanosystems operating on the molecular scale and
associates of the molecular assembler [7]. His ideas about the nanotechnology
can be quoted as; it is the principle of manipulation atom by atom, through
control of the structure of matter at the molecular level, which entails the
ability to build molecular systems with atom-by-atom precision, yielding a
variety of nanomachines. With respect to this reality, in 1998 National Science
and Technology Council (NSTC) of the USA created a working group of the
nanoscience, technology, and engineering. Succeeding this, they also
programmed the National Nanotechnology Initiative (NNI) in 2001 with the
objective of initiation to create a common platform for the academic institute,
industries, and private sector for working on this new technology. Following
this example, nowadays, the field of the nanoscience and nanotechnology are
being rapidly developed and getting heavily invested in most of the advanced
Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 1

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countries in the world and many developing countries including China and
India [8].
The word „Nano‟ originates from the Greek word „Nanos‟, which means
dwarf or extremely small. The nanotechnology and nanoscience are defined by
various means, which are often interchangeable. The nanotechnology could be
defined as, “The science involving designing, synthesis, characterization, and
application of materials which are characterized by at least one dimension in
the nanometer range where 1 nm = 10-9 m.” In the nanotechnology, two types
of perspectives named as, bottom-up and top-down are used. With the “bottom-
up” approach, nano scale materials are created by breaking down larger
materials physically or chemically. While in the “top-down” perspective, nano
scale objects are assembled atom-by-atom or molecule-by-molecule. The
nanotechnology includes the production and application of physical, chemical,
and biological systems at scales, which range from individual atoms or
molecules to submicron dimensions. It is also concerned with the integration of
the resulting nanostructures into larger system [9-11].
By merging the nanoscience with technology, new, improved, and
potential nanotechniques are developed, replacing older ones. The
nanotechnology has been tremendously interested since materials ranging in
nanoscale possess novel functionalities leading to have many imminent
technological applications. There is an expanding scope of creating new
knowledge, which explains the size dependent evolution of various physical
properties, and previously unnoticed features, and other things. Various basic
and applied scientific disciplines have been holed promptly within the
empirical discipline of nanotechnology. It has recently become one of the most
cardinal areas having potentiality to aid existing technologies with more
efficiency and better outcome [8]. The nanotechnology shows great promise for
providing us in the near future with many breakthroughs that will change the
direction of technological advances in a wide range of applications. Improved

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and more desirable products can be made by using nanosystems than

conventional materials. The nanotechnology is surely going to have a strong
impact in areas such as electronics, communication, medical device, medicine,
cosmetics, architectural, textile, agricultural, food, metallurgy, defense and
security, space, and many more [12]. Figure 1.1 shows the applications of the
nanotechnology in various disciplines.
Figure.1.1. Applications of the nanotechnology in various disciplines
1.2. Nano-bio technology

1.2.1. Biotechnology
The biotechnology is a broad scientific discipline in which biological
processes, organisms, cells or cellular components are exploited to develop
new technologies. Any high tech application that uses biological systems,
living organisms, or derivatives thereof, to make or modify products or
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processes for specific use can be encircled under the science of biotech. In the
field of biotechnology, the most efficient methods and techniques are studied
and developed. The biotechnology leads to get useful products for human by
using viable microorganisms, cells, and tissues of plants or animals, or even
certain functional components of organisms. The concept of biotechnology
encompasses a wide range of procedures for modifying living organisms
according to human purposes. New tools and products developed by
biotechnologists are useful in research, agriculture, industry, and many more.
Modern usage also includes genetic engineering as well as cell and tissue
culture technologies. The biotechnology also has an impact on the pure
biological sciences such as cell biology, molecular biology, microbiology,
embryology, genetics, biochemistry, and animal cell culture [13-15].
1.2.2. Fundamentals of nano-bio technology
Figure.1.2. The simultaneous interface of the nanoworld, bio systems, and

technology leading to the nano-bio technology

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The nano-bio technology is the simultaneous interface of the nanoworld,

bio systems, and technology as presented in figure 1.2. The nano-bio
technology is that branch of the nanotechnology that deals with biological and
biochemical applications or uses. The nano-bio technology takes most of its
fundamentals from the nanotechnology. The nano-bio technology often studies
existing elements of living organisms and nature to fabricate new nano-devices.
The nano-bio technology can be defined as the ways that nanotechnology is
used to create devices to study biological systems. On the other hand, Bio-nano
technology can be defined as the study of how the nanotechnology is
incorporated to study biological systems and to adapt these biological motifs
into improving existing nanotechnologies or creating new ones. It can be
stated, as the nano-bio technology is a specific application of the
nanotechnology, whereas the bio-nano technology is an essential branch of the
biotechnology. For example, many new medical technologies, which involve
nanoparticles as delivery systems or as sensors are categorized under the nano-
bio technology, since they use the nanotechnology to advance the goals of
biology. Conversely, DNA nanotechnology or cellular engineering would be
categorized under the bio-nano technology, since they involve working with
biomolecules on the nanoscale.
The molecular behavior at nanometer scales governs living systems,
where the disciplines of chemistry, physics, and biology all converge. Such
multidisciplinary approach will stimulate progress in the nano-bio technology.
The nano-systems in biology are the most complex and highly functional nano-
scale materials and machines, which occur in nature. Incredible precision
regarding regulation and control of biological systems is seen due to the
proteins and nucleic acids. The molecular building blocks of cells, such as
lipids, carbohydrates, and similar non-biological molecules are examples of
materials that possess unique properties due to their size, folding patterns at the
range of nanosize. Many nanotechnologists are approaching towards new

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materials and devices by getting inspired from nanosystems within biology [16-
18]. Understanding of biological processes on the nanoscale level is a strong
driving force behind the development of nano-bio technology [19]. The cells
are the fundamental unit of the living organisms, which are across round about
10 μm. Within the cell, the cell parts are much smaller and are in the sub-
micron size. The comparison of these cells constitutes with manmade
nanomaterials gives the idea to use the nanotechnology for nano-bio
applications (Figure.1.3). For example the nanosize protein of 5 nm, is
comparable with the dimensions of the smallest manmade nanoparticles, which
can be used as very small probes to spy at the cellular level [20].
Figure.1.3. Comparison of nanomaterials and bio systems of nanoscale [21]
1.3. Magnetic nanoparticles for nano-bio applications

Among the different types of nanomaterials, magnetic nanoparticles
(MNPs) are tremendously being focused due to their various exciting properties
that make them useful in a wide range of biomedical applications. The use of

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MNPs through the nanotechnology for biomedical applications is currently

gaining huge attention since the MNPs have the immense potential for
applications in diverse areas of biology and medicine. Scientific communities
are widely reporting the potentiality of MNPs for diagnostic and therapeutic
biomedical applications. Comparable size scales of MNPs and biological
materials promote their self to be used in biological and biomedical
applications. At the nanometer scale, the properties of materials seem to be
different from those at the macro scale. The distinctive physical and chemical
characteristics of nanomaterials like a high surface area to volume ratio,
tunable optical emission, unique electrical and magnetic behavior, and many
more other novel properties can be exposed to gain wide spectrum of nano-bio
applications. However, some drawbacks have limited the use of MNPs in nano-
bio applications, like potential toxicity, immunogenicity, decreased efficacy,
and cost of production. To overcome these drawbacks and enhance the
efficiency of MNPs, detailed studies have been undertaken to understand
fundamental interactions between nanomaterials and the biological
environment. For example, various nanoparticle synthesis techniques have been
developed for control over nanoparticle surface chemistry. These types of
perspectives enable to identify ways in which the MNPs interface can be
controlled to affect interactions with biomolecules for beneficial nano-bio
applications [22]. Essential aspects of MNPs regarding its nano-bio
applications are discussed in the following section.
1.3.1. Nano scaling laws of magnetic nanoparticles

Nano scaling laws for magnetic nanoparticles are critical, since they
play an important role in the designing of magnetic nanoparticles, so that they
gain optimized characteristics resulting in enhancing their use in nano-bio
applications. Regarding these, magnetic nanoparticles with precisely tuned
size, shape, and composition have been developed [23-26]. In magnetic

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nanoparticles, many desired phenomena have been observed that are distinct
from their bulk counterparts. It is found that the fundamental magnetic
properties such as coercivity (Hc) and susceptibility (x) are no more restricted
as material characteristics. These properties are adaptable to variations in their
size, shape, and composition. This results into utilization of these scaling
relationships accordingly to tune magnetism, from the ferromagnetic regime to
the superparamagnetic regime, which shows zero coercivity regarding
nanoscale [27-30]. Figure 1.4 represents how size affects on the magnetic
characteristics like coercivity, mass magnetization, and remanence of magnetic
nanoparticles.
Figure.1.4. Effect of size on the magnetism (e.g., coercivity, mass

magnetization, remanence) of magnetic nanoparticles [31]
1.3.1.1. Size effect

One of the interesting size-dependent phenomena of nanoparticles is
superparamagnetism. The volume of ferromagnetic material in which all
magnetic dipoles are present in groups and aligned in the same direction of the
exchange forces is referred as a magnetic domain. The ferromagnetism can be
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distinguished from paramagnetism regarding the concept of domains. When the

size of a ferromagnetic material is reduced below a critical value, it becomes a
single domain. The state of lowest free energy of ferromagnetic particles has
uniform magnetization for particles smaller than a certain critical size and has
non-uniform magnetization for particles larger than a certain critical size. The
former ones are referred to as a single domain particle, while the latter is called
multi-domain particles. Further, the particles show superparamagnetism when
the size of single-domain particles decreases below a critical diameter, so that
the coercivity (Hc) gains zero value and this impart superparamagnetism to the
particles. In superparamagnetic particles, zero coercivity and absence of
hysteresis are achieved due to thermal fluctuations, which are strong enough to
demagnetize spontaneously a previously saturated assembly. These results into
magnetization of the nanoparticles in the presence of an external magnetic
field, but the nanoparticles revert to a nonmagnetic state when there is no
magnetic field [32-34] (Figure.1.5).
Figure.1.5. Magnetic behavior of superparamagnetic nanoparticles under the

influence of external magnetic field
The principle behind this can be explained as; the main factor in
determining the superparamagnetic properties of the MNPs is magneto-
crystalline anisotropy, which originates from the spin–orbital (L–S) coupling at
crystal lattices. For MNPs with a cubic spinel structure, the great difference in

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L–S coupling strength between cations determines the relative magnitude of

magnetic anisotropy. The magnetic anisotropic energy barrier from a spin-up
state to spin-down state of the magnet is proportional to the product of the
magnetic anisotropy constant (Ku) and the volume (V) of the magnet. When the
thermal energy of the nanoparticle is sufficient to invert readily the magnetic
spin direction, it leads to magnetic fluctuation resulting in a net magnetization
of zero, and leading to superparamagnetism behavior. This transition
temperature from ferromagnetism to superparamagnetism is known as the
blocking temperature (Tb) [35].
1.3.1.2. Composition effect
Figure.1.6. (a) Ferrimagnetic spin structure of inverse spinel ferrites, (b) Spin
moments of various metal-doped MEIO nanoparticles (MeFe2O4, Me = Mn, Fe,
Co, Ni), (c) Mass magnetization values of various metal-doped magnetic-
engineered iron oxide nanoparticles (MeFe2O4, M) Mn, Fe, Co, Ni) [31]
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The anisotropic constant (Ku) is known to be strongly correlated with

various anisotropies such as shape, magneto crystalline, and composition [36-
38]. The magnetism of nanoparticles can be changed promptly by having a
compositional modification of nanoparticles by the adoption of magnetic
dopants [39]. For example, magnetism-engineered iron oxide (MEIO), Fe3O4,
nanoparticles possess a ferrimagnetic spin structure where Fe2+ and Fe3+
occupying Oh sites parallel to the external magnetic field and Fe3+ in the Td
sites of fcc (face centered cubic)-packed oxygen lattices parallel against the
direction of the field (Figure.1.6.a). Fe3+ possesses a d5 electron configuration
and Fe2+ has a d6 configuration with a high spin state. The total magnetic
moment per unit (Fe3+)Td(Fe2+Fe3+)OhO4 is found to be approximately 4μB. The
net magnetization gets changed by incorporating a magnetic dopant Me2+ (Mn,
Co, Ni) with electron configurations of d5, d4, and d3 respectively, to replace Oh
Fe2+ (Figure.1.6.b). The magnetic moment per unit MnFe2O4, CoFe2O4, and
NiFe2O4 is estimated as 5μB, 3μB, and 2μB, respectively because of
compositional change. Due to such compositional effects, the mass
magnetization value at 1.5 T gradually decreases from 110 emu to 101, 99, and
85 emu per mass of magnetic atoms (emu/g (Me+Fe)) for MnFe2O4, MEIO,
CoFe2O4, and NiFe2O4, respectively (Figure.1.6.c) [31].
Therefore, it can be stated as, to understand the behavior of existing
materials and to develop novel materials and to impart superior properties to
them, the nano scaling laws of engineered magnetic nanoparticles are important
factors. By considering these important factors, artificially engineered magnetic
nanoparticles can be used as promising tools for nano-bio applications.
1.3.2. Strategies to design magnetic nanoparticles

Magnetic nanoparticles have their own advantages that make them
useful for many applications in different kind of sectors. Tailoring and
conjugating them with other molecules and components make them more

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efficient for nano-bio applications. The combination of structural

characteristics of MNPs and integrated functions of multicomponent makes
MNPs more efficient. By this way, magnetic nanoparticles have the ability to
attract research interest and could lead to new opportunities in nano-bio
applications. Generally, there are two kinds of strategies to fabricate magnetic
nanoparticle-based multifunctional nanostructures. These are molecular
functionalization of magnetic nanoparticles and combination of magnetic
nanoparticles with other nanocomponents [40-41] (Figure.1.7).
Figure.1.7. The strategies to fabricate multifunctional magnetic nanoparticles

and their potential nano-bio applications (a) Molecular functionalization of
magnetic nanoparticles (b) Combination of magnetic nanoparticles with other
nano components [40]
1.3.2.1. Molecular functionalization of magnetic nanoparticles

The first strategy (Figure.1.7.a) one could adopt reliably is molecular
biofunctionalization. Magnetic nanoparticles can be forced to bind to specific

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biomolecules so that; they could detect or purify the biological entities after
being modified by biomolecules. By coating magnetic nanoparticles with
biofunctional molecules (e.g., antibodies, ligands, or receptors), it makes
magnetic nanoparticles enable to interact with a biological entity with high
affinity. Such biofunctional magnetic nanoparticles that respond to a magnetic
field confer features like high selectivity, specificity, sensitivity, and
magnetism, which make them efficient for many nano-bio applications. Most
of surface modification strategies for molecular functionalization of magnetic
nanoparticles are based on or derived from self-assembled monolayers [40, 42-
43].
1.3.2.2. Combination of magnetic nanoparticles with other

nanocomponents
The second strategy (Figure.1.7.b) is to combine magnetic nanoparticles
and other functional nanostructures by coating or sequential growth, resulting
in a single entity, which confers multiple functions of magnetic particles in
nanoscale. By doing this, an excellent platform could be provided to magnetic
nanoparticles for nano-bio applications. For example, magnetic nanoparticles
are used as seeds to grow the semiconducting chalcogenides, which produce
the core-shell or heterodimer nanostructures with both magnetic and
fluorescent properties. This can be used to demonstrate the intracellular
manipulation of nanoparticles and as a promising candidate for dual-functional
molecular imaging, which is a combination of magnetic resonance and
fluorescence imaging. Controlled drug delivery can be achieved by the
encapsulation of a potential anticancer drug by iron oxide nanoshells resulting
in the yolk-shell nanostructures as a promising novel nanodevice. Integration of
magnetic and metallic nanoparticles offers two distinct surfaces and properties.
It tallows different kinds of functional molecules to attach onto the specific
parts of the heterodimers, which may be bound to multiple receptors or, also

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could act as agents for multimodality imaging. Based on the magnetic

nanoparticles, one can combine QDs to exhibit magnetic and fluorescent
properties. It sequentially grows into metallic nanocomponents or form exotic
nanostructures, for example yolk-shell nanoparticles for the exploration of
nanomedicine [44-49].
1.4. Ferrites
Among various developed magnetic nanoparticles, ferrite nanoparticles
are the most explored magnetic nanoparticles up to date. Ferrites are a type of
ferrimagnetic ceramic compounds chemically composed of iron oxide such as
hematite (Fe2O3) or magnetite (Fe3O4) and oxides of other metals. They are
both ferrimagnetic electrically nonconductive in nature, so that, they can be
magnetized or get attracted to a magnet. Once the ferrite particles become
smaller in the nanosized range, they become superparamagnetic which prevents
self-agglomeration since they exhibit their magnetic behavior only when an
external magnetic field is applied. With the external magnetic field switched
off, the remanence falls back to zero. Ferrites can be divided into two families
based on their magnetic coercivity and bearing resistance to demagnetization.
These are soft ferrites and hard ferrites. Soft ferrites have low coercivity. The
low coercivity means the material's magnetization can be easily get reverse
direction without dissipating much energy (hysteresis losses); while the
material's high resistivity prevents eddy currents in the core, which results into
loss of energy. The examples of soft ferrites are manganese-zinc ferrite, nickel-
zinc ferrite etc. Hard ferrites have a high coercivity, remanence, and magnetic
permeability. The high coercivity leads the materials to be resistant to be
demagnetized, which is an essential characteristic for a permanent magnet. The
examples of hard ferrites are strontium ferrite, barium ferrite, cobalt ferrite etc.
[50]. Ferrites can be grouped into three classes, namely hexagonal ferrite,
garnet and spinel ferrite based on the crystal structure.

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1.4.1. Spinel ferrites

Among the different types of ferrites, spinel ferrite nanoparticles have
been a subject of interest in recent years due to their promising technology
applications, especially in nano-bio applications [51-52]. Extensive work on
structural and magnetic characterization of spinel ferrite in the form of nano-
size has been done by many workers [53-54]. The first systematic study
reported by Hilpert in 1909 regarding preparing of spinel ferrites was reviewed
by S. Amiri and H. Shokrollahi [55]. About 20 years after Hilpert's work, in
France, Forestier [56] started his chemical study on the preparation of various
ferrites, as well as the measurement of their saturation magnetization and Curie
temperature. In 1932, Japan's Kato and Takei [57-58] studied the formation of
solid solutions between some ferrites [59]. Efforts continued on ferrites until
researchers found some applications of these materials, especially for cobalt
ferrite in the world of medicine. The spinel ferrites are the important class of
magnetic materials because of their tremendous potentiality to have numerous
applications in several fields. Research on nano-size spinel ferrite has been
considerably increased due to their superior properties and applications in new
fields like magnetic drug delivery, catalyst, sensors etc. [60]. The combination
of magnetic and electrical properties makes spinel ferrite useful in many
technological applications. The basic electrical and magnetic properties of
ferrite can be modified to suit the desire applications. The modification of the
properties of ferrite can be brought by various ways. One of the important ways
is to use different synthesis methods to optimize the synthesis parameters. The
size and properties of spinel ferrite nanoparticles greatly depend on pH, fuel,
stirring time and speed, metal nitrates to fuel ratio, preparative parameters, etc.
[61-62]. The spinel ferrite is having the chemical formula MeFe2O4 (where M-
is a divalent metal ion such as Co, Ni, and Mn etc.). The wide variations in the
structural, electrical and magnetic properties are brought due to the cations of
different valence, which accommodate in the interstitial sites of spinel ferrites.

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1.4.1.1. Structural properties of spinel ferrites

The spinel ferrite structure MeFe2O4, where „Me‟ refers to the metal, can
be described as a cubic close-packed arrangement of oxygen atoms. Me2+ and
Fe3+ are two different crystallographic sites, which have tetrahedral and
octahedral oxygen coordination (termed as A and B-sites, respectively), so the
resulting local symmetries of both sites are different. The spinel structure
contains two cation sites for metal cation occupancy. There are 8 A-sites in
which the metal cations are tetrahedrally coordinated with oxygen, and 16 B-
sites, which possess the octahedral coordination. When the A-sites are occupied
by Me2+ cations and the B-sites are occupied by Fe3+ cations, the ferrite is
called a normal spinel. Structural formula of such ferrites is Me2+[Fe23+]O42−.
For example, this type of distribution takes place in zinc ferrites
Zn2+[Fe2+Fe3+]O42−. The structure is referred to as an inverse spinel when A-
sites are completely occupied by Fe3+ cations and the B-sites are randomly
occupied by Me2+ and Fe3+ cations. Structural formula of these ferrites is Fe3+
[Me2+Fe3+] O42−. For example, magnetite Fe3O4, nickel ferrites (NiFe2O4), and
cobalt ferrite (CoFe2O4) have an inverse spinel structure. In mixed spinels, the
cation distribution possesses an intermediate degree of inversion where both
sites contain a fraction of the Me2+ and Fe3+ cations. Structural formula of these
ferrite is Me1−δ2+Feδ3+ [Meδ2+Fe2−δ3+]O42−, where the degree of inversion is δ.
MnFe2O4 represent this type of structure, having inversion degree 0.2.
Magnetically, spinel ferrites display ferrimagnetic ordering. The magnetic
moments of cations in the A and B-sites are aligned parallel with respect to one
another. Between the A and B-sites, the arrangement is antiparallel and as there
are twice as many B-sites as A-sites, there is a net moment of spins yielding
ferrimagnetic ordering for the crystal. The choice of metal cation and the
distribution of ions between the A and B-sites, therefore, offer a tunable
magnetic system [63-64]. The structure of spinel ferrite is presented in figure
1.8.

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Figure.1.8. Spinel ferrite structure [65]
1.4.1.2. Magnetic properties of spinel ferrites

Neel explained magnetic properties of ferrites, postulating that the
magnetic moments of ferrites are a sum of magnetic moments of individual
sublattices. In the spinel ferrites, these are sublattice A consisting of cations in
tetrahedral positions and sublattice B with cations in octahedral positions.
Different values are seen, when there is an exchange interaction between
electrons of ions in these sublattices. Interaction between magnetic ions of
sublattices A and B (A–B interaction) is the strongest. A–A interaction is
almost ten times weaker and B–B interaction is the weakest. The complete or
partial antiferromagnetism (ferrimagnetism) is due to the dominant A–B
interaction [66].
1.4.2. Cobalt ferrites: A promising spinel ferrite for nano-bio applications

Among the different spinel ferrites cobalt ferrite (CoFe2O4) with inverse
spinel structure is promising magnetic nanoparticles for different kind of
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technological applications. Many researchers have reported magnetic properties

of cobalt ferrite nanoparticles with a view to understand magnetism at the nano
scale and their possible practical applications [67]. Cobalt ultra-fine powders
[68-69] and films [70-71] have attracted considerable attention for their wide
range of technological applications such as transformer cores, recording heads,
antenna rods, memory, ferrofluids, biomedical applications, sensors, and many
more [72-73]. In the case of CoFe2O4, the Co cation occupies one-half of the
octahedral coordination sites. Half the Fe3+ cations occupy the other half of the
octahedral coordination sites as well as all of the tetrahedral coordination sites
[74-76]. The crystal structure of CoFe2O4 is shown in figure 1.9.
Figure.1.9. Crystal Structure of CoFe2O4 where green atoms are Co, pink
atoms is Fe and blue atoms are O [77]
CoFe2O4 is a conventional magnetic material, with a Curie temperature
(Tc) of about 793 K, CoFe2O4 possess high magneto-crystalline anisotropy,
high coercivity, high electrical properties, moderate saturation magnetization,
good mechanical properties and greater physical and chemical stability, which
make it a good candidate for various applications. The anisotropy constant of

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bulk cobalt ferrite is between 1.8 x 106 and 3.0 x 106 erg/cm3. The saturation
magnetization of CoFe2O4 is 80.8 emu/g and 93.9 emu/g at room temperature
and 5 K respectively.
1.5. Nano-bio applications of magnetic nanoparticles

The use of magnetic nanoparticles for biological and clinical
applications is undoubtedly one of the most challenging research areas in the
field of nanomagnetism. In recent years, lots of attention has been focused on
the development of MNPs due to their fundamental properties. First, they have
size, which generally ranges from a few nanometers to tens of nanometers,
which places them at dimensions that are smaller than or comparable to those
of a cell (10–100 µm), a virus (20–450 nm), a protein (5–50 nm) or a gene (2
nm wide and 10–100 nm long). This means that they can „get closer‟ to a
biological entity of interest. Certainly, they can be coated with biomolecule to
make them interact with or bind to a biological entity, thereby providing a
controllable means of „tagging‟ or addressing it. Second, the MNPs can be
manipulated by using an external magnetic field, which provides a huge
advantage and opens up many nano-bio applications in vivo. Third, MNPs
exhibits interesting size dependent superparamagnetism. Such MNPs are highly
preferred due to their ability of being magnetized upon exposure of the
magnetic field, but they have no permanent magnetization (remanence) once
the field is turned off. MNPs specifically superparamagnetic nanoparticles,
based upon their unique physical, chemical and thermal properties, offer high
potential and have been proposed extensively for nano-bio applications. MNPs
are eligible for various biomedical applications, such as magnetic
hyperthermia, magnetic resonance imaging, target drug delivery, bacteria
detection, cell labeling, magnetic separation, and enrichment of DNA [78-81]
(Figure.1.10). Brief descriptions of these bioapplications of magnetic
nanoparticles are given in following section.

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Figure.1.10. Different nano-bio applications of magnetic nanoparticles
1.5.1. Magnetic hyperthermia

Hyperthermia is one of the modes of cancer treatment, where cancer
cells are killed by exposing it to high temperatures (42-46 ºC). Hyperthermia
offers an attractive approach for the treatment of cancer because, as a local
therapy, it is associated with fewer side effects in comparison with chemo and
radiotherapy. Additionally, it can be used in combination with all conventional
modes of treatment. Various clinical trials have demonstrated the efficacy of
such combinations [82-85]. When Ferro fluids, a mixture of nanoscale
magnetic particles and water, are infused into a tumor and exposed to an
external alternating magnetic field, it can become a heat source capable of
raising temperatures high enough to weaken or destroy the cancerous cells as
illustrated in figure 1.11.

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Figure.1.11. Illustration of magnetic hyperthermia [86]

The absorption efficiency of any material to generate heat due to an
applied magnetic field is measured in terms of specific absorption rate (SAR)
or specific loss power (SLP). These terms are generally used to define the
transformation of magnetic energy into heat [87]. MNPs based hyperthermia
involves distribution of particles throughout the targeted tumor site followed by
the generation of heat to the tumor using an external AC magnetic field. A
mechanism of heating by magnetic nanoparticles includes three types of
processes, which are hysteresis losses, Neel and Brownian relaxation. The
crystal size and composition of the particles play an important role in the
relative contribution of each process. Magnetic nanoparticles with core
diameters of less than 20 nm are single-domain particles, which mean they
consist of only a single organized crystal. The external rotation (Brownian) and
internal (Neel) diffusion of the particle‟s magnetic moment combinely govern
magnetization relaxation in such small magnetic nanoparticles with a negligible
contribution of hysteresis loss [88]. The characteristics of well-dispersed
magnetic nanoparticles like high magnetic crystalline anisotropy, moderate

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saturation magnetization, and small superparamagnetic size play an important

role in the controlled heat generation, which make them the appropriate choice
for hyperthermia [84, 89].
1.5.2. Magnetic resonance imaging

For proper medical diagnosis and treatments, proper imaging of internal
organs with high spatial resolution is one of the essential concerns. Magnetic
resonance imaging (MRI) is a very important imaging technique, since it
enables to achieve extraordinarily high temporal and spatial resolution without
any invasive procedure. MRI is the standard clinical test to determine the
presence, location and size of a tumor. However, this technique has not been
able to reach the resolution required to diagnose metastatic cancer or individual
cancer cells. Hence, nanoscale materials have been focused for better imaging
[90]. In MRI, computer-assisted imaging is done by achieving relaxation
signals of proton spins within the human body excited by radiofrequency
waves in a strong magnetic field. Although MRI itself gives detailed images, it
may not be accurate, since normal tissues and lesions often show only small
differences in relaxation time. To overcome this, MRI contrast agents can be
used to clarify images and allow better interpretation. Increased recognition of
magnetic nanoparticles as MRI contrast agents is resulting into visualization of
the disease-specific biomarkers at the molecular and cellular levels. There are
two independent relaxation times, known as longitudinal and transverse
relaxation (T1 and T2), which are used to generate the magnetic resonance
image [91]. Gd-based contrast agents enhance the signal in T1-weighted images
[92]. On the other hand, superparamagnetic iron oxide (SPIO) provides a
strong contrast effect in T2-weighted images, due to its different contrasting
mechanism [93]. It was the first nanoparticulate MRI contrast agent, which has
received great attention since its development as a liver contrasting agent and is
still used clinically [94]. Addition to this, nanoparticulate properties of these

Chapter-1
Introduction
materials is advantageous over the conventional imaging agents regarding

different bio distribution. r1 and r2 are the longitudinal and transverse relaxation
rate per millimolar of agent respectively, which can be determined
experimentally from the slope of the relaxation rate curve, which are the
contrast agent relaxivities. The r2 relaxivity remains proportional to the
saturation magnetization and the concentration of the MNPs. For a T2 contrast
agent, higher value of r2/r1 ratio leads to better T2 contrast efficacy. Among the
r2/ r1 values for MnFe2O4, Fe3O4 and CoFe2O4, the cobalt ferrites MNPs shows
higher value and offer the excellent contrast. Among these three compositions,
the cobalt ferrite nanoparticles have the highest magnetization, initial
susceptibility, and anisotropy energy, which lead to high r2 proton relaxivity.
The cobalt nanoparticles have a larger effect on proton relaxation, resulting in
improved magnetic resonance contrast and allowing smaller particle cores to be
used without the compromising sensitivity [95]. Figure 1.12 illustrates T2 and
T1 weighted MRI of tumor cells by using magnetic nanoparticles.
Figure.1.12. MRI of tumor cells by using magnetic nanoparticles [96]

Chapter-1
Introduction
1.5.3. Target drug delivery

Paul Ehrlich, the Nobel Prize winner in medicine, proposed that if an
agent could selectively target a disease-causing organism, then a toxin for that
organism could be delivered along with the agent of selectivity. He also
proposed a „magic bullet‟ which able to kill exclusively the targeted organism
[97]. Targeted Drug delivery indicates a pharmaceutical dosage form to
transport or deliver the drugs to a desired place in the body. Effective, targeted
drug delivery systems require four key requirements: retain, evade, target and
release. For example, formulations intended for intravenous administration
requires efficient drug loading into some type of delivery vehicle (carrier), so
that drug sufficiently resides in the circulation to reach the sites of the body,
where it is intended to act. Further drug should be retained by specific
characteristics within intended sites i.e. targeting. The drug should be released
within a time for effective function, in response to an external trigger such as a
change in pH, ionic strength, or temperature at the intended site. Ideally, the
carrier system, whether it is synthetic or natural, should be able to provide a
nontoxic support system that can be manufactured on an industrial scale and
can be translated into a cost-effective delivery system. Target drug delivery
leads to a reduction in the dosages and side effects of drugs, having a uniform
effect of the drug, and reduced fluctuation in circulating drug levels clinically.
Thus, development of an effective drug delivery system is a great deal of
challenge faced by pharmaceutical industries. In this regard, nanoparticles are
the ideal candidates for drug delivery because they do not only act as a
nanocarrier for passive targeting but also coating makes them useful for active
targeting. In the case of magnetic nanoparticles (MNPs), it provides the dual
advantages. First, the drug can be entrapped or loaded on the surface of MNPs
as it allows modifying the surface by using different surface functionalization
strategies. Second, the drug can be delivered to tissues by magnetic targeting
[98]. The morphological difference between normal and cancer tissue

Chapter-1
Introduction
surroundings helps the selective penetration of MNPs in tumor region. Usually

solid tumors are characterized by leaky vasculature, which promotes
extravasation of the MNPs from the blood and their subsequent retention in the
tumor interstitium. In the contrast, the healthy tissue forms an effective barrier
that prevents extravasation of MNPs. Hyperthermia-based controlled drug
delivery system can be also a prominent alternative for cancer treatment. In this
case, the therapeutic effect can then be triggered by local overheating of the
cancer cells via hyperthermia [99] and/or upon the MNPs gated-delivery of
cancer drugs. This approach is represented in figure 1.13.
Figure.1.13. Representation of two types of mechanism used in hyperthermia

mediated drug delivery system (a) The drug delivery by using a bond
mechanism (b) The drug delivery by using polymeric matrix [100]
For targeted delivery, different kinds of methods are used to attach a
drug to the surface of MNPs. For example, the drug molecule can be loaded in
the magnetic nanoclusters [101], encapsulated in stimuli responsive polymer

Chapter-1
Introduction
shell [102], covalently linked via cleavable bond [103] or trapped in the
magneto liposome [104-105] (Figure.1.14). Various drugs have been
successfully incorporated into MNPs, such as paclitaxel, doxorubicin,
epirubicin, and methotrexate on the surface by using different methods.
Figure.1.14. Representation of magnetic nanoparticles-drug nanosystem for

targeted drug delivery [105]
1.5.4. Bacteria detection

Interest in magnetic nanoparticles for detection of bacteria arises due to
beneficiary factors like, the similar size of nanoparticles, magnetic behavior,
and ability to attach biomolecules such as proteins and nucleotide probes. It
takes usually long time to detect and capture bacteria at low concentrations by
conventional techniques. An important advantage of using magnetic

Chapter-1
Introduction
nanoparticles to capture bacteria is the simple separation of bacteria from

biological samples using magnets so that rapid detection could be possible.
Magnetic nanoparticles have been utilized to magnetic capture of
pathogens in the sample preparation, giving aid to detection and identification,
and are promising candidates for microbial decontamination. The wonderful
properties of MNPs help to control the location and motion of bacteria with
externally applied magnetic fields. Since, biological media lacks in naturally
occurring magnetic components, by using magnetic nanoparticles pathogens
can be selectively controlled and detected with excellent specificity and low
background noise. Integration of bioconjugated magnetic nanoparticles with
different analytical methods has opened new opportunities for bacteria analysis,
[106] as showed in figure 1.15.
Figure.1.15. Schematic representation showing the bioconjugated

multifunctional magnetic nanoparticles to selectively capture and separate
bacteria from the mixture, which can be detected by different techniques [107]

Chapter-1
Introduction
For a specific interaction to occur, magnetic nanoparticles are usually

functionalized with bio recognition molecules, such as an antibody, aptamers,
bio protein, and carbohydrates, which allow bacteria recognition and targeting
[108]. Several groups have confirmed superior performances of magnetic
nanoparticles (MNPs) and demonstrated exciting and promising applications
for capturing bacterial pathogens. For example, Gu et al. [109-110] showed
that E. coli at ultra-low concentrations was effectively captured by
vancomycin-conjugated FePt nanoparticles. Kell et al. [111] also developed a
series of vancomycin-modified nanoparticles and employed them in magnetic
confinement assays to isolate a variety of bacteria from aqueous solution.
1.5.5. Enzyme immobilization

The definition of immobilization is to fix something so that it cannot
move. The process of immobilization leads to the physical confinement or
localization of the enzyme in a certain region of space with retention of their
catalytic activities. By immobilizing an enzyme, some structural changes can
occur, thereby changing its properties and activity to some extent. Since
enzymes are highly sensitive, their immobilization requires a suitable
methodology for keeping or eventually improving their properties such as
stability and activity, thus increasing the possibility of reusing it.
The best results have been seen, when several magnetic particles and
magnetic supports are used. For example, microspheres of various biomaterials
encapsulating the magnetic particles and copolymers with magnetic particles
were used for enzyme immobilization [112]. As a very promising strategy,
superparamagnetic nanoparticles based on magnetite (Fe3O4) and maghemite
(γ-Fe2O3) have been recently employed. These are used as supporting materials
for enzymes, which exhibit characteristics like large surface area, mobility and
high mass transfer. The bulk material exhibits a direct response to the field,
while nanoparticles coherently rotate their single vector to overcome particle

Chapter-1
Introduction
anisotropy. In addition, the high surface to volume ratio provided by the

magnetic nanoparticles favors high binding capacity and high catalytic
specificity of the conjugated enzyme. Magnetic field susceptibility also
revealed a mechanism for the efficient recovery of the enzyme complex,
thereby preventing the enzyme contamination of the final product. Enzyme
stability is maximized with nanoscale support with the additional advantages of
possible modulation of the catalytic specificity, lower transfer resistance to
solve the diffusion problem and lower operational cost [113-114].
However, since the proteins have been immobilized directly on the
surface of such particles, their stability and activity will be greatly influenced
by environmental factors, such as the temperature, pH, protease, and
immunological responses. Furthermore, pure magnetic particles may not be
very useful in practical applications, since they are likely to form a large
aggregation, alter magnetic properties, and may undergo quick biodegradation
when they are directly exposed to the biological system. Therefore, a suitable
coating is essential to prevent the occurrence of such limitations [115]. In
addition, due to anisotropic dipolar attraction, unmodified nanoparticles of
ferrites tend to form agglomerates, and lose their single-domain associated with
specific properties, when subjected to a biological system. It has been shown
that aggregation of particles can be prevented and chemical stability can be
improved by coating the surface of nanoparticles. Another advantage of coating
relates to terminal groups present on the surface, which can be modified with
coupling agents so that it covalently bind to multiple ligands specific to the
surfaces of these magnetic nanoparticles [116].
Ansari and Husain reviewed for potential applications of enzymes immobilized
on/in nanomaterials. They described how magnetic nanoparticles were used for
bioapplications by immobilized enzymes, which are mentioned in table 1.1
[117].

Chapter-1
Introduction
Table.1.1. Use of MNPs for bioapplications of immobilized enzymes
Enzyme Nanoparticle utilized Applications
Cellulose-coated
α-Amylase Starch degradation
magnetite
Cholesterol Analysis of total cholesterol

Fe3O4
oxidase in serum
Silica coated Production of fusion proteins

Haloalkane
iron oxide Containing dehalogenase
dehalogenase
sequences
Keratinase Fe3O4 Synthesis of keratinase
Bioremediation of
Laccase Chitosan-MNPs
environmental pollutants
Hydrolysis of para-
Lipase Fe3O4
nitrophosphophenol (pNPP)
Netto and Toma did an interesting review on the use of

superparamagnetic particles for the immobilization of enzymes belonging to
the categories of hydrolases, oxidoreductases, and transferases. He found that
almost all enzymes could be successfully immobilized onto magnetic
nanoparticles, which can be used further, and many progresses are going on in
this field [118].
Figure 1.16 shows an example for schematic immobilization of enzyme
by functionalized magnetic nanoparticles. It illustrates a two-step method of
polydopamine surface modification and lipase immobilization. In first step iron
oxide MNPs prepared by alkaline co-precipitation of Fe (II) and Fe (III) have
been first incubated in an alkaline dopamine solution for several hours to create
an adherent polydopamine film on MNPs (PD-MNPs). In second step enzyme

Chapter-1
Introduction
was immobilized by exposure of PD-MNPs to a lipase containing solution

[119].
Figure.1.16. Schematic immobilization of lipase enzyme by polydopamine

functionalized magnetic nanoparticles [119]
1.5.6. Cell labeling and Magnetic separation

In biomedicine, it is often advantageous to separate out specific
biological entities from their native environment in order to prepare
concentrated samples for the subsequent analysis or other use. Magnetic
separation using biocompatible nanoparticles is one of the ways to achieve this.
It is a two-step process, involving (i) the tagging or labeling of the desired
biological entity with magnetic material, and (ii) the separating out of these
tagged entities via a fluid-based magnetic separation device. Tagging is made
possible through chemical modification of the surface of the magnetic
nanoparticles. It is usually done by coating MNPs with biocompatible
Chapter-1
Introduction
molecules (e.g. dextran, phospholipids, polyvinyl alcohol, etc.) as well as by

providing a link between the particle and the target site on a cell or molecule.
In addition, coating increases the colloidal stability of the magnetic fluid.
Functionalizing magnetic nanoparticles by using antibodies or other biological
macromolecules such as hormones or folic acid enables them for specific
binding towards enzyme. For example, as antibodies specifically bind to their
matching antigen this provides a highly accurate way to label cells. Magnetic
particles coated with immune specific agents have been successfully bound to
red blood cells, golgi vesicles, urological and lung cancer cells, bacteria, etc.
[93].
Magnetic separation has been successfully applied to many aspects of
biomedical and biological research. The increasing use of magnetic carriers in
biochemical and molecular biology processes has many advantages, as
compared to other non-magnetic separation processes [120]. It has proven to
be a highly sensitive technique for the selection of rare tumor cells from blood,
and is especially well suited for the separation of low numbers of target cells
[121]. For example, this has led to the enhanced detection of malarial parasites
in blood samples either by utilizing the magnetic properties of the parasite
[122] or through labeling the red blood cells with an immunospecific magnetic
fluid [123]. The DNA of a sample is amplified and identified through
polymerase chain reactions by using this as a pre-processing technology [124].
Cell counting techniques have also been developed. One method estimates the
location and number of cells tagged by measuring the magnetic moment of the
microsphere tags [125], while another uses a giant magneto resistive sensor to
measure the location of microspheres attached to a surface layered with a
bound analyte [126]. In another application, magnetic separation has been used
in combination with optical sensing to perform „magnetic enzyme linked
immunosorbent assays‟ [127-128]. These assays use fluorescent enzymes to
determine optically the number of cells labeled with the assay enzymes, where

Chapter-1
Introduction
the target material must bind to a solid matrix. To modify the procedure the
magnetic microspheres can be used to act as the surface for initial
immobilization of the target material and magnetic separation can be used to
increase the concentration of the material. The mobility of the magnetic
nanoparticles allows a shorter reaction time and a greater volume of the reagent
to be used than in standard immunoassays where the antibody is bound to a
plate. Figure 1.17 represents a procedure for immunoassay by using MNPs,
where the captured analytes can be isolated by precipitation of the magnetic
nanoparticles in an external magnetic field. The nanoparticles are redispersed
as soon as this magnetic field is removed. The analyte labeled nanoparticles
remained immobilized on surfaces, which are modified with a secondary
antibody and thus quantified by diagnostic magnetic resonance (DMR)
technique.
Figure.1.17. Immunoassay of labeled analyte and its separation, by using

magnetic nanoparticles [129]

Chapter-1
Introduction
1.5.7. Enrichment of DNA

Highly sensitive and selective DNA sensors are of prior requirements
regarding genetic engineering. These have a wide role in the diagnosis of
genetic diseases, detection of infectious agents, gene therapy, and identification
in forensic and environmental cases. In recent years, many studies have been
devoted to develop DNA sensors due to the simplicity, specificity, exceptional
sensitivity and selectivity for the detection of specific genes. Some
conventional techniques like PCR and real-time PCR have the ability of
amplification from small amounts of DNA into readable quantities and being
applied mostly in the fields of biomedicine. However, these techniques demand
more time, sample preparation, high-tech equipped apparatus and well-
educated operators. Unlike to this, nano-bio sensors have opened a perspective
to overcome most of the disadvantages of the conventional detection methods
for the applications in agriculture, clinics, drugs, environment, and food.
Magnetic nanoparticles-DNA interaction can open up new possibilities in
various biomedical applications, which are based on fundamental properties of
DNA and magnetic nanoparticles (Figure.1.18).
Figure.1.18. DNA-magnetic nanoparticles interaction and its biomedical

applications

Chapter-1
Introduction
Nucleic acid - MNP assembly holds great potential for genetic tests and
nanomedicine at the molecular level. For successful achievement of the
genetically engineered biomedical applications, the prior requirement is the
enrichment of DNA, which can be effectively done by using magnetic
nanoparticles and specifically tailoring it for DNA [130]. For example, both
modified and unmodified cobalt ferrite nanoparticles are suitable for the
isolation and purification of genomic DNA. It is possible to achieve DNA
adsorption and desorption on modified cobalt ferrite surfaces, which is similar
to those for silica-based supports [131]. Some key factors play an important
role in the enrichment of DNA by using magnetic nanoparticles. These are type
of interaction between DNA and magnetic nanoparticles, surface modification
of magnetic nanoparticles, functional group, and mode of adsorption of DNA
on the surface of magnetic nanoparticles. By altering these factors, many types
of literatures have reported potential nano-bio applications, where nucleic
materials are enriched in such a fashion so that they can possess integrity and
the potentiality for a variety of applications. From various reported
applications, some are listed in table 1.2.

Chapter-1
Introduction
Table.1.2. List of nano-bio applications by enrichment of nucleic acid using

MNPs
Surface Functional Nucleic

MNPs Applications Ref
modifier group acid
Genomic
Tetraethylo DNA of
Pathogen 132
Fe3O4 rthosilicate Si-o hepatitis
detection
(TEOS) virus type
B (HBV)
2-
Isolation
carboxyeth
mRNA, and
Fe3O4 yl –COOH 133
pDNA extraction
phosphonic
acid
Plasmid DNA
Fe3O4 Chitosan -NH2 134
DNA isolation
Human
TEOS and DNA
Fe3O4 -NH2 Genomic 135
APTMS separation
DNA
Fe2O3, 2- λ phage MRI contrast
- 136
CoFe2O4 pyrollidone DNA agent
Polyacrylic Plasmid Gene
Fe3O4 -NH2 137
acid, PEI DNA therapy
antisense
oligo Poly- In vitro
Fe3O4 Hemin 138
(deoxy) nucleotide diagnosis
nucleotides

Chapter-1
Introduction
1.6. Statement of problem

Spinel ferrite nanoparticles are of greater interest for nano-bio
applications. Among the high potential spinel ferrite nanoparticles, the class of
cobalt ferrite has specific properties, which have made them suitable candidates
for different nano-bio applications. Cobalt ferrite has been preferred due to its
high Curie temperature, relatively high saturation magnetization, high
magneto-crystalline anisotropy, and excellent chemical stability. Additionally,
it shows the ease of synthesis, wear resistance, mechanical hardness, and
electrical insulation. These mentioned properties make cobalt ferrite as one of
the most promising candidates for different nano-bioapplications. By reviewing
literatures, for present thesis work, cobalt ferrite was selected as a choice of
magnetic nanoparticles for different types of nano-bio applications like bacteria
detection, enzyme immobilization, and magnetic fluid hyperthermia for cancer
cell treatment.
In most of the cases, synthesis and functionalization strategies are
carried out in two different steps, which are always tedious, time consuming
and required multiple process. Typically, in a multistep post synthetic grafting
process, costly organic chemicals are used to provide the desired functionality,
which makes the process inefficient for large-scale synthesis. In addition,
frequent exposure of magnetic particles to harsh reaction conditions may affect
magnetic properties. Hence, to get rid from these drawbacks, one pot synthesis
method including in-situ functionalization of magnetic nanoparticles was
preferred and aimed to synthesize in a single step.
It is well established that prior to the use of magnetic nanoparticles for
nano-bio applications, they must satisfy certain criteria such as
biocompatibility, improved colloidal stability in physiological media and
ability to carry payloads, such as drug, enzyme, proteins, carbohydrates, etc.
For all this, chemical modification of the surface of magnetic nanoparticles
through specific functionalization strategy is necessary. Hence, the different

Chapter-1
Introduction
techniques of in-situ functionalization with different materials were selected to

prepare functionalized magnetic nanoparticles. An amine and triethylene glycol
functionalization was preferred for desired nano-bio applications.
However, considering different nano-bioapplications aimed to be
achieved, it was also found that for the magnetic hyperthermia (MFH) therapy,
magnetic nanoparticles should fulfil some different prerequisites unlike to other
bioapplications. Regarding this, CoFe2O4 nanoparticles lack themselves for
some of these prerequisites. In 2008, D. Kim et al. investigated the heat
generation of cobalt ferrite nanoparticles when dispersed in aqueous media, as
heating agents for magnetically activated drug delivery and hyperthermia
[139]. In 2012, I. Sharifi et al. investigated the effect of the cation distribution
and particle size on the magnetic and structural properties of Cobalt ferrite
synthesized by chemical methods. Furthermore, they summarized the ferrite-
based magnetic ferrofluids used in hyperthermia applications [140-141]. Chen
et al. showed that suspensions of CoFe2O4 nanoparticles with a value of 2
mg.mL-1 under the alternating magnetic field showed large output power and
stable magnetic intensity, leading to a rapid increase in temperature [142]. The
self-heating characteristics of Cobalt ferrite nanoparticles for hyperthermia
application were investigated by Lee et al. They studied that, CoFe2O4 NPs are
not suitable for hyperthermia application, because the H0.f for the maximum
temperature rising is beyond the biological and physiological, safety, range
(H0.f ≤ 4.85×108Am-1s-1) in bio-medical applications [143].
For MFH application, a well-designed heating mediator with correctly
adjusted magnetic properties is necessary. The magnetization and coercivity of
Cobalt ferrite MNPs appear to be crucial for the desired heating capability.
Also for safety purpose, it is required to preset the Curie temperature (Tc)
slightly above the therapeutic range in order to reach the self-controlled heating
regime, so that the danger of local overheating and damage of the surrounding
healthy tissue could be ruled out. All these requisites of MFH therapy slightly

Chapter-1
Introduction
get apart from properties of bare Cobalt ferrite nanoparticles. Therefore, based
on the literature reviewed, it was observed that more attention has been
recently paid to magnetic cores of Cobalt zinc ferrite (Co1-XZnXFe2O4) (CZF)
NPs, which exhibits moderate heating efficiency and shows optimum Tc for
MFH therapy. Hence, along with Cobalt ferrite (CoFe2O4), an attempt was also
made to evaluate the cobalt zinc ferrite NPs, which offers variation of the
magnetic properties by the compositional changes.
The Zn-substitution in CoFe2O4 leads to some distorted spinel structures
depending upon the concentration of the precursor solutions. It was analysed
that when ferrites are sufficiently diluted with non-magnetic atoms, they can
show a wide spectrum of magnetic structures, ferrimagnetic order, etc. The
addition of foreign elements modifies the magnetic properties, such as
coercivity and saturation magnetization. Zn2+ ions commonly substitute for
Co2+ at the tetrahedral sites, resulting in an increase in the lattice parameter
because of the larger ionic radius of Zn2+ (0.74 Å), as compared to the Co2+
(0.72 Å). Zn2+ ions in high amounts can reduce the Curie point, the coercivity
and the saturation magnetization. Recently Nikam et al. have studied and
suggested that composition of Co0.5Zn0.5Fe2O4 shows high magnetization value
and minimum coercivity [144]. Duong et al. shown that when the zinc
substitution increases from x = 0 to 0.4, the saturation magnetization increases
from 72.1 to 99.7 emu/g [145]. Hou et al. in the investigation had shown that
the maximum saturation magnetization value of the prepared sample was with
concentration of Co0.5Zn0.5Fe2O4 NPs [146]. Also, Cobalt zinc ferrite
(Co0.5Zn0.5Fe2O4) NPs was reviewed as the material of high resistivity, high
magnetization, high permeability, minimum coercivity, suitable Curie
temperature, chemical stability and low eddy current loss, which are suitable to
produce hyperthermia. Therefore, aiming to achieve the hyperthermia
application, an appropriate composition as Co0.5Zn0.5Fe2O4 was selected to
achieve the desired hyperthermia performance.

Chapter-1
Introduction
Nevertheless, it was found that besides heating induction ability, the in

vitro study for magnetic fluid hyperthermia on different cell lines are less
explored. In addition, possible mechanisms of cell death are yet required to
study deeply. Hence, the present thesis work was also aimed to study possible
cell death mechanisms and to carry out in vitro study on different types of cell
lines so that it could get further direction for in vivo applications for cancer
therapy.
Based on the above considerations, objectives of the present thesis were
designed as follows,
 To characterize the amine functionalized Cobalt ferrite and amine

functionalized Cobalt zinc ferrite nanoparticles for the desired physical,
chemical, and biological properties required for nano-bio applications.
 To synthesize amine functionalized Cobalt ferrite (AF-CoFe O ) 2 4

amine functionalized Cobalt zinc ferrite (AF-Co0.5Zn0.5Fe2O4) (AF-CZF)
nanoparticles by the modified high thermal decomposition method.
 To compare and evaluate the amine functionalized Cobalt ferrite

and amine functionalized Cobalt zinc ferrite nanoparticles, to lead to
nano-bio applications.
 To study the potential nano-bio applications of selected amine

functionalized nanoparticles.
 To synthesize triethylene glycol functionalized Cobalt zinc ferrite

nanoparticles (TEGF-Co0.5Zn0.5Fe2O4) (TEGF-CZF) by using the polyol
method.

Chapter-1
Introduction
 To study the physical, chemical, and biological properties of

triethylene glycol functionalized Cobalt zinc ferrite nanoparticles.
 To explore the heat rising capacity of functionalized triethylene

glycol functionalized Cobalt zinc ferrite nanoparticles.
 To study in vitro magnetic fluid hyperthermia of triethylene glycol

functionalized Cobalt zinc ferrite nanoparticles and possible mechanism
of cell death.
With respect to these objectives the outline of the thesis work is

presented in figure 1.19.
Figure.1.19. The outline of the thesis work

Chapter-1
Introduction
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Chapter-1

1.1. Introduction to nanotechnology

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 1

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 2

and more desirable products can be made by using nanosystems than

Figure.1.1. Applications of the nanotechnology in various disciplines

1.2. Nano-bio technology

1.2.2. Fundamentals of nano-bio technology

Figure.1.2. The simultaneous interface of the nanoworld, bio systems, and

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 4

The nano-bio technology is the simultaneous interface of the nanoworld,

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 5

Figure.1.3. Comparison of nanomaterials and bio systems of nanoscale [21]

1.3. Magnetic nanoparticles for nano-bio applications

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 6

MNPs through the nanotechnology for biomedical applications is currently

1.3.1. Nano scaling laws of magnetic nanoparticles

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 7

Figure.1.4. Effect of size on the magnetism (e.g., coercivity, mass

1.3.1.1. Size effect

distinguished from paramagnetism regarding the concept of domains. When the

Figure.1.5. Magnetic behavior of superparamagnetic nanoparticles under the

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 9

L–S coupling strength between cations determines the relative magnitude of

1.3.1.2. Composition effect

The anisotropic constant (Ku) is known to be strongly correlated with

1.3.2. Strategies to design magnetic nanoparticles

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 11

efficient for nano-bio applications. The combination of structural

Figure.1.7. The strategies to fabricate multifunctional magnetic nanoparticles

1.3.2.1. Molecular functionalization of magnetic nanoparticles

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 12

1.3.2.2. Combination of magnetic nanoparticles with other

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 13

could act as agents for multimodality imaging. Based on the magnetic

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 14

1.4.1. Spinel ferrites

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 15

1.4.1.1. Structural properties of spinel ferrites

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 16

Figure.1.8. Spinel ferrite structure [65]

1.4.1.2. Magnetic properties of spinel ferrites

1.4.2. Cobalt ferrites: A promising spinel ferrite for nano-bio applications

technological applications. Many researchers have reported magnetic properties

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 18

1.5. Nano-bio applications of magnetic nanoparticles

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 19

Figure.1.10. Different nano-bio applications of magnetic nanoparticles

1.5.1. Magnetic hyperthermia

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 20

Figure.1.11. Illustration of magnetic hyperthermia [86]

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 21

saturation magnetization, and small superparamagnetic size play an important

1.5.2. Magnetic resonance imaging

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 22

materials is advantageous over the conventional imaging agents regarding

Figure.1.12. MRI of tumor cells by using magnetic nanoparticles [96]

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 23

1.5.3. Target drug delivery

Centre for Interdisciplinary Research, D. Y. Patil University, Kolhapur. 24

surroundings helps the selective penetration of MNPs in tumor region. Usually

Figure.1.13. Representation of two types of mechanism used in hyperthermia