58 Chapter 2 Nanomaterials in biomedical diagnosis

2.1 Introduction
Nanoscience is a young and flourishing field that crosses
almost every discipline of science and engineering. It focuses
on molecular events encompassing research into the mechan-
isms of assembly, structure and function of nanomaterials,
along with a focus on their atomic and molecular composition.
Nanomaterials exhibit potential applications in a wide range of
fields including medicine, chemistry, physics, biology, and com-
puter science. In last two decades research has focused the
applications of nanomaterials in biomedicine.
Nanomaterials range from 1 to 100 nm in dimension and are
used for the fabrication of new processes, devices, and materi-
als [1]. Progress in the field of medical nanotechnology with
clinical relevance provides a fertile ground for the development
of nanomaterials for patient benefit. Nanomaterials may have
wider applications ranging from monitoring molecule signals in
the biological system to rectifying image processing or as car-
riers for drugs to target specific locations in the body (Fig. 2.1).
They are making a major impact on human health by their
comprehensive and progressive applications in diagnostics and
therapeutics [2,3].
Nanomaterials exhibit novel and unique characteristics at a
nanometer scale with zero, one, two, and three dimensions. In
general nanomaterials are inorganic, organic, or inorganic/
organic composite nanostructures. They include nanoparticles,
nanowires (NWs), and nanopatterns. They possess attractive

Figure 2.1 Scope of nanomaterials in medical sciences.

 Chapter 2 Nanomaterials in biomedical diagnosis 59

physicochemical properties such as ultrasmall sizes, high sur-

face area to volume ratios, tunable optical emissions, enhanced
mechanical properties, and superparamagnetic behaviors [4,5].
Nanomaterials are gaining attention due to their cost effective-
ness, selectivity, and sensitivity along with their potential for
rapid diagnosis and mitigation of pathogenic and genetic dis-
eases (e.g., HIV and cancer) in humans and animals [6,7]. In
addition nanomaterials have also been used as advanced con-
trast agents in clinical imaging technologies such as MRI, com-
puter tomography, and ultrasound. The fabrication of various
medical devices like pacemakers, defibrillators, wireless pulse
oximeters, and ECG monitors with communication capabilities
are also composed of nanoscale materials [8,9].
This chapter focuses on various types of nanomaterials,
approaches of their synthesis, characterization techniques,
applications, and their future possibilities in prominent diag-
nostic fields.

2.2 Classification
Nanomaterials are nanostructured objects 100 nm or less in
size. Depending on the nanostructure of these materials, they
are called nanoparticles, nanopores, nanorods, NWs, nanorib-
bons, nanotubes, and nanoscaffolds. A variety of nanostructures
can be created by altering dimensions, pore size, and composi-
tion of the materials. Nanomaterials can be used for diagnostic
purposes due to their existence in various forms like single and
multiple merged aggregates or combined forms with different
shapes. The desirable electrical, optical, magnetic, and mechan-
ical properties of nanomaterials have led to considerable atten-
tion being paid to them.
The characteristic features of nanomaterials are based upon:
1. size and size distribution of materials (,100 nm);
2. chemical composition;
3. interfaces present—specifically grain boundaries, different
phase interface; and
4. interaction between constituents dormant [10].
The nanomaterials can be further classified based on dimen-
sions, pore size, and composition, as discussed in the following.

2.2.1 Dimensions
1. Zero dimension (clusters, filaments, cluster assemblies)
2. One dimension (nanotubes, NWs, nanofibers)
60 Chapter 2 Nanomaterials in biomedical diagnosis

3. Two dimensions (nanoplates, nanofilms)

4. Three dimensions (nanoparticles, quantum dots [QDs])

2.2.2 Pore size

1. Microporous (naturally occurring clay materials)
2. Mesoporous (MCM-41, MCM-48, SBA-15, carbon mesoporous)
3. Macroporous (carbon microtubes, porous gels, porous glasses)

2.2.3 Composition
1. Carbon based
a. Carbon nanotubes (CNTs)
i. Single-celled nanotube
ii. Multicelled nanotube
b. Fullerenes
2. Metal based
3. Hybrid
a. Three-dimensional metal matrix
b. Two-dimensional lamellar
c. NWs
i. Single-dimension core shell
ii. Zero-dimensional core shells

2.2.4 Types of nanomaterials

Various types of nanomaterials, along with their characteris-
tic features, are explained as follows.

2.2.4.1 Clusters
Clusters are smaller atomic and molecular agglomerates
consisting of molecules with diameters on a nanometer scale.
Metal nanoclusters are composed of smaller atoms of either
single or multiple elements of a diameter less than 2 nm.
Nanoclusters exhibit good physical and chemical properties due
to their nanometric size [11]. They can be classified as bulk par-
ticles, nanoparticles, nanostructures, and atomic clusters. Bulk
metals show efficient electrical conductivity and optical reflec-
tion, whereas metal nanoparticles show strong color due to sur-
face plasmon resonance. The decrease in size of the metal
nanoclusters leads to decrease in distinct energy levels similar
to the energy levels of molecules. Nanoclusters can be further
used for bioimaging, in vitro diagnostic, cancer therapies, and
surgeries, and also in drug delivery systems [12].
 Chapter 2 Nanomaterials in biomedical diagnosis 61

2.2.4.2 Nanotubes
In the field of nanotechnology, carbon atoms can form tiny
tubes (nanotubes) or hollow balls (fullerenes or buckyballs). In
nanotubes carbon molecules are arranged in a hexagonal struc-
ture by forming covalent bonds with three other carbon atoms.
CNTs have smaller diameters but are several centimeters long.
They are stronger in nature and can be easily bent but recover
their position if released. CNTs can be either multiple concen-
tric cylinders of carbon atoms, called multiwalled CNTs, or
nanotubes with only one cylinder, called single-walled CNTs.
Ligand binding to CNTs is commonly utilized for solubility
enhancement as well as targeting. However, nanotubes are sus-
ceptible to oxidation by strong acids [13]. Owing to high surface
area for conjugation, limited solubility, stability at physiological
pH makes them suitable for biochemical and biomedical appli-
cations. The entry of CNTs into cells is initiated either by the
process of endocytosis or insertion through the cell membrane.
Still, the role of CNT in the diagnosis and treatment of malig-
nant melanoma is an emerging area of research [14].

2.2.4.3 Nanowires
NWs are nanosized channels forming a path for electrical
currents at a very low amplitude and are formed from CNTs,
metal oxides, or silicon. They have a smaller size and diameter,
which facilitates the detection of even a small change in electri-
cal properties [15]. NWs can be used as platform for multi-
plexed chemical/biological recognition like determination of
conformational changes during antigen-antibody interactions.
The perturbation of electrical potential at NW surface associ-
ated with binding events of charged biomolecules (sign and
quantities of their charges depend on the isoelectric point of
biomolecules and solution pH) leads to a change of the charge
carrier density and conductance of NW devices in real time.
Thus several NWs attached to antibodies and incorporated into
a single device can be used to detect diseases like cancer. For
example, silicon NWs ion sensitive field effect transistor arrays
can detect several prostate cancer biomarkers. Furthermore NW
can be used in measuring ribonucleic acid (RNA) expression
levels of cancer-testis antigens and as biomarkers for aggressive
prostate cancer. Silicon and zinc oxide NW are also used to
detect ssDNA and corresponding mutations in ssDNA. For
example, a NW platform functionalized with ssDNA detected
the gene mutation for breast cancer [16,17].
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2.2.4.4 Nanofibers
Nanofibers are formed by a network of porous mesh with
remarkable inter-connectivity between the pores [18]. Natural
polymers, synthetic polymers, carbon-based polymers, semi-
conducting polymers, and composite materials are commonly
utilized to prepare nanofibers [19]. Nanofibers have been widely
used in tissue engineering and regenerative medicine, wound
dressing, delivery of therapeutic agents, and development of
biological sensors due to their distinctive structural design and
physicochemical properties [20].

2.2.4.5 Nanogels
Nanogels (NGs) are hydrogels consisting of polymeric chains
cross-linked with covalent bonds or noncovalent interactions.
NGs are highly sensitive to external stimuli leading to changes
in volume and shape much more rapidly than macroscopic
gels. A number of molecules can be accommodated in their
inner free spaces [21]. The biodistribution is affected by the
elasticity of NGs. NGs used to target human umbilical vein
endothelial cells (HUVECs) in a model cell system have been
described. The capacities of zwitter ionic poly(carboxybetaine
methacrylate) (pCBMA) NG conjugated to cyclo[Arg-Gly-Asp-D-
Tyr-Lys] (cRGD) was evaluated for targeting HUVECs. The
results of the study established the selective binding affinity of
pCBMA to HUVECs expressing Rv_3 or Rv_5 integrins [22].

2.2.4.6 Nanoshells
Nanoshells are gold coated, miniscule structures capable of
absorbing specific wavelengths of light. They exhibit the charac-
teristic property of concentrating at inflammatory or cancer
lesion sites via an enhanced permeation and retention phenom-
enon. Thus nanoshells can be used as an alternative nanocarrier
for drug delivery systems like molecular conjugates due to their
affinity for binding to the antigen’s revealed surface of a cancer
cell or in a tumor microenvironment [23]. Under in vivo condi-
tions, energy via mechanical, radio frequency, or optical source
to cancerous cells facilitates excitation of nanoshells. Upon
relaxation to ground state, nanoshells produce fierce confined
heating with selective killing of cancer cells without injuring
healthy cells. The most functional nanoshells absorb near-
infrared light and convert light energy to heat energy to mimic
their cancer cell killing potential [24].
 Chapter 2 Nanomaterials in biomedical diagnosis 63

2.2.4.7 Quantum dots

QDs are spherical, fluorescent nanocrystals of semiconductor
materials with a diameter of 2 8 nm. They exhibit distinctive
properties like narrow and broad emission spectra, negligible
photo bleaching, high sensitivity, and stable fluorescence with
simple excitation without the need for laser [25]. QDs in an
excited state produce distinguishable highly sensitive fluores-
cence emission helpful for genotype determination, image-
guided surgery, and molecular diagnostics. QDs serve as simple
fluorescent reporters in immunoassays, microarrays and fluores-
cent imaging applications. In addition, QD probes such as
quenching sensors and barcoding systems are paving the way
for highly-sensitive genetic and epigenetic detection of diseases,
multiplexed identification of infectious pathogens, and tracking
of intracellular drug and gene delivery. QD are inorganic fluoro-
phores that offer significant advantages over conventionally
used fluorescent markers. Usually QDs are attached to multiva-
lent bioconjugates having a polymer coat, or they remain fixed
into microbeads. Some QDs like CdSe ZnS cannot be used for
hemolytic analysis since their emission is far from infrared. QDs
have shown good efficacy for cancer diagnosis as well as for
therapeutic purposes. QDs, in combination with fluorescence
microscopy, show higher resolution of cells in living animals.
QDs coated with an polyacrylate cap and linked covalently with
antibodies can be used as the breast cancer marker Her2, when
carbohydrate coated they can be used for cancer imaging, and
when multicolored optical coding is applied they can be used
for biological assay. QDs are widely used as an alternative to
conventional flurophores and for developing biosensors for
detecting biomolecules such as proteins, neurotransmitters,
enzymes, and amino acids [26].

2.2.4.8 Fullerenes
These are the carbon allotropes with 60 atoms of carbon and
arranged in truncated icosahedron shapes [27]. There are a
number of fullerenes such as alkali-doped fullerenes, endohe-
dral fullerenes, endohedral metallofullerenes, exohedral fuller-
enes, and heterofullernes. The alkali-doped fullerenes are the
carbon structures containing alkali metal ions between fuller-
enes, contributing valence electrons to neighboring fullerenes
[28]. Similarly, the endohedral fullerenes have atoms enclosed
inside the fullerenes, and when these are enclosed within the
metallic atom they are known as metallofullerenes. The exohe-
dral fullerenes, also known as functional fullerenes, are
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synthesized with chemical reactions. The hetero fullerenes are

the fullerenes containing one or more carbon atoms, which are
replaced by other atoms such as nitrogen, boron, etc.

2.2.4.9 Metal-based nanomaterials

Metal-based nanoparticles are used in many applications
such as sunscreens and cosmetics, as antibacterial agents, and
for diagnostic and therapeutic purposes. Metal-based nanoma-
terials include silver nanomaterials, gold nanoparticles, and
metallic and nonmetallic nanoparticles [29]. In the bioanalytical
field for medical diagnosis, gold nanoparticles are used at a
higher rate because they can be easily synthesized, show higher
biocompatibility, and are nontoxic. These nanomaterials can
also be used as labeling agents as well as biosensing agents.
Gold nanomaterials can be used for light-based techniques
because they have higher scattering properties, strong absorp-
tion, and plasma resonance. The ease of surface modification of
gold nanoparticles by using a variety of biomolecules, like oligo-
nucleotides and proteins, to create specific/selective binding,
enables for the development of new biosensor platforms with
enhanced capabilities in the detection of various biological/
chemical analytes. Silver nanoparticles are helpful in the sepa-
ration of bacteria, viruses, and other blood components, facili-
tating clearer Raman spectroscopy signals of the components.
This method further helps in identifying the bacteria and
viruses in a much shorter time span [30].

2.3 Approaches to nanomaterial production

The nanomaterials have opened, and will continuously
unlock, synchronous and unpredictable domains of their appli-
cations with the development of newer technologies.
The development of nanofunctional devices depends on the
controlled assembly of nanoscale objects for particular applica-
tions. The two general approaches to the synthesis of nanoma-
terials and fabrication of nanostructures are top-down and
bottom-up approaches (Fig. 2.2).

2.3.1 Top-down approach

The top-down approach involves successive scaling down of
larger (macroscopic) complex bulk materials to smaller, nano-
sized materials using externally controlled physical processes
like etching through the mask, crushing, milling, or grinding.