Polymer Nanocomposites for Biomedical

and Biotechnology Applications 4

Jasaswini Tripathy

4.1 Introduction

Polymer matrix–based nanocomposites have emerged as a new class of hybrid

materials since last two decades and have attracted significant interest of researchers.
Polymer composites represent efficient strategies to upgrade the structural and
functional properties of polymers. The nanoparticles-reinforced polymer composites
often show significant improvements in properties that cannot be achieved by using
the polymer alone. Polymer nanocomposites are formed by combination of polymers
and inorganic/inorganic nanofillers. Nanofillers are nanostructures which have
dimension less than 100 nm and possess high interfacial area per volume
[12Ray]. Most commonly, nanofillers are carbon nanotubes, silicates, metals,
metal oxide, and ceramics. These fillers consist of different properties such as
mechanical, chemical, thermal, electrical, etc. In this regard, the fundamental under-
standing and knowledge of the nanostructures are required for fabrication of mate-
rials for desired applications. The interaction between nanofillers and polymer
matrix is the basis of mechanical and functional properties of polymer
nanocomposites and by tuning various parameters and controlling the interaction
between nanostructures and polymers, unique properties combinations can be
achieved [07Lif, 11Gah].
By exploiting the inherent properties of nanomaterials and selecting suitable
polymer matrix, a variety of nanocomposites materials have been generated with

J. Tripathy (*)
Department of Chemistry, School of Applied Sciences, KIIT University, Bhubaneswar,
Odisha, India
e-mail: [email protected]

# Springer-Verlag GmbH Germany 2017 57

D.K. Tripathy, B.P. Sahoo (eds.), Properties and Applications of Polymer
Nanocomposites, DOI 10.1007/978-3-662-53517-2_4
58 J. Tripathy

improvised properties. Current opportunities for polymer nanocomposites arise from

the multitude of applications based on different functional requirements
[08Pau]. The polymer-based nanocomposites have generated huge interest in
many biomedical and biotechnological applications [14Goe]. Biomaterials based
on polymer nanocomposites are field of interdisciplinary research which brings
together collaborative efforts of materials science, nanotechnology, and biological
science. A synergistic combination of physical, chemical, and biological properties
of nanocomposites provides an exciting platform for designing and development of
improved materials for biomedical applications [09Mit, 10Sat].
Polymer nanocomposites show improved mechanical properties, and thus can be
used to replicate high performance materials such bone and silk [08Lee]. Nature
offers strategies to mimic biological tissues containing hard and soft components.
Biologically inspired materials can be fabricated by unique combination of soft
polymer matrix and hard nanostructure; thus, these new composite materials are
being developed by dispersing hard inclusions in polymer matrix [08Vai,
09Mit]. However, one of the key challenges faced during fabrication of polymer-
based nanocomposites is uniform dispersion of nanofillers into polymer matrix for
which different processing technologies have been used by researchers
[05Dun]. Another key issue which needs to be addressed while fabricating
nanocomposites for biomedical applications is biocompatibility factor. Thus by
balancing mechanical properties, functionalities, and biocompatibility factors, poly-
mer nanocomposites are finding many interesting applications in the emerging
technologies such as tissue engineering, biomedical imaging, sutures, surgical
implants, drug delivery-based applications, etc. [14Goe].
In tissue engineering, scaffold serves as a template and structural support for
cell adhesion, proliferation, differentiation, and extracellular matrix formation
[10Arm]. Replicating some physical properties of natural tissue and reproducing
their complexity and efficiency are very challenging; thus, scaffold matrices have
to be designed which not only resemble the structural and mechanical properties
of a natural tissue but also imitate the signaling environment of the natural
extracellular matrix (ECM). In this regard, polymer-based nanocomposites
[14Ser] have been widely explored. These materials are applicable for hard and
soft tissue engineering due to their unique mechanical and electrical properties.
Apart from tissue engineering, polymer nanocomposites are also considered as
potential candidates for drug delivery as they provide possibility to deliver large
and controlled doses of therapeutic agents at the action site. The polymer-based
polymer composites have been utilized in a myriad of bioapplications
(Scheme 1). To achieve different functional requirements, these polymer-based
nanocomposites are tailored with functionalities which open new possibilities in
developing advanced biomaterials for various biomedical and biotechnological
applications.
Here, in this chapter we focus on biomedical applicability of carbon nanotube
(CNT), graphene, and nanoclay-based polymer nanocomposites. This chapter high-
lights applications of these polymer nanocomposites in emerging biomedical fields
such as tissue engineering, drug delivery, etc.
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 59

Scaffold for
tissue
engineering
Wound Medical
Healing implants

Dental Drug
Polymer
applications release
Nanocomposites

Gene
Bio delivery
imaging

Biosensors

Scheme 1 Biomedical and biotechnological applications of polymer nanocomposites

4.2 Nanofillers for Polymer-Based Nanocomposites

Polymer-based nanocomposites for bioapplications are fabricated by reinforcing

nanofillers into biocompatible polymers which include both natural and synthetic
polymers. This section presents three different nanofillers which are extensively used
for the preparation of polymer-based nanocomposites:

1. Carbon nanotubes as fillers

2. Graphene as fillers
3. Nanoclay as fillers

4.2.1 Carbon Nanotubes as Fillers

Carbon nanotubes (CNTs) are allotropic forms of carbon discovered in late 1950s;
however, the synthesis was first reported by Lijima [91Lij] and Bacon [60Bac]. The
major methods adopted for synthesis of carbon nanotubes include laser ablation
[96The], arc-discharge [97Jou], template-directed synthesis, chemical vapor depo-
sition (CVD) [99Cas], and catalyst chemical vapor deposition (CCVD). However,
60 J. Tripathy

CVD is the most commonly used method due to its low setup cost and high
production yield. CNTs are either composed of monolayered graphene sheet
(Single-walled carbon nanotubes, SWCNTs) or several concentric graphitic layers
with multiwalled structure (multiwalled carbon nanotubes, MWCNTs). CNTs have
fascinating electrical, mechanical, as well as thermal properties [14Ser]. They are
known to have exceptional mechanical properties such as high Young’s modulus and
tensile strengths up to 63 GPa (10–100 times higher than steel) [96Tre, 00Yu], as
well as thermal stability up to 2800  C under vacuum [01Tho]. Due to their unique
properties, these nanosized structures find wide application in many areas of material
science including nanocomposites [12Wan].
CNTs have been recognized as effective reinforced materials for polymer
nanocomposites because of their excellent mechanical, electrical, and surface prop-
erties such as high aspect ratio and enormous surface area. Incorporation of CNT
significantly increases the mechanical strength of polymer matrix. In addition to the
role of carbon nanotubes, the mechanical behavior of polymer nanocomposites are
also dependent on polymer nature (amorphous/crystalline/semicrystalline). Also the
adhesion between carbon nanotubes and the polymer matrix, and the homogenous
dispersion of nanotubes in the polymer matrix are equally important in improving
the properties [02Coo, 03Won] which is influenced by fabricating methods (melt
blending/latex technology/in situ polymerization/solvent casting/). Integration of
CNT as nanofiller in polymer matrix not only improves the physicochemical prop-
erties such as strength and flexibility [10Spi] but also adds new functionalities due to
which they find diverse applications [14Ser]. Sometimes CNTs are functionalized to
explore their potential in chemical and biological applications [12Mac]. They have
been exploited for various biological applications such as the preparation of tips for
atomic force microscopy, fuel-powered artificial muscles, and nanosurgical needles
and biosensors, etc. [03Wan, 14Ser].

4.2.2 Graphene as Fillers

Graphene is a single layer two-dimensional material with a hexagonal-packed lattice

having many unique properties such as high carrier mobility at room temperature
[04Nov], high Young’s modulus [08Lee], and excellent conductivity
[08Bal]. Graphene can be synthesized by chemical vapor deposition (CVD) growth,
mechanical exfoliation of graphite, or exfoliation of graphite oxide
[10Dre]. Currently, graphene-based polymer nanocomposites are an exciting area
of research due to the superior properties of graphene compared to polymers
[09Xu]. Due to its high thermal and electrical conductivity, it is reported to be better
nanofiller than other carbon-based polymer composites. The distribution of graphene
layers in the polymer matrix and interfacial bonding between the graphene layers
and polymer matrix are very important in determining the physicochemical proper-
ties of the nanocomposite. Since pristine graphene is not compatible with organic
polymers and does not form homogeneous composites, graphene oxide (GO) sheets
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 61

are more preferred [07Sta] as nanofiller for polymer nanocomposites. The remark-
able property of graphene provides possibilities for various fields.
One such interesting area of research is biomedical applications of graphene-
based polymer nanocomposites. Indeed, the number of publications on
bioapplications of graphene-based nanocomposites has grown exponentially in
recent years. Graphene-based polymer composites have been utilized in a myriad
of bioapplications.

4.2.3 Nanoclay as Fillers

Nanoparticles derived for clay minerals have been considered as very effective
reinforcing materials for designing polymeric nanocomposites. Due to their abun-
dant nature, low cost, and interesting properties such as high strength, stiffness and
high aspect ratio of individual platelets, clay-based polymer composites are used in
different fields. Clay minerals include both natural clays (e.g., montmorillonite,
hectorite, and saponite) and synthesized clays. Depending upon the nature of the
components and processing conditions, layered clays filled into a polymer matrix
form either conventional composite or nanocomposite. In case of conventional
composite, polymer cannot intercalate into the galleries of clay minerals, whereas
nanocomposites can further be divided into two types called as intercalated
nanocomposites and exfoliated or delaminated nanocomposite.
In case of intercalated nanocomposites, monolayer of extended polymer chains is
impregnated into the gallery of clay minerals resulting in an ordered multilayer
structure with alternate polymer layers and clay platelets, whereas in exfoliated
nanocomposites clay platelets are completely and uniformly dispersed in a contin-
uous polymer matrix resulting in enhanced polymer–clay interaction [10Kil]. Clay-
based nanocomposites exhibit strong interfacial interactions between the dispersed
clay layers and the polymer matrix which leads to enhanced mechanical, thermal,
and barrier properties over the virgin polymer.
The incompatibility between hydrophilic clay and hydrophobic polymer is one of
the shortcomings, which often causes agglomeration of clay mineral in the polymer
matrix. Therefore, surface modification of clay minerals is the most important step to
achieve polymer nanocomposites. The unique layered structure and high intercala-
tion capabilities of clay minerals allow them to be chemically modified to be
compatible with polymers which are essential for development of clay-based poly-
mer nanocomposites The selection of clay for polymer nanocomposites depends
upon on their targeted applications.
One of the promising applications of these nanocomposites is in the field of
biomedical and biotechnological arenas. Clay-based polymer nanocomposites have
been investigated in many biomedical and biotechnological applications such as
tissue engineering, drug delivery, biosensors, and biomedical devices. However,
some of the major challenges to consider while developing materials for biomedical
relevance include long-term biocompatibility and biodegrability issues.
62 J. Tripathy

4.3 Bioapplications of Polymer Nanocomposites

4.3.1 Biomedical Applications of CNT Polymer Nanocomposites

Recently carbon nanostructures-reinforced polymer nanocomposites have increas-

ingly drawn attention of researchers for their use in biotechnology and biomedical
field. They have been exploited for variety of biomedical applications. Particularly
CNT-based polymer composites are widely investigated [14Ser]. A significant
amount literature is available on nanocomposite made from polymers and nanotubes
as biomaterials.

4.3.1.1 Tissue Engineering

The use of CNTs/polymeric composites as scaffolds for bone engineering has
recently become a subject of interest. The scaffolds used for tissue engineering serves
as substrates for cell adhesion, proliferation, differentiation, extracellular matrix
(ECM) formation, and to guide tissue regeneration. The ideal scaffold for tissue
regeneration should possess sufficient mechanical properties. In this regard, carbon
nanotubes have the potential in providing the needed structural reinforcement for
tissue scaffold [10Arm]. The much anticipated use of CNTs in these scaffolds is
mainly to improve their overall mechanical properties and to promote and guide bone
tissue regeneration. By integrating a small fraction of carbon nanotubes into polymer
matrix, significant improvements in the mechanical strength of the composite is
observed [05Che]. Wang et al. have shown that MWCNTs blended with chitosan
significantly improved the mechanical properties compared with those of chitosan
[05Wan]. The biological properties of CNT scaffolds have been confirmed by in vitro
studies. Several cells types have been successfully grown on carbon nanotubes-based
polymer composites. Jell et al. have reported synthesis of porous thermoplastic
polyurethane–multiwalled carbon nanotubes (CNTs) foams by thermally induced
phase separation (TIPS) method [08Jel]. They have shown that CNT incorporation
significantly improved the compression strength and stiffness of the nanocomposite
scaffold. Through in vitro studies, they found that osteoblast production of the potent
angiogenic factor VEGF (vascular endothelial growth factor) increased in proportion
to CNT loading which verifies the potential influence of the nanocomposite scaffolds.
Shi et al. fabricated porous nanocomposite scaffolds using a thermal-cross-linking
and particulate-leaching technique) [07Shi]. Through in vitro cultures, they con-
firmed that mesenchymal stem cells (MSCs) adhere and proliferate on all the
PPF-SWNT scaffolds. Sitharaman et al. [08Sit] tested the biocompatibility of porous
PPF-SWNT scaffolds in a rabbit model. They found that implants made of
PPF-SWNTs displayed only mild inflammatory responses. The PPF-SWNT
nanocomposite scaffolds showed significant bone ingrowth after 12 weeks of implan-
tation with increased collagen matrix production (Figs. 4.1 and 4.2).
One of the careful considerations while fabricating carbon nanotube scaffold is
uniform dispersion of carbon nanotube in polymer matrix to achieve desired elec-
trical conductivity and mechanical strength. The uniform distribution of CNT in
polymer matrix transfers the load from the matrix to the nanotube which is essential
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 63

Fig. 4.1 Representative micro CT images of the rabbit femoral condyle 12 weeks after the
implantation of either a (a) PPF or (b) US-tube/PPF scaffold (Adapted from Sitharaman (2008))

40
*
35

30
*
% Bone Volume

25

20

15

10

0
PPF US-tube PPF US-tube Trabecular Bone
(4 Weeks) (4 Weeks) (12 Weeks) (12 Weeks)

Composites

Fig. 4.2 Percent bone volume at the initial defect site after 4 and 12 weeks for each of the study
groups, as measured from reconstructed three-dimensional micro CT images (Adapted from
Sitharaman (2008))

for enhancing the mechanical and electrical properties of the composite. Different
techniques have been explored to improve the CNT dispersion in different polymer
matrix [10Arm]. Functionalization of CNT is also an effective route to increase the
dispersing ability. Furthermore, the functionalization of CNTs increases water
64 J. Tripathy

Fig. 4.3 SEM images of MSCs cultured on (a, d) smooth PLGA, (d, e) PLGA/MWCNTs, and
(c, f) PLGA/c-MWCNTs films for 24 h at 100 magnification (a–c), and at 1000  magnification
(d–f) (Adapted from Lin (2011))

miscibility and biocompatibility of CNT. Covalent or noncovalent addition of

various chemical functional groups can be done to the side walls and tips of CNTs
to control the interaction between polymer and carbon nanotube. The nature of the
functional group at the CNT surface plays a determinant role in the mechanism of
interaction with cells. Lin et al. fabricated nanocomposites by incorporating
carboxyl-functionalized multiwalled carbon nanotube (c-MWCNT) into poly(lac-
tic-co-glycolic acid) (PLGA) matrix [11Lin]. Their observations revealed that
c-MWCNTs gave a better dispersion than unmodified MWCNTs in the PLGA
matrix with increased mechanical properties of the nanocomposites. They have
shown that 7-week period in vitro degradation test accelerated the hydrolytic
degradation of PLGA. In addition, the cells could adhere to and spread on films
via cytoplasmic processes. Their results demonstrated that c-MWCNT-modified
PLGA films were beneficial for promoting cell growth and inducing MSCs to
differentiate into osteoblasts (Fig. 4.3).
Shi et al. fabricated nanocomposite scaffolds by incorporating single-walled
carbon nanotubes (SWNTs) in poly(propylene fumarate) (PPF) matrix. They showed
that the functionalization of single-walled carbon nanotubes (SWNTs) increases the
interaction between nanotubes and polymer matrix which in turn enhances mechan-
ical properties [06Shi].
The electrical conductivity of CNS-based nanocomposites is used to direct cell
growth due to their ability to conduct electricity stimulus into the tissue healing
process. Khang et al. used the electrical conductivity of CNTs to increase the cell
density on the on CNTs–polycarbonate urethane nanocomposite surface via electri-
cal stimulation [08Kha]. Supronowicz et al. have provided evidence that electrical
stimulation delivered through nanocomposites of poly(lactic acid) and MWCNTs
promotes osteoblast functions [02Sup].
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 65

For the preparation of 2D CNT films and fibers, the techniques involved are
mainly layer-by-layer (LbL) deposition and solution–evaporation technique,
[05Wan, 10Byr], whereas for the preparation of 3D CNT-based scaffolds the
methods such as freeze-casting and electrospinning [14Ser] have been opted which
generally produce scaffolds with controlled and interconnected porosity. Concerning
nerve tissue repair, Kabiri et al. reported CNTs/PLLA fiber scaffolds as a as potential
candidates for neural tissue regeneration [12Kab].

4.3.1.2 Other Bioapplications

Other biomedical and biotechnological applications of CNT-based polymer
nanocomposite include drug delivery, wound healing development of chemiresistors
[10Cho], dental composites [13Bor], and microcatheters [05End].

4.3.2 Graphene-Based Polymer Nanocomposites

Graphene has been exploited for diverse applications due to its unique properties. Of
particular interest is a biological application of graphene which is well documented.
Graphene oxide (GO) is more widely used as compared to graphene due to the
presence of carboxylic, epoxy, and hydroxide groups, which provide opportunity for
functionalization. GO has been functionalized with many biocompatible polymers
for requisite biological applications. Graphene-based polymer composites have been
investigated for various biomedical applications. They are anticipated to be promis-
ing materials to be used for tissue-engineered scaffolds, drug delivery vehicles,
biosensors, etc.

4.3.2.1 Tissue Engineering

Due to enhanced mechanical and electrical properties, graphene-reinforced polymer
composites hold immense potentials for tissue engineering scaffolds. The graphene-
based polymer nanocomposites have been reported to be biocompatible and bene-
ficial for the growth of the cells [11Par]. Sayyar et al. prepared graphene/
polycaprolactone composites by covalently linking the polymer to the graphene
chains which resulted in improved conductivity and mechanical properties. The
growth of Fibroblast (L-929), neural (PC-12) and muscle (C2C12) cell lines on
cPCl–CCG materials were assessed by comparing with growth of these cell types on
pristine PCl [13Say]. They found that all cell lines proliferated on PCl, cPCl–CCG
0.5% and 5 % in a similar way to tissue culture plastic suggesting the potential
applications of fabricated nanocomposite for tissue engineering (Fig. 4.4). In a
similar approach GO–chitosan hydrogel scaffolds prepared by covalent linking
chitosan with graphene exhibited better mechanical properties and lower degrada-
tion rate. In addition, there was significant improvement in cell adhesion, differen-
tiation, proliferation, and calcium phosphate deposition of mouse preosteoblast
MC3T3-E1 cells on the hydrogel [11Dep]. Zhang et al. reported that the tensile
strength and compressive strength of PVA-based hydrogels were significantly
enhanced by incorporating GO without affecting their cytocompatibility [11Zha].
66 J. Tripathy

Fig. 4.4 Growth curves of three cell lines on PCl, tissue culture plastic, and cPCl–CCG materials.
Fibroblasts (a, L-929 cell line), muscle cells (b, C2C12 cell line), and neural cells (c, PC12 cell line)
all adhered to and proliferated on the materials for 72–96 h (Adapted from Sayyar (2013))

In another study, graphene-reinforced chitosan films did not show any toxicity
when tested on murine fibrosarcoma L929 cell culture. The mechanical properties
were also enhanced [10Fan]. Recently, Shin et al. fabricated 3D composite scaffolds
from gelatin methacrylate and GO [13Shi]. The incorporation of GO into hydrogels
enhanced their mechanical and electrical properties without affecting encapsulated
fibroblast cells. Lu et al. [12Lu] fabricated chitosan–PVA nanofibrous scaffolds
containing graphene. Further, they explored wound healing property of composite
with and without graphene along with control (no scaffold) and found that the
samples containing graphene healed completely and at a faster rate in both mice
and rabbit. The wound healing property of graphene composite was attributed to free
electron in graphene which inhibits the prokaryotic cell multiplication. This was
further confirmed by performing antibacterial study on E. coli.

4.3.2.2 Drug Delivery

In addition to tissue engineering, the graphene-based polymer composites were
found to be potential candidates for other biomedical applications including drug
delivery. They are exploited for loading poorly soluble drugs due to their high
surface area, π–π stacking, and hydrophobic interactions of graphene.
A number of studies have been reported on applications of chitosan and graphene
nanocomposite for loading various drugs like 5-fluorouracil [11Ran] and camptothecin
(CPT) [11Bao]. Liu et al. successfully loaded doxorubicine (DOX), an anticancer drug,
into graphene nanosheets (GS) using gelatin as a reducing and functionalizing agent
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 67

[11Liu]. Gelatin–GS–DOX exhibited higher drug loading capacity due to large surface
area and relatively higher π interactions. The Gelatin–GS–DOX complex also showed
high toxicity towards MCF-7 cells through endocytosis. Sun et al. [08Sun] fabricated
targeted delivery system by conjugating rituxan (CD20+ antibody) with polyethylene
glycol–nanographene oxide (PEG–NGO). Loading of doxorubicin (DOX) onto
PEG–NGO conjugate was favored due to the noncovalent π–π stacking and in vitro
pH-dependent drug release was studied.
Kim et al. developed stimuli-responsive nanocarrier for intracellular cytosolic
delivery of DOX by functionalizing rGO with PEG and branched polyethylenimine
(BPEI) [13Kim]. DOX was released in response to near infrared (NIR), acidic pH, and
high intracellular levels of glutathione (GSH). In another study [12Dem], DOX-loaded
PEG–GO nanocomposites were developed and released in response to GSH.
Miao et al. [13Mia] demonstrated successful codelivery of anticancer drug
doxorubicin (DOX) and photosensitizer (Ce6) using polyethylene glycol-grafted
graphene oxide (pGO). They revealed that pGO nanosheets increased the cellular
uptake as well as tumor tissue accumulation of Ce6, compared to treatment with free
drugs (Fig. 4.5).

4.3.2.3 Gene Delivery and Bioimaging

Graphene-based polymer nanocomposites have also been explored for other bio-
medical and biotechnological applications such as gene delivery. For the purpose of
gene delivery, Chen et al. functionalized graphene with cationic polymer polyethy-
lenimine (PEI) [11Che]. PEI acts as a nonviral gene vector due to its strong
electrostatic interactions with negatively charged phosphates of RNA and DNA.
However, it shows low biocompatibility and high toxicity which limits its use. It was
revealed that PEI–GO exhibited gene delivery high transfection efficiency and lower
cytotoxicity compared to PEI alone. Kim et al. complexed PEG–BPEI–rGO with
plasmid DNA for photothermally controlled gene delivery. The complex did not
show any cytotoxicity to PC3 and NIH/3 T3 cells. The presence of rGO accelerated
endosomal membrane disruption led to higher transfection efficacy when subjected
to NIR irradiation [13Kim].
Graphene-based nanocomposites have also been fabricated for bioimaging pur-
poses. Shen et al. [12She] prepared a multifunctional nanocomposite as MRI probe
by combining PEG–functionalized GO complex and gadolinium–diethylenetriami-
ne-pentaacetic acid–poly(diallyldimethylammonium)chloride (Gd–DTPA–PDDA).
GO–IONP (superparamagnetic iron oxide nanoparticles) functionalized with PEG
have been successfully used for drug delivery and bioimaging applications. [12Ma].

4.3.3 Clay-Based Nanocomposites

Silicate-based polymer nanocomposites have been anticipated as the next-generation

materials for various biomedical applications due to the enhanced surface interac-
tions of silicate nanoparticles and polymer chains [12Gah].
68 J. Tripathy

Fig. 4.5 In vivo biodistribution of pGO nanophysisorplexes. SCC7-bearing mice were systemi-
cally treated with pGO, free Ce6, Ce6/pGO, or with Ce6/Dox/pGO (Ce6 10 mg/kg and Dox 5 mg/
kg). After 1 h (a), 24 h (b), and 48 h (c), the in vivo distributions of Ce6 fluorescence were
visualized using a molecular imaging system. (d) Optical images are provided for location of tumors
(Adapted from Miao 2013)

4.3.3.1 Tissue Engineering

Nanoclays (synthetic silicates) are widely used to reinforce polymer to improve
physical and mechanical properties of polymeric matrix [10Wu] due to their aniso-
tropic and high aspect ratio morphology. The physical and chemical properties of
nanocomposite matrix can be controlled by addition of nanoclay to polymeric
matrix. Nitya et al. have shown that the incorporation of halloysite nanoclay within
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 69

Fig. 4.6 SEM images of mineralized PCL nanocomposite scaffolds: SEM images showing
mineralization after 21 days in SBF on a PCL, b PCL/4 % NC, and c PCL/6 % NC composite
scaffolds (Adapted from Nitya (2012))

the PCL scaffolds not only increased the mechanical strength but also the protein
adsorption and cell adhesion of the nanocomposite [12Nit]. Their results indicated
that the human mesenchymal stem cells (hMSCs) seeded on these scaffolds prolif-
erated faster than in PCL scaffolds (Fig. 4.6).
Gaharwar et al. have they have shown that silicate nanoparticles (Laponite RD)
can be used to effectively control the adhesion, spreading, and proliferation of
fibroblast and preosteoblast cells on silicate cross-linked PEO surfaces [10Gah]. In
a similar study, they have revealed through in vitro cell culture studies that increase
in silicate concentration in silicate cross-linked poly(ethylene oxide) (PEO)
nanocomposites enhanced the attachment and proliferation of human mesenchymal
stem cells significantly [12Gah].
In a recent study, nanoclay has been shown to induce osteogenic differentiation in
hMSCs without using any growth factors [13Gah]. Ambre et al. have shown that
MMT clay modified with 5-aminovaleric acid increases interlayer spacing and
improves biocompatibility with human osteoblasts. They used MMT clay modified
with 5-aminovaleric acid for preparing chitosan/polygalacturonic acid (ChiPgA)
composite scaffolds [10Amb] In a similar approach, in another study they have
reported fabrication of biopolymer (ChiPgA) scaffolds and films by intercalated
nanoclays containing organomodified MMT clay with HAP (in situ HAPclay) for
70 J. Tripathy

Fig. 4.7 (a–d) SEM micrographs of human MSCs on ChiPgA/in situ HAP clay scaffolds after
18 days of culture (Adapted from Ambre (2013))

bone tissue engineering. These HAPclay scaffolds were able to promote osteogenic
differentiation of hMSCs [13Amb] (Fig. 4.7).

4.3.3.2 Drug Delivery

One of the biggest challenges in developing polymer-based drug delivery system is
to control release of encapsulated or entrapped drugs. The therapeutic effects of the
drugs and their biological activity can be optimized by controlling release kinetics of
drugs. Silicate-based polymer nanocomposites show good barrier properties for
diffusion of small molecules and thus can be applied for sustained drug release
applications. Various drug delivery systems have been designed based on clay-based
polymer nanocomposites for drug delivery-based applications.
Saha et al. [14Sah] fabricated film as well as nanofibrous web form of polyure-
thane/MMT clay nanocomposite as a delivery system for chlorhexidine acetate
(CA), a bactericidal agent. Sustained release of drug was attributed to the presence
of bulky and immobilized drug cation in the clay interlayer spacing which hinders
the exchange of the cationic species present in the buffer media (Fig. 4.8).
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 71

90 1% CA
1% CAMt
80 5% CA
5% CAMt
70
Drug released (%)

60

50

40

30

20

10

0 20 40 60 80 100 120 140 160 180 200

Time (hrs)

Fig. 4.8 Release profile of CA and CAMt-loaded film samples in PBS media at 37  C (Adapted
from Saha (2014))

Montmorillonite clay-based polyurethane nanocomposites have been explored as

implantable drug delivery system. The therapeutics loaded nanocomposites were
implanted in specific organs to control the release of a therapeutic agent to a specific
target and to prevent different types of pathological processes [11Sil].
Lee and coworker [03Lee, 04Lee] fabricated clay-based poly (N-isopropyla-
crylamide) nanocomposite hydrogels and studied their drug release as well as
swelling behaviors. It was concluded that clay in nanocomposites led to the
decrease in swelling ratio and increase in strength of the nanocomposite hydrogels.
In addition, they found that the release behavior of model drugs was largely
depended on various factors including the content of clay and its intercalated
agents.
Mishra et al. [14Mis] prepared nanocomposites of polyurethane by dispersing
organically modified 2-D nanoclay followed by prepolymerization and subsequent
chain extension using various chain extenders. These nanocomposites showed
sustained release of drug as compared to the pristine PU. They have shown that by
increasing the length of chain extender larger crystallites were formed which
restricted drug diffusion due to barrier effect (Fig. 4.9).
A sustained release of dexamethasone drug was observed when organic modified
silicate nanoparticles (Cloisite clay) were added to poly (ethylene-co-vinyl acetate)
[56]. The drug release kinetics was suggested to be dependent on degree of disper-
sion as well as the aspect ratio of the silicate nanoparticles [03Cyp]. Li et al. have
proposed Laponite nanoparticle-enriched alginate gels for the controlled delivery of
cationic drugs [11Li] [14Mis].
72 J. Tripathy

Fig. 4.9 Microstructure-controlled drug delivery. Crystallite size increases with the increase in
aliphatic chain length of the chain extender. The formed with the chain extenders EG, BD, HD, and
DD were designated as EG-PU, BD-PU, HD-PU, and DD-PU, respectively. BD-PU, which has a
small crystallite size, shows prompt release, whereas DD-PU, which has larger crystallites, shows
lower drug release. Moreover, in the case of DD-NC, the delayed diffusion of drug is controlled by
crystallite size and 2D nanoclays as compared to that of BD-NC (Adapted from Mishra (2014))

4.3.3.3 Other Biomedical Applications

Clay-based polymer nanocomposites find diverse applications in biomedical field.
For example, poly vinyl alcohol-clay-based polymer hydrogels have been exploited
for wound dressing [07kok]. Bionanocomposites based on montmorillonite com-
bined with HAP and chitosan have been evaluated as implants [08Kat]. Clay-based
polymer composites have been also explored for food packaging applications
[07Sor].

4.4 Concluding Remarks

There has been tremendous progress in synthesis of polymer-based nanocomposites

due to the unique properties of nanostructured reinforcing materials. The synthesis
and functionalization of nanofillers open new avenues for exploring their use in
fabrication of polymer-based nanocomposites for bioapplications. These polymer
nanocomposites have demonstrated improved properties significantly compared to
the virgin polymer; thus, remarkable progress has been achieved in exploiting their
use for emerging technologies such as drug/gene delivery and tissue engineering.
This chapter provides insights to multitude of bioapplications of polymer
nanocomposites, thus expanding the range of applications from scaffolds for cell
growth to biosensors. In spite of numerous efforts that have been taken to prepare
polymer-based nanocomposites for bioapplications, there are still many challenges
that need to be addressed to reach their full potential. While using these
nanocomposites as biomaterial, the requisite criteria include biocompoatibilty and
biodegradabilty and a host of other parameters which must be assessed properly with
relevant preclinical studies to avoid false expectations. A careful selection of mate-
rials for developing nanocomposites and investigating their cellular level interac-
tions will be instrumental for their biological applications.
4 Polymer Nanocomposites for Biomedical and Biotechnology Applications 73

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