Encyclopedia of Polymeric Nanomaterials

DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Biocomposites
Kazutoshi Haraguchi*
Department of Applied Molecular Chemistry College of Industrial Technology, Nihon University, Narashino, Chiba,
Japan

Synonyms
Biomaterials; Inorganic/organic hybrid materials

Definition
Biocomposites are defined as biocompatible and/or eco-friendly composites. They consist of a large
variety of organic and/or inorganic components, such as natural and synthetic polymers, poly-
saccharides, proteins, sugars, ceramics, metals, and nanocarbons. Biocomposites are present in
various forms, such as films, membranes, moldings, coatings, particles, fibers, and foams. In
addition to the studies aimed at improving basic mechanical properties and functionalities of the
materials, a large number of studies have been conducted to develop eco-friendly composite and/or
biomedical materials for use in the fields of sensors, tissue engineering, implants, and scaffolds.

Introduction
Biocomposite is a category of biocompatible and/or eco-friendly (i.e., green) composites in a broad
sense [1, 2]. High-performance biocomposites are often used in contact with living tissues [3]. A
wide variety of components, such as polymers (e.g., natural and synthetic polymers) [4–6], ceramics
(e.g., hydroxyapatite (HAp), silica, alumina, zirconia, clay) [7–9], carbons (e.g., graphene, fullerene,
carbon nanotube (CNT)) [10, 11], and metals (e.g., ferrotitanium, titanium alloys, silver, gold,
stainless steel) [12, 13], have been used for the preparation of biocomposites. Among the various
organic/organic, organic/inorganic, and inorganic/inorganic composites, an organic/inorganic
nanocomposite (NC) consisting of organic polymers and inorganic nanoparticles or nanofibers is
one of the most promising biocomposites for biomedical or biomaterial research. This is because of
their nanostructured surface morphology and distinguished properties derived from the combination
of characteristics of distinct constituent elements. For example, bone is a typical organic/inorganic
nanostructured composite consisting of collagen/HAp formed in vivo. Because biocomposites are
often used in contact with living tissues, cells, or blood, bio-related characteristics such as
non-toxicity, biocompatibility, anti-biofouling performance, and cell cultivation capability are
required. In this section, recent studies on the development of biocomposites are briefly discussed.
Then, examples of biocomposites, i.e., organic (polymer)/inorganic (clay) nanocomposite hydrogel
(NC gel) and soft nanocomposite (M-NC), are presented, and their application safety and charac-
teristics are examined based on their interactions with cells and living tissues.

*Email: haraguchi.kazutoshi@nihon-u.ac.jp

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Recent Studies on Biocomposites

Biocomposites generally consist of two or more organic and/or inorganic components. Various
kinds of natural (i.e., biological) polymers, for example, polysaccharides (e.g., starch, cellulose,
alginate, chitosan), proteins (e.g., keratin, silk fibroin, collagen), sugars (e.g., glucose), and lignins,
and synthetic polymers, for example, poly(vinyl alcohol) (PVA), poly(lactic acid), poly(aniline), and
polypropylene, are used as organic components [1, 2]. The compositions of biocomposites are
selected based on organic/inorganic, organic/organic, and inorganic/inorganic combinations
according to the purpose of the application. Some examples of organic/inorganic biocomposites
are chitosan/HAp, alginate/HAp, glucose oxidase/CNT/graphene oxide, glucose oxidase/poly(ani-
line)/Au/graphene, PVA/graphene, calcium alginate/CNT, PVA/gelatin/clay, starch/clay, and bioac-
tive glass/PVA/sodium alginate. Examples of organic/organic biocomposites include sodium
alginate/silk fibroin, starch/lignin, poly(lactic acid)/lignocellulosic fiber, starch/PVA/cellulose, and
chitosan/sisal cellulose. Further, inorganic/inorganic ones include HAp/CNT/Ag, HAp/titania rods,
and HAp/alumina/zirconia, etc.
Various methods are adopted for the preparation and processing of biocomposites. These methods
include electro-spinning, layer-by-layer deposition, electrodeposition, thermo-molding,
compression-molding, sintering, and isothermal crystallization. Biocomposites are prepared in
a variety of forms, such as films, membranes, moldings, fibers, coatings, foams, and hydrogels.
The major objectives of biocomposite studies are to reveal their structure/composition-property
relationships, safety, eco-friendliness, and biocompatibility (e.g., the interactions with tissues, cells,
and blood), in addition to the development of practical bio-devices [3], such as glucose sensors,
implants, scaffolds, adsorbents, and tissue replacements.

Fundamental Properties of NC Gels and M-NCs

Polymer hydrogels, which consist of a three-dimensional polymer network and a large amount of
water, have been utilized in many bio-related research fields, such as biology, biochemistry,
medicine, pharmacy, food chemistry, and analytical chemistry. However, these materials are
mechanically fragile because of their randomly arranged chemically cross-linked networks.
Recently, this limitation has been overcome by the development of a new type of hydrogel, i.e.,
organic/inorganic NC gel, with a unique polymer/clay network structure [14–17]. Further, based on
the study of NC gel, a new transparent and soft nanocomposite (M-NC) consisting of hydrophobic
polymer and inorganic clay was developed [18]. Both NC gels and M-NCs were synthesized by in
situ free-radical polymerization in the presence of exfoliated clay platelets (hectorite) in aqueous
systems and were present in various forms and sizes. Here, the disklike clay nanoparticles func-
tioned as multifunctional cross-linkers for the formation of new network systems [19]. The NC gels
and M-NCs obtained by this approach are shown in Fig. 1a, b, respectively. The NC gels showed an
extraordinarily high mechanical toughness, i.e., high elongation at break (>1,000 %), and control-
lable strength and modulus over a wide range, in addition to excellent swelling and stimulus
sensitivities [17]. Moreover, M-NCs exhibited dramatic improvements in optical and mechanical
properties, including high transparency, high reversible extensibility, and well-defined yielding,
despite their high clay contents [18]. Thus, those serious disadvantages, such as intractability,
mechanical fragility, optical turbidity, poor processing ability, and low stimulus sensitivity, which
were associated with the conventional chemically cross-linked polymeric materials, were overcome
in NC gels and M-NCs. Further, uniform aqueous dispersions of polymer/clay NC microspheres

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Fig. 1 Optical images showing (a) NC gel film, (b) M-NC film [8], and (c) aqueous dispersion of polymer/clay NC
microspheres, whose transparency was changed by altering the temperature across the LCST

(Fig. 1c), which can be utilized for coating materials, were obtained on the basis of NC gels and
M-NCs [20]. Here, the transparency and particle size were reversibly changed by altering temper-
ature across the transition temperature (lower critical solution temperature: LCST) due to the coil-to-
globule transition of polymer chains.

Cell Harvests on NC Gels and M-NCs

It was found that a confluent culture of cells, such as normal human dermal fibroblast (NHDF) cells
(Fig. 2a (i)), human hepatoma (HepG2) cells, and human umbilical vein endothelial cells
(HUVECs), could be obtained on the surfaces of NIPA-NC gels consisting of poly(N- isopropyla-
crylamide) (PNIPA)/clay networks, regardless of gel thickness, although cell culture on conven-
tional chemically cross-linked PNIPA hydrogels was almost unachievable [21]. This was because of
the combined effects of clay nanoparticles, which showed negative surface charges, and hydropho-
bic PNIPA chains, which exhibited coil-to-globule transition at a lower critical solution temperature
(LCST ¼ 32  C). It was also observed that NHDF cells cultured on PNIPA-NC gels could be
spontaneously detached in the form of cell sheets without trypsin treatment by simply decreasing the
temperature to 20  C (<LCST), due to the thermoresponsive transition of the NIPA-NC gel (Fig. 2a
(ii)) [21].
It was found that NHDFs could be attached to a polystyrene dish coated with a photo-polymerized
NIPA-NC gel or aqueous dispersion of PNIPA/clay microspheres, and a confluent cell culture could
be achieved (Fig. 2b(i)) [22]. Further, the cell sheet could be subsequently detached from the dish by
simply decreasing the temperature without using trypsin treatment (Fig. 2b(ii)) [22].
In the case of M-NC films consisting of poly(2-methoxyethyl acrylate) (PMEA) and clay
(hectorite), various cell types, such as NHDF, Balb3T3, HUVEC, and primary normal bovine aortic
endothelial cell (BVAEC), could be cultivated to confluence (Fig. 3a) [23], although they could
hardly be cultivated on the surface of chemically cross-linked PMEA. Further, the M-NC films could
be used to harvest living cells, which might be detached by simply decreasing the temperature of the
medium and/or simultaneously using gentle pipetting without any enzymatic digestion (Fig. 3b)

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Fig. 2 (a) (i) NHDF cells cultured on NIPA-NC gel and (ii) detachment of cell sheet by decreasing the temperature to
20  C. (b) (i) NHDF cells cultured on photo-NIPA-NC gel coated on a polystyrene dish and after the detachment of the
cell sheet by reducing the temperature to 10  C for 5 min. (c) Harvested MSC sheet

[23]. The detached cells were obtained as single cells or a contiguous cell sheet, both of which were
viable and reculturable.
Furthermore, effective stem cell culture in vitro is necessary for tissue engineering and regener-
ative medicine. A thermoresponsive MD-NC gel [24] consisting of MEA-DMAA (N,
N-dimethylacrylamide) copolymer-clay networks can be applied for culturing human mesenchymal
stem cells (MSCs). It was found that MSCs were effectively cultured, harvested without
trypsinization, and differentiated on the MD-NC gel (Fig. 2c) [25].

Biomedical Properties and Safety

To investigate biocompatibility, film and rod samples of NC materials (NIPA-NC and DMAA-NC
gels and M-NCs) were implanted into rabbits (intramuscular) and goats (hypodermal) for 6 months.
It was found that neither inflammation nor concrescence occurred around any of the materials
(Fig. 4a(i)) except for the formation of a thin uniform encapsulation (Fig. 4a(ii)) after 3 months. In
contrast, chemically cross-linked hydrogels broke into pieces (and missing) during the implantation
because of their low strength and fragility. Further, NC gels used as wound dressings for various
types of traumas, including epidermis wounds, full-depth skin wounds (Fig. 4b) [26], and decubitus
(i.e., pressure ulcers), were tested in animal models. It was concluded that NC gels, which
simultaneously absorbed the exudate and prevented the formation of intractable granulomatous
tissue, could comfortably be used as dressing materials for wound healing [26]. In addition, it was

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Fig. 3 (a) Phase-contrast photomicrographs of 3T3, HUVEC, and BAEC cells cultured on the surface of M-NC film.
(b) (a–1)–(a–3) Cell detachment of 3T3 cultured on the surface of M-NC11 film, by reducing the medium temperature to
5–20  C and using gentle pipetting. (b–1)–(b–3) Models depicting cell-detachment behavior corresponding to
(a–1)–(a–3) [15]

found that the modified NC gels (Fig. 4c) with extremely low density (0.3 g/cm3) and well-
controlled surface adhesion might be used as wheel cushions, due to pressure dispersion effects,
to reduce the shear force, and as adhesive skin sheets (or patches) without causing irritation.
The safety of NC gels and M-NCs was evaluated by using biological tests for medical devices.
The safety of NC gels and M-NCs was confirmed in all tests, including the sensitization test,
irritation test, intracutaneous test, and in vitro cytotoxicity test. Figure 5 shows the results of in vitro
cytotoxicity tests for MD-NC gels [24].

Summary
First, recent examples of nanocomposites used as biocomposites were presented. Next, the charac-
teristics of NC gel, aqueous dispersion of NC gel, and M-NC were discussed by considering cell

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Fig. 4 (a) Optical images showing (i) a goat (hypodermal) specimen after the implantation of NC gel rod for 6 months,
(ii) encapsulation formed in the surrounding NC gel, (b) healing of full-depth skin wound by NC gel sheet as a wound
dressing, and (c) lightweight porous NC gel foam
Number of colonies (% against the control)

100

80

60

40 M-NC2
MD30-NC2
MD10-NC5
20
MD10-NC10

0
0 20 40 60 80 100
Concentration of extract (%)

Fig. 5 In vitro cytotoxicity tests of MD-NC gels (MD30-NC2, MD10-NC5, and MD10-NC10) and M-NC (MD-NC2)
using V79 cells. The change in the number of colonies by altering the concentration (0–100 %) of extract added to
culture medium is represented in percentage against the number of colonies obtained in the culture medium without
sample extract [12]

culture, in vivo implantation, and biological safety. These biocomposites not only showed distin-
guished mechanical, optical, and thermosensitive properties but also exhibited excellent bio-related
properties that allowed for effective cell culture and subsequent thermoresponsive harvest of various
types of cells, e.g., human MSCs, on the surfaces of NIPA-NC gel, MD-NC gel, and M-NC.

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Encyclopedia of Polymeric Nanomaterials
DOI 10.1007/978-3-642-36199-9_316-1
# Springer-Verlag Berlin Heidelberg 2014

Furthermore, the safety of these NC materials was confirmed by biological tests and results of
in vivo implantation.

Related Entries
▶ Application of CL/P Nanocomposites
▶ Polymer Gels

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# Springer-Verlag Berlin Heidelberg 2014

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