Anda di halaman 1dari 23

Progress in Polymer Science 35 (2010) 10931115

Contents lists available at ScienceDirect

Progress in Polymer Science


journal homepage: www.elsevier.com/locate/ppolysci

Composite materials based on silk proteins


John G. Hardy, Thomas R. Scheibel
Lehrstuhl fr Biomaterialien, Universitt Bayreuth, Universittsstr. 30, 95440 Bayreuth, Germany

a r t i c l e i n f o a b s t r a c t

Article history: A number of animals have evolved to produce silk-based composite materials for a variety
Received 24 September 2009 of task-specic applications. The review initially focuses on the composite structure of
Received in revised form 5 March 2010
silk bers produced naturally by silkworms and spiders, followed by the preparation and
Accepted 14 April 2010
applications of man-made composite materials (including bers, lms, foams, gels and
Available online 22 April 2010
particulates) incorporating silk proteins in combination with other polymers (both natural
and synthetic) and/or inorganic particles.
Keywords:
2009 Elsevier Ltd. All rights reserved.
Biomimetic
Biomaterials
Composite
Proteins
Silk

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1094
1.1. Fibers produced by silkworms are natural composites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1094
1.2. Fibers produced by spiders are natural composites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1094
1.3. Sources of silk proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1096
1.4. Processing silk proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1096
2. Man-made composite materials based on silk proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1096
2.1. Man-made composites based on silk proteins and synthetic polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1096
2.1.1. Man-made composites based on silk proteins and non-biodegradable synthetic polymers . . . . . . . . . . . . . . . . . . 1096
2.1.2. Man-made composites based on silk proteins and biodegradable synthetic polymers . . . . . . . . . . . . . . . . . . . . . . . 1100
2.2. Man-made composites based on silk proteins and biopolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1101
2.2.1. Man-made composites based on silk proteins and other proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1101
2.2.2. Man-made composites based on silk proteins and polysaccharides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1103
2.3. Man-made composites based on silk proteins and inorganic particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1105
2.3.1. Man-made composites based on silk proteins and metal nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1105
2.3.2. Man-made composites based on silk proteins and biominerals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1106
3. Applications of composite materials based on silk proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1107
3.1. Textiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1107
3.1.1. Composites based on silk proteins and synthetic polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1107
3.1.2. Composites based on silk proteins and biopolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1107
3.1.3. Composites based on silk proteins and metal nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108

Corresponding author. Tel.: +49 921557360; fax: +49 921557346.


E-mail addresses: john.hardy@bm.uni-bayreuth.de (J.G. Hardy), thomas.scheibel@bm.uni-bayreuth.de (T.R. Scheibel).

0079-6700/$ see front matter 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.progpolymsci.2010.04.005
1094 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

3.2. Sutures and dressings for wounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108


3.2.1. Sutures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108
3.2.2. Wound dressings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108
3.3. Tissue scaffolds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108
3.3.1. Composites based on silk proteins and other proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108
3.3.2. Composites based on silk proteins and polysaccharides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1108
3.3.3. Composites based on silk proteins and synthetic polymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.3.4. Composites based on silk proteins and inorganic particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.4. Biocompatible coatings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.4.1. Anticoagulant coatings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.4.2. Antibacterial coatings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.4.3. Improved cell adhesion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1109
3.5. Drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1110
3.5.1. Low molecular weight drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1110
3.5.2. High molecular weight drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1110
3.6. Materials with novel electronic, magnetic and optical properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1110
3.7. Enzymatically active biomaterials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1111
4. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1111
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1111
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1111

1. Introduction The broin nanobrils are composed of a complex of


three proteinaceous components: a large protein, known as
A number of animals (best known being arthropods) heavy chain (H-chain) broin (of ca. 350 kDa) that is linked
have evolved to produce a variety of task-specic silk to a second small protein, known as light chain (L-chain)
protein-based composite materials (see Table 1) [1]. Of the broin (of ca. 25 kDa) via disulde bonds; and a third small
greatest economic importance are Bombyx mori (B. mori) glycoprotein, known as the P25 protein (of ca. 30 kDa) is
silkworms that produce cocoons from silk composite bers associated via non-covalent hydrophobic interactions. The
to protect them from predators during their metamor- molar ratios of H-chain:L-chain:P25 are 6:6:1; the H-chain
phosis into moths. Web-weaving spiders (such as Araneus is hydrophobic and contains blocks of (Gly-Ala-Gly-Ala-
diadematus or Nephila clavipes) produce a number of dif- Gly-Ser)n that are known to form anisotropic -sheet-rich
ferent silk composite bers to capture prey (in webs), nanocrystals, whereas the L-chain is more hydrophilic and
to protect/preserve their offspring/prey (in cocoons), and relatively elastic, and the P25 protein is believed to play a
as lifelines to escape from predators. Certain silk bers role in maintaining the integrity of the complex [15].
have mechanical properties superior to Nylon, Kevlar and The glue-like glycoproteins are known as sericins (a set
high-tensile steel. The mechanical properties and biocom- of serine-rich glycoproteins) that coat the broin laments
patibility of naturally occurring silkworm and spider silk and ensure the cohesion of the cocoon by sticking the twin
composite bers have allowed humans to use such bers laments together and constitute 2530% of the weight of
for millennia for applications as diverse as currency, hunt- the ber (see Fig. 1B and C) [16]. Finally the ber is coated
ing (bow strings, cross-hairs, shing lines or nets), paper, with a variety of other proteins postulated to protect the
textiles and wound dressings [24]. cocoon against microbes and predators (see Fig. 1C) [17,18].
This article begins with a brief explanation of the com-
posite structure and function of natural bers based on
1.2. Fibers produced by spiders are natural composites
silk proteins [510], followed by the preparation of man-
made composite materials based on silk proteins and their
Silk bers made of proteins produced in the major
prospective applications.
ampullate (MA) silk gland (of ca. 250350 kDa) have a very
high-tensile strength (comparable to Kevlar) and moder-
1.1. Fibers produced by silkworms are natural composites ate elasticity. MA silks are used as a scaffold upon which
to attach other silks during the construction of a web and
Mankind has farmed B. mori silkworms for thousands of as a lifeline when it is necessary to escape from a predator.
years, which has facilitated an understanding of its com- MA silks have diameters between 1 and 20 m (depending
posite structure (see Table 1 and Fig. 1), and more recently upon spider species) and have a coreshell type struc-
its potential for biomedical applications [11]. Silkworm ture (see Fig. 2 and Table 1) [6,7]. The core contains two
silk bers are composed of two protein microlaments major proteins (e.g. MaSp1 and MaSp2 produced by N.
(known as brins) embedded in a glue-like glycoprotein clavipes spiders or ADF-3 and ADF-4 produced by Araneus
coating (see Fig. 1B and C) [12]. The brins are broin l- diadematus spiders) that are composed predominantly of
aments composed of bundles (of ca. 100 nm in diameter) glycine, alanine and proline (although the quantity of the
of nanobrils (individually of ca. 5 nm in diameter) that latter varies signicantly between individual proteins and
are preferentially aligned with the long axis of the ber spider species) [19]. Major ampullate spidroins are rem-
[13,14]. iniscent of block copolymers containing blocks of (Ala)n ,
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1095

Table 1
Examples of natural composite materials incorporating silk proteins.

Example Components Role References

Silkworm cocoons Fibroins Mechanical properties [1118]


Sericins Glue-like glycoproteins
Peptides Antimicrobial
Lipids Camouage

Spider webs (e.g. lifeline) Major ampullate spidroins Mechanical properties [6,7,1925]
Minor ampullate-like spidroin Mechanical properties
Glycoproteins Unknown
Lipids Unknown

Mussel byssus bers Collagens with specic anks: Mechanical properties [810]
preCol-D Spider silk-like (stiff)
preCol-P Elastin-like (elastic)

Mollusc shells Silk-like protein Energy distribution [140,166168]


Mechanics
Chitin Scaffold
Calcium carbonate Hard mineral

Fig. 1. A) Photograph of a Bombyx mori silkworm. B) Electron micrograph of partially degummed B. mori silkworm cocoon bers, in which the two brins
of broin and the coating of sericins and other proteins postulated to protect the cocoon against microbes and predators are pointed out. C) Schematic
illustration of the composite structure of a cocoon ber, in which the two brins of broin and the coating of sericins and other proteins postulated to protect
the cocoon against microbes and predators are pointed out [2]. Adapted with the permission of both the authors and publisher, Copyright 2008, Elsevier
Science Ltd., Oxford, UK.

(Gly-Ala)n , (Gly-Gly-X)n or (Gly-Pro-Gly-X-X)n where X is due to the difference in primary amino acid sequence of
typically tyrosine, leucine or glutamine. The alanine-rich the two proteins [24,25]. MaSp1 is distributed more or less
blocks (Ala)n and the (Gly-Ala)n anking them are known homogeneously throughout the core of the lifeline ber
to form -sheet structures that are responsible for the forming a -sheet crystal reinforced matrix, with elon-
high-tensile strength of MA silks; whereas the blocks of gated island-like structures of MaSp2 in the core of the
(Gly-Gly-Ala) form 31 -helices, and the blocks of Gly-Pro- ber that are believed to act as deformable shock absorbers,
Gly-X-X form -turn spirals imparting elasticity/exibility improving the elasticity of the bers. These shock absorbers
to the proteins [2023]. further reinforce the ber, thereby reducing the likelihood
Of particular note is that the two major proteins com- of breaking upon the impact of prey into the web. This
prising the core lament (of bers spun by N. clavipes core lament is coated in a layer of minor ampullate (MI)
spiders) are inhomogeneously distributed [6,7]. The ori- spidroin-like protein (known as the skin), a glycoprotein
gin of the inhomogeneous distribution is believed to be coat, and nally a lipid coat (see Fig. 2C) [6,7].

Fig. 2. A) Photograph of an Araneus diadematus spider. B) Electron micrograph of major ampullate bers. C) Schematic illustration of the composite structure
of a major ampullate ber. The core ber is composed of major ampullate (MA) spidroins, coated by a layer of minor ampullate-like (MI-like) spidroin
known as the skin, a layer of glycoprotein and nally a layer of lipids [2]. Adapted with the permission of both the authors and publisher, Copyright 2008,
Elsevier Science Ltd., Oxford, UK.
1096 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

1.3. Sources of silk proteins prepared by freeze-drying hydrogels, gas foaming or salt-
leaching; silk spheres can be prepared by electrospraying or
Although many animals produce silk proteins for vari- precipitation upon addition of a poor solvent to a solution
ous different applications [1], in this review we will focus of silk, and silk capsules can be prepared by adsorption of
on the silkworm and spider silk proteins as they are the the protein at the interface of a water-in-oil emulsion. The
most extensively studied and readily abundant. materials formed by the above processes are often treated
Humans domesticated B. mori silkworms thousands of with alcohols (typically methanol) or aqueous solutions of
years ago in order to be able to produce silk on an indus- kosmotropic salts (such as potassium phosphate) in order
trial scale for a variety of applications. A simple washing to induce -sheet formation in the material, which reduces
procedure after harvesting the cocoons removes the sericin the solubility in water and changes the mechanical prop-
coating on the silk bers yielding clean B. mori silk broin erties (Fig. 3) [3,34,35]. We refer interested readers to a
[16]. A similar process is applicable to silk bers produced selection of recent reviews including sections on the pro-
by wild silkworms (such as Antheraea pernyi, Antheraea cessing of silk-based materials [3,11,34,35].
mylitta, or Samia cynthia ricini silkworms [16]) providing
us with a source of silk proteins with differing amino acid 2. Man-made composite materials based on silk
sequences in the protein (therefore differing biocompati- proteins
bility, biodegradability and solubility in various solvents).
Although it is possible to harvest milligrams of spider As noted in Section 1, the mechanical properties and
silk bers, either at their point of application (e.g. from their biocompatibility of naturally occurring silkworm and spi-
egg cocoons or webs) or directly from the animal (via forced der silk protein-based bers have allowed humans to use
silking), this is an incredibly time consuming and expensive such bers for millennia for a variety of applications. The
process (e.g. a 3.4 m rug produced from the major ampul- development of new composite materials based on silk pro-
late silk bers of Nephila madagascarenis spiders, took 70 teins has been driven in part by the necessity to address the
people 4 years to prepare from the silk collected from over shortcomings of natural silk bers for certain applications
1 million spiders at a cost of over half a million US dollars (e.g. in textiles) and our ability to utilise the proteins as
[26]). Unfortunately attempts to farm spiders on an indus- building blocks of materials with interesting new applica-
trial scale have been fruitless owing to their cannibalistic tions.
nature, yet the determination of the primary structures
of natural spider silk proteins has allowed the produc- 2.1. Man-made composites based on silk proteins and
tion of spider silk-like proteins using recombinant DNA synthetic polymers
technology on an industrial scale [27]. Recombinant DNA
technology has also been used to prepare chimaeric/hybrid 2.1.1. Man-made composites based on silk proteins and
proteins incorporating silkworm silk-like or spider silk-like non-biodegradable synthetic polymers
sequences and other sequences that enhance the proteins Our ability to synthesize diverse structures with tunable
solubility, or improve biomineralization/cell adhesion of properties has made synthetic polymer based materials
materials prepared from the proteins [28]. absolutely ubiquitous in our daily lives. Indeed the rst
reports of silk-based composite bers in 1956 utilized non-
1.4. Processing silk proteins biodegradable synthetic polymers (poly(acrylonitrile) [36]
in this case) to improve their properties for textiles appli-
Natural silkworm silk bers require very little process- cations, and since this time a variety of other materials
ing prior to using them in the textiles industry (e.g. dyeing morphologies have been prepared based upon silk proteins
[2931], or chemical modication to render them water- in combination with non-biodegradable synthetic poly-
proof [32,33]). However, in order to prepare alternative mers. Table 2 highlights a small selection of interesting
material morphologies (e.g. lms, foams, hydrogels, bers examples.
with nanometer scale diameters, spheres or capsules) or
composite materials, it is typically necessary to dissolve 2.1.1.1. Carbon nanotubes. Carbon nanotubes are a
the proteins in a solvent capable of denaturing the protein recently discovered allotrope of carbon with nanometer
(by breaking the strong intermolecular hydrogen bonds scale diameters and lengths that can be many thousands of
stacks of -sheets are particularly important for silk pro- times longer. Carbon nanotubes are typically categorized
tein interaction), such as concentrated aqueous solutions of as either single-walled nanotubes (SWNT) or multi-walled
inorganic/organic salts, uorinated solvents, ionic liquids nanotubes (MWNT), both of which exhibit extraordinary
or strong acids [3,34,35]. Once the silk protein is dissolved strength and heat conduction [37,38].
it can be processed into a variety of different material mor- Fibers composed of B. mori broin and SWNT were pro-
phologies; for example silk bers with m scale diameters duced by electrospinning blends of 12 wt% B. mori broin
can be prepared by hand-drawing or dry-/wet-spinning, and between 0.5 and 5 wt% SWNT in formic acid solution,
and silk bers with nm scale diameters can be prepared by resulting in smooth bers with diameters of 20250 nm.
electrospinning. Silk lms can be prepared by casting and There was evidence for both random coil and -sheet sec-
dip-/spin-coating; silk hydrogels can be prepared by expo- ondary structures in the broin which acted as a matrix for
sure of aqueous solutions of silk proteins (after dialysis to the dispersion of SWNT that were roughly aligned with the
remove any denaturant) to various stimuli including salt, long axis of the ber. It was found that by including 1 wt%
shear, sonication or up-concentration; silk foams can be of SWNT it was possible to improve the Youngs modulus
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1097

Fig. 3. Processing conditions of naturally/recombinantly produced silk proteins.

of the resultant bers by up to 460% at the expense of the by electrospinning from formic acid solution, resulting in
strength and strain to failure; all other ratios of broin and bers with improved Youngs modulus and tensile strength
SWNT were observed to worsen the mechanical proper- at the expense of the elongation at break [40].
ties of the bers due to poor alignment and dispersion of Films composed of B. mori broin and MWNT were cast
the SWNT within the broin matrix [39]. Likewise bers from aqueous solution, and there was evidence for ran-
composed of B. mori broin and MWNT can be produced dom coil, -helix and -sheet secondary structures in the
1098 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

Table 2
Examples of man-made composite materials based on silk proteins and synthetic polymers.

Morphologies Components Improvement vs. silk References

Fibers, lms or spheres Either A) B. mori broin or B) N. Electrical conductivity, [3749]


clavipes spidroin with carbon mechanical properties
nanotubes
Fibers B. mori broin and Electrical conductivity [69]
poly(pyrrole)
Films B. mori broin and poly(vinyl Water permeability [74]
alcohol)
Films B. mori broin and pellethane Biocompatible coating with [94]
heparin anticoagulant properties
Foams B. mori broin and Biomineralization (improved [7880]
poly(aspartic acid) mech. properties) needed for
tissue engineering
Foams B. mori broin and Cell adhesion and proliferation, [85]
poly(-caprolactone) i.e. tissue engineering

broin. There were notable improvements in tensile mod- other than 50:50 broin:nylon [50]. It was also possible to
ulus and tensile strength at the expense of elongation at prepare particles composed of B. mori broin and nylon 66
break [41]. Films composed of recombinantly produced by precipitation from formic acid solutions of the polymers
proteins based on the major ampullate spidroins of N. upon the addition of acetone, followed by ltration, wash-
clavipes with SWNT functionalized with octadecylamine ing with acetone and water drying; yielding particles in
were cast from hexauoroisopropanol solution and sub- which the polymers were phase separated [50].
sequently vacuum dried. The as-cast lms had very low
-sheet content (as hexauoroisopropanol induces -helix 2.1.1.3. Poly(acrylamide). Poly(acrylamide) is a highly
formation in proteins [42]) and consequently had poor water absorbent polymer used in both bulk applications
mechanical properties (stiffness, strength and ductility), such as waste water treatment and if highly puried
however, including small quantities (volume fraction of ca. (to remove the neurotoxic monomer acrylamide) as soft
0.1%) of SWNT improved these properties by 1445% [43]. contact lenses. Films composed of B. mori broin and
Fibers of B. mori broin with a MWNT coating were used to poly(acrylamide) were cast from aqueous solution, incu-
reinforce lms cast from polymer melts of poly(butylene bated in a humid atmosphere to induce -sheet formation
succinate). It was shown that a small amount of silk bers in the broin, and nally vacuum dried. The polymers
(ca. 3 wt%) dramatically improved the tensile strength and were phase separated, with spherical poly(acrylamide)
modulus of the composites [44]. particles dispersed in a continuous phase of broin. Films
Fibers and microspheres of B. mori broin have been with a poly(acrylamide) content of up to 25 wt% showed
coated with MWNTs by immersion of the bers or micro- improved mechanical stability compared to lms of broin
spheres in aqueous suspensions of the MWNTs, followed alone [51].
by washing and drying. Such coatings were stable to both
extensive washing and sonication due to the formation of 2.1.1.4. Poly(acrylonitrile). Fibers composed of poly
hydrogen bonds between the broin and the carboxylic (acrylonitrile) and its copolymers are cheap, antibacterial
acids on the surface of the nanotubes (introduced during and stable to photo-oxidation, and are commonly used
the nanotube purication procedure) [40,45,46]. for textiles and the reinforcement of composite materials;
Hydrogels composed of B. mori broin and MWNT can however, these polymers absorb little moisture (due
be prepared by sonication of the nanotubes in 5 wt% solu- to their hydrophobicity) and collect static electricity.
tions of B. mori broin at pH 12 followed by reduction of the Fibers composed of B. mori broin and poly(acrylonitrile)
pH of the solution to pH 4.5. Interestingly this gelation pro- were produced by wet-spinning solutions of 8.5 wt%
cess could be reversed by increase of the pH of the solution broin/poly(acrylonitrile) in aqueous ZnCl2 (56 wt%) into
to pH 12, although the authors do not mention if the broin an aqueous coagulation bath [36]. The bers were air-dried
is hydrolyzed under such conditions [47,48]. Freeze-drying for a few hours, drawn to ca. six times their original length,
such hydrogels yielded foams in which aggregated MWNT washed rst with aqueous Na2 CO3 (10 wt%) (neutralizing
were embedded in a continuous broin matrix. The MWNT the ZnCl2 ), then with water and nally air-dried. The resul-
interacted with the broin via hydrogen bonding interac- tant bers were transparent with circular cross-sections
tions between the broin and the carboxylic acids on the and were smooth (except at broin:poly(acrylonitrile)
surface of the nanotubes [49]. ratios of 20:80). X-ray diffraction studies showed that
the polymers were neither highly crystalline nor strongly
2.1.1.2. Nylon. Nylon 66 is a cheap and strong polyamide oriented, and scanning electron microscopy (SEM) studies
that is widely used in the plastics and textiles industries. carried out later indicated the polymers to be phase sep-
Films composed of blends of B. mori broin and nylon 66 arated [52]. Electrical resistivity measurements indicated
were cast from formic acid solution and vacuum dried. that the bers were contaminated with residual Zn; this
The polymers in the resulting lms were crystalline and problem could be overcome using sodium thiocyanate as
appeared to be homogenously distributed at all blend ratios the protein denaturant in the spinning dope. Subsequent
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1099

studies showed that the bers had a core-sheath structure, of applications. Fibers composed of B. mori broin and
with a broin-rich sheath, and a poly(acrylonitrile) rich poly(ethylene oxide) (900 kDa) were produced by elec-
core containing long brils of broin aligned with the trospinning aqueous solutions of 28 wt% B. mori broin
direction of the ber (caused by elongational ow forces with up to 1.8 wt% poly(ethylene oxide), yielding smooth
during the ber spinning process). The resulting bers had bers with diameters of 590910 nm. The polymers in the
mechanical properties usable in the textiles industry. as-electrospun bers were phase separated, with island-
Compatibilization of B. mori broin and poly(acrylo- like regions of crystalline poly(ethylene oxide) (elongated
nitrile) can be achieved through the addition of small along the long axis of the ber) and an unstructured silk
amounts (ca. 2 wt%) of broin-graft-poly(acrylonitrile) broin matrix (evaporation of the solvent during elec-
copolymers to mixtures of broin and poly(acrylonitrile). trospinning is fast, and silk broin crystallizes slowly in
The best compatibilization was achieved with a broin- water relative to poly(ethylene oxide)). Treatment of these
graft-poly(acrylonitrile) copolymer bearing (on average) bers with 90/10 (v/v) methanol/water rendered the sur-
one long poly(acrylonitrile) chain (of ca. 106 kDa), which face of the bers rough, due to partial dissolution of the
almost eliminates phase separation. When preparing bers poly(ethylene oxide) and simultaneous induction of -
based on mixtures of B. mori broin and poly(acrylonitrile) sheet formation in the broin [58,59]. Almost all of the
for textiles applications, complete compatibilization is poly(ethylene oxide) could be removed by subsequently
undesirable as it reduces the handle and luster of the bers washing the bers with water.
imparted by the silk sheath. A copolymer bearing (on aver- It is also possible to electrospin bers with a core-sheath
age) two short poly(acrylonitrile) chains (of ca. 65 kDa) was morphology by a two-uid electrospinning technique (one
used to successfully improve the tensile strength by 50% uid containing B. mori broin and the other poly(ethylene
(relative to the ber in the absence of the compatibilizer) oxide)) using water as the common solvent. The two-uid
whilst maintaining a broin-rich sheath [53,54]. electrospinning process is the key to producing a core
Particles composed of B. mori broin and poly lament of unblended, unstructured broin and allowed
(acrylonitrile) were prepared by precipitation from aque- a variety of core-sheath diameters to be prepared [60].
ous solutions of the polymers upon the addition of Exposure of the as-spun bers to a humid atmosphere
methanol (inducing -sheet assembly in the broin), fol- plasticized the bers and allowed -sheet formation in
lowed by ltration, dialysis (to remove methanol) and vac- the broin, and the poly(ethylene oxide) sheath could
uum drying, yielding particles in which the polymers were be removed by subsequently washing the bers with
phase separated. In contrast, the polymers in particles com- water.
posed of B. mori broin and poly(acrylonitrile-co-methyl Films composed of B. mori broin and poly(ethylene
acrylate) were more homogeneously distributed due to the oxide) were cast from aqueous solution, treatment with
existence of hydrogen bonds between the polymers [55]. methanol induced -sheet formation in the broin. At a
blend ratio of 98:2 broin:poly(ethylene oxide) a homoge-
2.1.1.5. Poly(allylamine). Poly(allylamine) is a polycationic nous lm was formed with improved tensile strength and
polymer that is highly soluble in water and utilized widely elongation at break in comparison with lms composed
in biomedical applications. Films composed of B. mori solely of broin. In lms with a higher poly(ethylene oxide)
broin and poly(allylamine) were cast from aqueous solu- content the polymers were phase separated, with broin
tion and found to be more stable in water than lms particles homogeneously dispersed in a poly(ethylene
composed only of poly(allylamine). The polymers were oxide) matrix. Films with a poly(ethylene oxide) con-
compatible, yet Fourier transform infrared (FTIR) spectro- tent of greater than 30 wt% had improved elongation
scopic studies indicated only weak interactions between at break at the expense of tensile modulus and ten-
the polymers [56]. sile strength. As previously demonstrated with the bers,
the poly(ethylene oxide) matrix could be washed away
2.1.1.6. Poly(epoxides). Poly(epoxides) are commonly used in water, yielding a porous lm composed of broin
as adhesives, coatings and structural materials. Incor- [61,62].
poration of waste silk fabric (1 wt%) into lms cast of Hydrogels composed of B. mori broin and
Araldite HY960 epoxy resin (with a monomer:hardener poly(ethylene oxide) were prepared simply by mix-
ratio of 5:1) yielded resins with improved stiffness at the ing aqueous solutions of the polymers. Hydrogelation
expense of tensile strength, ductility and elongation at occurred faster as the poly(ethylene oxide) content
break [44]. In contrast, the incorporation of waste silk fabric increased due to restriction of the mobility of the broin,
(degummed B. mori broin bers) (up to 25 wt%) into lms encouraging the formation of -sheets between the
cast from polymer melts of an epoxy resin (synthesized proteins [63]. Cross-linking of the poly(ethylene oxide)
from Vanticos LY556 monomer and HY951 hardener) led polymers was demonstrated to signicantly improve the
to increases in tensile strength, tensile modulus and elon- mechanical properties (tensile strength and elongation at
gation at break. The toughness could be further increased break) and water swellability of the hydrogels [6466].
by addition of either poly(methyl methacrylate) (4 wt%) or Hydrogels composed of B. mori broin and a
poly(carbonate) (6 wt%) [57]. poly(ethylene oxide)-poly(propylene oxide) copoly-
mer (Pluronic F-127) were prepared simply by mixing
2.1.1.7. Poly(ethylene oxide). Poly(ethylene oxide) (also acidied (pH 4) aqueous solutions of the polymers. The
known as poly(ethylene glycol)) polymers are cheap, viscosity of the hydrogels was found to increase as the
water-soluble and biocompatible, with a wide variety concentration of Pluronic F-127 increased [67].
1100 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

2.1.1.8. Poly(pyrrole). Poly(pyrrole) is a biocompatible (a consequence of which is that they swell more in water),
conducting polymer discovered in the 1960s currently the degree of swelling could be reduced by annealing the
investigated for a wide variety of biomedical applications lms at 7080 C which induces -sheet formation in the
[68]. Forcibly silked major ampullate silk bers (pro- broin (as shown by FTIR) due to the slow removal of resid-
duced by Nephila edulis spiders) were dip-coated with ual water from the lm (as shown by differential scanning
poly(pyrrole) followed by air drying, with almost no detri- calorimetry (DSC)), thereby increasing their water resis-
mental effect upon the mechanical properties of the bers tance [74].
[69]. Such composite bers may nd application in elec- Hydrogels composed of B. mori broin and poly(vinyl
trically stimulated tissue scaffolds or drug delivery devices alcohol) were prepared by mixing aqueous solutions of the
[68]. polymers, incubation at 50 C for 2 h, freeze-drying and
re-hydration. The elongation at break of the hydrogels was
2.1.1.9. Poly(styrene). Poly(styrene) is cheap and utilized demonstrated to increase as the poly(vinyl alcohol) con-
in a wide variety of applications. Films composed of B. mori tent increased, yet the strength was unaffected [75]. Foams
broin and ordered arrays of poly(styrene) microspheres composed of B. mori broin and poly(vinyl alcohol) have
were prepared by casting aqueous solutions of broin been prepared by either air or freeze-drying the aforemen-
onto the microspheres, followed by air drying and subse- tioned hydrogels. Deeper freezing increased the -sheet
quent treatment with ethanol to induce -sheet assembly content of the foams (as did air drying) and consequently
in the broin. Fibroin foams with three-dimensionally the mechanical properties (strength and elongation at
ordered pores could be produced by washing these lms break) were improved relative to the freeze-dried foams.
with toluene which selectively dissolved the poly(styrene) Moreover, the strength and elongation at break increased
microspheres, yielding foams that were superhydrophobic as the poly(vinyl alcohol) content increased [75,76].
and mechanically stable [70].
2.1.2. Man-made composites based on silk proteins and
2.1.1.10. Poly(vinyl alcohol). Poly(vinyl alcohol) is a cheap, biodegradable synthetic polymers
water-soluble, non-toxic, biocompatible and degradable Biodegradable synthetic polymers are more attractive
polymer. Fibers composed of blends of B. mori broin and than their non-biodegradable brethren for many materials
poly(vinyl alcohol) were produced by wet-spinning solu- applications, particularly when considering their disposal
tions of 19 wt% broin/poly(vinyl alcohol) in formic acid at the end of their useful lifetimes as they pose fewer envi-
into a methanol coagulation bath where they were left ronmental hazards. In this context a number of researchers
overnight to allow solidication of the ber and -sheet have investigated the properties of composite materials
formation in the broin [71]. Once solid, the bers were composed of silk proteins and biodegradable polymers for
plasticized in hot water (70 C), drawn to ca. 4.5 times a variety of applications [77]. Table 2 contains a small selec-
their original length, and nally air-dried under tension. tion of interesting examples.
The resultant white bers had circular cross-sections (with
diameters of between 220 and 270 m) and were smooth. 2.1.2.1. Poly(aspartic acid). The poly(aspartic acid)
X-ray diffraction studies indicated that the broin was crys- domains of certain natural proteins are known to be
talline and that the poly(vinyl alcohol) was amorphous, involved in biomineralization processes. Foams composed
and SEM studies demonstrated that the polymers were not of B. mori broin and poly(aspartic acid) were prepared
macroscopically phase separated. Wet-spun bers com- by casting mixtures of the polymers in aqueous solution
posed solely of B. mori broin had reasonable breaking into a container packed with sodium chloride, incubation
strength, but were brittle and inexible. Blending up to for 24 h at room temperature (allowing the formation of
50 wt% of poly(vinyl alcohol) was shown to increase the -sheets between the broin), followed by salt-leaching
tenacity and elongation at break of the bers (at the by washing with water. The foams were -sheet rich,
expense of the breaking strength). and had a continuous network of pores of approximately
It is possible to prepare lms by casting aqueous solu- 750 m [7880]. Inclusion of poly(aspartic acid) in such
tions of B. mori broin and poly(vinyl alcohol). The lms composite materials is benecial for their subsequent
were annealed at 200 C which crystallized the poly(vinyl biomineralization, as discussed later in this review.
alcohol) without affecting the conformation of broin (as
shown by FTIR), followed by immersion in methanol to 2.1.2.2. Poly(-caprolactone). Poly(-caprolactone) is a
induce -sheet formation in the broin (also shown by biodegradable polyester approved by the US Food and
FTIR). The polymers in the lms were phase separated, and Drug Administration (FDA). Films composed of poly(-
the mechanical properties were consequently non-additive caprolactone) reinforced with naturally spun B. mori
[72,73]. It is also possible to prepare transparent lms by broin bers were prepared by melt-mixing of the poly(-
casting mixtures of B. mori broin and poly(vinyl alcohol) in caprolactone) and broin bers followed by hot pressing
hexauoroisopropanol solution. At a weight ratio of 80:20 (at 140 C) into the desired form. The lms with 3545 wt%
broin:poly(vinyl alcohol) the lm was homogenous, with ber content had optimal mechanical properties [8183].
a higher -sheet content than lms cast from broin alone. Subsequent studies on such systems showed that exposure
At higher weight ratios the polymers were phase separated, to electron beams generates radicals on both the poly(-
with broin particles (of 17 m) dispersed in a continuous caprolactone) and broin leading to both cross-linking
phase of poly(vinyl alcohol). An increase in the poly(vinyl and degradation of the polymers, which resulted in mod-
alcohol) content increases the hydrophilicity of the lms erate increases in tensile strength and exural strength
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1101

below 150 kGy, and notable decreases above 150 kGy [84]. good mechanical properties of the homopolymers make
Composite materials in which a B. mori broin foam lled them ideal for load bearing applications, whereas the
the pores of a poly(-caprolactone) foam were prepared copolymers are preferred for drug delivery applications.
by freeze-drying poly(-caprolactone) foams lled with Films composed of B. mori broin and poly(lactic acid)
aqueous solutions of broin [85]. were cast from aqueous dioxane solution and dried at 30 C
and 50% relative humidity for 48 h. The polymers were
2.1.2.3. Poly(-caprolactone-co-d,l-lactide). Poly(- phase separated with poly(lactic acid) particles dispersed
caprolactone-co-d,l-lactide) is a biodegradable polymer in a continuous phase of broin, and they were shown to
of great popularity that degrades relatively quickly. Films interact via hydrogen bonding interactions. Furthermore,
composed of B. mori broin and poly(-caprolactone-co- mixing up to 10 wt% of poly(lactic acid) was demonstrated
d,l-lactide) were prepared by casting suspensions of B. to moderately improve the tensile strength and elongation
mori broin particles in solutions of poly(-caprolactone- at break of the lms and decrease the hydrophilicity of the
co-d,l-lactide) in acetone, followed by vacuum drying. lms [91].
The -sheet rich broin particles were homogeneously Foams composed of B. mori broin and poly(l-lactide)
distributed within the poly(-caprolactone-co-d,l-lactide) were prepared by freeze-drying dioxane solutions of
matrix, and the lms became more opaque as the broin blends of soluble poly(l-lactide) and insoluble -sheet
content increased. Furthermore, the mechanical properties rich broin particles (550 m). The broin particles were
(storage modulus and hardness) of the lms were observed homogeneously distributed throughout the poly(l-lactide)
to improve as the broin content increased, indicative of foam and were shown to interact with the poly(l-lactide)
hydrogen bonding between the broin particles dispersed matrix via hydrogen bonding interactions; furthermore,
in the poly(-caprolactone-co-d,l-lactide) matrix [86]. the presence of broin increased the hydrophilicity of the
foams [92].
2.1.2.4. Poly(carbonate-urethane). Poly(urethane) elas- Fibers composed of Agelena labyrinthica spidroin
tomers have been used extensively in biomedical and poly(d,l-lactide) were produced by electrospinning
applications due to their biocompatibility and mechan- emulsions of (A. labyrinthica spidroin in formic acid)-in-
ical properties. Poly(ester urethanes) are unsuitable for (poly(d,l-lactide) in acetone), followed by drying under
long-term implantation as they are hydrolyzed relatively vacuum for 4 days, yielding bers with diameters of
quickly in vivo; poly(ether urethanes) are more stable to 891050 nm. The polymers in the as-electrospun bers
hydrolysis, but observed to oxidize in vivo after extended were phase separated, and the proteins adopted pre-
periods; poly(carbonate urethanes) by contrast are more dominantly -helical/random coil conformations (with
stable to both hydrolysis and oxidation in vivo and have some -sheet content). At low weight ratios (of <15 wt%
more potential for use as long-term implants. Bilayer spidroin:poly(d,l-lactide)) this resulted in granular regions
lms composed of B. mori broin and medical grade of silk spidroin within a poly(d,l-lactide) matrix, whereas at
poly(carbonate-urethane), were prepared by: rst cast- higher weight ratios (of >15 wt% spidroin:poly(d,l-lactide))
ing lms of poly(carbonate-urethane) from solution in the bers had a core-sheath morphology (in which the core
tetrahydrofuran/dioxane, drying, and applying a second was spidroin). As the spidroin content increased, so did the
layer of broin (cast from aqueous solution), drying and mechanical properties (tensile strength and elongation at
nally treating with methanol (inducing -sheet assembly break) of the resultant bers [93].
in the broin). The broin surface was slightly rough and
stable to prolonged incubation in phosphate buffered 2.1.2.7. Poly(urethane) pellethane. Pellethane is the trade
saline solutions [8789]. name of a family of polyurethanes that are biodegradable,
thermoplastic elastomers. Films composed of pellethane
2.1.2.5. Poly(lactic-co-glycolic acid). Poly(lactic-co-glycolic (2363-80A), B. mori broin particles (of ca. 3.6 m) and
acid) copolyesters have been developed commercially for the sodium salt of heparin (10 wt%) were cast from DMF
a wide range of biomedical applications. Poly(lactic-co- solution, and the solvent removed at room temperature,
glycolic acid) microspheres (with diameters of ca. 53 m) followed by a nal drying step at 80 C for 24 h, resulting
were coated with silk by adsorption (due to hydrophobic in lms that were relatively smooth in which the broin
interactions) from an aqueous solution of B. mori broin and heparin were homogenously dispersed in a matrix of
(0.1% w/v), prior to repetitive centrifugation/washing pellethane [94].
steps, exposure to aqueous methanol (inducing -sheet
assembly in the broin), and drying under a stream of 2.2. Man-made composites based on silk proteins and
nitrogen (two further coats of broin were applied using biopolymers
the same procedure). The silk coating was approximately
1 m thick and stable to prolonged incubation in phos- 2.2.1. Man-made composites based on silk proteins and
phate buffered saline solution [90]. other proteins
As described in Section 1, naturally occurring silk bers
2.1.2.6. Poly(lactic acid). Poly(lactic acid) which is also are composite materials based upon the combination of a
known as poly(lactide), has been developed for a variety of variety of proteinaceous components (broins and sericins
biomedical applications; homopolymers of l-lactide or d- in the case of silkworm cocoon bers, or major ampul-
lactide tend to be crystalline and degrade slowly, whereas late spidroins 1/2 (or analogues), minor ampullate silk-like
copolymers tend to be amorphous and degrade faster. The proteins, glycoproteins and lipids in the case of spiders
1102 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

Table 3
Examples of man-made composite materials based on silk proteins and biopolymers.

Morphologies Components Improvement vs. silk References

Fibers or spheres B. mori broin and either bone Tissue engineering: growth [118,119]
morphogenic protein-2 or factor determined
insulin-like growth factor cell-differentiation
Films B. mori broin and either A) Catalytic activity of the [104108,161,218]
glucose oxidase or B) respective enzyme
horseradish peroxidase or C)
-galactosidase
Films B. mori broin and green Non-linear optical properties [117]
uorescent protein
Microspheres embedded in foams or hydrogels Either A) A. mylitta broin or B) Drug delivery without burst [119,130]
B. mori broin and alginate prole
Fibers B. mori broin cellulose Cheap textiles from waste [131,137]
materials

dragline bers). In vitro a variety of other composite materi- The water swellability of the foams increased as the col-
als have been prepared based upon mixtures of silk proteins lagen content increased. The mechanical properties (yield
and other proteins/enzymes that have a number of exciting stress and compressive moduli) increased with increas-
applications. Table 3 shows some interesting examples. ing collagen content [100102]. Subsequent studies with
recombinantly produced human-like collagen yielded sim-
ilar results [103].
2.2.1.1. Collagen. Collagen is the most abundant protein
in mammals, and is particularly important in the extra-
cellular matrix. Fibers composed of B. mori broin and 2.2.1.2. Enzymes. Enzymes are biomolecules that catalyze
collagen were produced by electrospinning solutions of B. certain chemical reactions, and their incredible catalytic
mori broin and collagen in hexauoroisopropanol. Incuba- efciency has resulted in numerous biomedical and tech-
tion in an atmosphere of water and glutaraldehyde vapor, nical applications of enzymes that are easy to isolate. Films
followed by immersion in an aqueous solution of glycine (to composed of B. mori broin and the enzymes Glucose oxi-
block unreacted aldehyde groups), induced -sheet forma- dase or Horseradish peroxidase were prepared by casting
tion in the broin and chemically cross-linked the proteins. from aqueous solutions, followed by exposure to aqueous
The samples were nally vacuum dried, resulting in smooth methanol (inducing -sheet assembly in the broin), and
bers with average diameters of 320360 nm [95,96]. air drying (Table 3) [104108]. The enzymes were homo-
Films composed of B. mori broin and recombinant geneously distributed and remained trapped within the
human-like collagen were prepared by casting from warm -sheet cross-linked broin matrix even after long periods
(5060 C) aqueous solutions, followed by drying at 70 C. of exposure to water.
The lms cast from broin alone contained some -
sheet crystals, probably due to the high temperature 2.2.1.3. Fibroins of different species. As outlined in Section
drying/annealing process. The presence of collagen in the 1, mankind has harvested B. mori silkworm broin for
lms resulted in the formation of hydrogen bonds between thousands of years for use in diverse applications. Other
the two proteins (at the expense of -sheet content), and species of silkworms produce different broins for protec-
the lms were consequently slightly less mechanically tive cocoons that have in recent years been investigated
stable than lms cast from broin alone. Recombinant with a view to understanding the molecular basis for the
human-like collagen is more hydrophilic than broin, so different mechanical properties of their silks, and their
the surfaces of the lms were more hydrophilic than those potential use in various medical applications.
cast from broin alone [97]. Films composed of B. mori Films composed of B. mori broin were prepared by cast-
broin and bovine collagen cast under analogous condi- ing from aqueous solutions (of soluble broin and insoluble
tions showed moderate improvements in the wet-state -sheet rich broin nanoparticles) followed by drying at
exibility of the lms at the expense of strength and stiff- 45 C. It was demonstrated that the presence of -sheet
ness [98]. Films composed of B. mori broin and rat-tail rich broin nanoparticles induced -sheet formation in the
collagen were prepared by casting from aqueous acetic acid lms, yielding smooth lms that were insoluble in water
solutions, air drying, treating with methanol to induce - [109].
sheet formation in the broin, and air drying again. The B. mori and A. pernyi broins have notably different pri-
lms were optically transparent, smooth and hydrophilic mary structures. The glycine content being higher in B. mori
(water uptake of ca. 30% dry weight) [99]. broin, and the content of acidic and basic amino acids
Foams composed of B. mori broin and bovine col- being higher in A. pernyi broin; moreover, the -sheet
lagen were prepared by freeze-drying aqueous solutions forming segments of B. mori broin are composed mainly
of the polymers. The foams were subsequently treated of (Gly-Ala)n repeats, whereas in A. pernyi broin they are
with methanol (inducing -sheet assembly in the broin) composed mainly of (Ala)n repeats. Films composed of B.
and air-dried. The polymers were homogeneously dis- mori and A. pernyi broins were prepared by casting from
tributed in the foams and interacted via hydrogen bonds. aqueous solutions (of native proteins extracted from the
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1103

lumen of the silkworms), followed by incubation in a humid incorporated in B. mori broin microspheres which were
atmosphere (allowing the formation of -sheets in the subsequently incorporated within either an alginate gel or
broins) and vacuum drying. Various techniques demon- a B. mori broin foam [119].
strated that the broins were phase separated in the lms
[110]. 2.2.1.7. Keratin. Keratins are tough structural proteins
Films composed of B. mori broin and an engineered produced by animals. Fibers composed of B. mori broin
hybrid protein-based on the repeat sequence of N. clavipes and oxidized merino wool keratin were produced by elec-
spidroin and an integrin-binding Arg-Gly-Asp motif (more trospinning solutions of B. mori broin and keratin in
commonly referred to as an RGD motif), were prepared formic acid. The as-spun bers were smooth with diam-
by spin-coating from hexauoroisopropanol solutions, and eters between 160 and 900 nm, and the proteins adopted
either left to dry at room temperature or annealed by incu- predominantly -helical/random coil conformations [120].
bation in a hot (70 C) humid atmosphere. The proteins Immersion of the bers in methanol induced -sheet for-
in unannealed lms were predominantly randomly coiled, mation in the broin, and incubation in an atmosphere of
whereas those in the annealed lms were -sheet rich with formaldehyde vapor chemically cross-linked the proteins,
-sheet content increasing with spidroin content [111]. further improving their mechanical stability [121].
Films composed of B. mori broin and merino wool ker-
2.2.1.4. Gelatin. Gelatin is produced by hydrolysis of colla- atin were prepared by casting from formic acid solution.
gen, and used as a gelling agent in drug formulations, food, The presence of the keratin promoted -sheet formation in
and photography. It was possible to induce hydrogel for- the broin (due to hydrogen bonding interactions between
mation in aqueous solutions containing B. mori broin and the proteins), as did treatment of the lms with methanol
gelatin by exposure to methanol (inducing -sheet forma- [120,122,123].
tion in the broin) [112,113]. It is also possible to prepare
tubes of gelatin-based hydrogel reinforced with naturally 2.2.1.8. Sericin. As described in Section 1, sericins are gly-
spun B. mori broin bers, yielding tubes with mechanical coproteins produced by silkworms that are used as a
properties similar to those of arteries [114]. Films com- protective glue coating on broin bers [16]. Films com-
posed of B. mori broin and gelatin were cast from aqueous posed of B. mori broin and sericin were prepared by
solutions, followed by treatment with aqueous methanol casting from aqueous solutions, followed by drying at 80 C.
and vacuum drying. The proteins were homogeneously dis- The proteins in the as-cast lms were amorphous and
tributed and were demonstrated to interact via hydrogen phase separated. Exposure of the lms to methanol induced
bonds, and blend lms were more thermally and mechan- -sheet formation in the broin, but this was retarded
ically stable than the lms cast from either protein alone (although not prevented) by the presence of sericin in
[115]. Subsequent studies demonstrated it is possible to the lms, suggesting the interaction of the polymers via
prepare microporous broin lms by dissolution of the hydrogen bonding interactions at the interface of the phase
gelatin in aqueous phosphate buffered saline solution at boundary [124].
37 C [116].
2.2.1.9. Spidroins of different species. As described in Sec-
2.2.1.5. Green uorescent protein. Green uorescent pro- tion 1, spidroins are brous proteins produced by spiders
tein uoresces green when exposed to blue light, and is for a number of task-specic applications. Natural major
used extensively in cell and molecular biology. Films com- ampullate silk bers are composed of multiple proteins (see
posed of B. mori broin and green uorescent protein were Table 1) [6,7], and in an effort to mimic the natural spin-
cast from aqueous solution and air-dried (Table 3). The ning process [125] we have used a microuidic device (as
lms had rough surfaces and the green uorescent protein its dimensions are similar to a spiders spinning duct) to pre-
was inhomogeneously distributed throughout the broin pare bers composed of recombinantly produced spidroins
matrix [117]. Such composite materials have potentially (eADF-3 and eADF-4 [24]) that are based upon the con-
interesting nonlinear optical applications. sensus sequences of the major ampullate silks ADF-3 and
ADF-4 of A. diadematus spiders. Using biomimetic condi-
2.2.1.6. Growth factors. Bone morphogenic proteins are tions (combining increasing the phosphate concentration,
a group of growth factors and cytokines that induce pH decrease and elongational ow) we successfully pre-
the formation of bone and cartilage; such proteins can pared bers with m scale diameters in which the proteins
be produced recombinantly and are currently under were -sheet-rich [126].
development for medical and dental applications. Fibers We have also cast lms from mixtures of such recom-
composed of B. mori broin, poly(ethylene oxide) and bone binantly produced spidroins in hexauoroisopropanol
morphogenic protein-2 were electrospun from aqueous solution. The as-cast lms were -helix rich and water-
solution, immersed in methanol (rendering the bers insol- soluble, whereas those subsequently treated with either
uble in water due to -sheet assembly in the broin) then methanol or potassium phosphate were -sheet rich and
dried, yielding smooth bers with diameters of approxi- consequently insoluble in water [127].
mately 570 nm (Table 3). The polymers in the bers were
phase separated with the proteins in one phase and the 2.2.2. Man-made composites based on silk proteins and
poly(ethylene oxide) in another [118]. polysaccharides
More recently, bone morphogenic protein-2 and Polysaccharides are ubiquitous in nature and are typi-
recombinant human insulin-like growth factor-1 were cally utilized for energy storage or as structural elements in
1104 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

plant and cell walls that are often used in composite mate- studies demonstrated the potential to form porous lms
rials owing to their high natural abundance and relatively (composed of cellulose) via casting Cuoxam solutions of the
low cost. Table 3 shows some interesting examples. polymers, followed by sequential treatment with: aqueous
sodium hydroxide (removing the broin), aqueous sulfu-
2.2.2.1. Alginate. Alginate is a linear polysaccharide found ric acid, acetone/acetic acid, aqueous glycerine, and water,
in the cell walls of algae, and is utilized in the biomed- before nally air drying [133,134].
ical, food and textiles industries. Films composed of B. It is also possible to prepare lms composed of B. mori
mori broin and sodium alginate were prepared by casting broin and cellulose by casting from N-methylmorpholine
from aqueous solution, followed by incubation in a humid oxide solution, followed by treatment with aqueous alco-
atmosphere (plasticizing the protein, allowing the forma- hol, washing with water and air drying. DSC demonstrated
tion of -sheets in the broin). The mechanical properties the formation of intermolecular hydrogen bonds between
of the lms were optimal when the alginate content was the broin and cellulose, and that the broin was -sheet
30 wt%, and the water absorbance of the lms was notably rich [135].
improved by mixing alginate with broin [128]. Foams pre-
pared by freeze-drying aqueous solutions of B. mori broin 2.2.2.3. Cellulose xanthate (viscose). Cellulose xanthate
and sodium alginate had compressive moduli that were (viscose) is a polymer popular in the textiles industry.
better than the foams composed of broin alone [129]. Fibers composed of B. mori broin and viscose were
Alginate microspheres (with diameters of ca. 700 m) produced by wet-spinning solutions of broin/viscose in
were prepared by precipitating alginate in aqueous calcium aqueous sodium hydroxide solution into an acidic coagu-
chloride solution, and puried by shaking, ltration and lation bath (aqueous sulfuric acid and ammonium sulfate)
washing in water. The resulting microspheres were coated where they were left overnight to allow complete solidi-
with silk by adsorption (due to hydrophobic interactions) cation of the ber. The bers were subsequently washed in
from an aqueous solution of B. mori broin (0.1% w/v), various methanolic solutions before being air-dried under
prior to repetitive centrifugation/washing steps, expo- tension. The resultant white bers had circular cross-
sure to aqueous methanol (inducing -sheet assembly in sections, and the surface of the bers had very ne grooves
the broin), and drying under a stream of nitrogen (two aligned with the ber axis. The polymers were phase sep-
further coats of broin were applied using the same pro- arated, with silk dispersed in a viscose matrix [136], and
cedure). The silk coating was approximately 10 m thick the bers had mechanical properties suitable for certain
and slightly rough [90]. It is also possible to prepare micro- textiles applications.
spheres composed of alginate and A. mylitta broin by
an analogous method (Table 3) [130], and more com- 2.2.2.4. Chitin. Sodium N-acetylchitosan (chitin) is the
plex composite materials composed of such microspheres main component of the cell walls of fungi, and the
embedded in A. mylitta broin foams [130]. exoskeletons of arthropods and insects, that is utilized in
adhesives, foods and various biomedical/ltration tech-
2.2.2.2. Cellulose. Cellulose is the most common organic nologies. Fibers composed of B. mori broin and chitin
compound on Earth, which is found in the cell walls of many were produced by wet-spinning solutions of broin/chitin
plants, and is widely used in the textile industry (in the in aqueous sodium hydroxide solution into an acidic coag-
form of cotton, hemp or jute) as it is cheap, biodegradable ulation bath (aqueous sulfuric acid and ammonium sulfate)
and moisture absorbent. Fibers composed of B. mori broin where they were left overnight to allow complete solidi-
and cellulose were produced by wet-spinning solutions cation of the ber. The bers were subsequently washed
of broin/cellulose in N,N-dimethylacetamide (containing in various methanolic solutions before being air-dried. The
7 wt% LiCl) into an ethanol coagulation bath, and were resultant white bers had circular cross-sections, and the
washed for 34 days to assure extraction of all of the LiCl surface of the bers was slightly rough. The mechanical
(Table 3) [131]. The resultant bers had circular cross- properties of the bers were poorer than those of either
sections (with diameters of between 20 and 36 m) and silk or chitin alone, but potentially usable in the textiles
were smooth. SEM studies demonstrated that the poly- industry [137]. It is also possible to prepare bers com-
mers were not macroscopically phase separated, and X-ray posed of B. mori broin, viscose and chitin in an analogous
diffraction studies indicated that the crystalline broin was way [136], or to prepare bers by electrospinning from
dispersed in an amorphous matrix of cellulose. hexauoroisopropanol solutions [138,139].
Films composed of B. mori broin and cellulose were Films composed of blends of B. mori broin and either
prepared by casting aqueous cuprammonium hydroxide chitin [140] or carboxymethyl chitin [141] were prepared
(Cuoxam) solutions of the polymers, followed by treat- by casting from aqueous solution, followed by treatment
ment with acetone/acetic acid, and washes with aqueous with aqueous methanol (inducing -sheet assembly in the
glycerine and water, before air drying. Infrared absorption broin), and vacuum drying; yielding lms in which the
spectra showed intermolecular hydrogen bonds between polymers were homogeneously distributed and interact via
the broin and cellulose, and peaks characteristic of the hydrogen bonding.
crystallinity of the polymers (-sheet rich broin and cel- It is possible to deposit a layer of B. mori broin on
lulose II). The mechanical properties (tensile strength and porous -chitin bers by dip coating the -chitin bers in
elongation at break) were distinctly improved relative to an aqueous solution of B. mori broin followed by drying
the lms cast from broin alone, despite minor degrada- and treatment with aqueous methanol (inducing -sheet
tion of the broin in the Cuoxam solution [132]. Subsequent assembly in the broin) [142,143]. Such chitin bers can
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1105

also be used to reinforce B. mori broin foams prepared analogous foams including chondroitin-6-sulfate (which
by freeze-drying suspensions of the chitin bers in broin is also found in the extracellular matrix) [155].
hydrogels [144].
2.3. Man-made composites based on silk proteins and
2.2.2.5. Chitosan. Chitosan is a linear polysaccharide pro- inorganic particles
duced by deacetylation of chitin, and is utilized in
agriculture, biomedicine and ltration technologies. Fibers 2.3.1. Man-made composites based on silk proteins and
composed of B. mori broin and chitosan were electro- metal nanoparticles
spun from formic acid solution, immersed in aqueous A focus of intense current research interest is the
methanol (to induce -sheet assembly in the broin) and immobilization of biomolecules on nanoscale/colloidal
then vacuum dried. When the chitosan content was below substrates owing to their exciting applications in biochem-
30 wt% the polymers were homogenously distributed and istry, biotechnology and medicine. Many proteins can be
the bers were smooth with diameters of 130780 nm. conjugated to metal colloids via simply mixing the pro-
However, in bers with higher chitosan content there was teins with a metal colloidal sol (formed prior to mixing),
evidence of phase separation with chitosan-rich beads on and the protein typically binds the metal via amines or thi-
the surface of the bers [145]. ols displayed on their backbones. Table 4 highlights a small
Films composed of blends of B. mori broin and chi- selection of interesting examples.
tosan were prepared by casting from aqueous acetic acid
solution, followed by air drying [146]. At all blend ratios
2.3.1.1. Silver nanoparticles. Silver nanoparticles have
the polymers were homogeneously distributed and the
interesting antibacterial and electronic properties, and
broin was -sheet rich. Furthermore, a chitosan content
are currently the subject of much debate due to poten-
of 30 wt% signicantly improved the mechanical properties
tial environmental risks. B. mori broin bers (either
(tensile strength and initial tensile modulus) of the lms
naturally spun or electrospun) have been coated with
relative to lms cast from broin alone [147]. By contrast,
silver nanoparticles by (1) simply immersing the bers
lms composed of A. pernyi broin and chitosan exhibited
in a warm aqueous solution of the nanoparticles [156]
phase separation in lms with chitosan contents >40 wt%,
or (2) the tyrosine mediated reduction of Ag(I) to Ag(0)
with chitosan particles dispersed in a broin-rich matrix
under basic conditions [157,158], or (3) the photochemical
[148].
reduction of Ag(I) to Ag(0) (under irradiation with UV
Hydrogels composed of B. mori broin and chitosan
light) (Table 4) [159]. The nanoparticles were shown to
were prepared simply by mixing acidied (pH 4) aqueous
interact with some nitrogen and oxygen atoms present in
solutions of the polymers. The broin hydrogels inter-
broins.
acted with the chitosan via both physical entanglement
and hydrogen bonding interactions, and the viscosity of the
hydrogels was found to increase as the concentration of 2.3.1.2. Gold nanoparticles. Gold nanoparticles have inter-
chitosan increased [67]. Foams composed of B. mori broin esting electronic and optical properties and are utilized
and chitosan were prepared by freeze-drying aqueous solu- widely in biological and biomedical applications. Solutions
tions of the polymers. The foams were subsequently treated of B. mori broin have been used as an active scaffold for
with mildly basic aqueous methanol (inducing -sheet growing gold nanoparticles stabilized by a broin shell via
assembly in the broin and neutralizing the chitosan), and an in-situ redox reaction between the tyrosine residues on
washed with basic water and then phosphate buffered the silk protein and Au(III) (which is reduced to Au(0))
saline solution. The mechanical properties (tensile strength during which the pH 9 solution changed from yellow
and elastic modulus) increased with increasing broin con- to purple (Table 4). The gold cores were ca. 15 nm in
tent (as the gel was more highly cross-linked by -sheets), size and the core/shell particles (of ca. 45 nm) were sta-
and consequently their swellability in water was observed ble for months at room temperature [160,161]. Naturally
to decrease [149,150]. spun P. phalangoides spider silk bers have been coated
in gold nanoparticles via incubation of the ber in aque-
2.2.2.6. Hyaluronic acid. Hyaluronic acid is a gly- ous chloroauric acid, followed by washing with water, and
cosoaminoglycan that is found in the extracellular drying (Table 4) [162]. It is also possible to dip-coat natural
matrix, and has been investigated for cosmetic and spider silk bers with hydrophobically functionalized gold
medical applications. Foams composed of B. mori broin nanoparticles with almost no detrimental effect upon the
and hyaluronic acid were prepared by freeze-drying mechanical properties of the bers [69].
aqueous solutions of the polymers. The foams were subse-
quently treated with aqueous methanol (inducing -sheet 2.3.1.3. Transition metal oxides/suldes. Transition metal
assembly in the broin) and air-dried. The foams were oxides and suldes have a vast number of applications
demonstrated to be phase separated with a hyaluronic owing to their interesting electronic, magnetic and optical
acid rich phase and a broin/hyaluronic acid phase. The properties. Magnetic nanoparticles (such as superparam-
breaking strength moderately increased at the expense agnetic magnetite (Fe3 O4 )) have been used in numerous
of compressive modulus with increasing hyaluronic acid biomedical and technical applications, and semiconducting
content [151154]. It is possible to generate materials cadmium sulde nanoparticles are utilized in photoresis-
mimicking the natural extracellular matrix by incorpo- tors and solar cells. It has proven possible to dip-coat
ration of collagen coated B. mori broin bers within natural spider silk bers with such nanoparticles with
1106 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

Table 4
Examples of man-made composite materials based on silk proteins and inorganic particles.

Morphologies Components Improvement vs. silk References

Fibers or lms B. mori broin and silver Antibacterial properties [156159]


nanoparticles
Fibers B. mori broin and gold Electrical conductivity [69,160162]
nanoparticles
Fibers B. mori broin and magnetite Response to magnetic elds [69]
nanoparticles
Various B. mori broin and either A) Biomineralization (improved [79,80,118,182,184,187,193,195,207]
calcium carbonate or B) mechanical properties) needed
calcium phosphate or C) silica for tissue engineering

almost no detrimental effect to their mechanical properties mation of aragonite crystals [169], as was the presence of
(Table 4) [69]. -sheets in the broin lm [170]. Interestingly, biominer-
Titanium dioxide is used as a white pigment in foods, alization of naturally spun P. phalangoides spider silk bers
paints and sunscreens. Films composed of B. mori broin in the same fashion yielded only calcite crystals [171].
and titanium dioxide nanoparticles (of ca. 80 nm) were pre- Biomineralization of porous -chitin bers coated with
pared by air drying aqueous ethanol solutions that had been B. mori broin and macromolecules extracted from calcitic
sonicated for 30 min. The titanium dioxide nanoparticles biominerals yielded calcite, as did control samples with-
were evenly dispersed within a broin matrix that was out the broin. In contrast, biomineralization of porous
-sheet rich due to exposure to ethanol and sonication, -chitin bers coated with B. mori broin and macro-
and various techniques demonstrated the interaction of the molecules extracted from aragonitic biominerals yielded
nanoparticles with nitrogen and oxygen atoms displayed aragonite, whereas control samples without the broin
upon the backbone of the protein. The mechanical proper- yielded vaterite, suggesting that the silk protein was
ties (maximum load, maximum stress and elasticity) of the involved in specic polymorph determination [142].
broin lms were improved by including titanium dioxide Particles composed of B. mori broin and calcium car-
nanoparticles, and found to be optimal when the titanium bonate were prepared by precipitation from basic aqueous
dioxide content was between 0.2 and 0.4 wt% [163165]. solutions of broin micelles, calcium chloride and sodium
bicarbonate, in which the broin acted as a scaffold for
2.3.2. Man-made composites based on silk proteins and the nucleation and crystallization of calcium carbonate
biominerals microparticles via hydrogen bonding interactions. In the
Biominerals are natural composite materials based absence of the broin, a mixture of plate-like vaterite crys-
upon biomolecules (such as proteins) and minerals pro- tals (majority) and rhombic calcite crystals (minority) are
duced by living organisms via processes known as formed [172]; in the presence of broin at pH 8.6, a mixture
biomineralization, yielding materials with impressive of spherical and rice-like crystals are formed (composed
mechanical properties such as bones, shells and teeth. solely of calcite) probably because complexes of broin
During biomineralization the biomolecule acts as a tem- and CaCO3 clusters are thermodynamically stable; whereas
plate for the formation of the mineral phase (sometimes at pH 9.6, spherical crystals are formed (composed of a
even controlling which polymorph is produced) and such mixture of calcite and vaterite) due to the lower bind-
natural composite materials have been mimicked in vitro ing strength of complexes of broin and CaCO3 [173175].
utilizing silk proteins as templates for the biomineral- Interestingly, incubation of degummed naturally spun B.
ization process (Table 4 shows a selection of interesting mori broin bers in a solution at pH 8.6 led to the deposi-
examples). tion of aragonite crystals on the broin template, that were
aligned with the long axis of the bers [176].

2.3.2.1. Calcium carbonate. Calcium carbonate is found in


the shells of various marine organisms which are natu- 2.3.2.2. Calcium phosphate. Calcium phosphate is found in
ral protein-based biomineral composite materials; mollusc bone and tooth enamel which are both natural biomineral
shells are a particularly pertinent example composed of composite materials (protein/calcium phosphate). In vitro
chitin, a silk-like protein, and calcium carbonate [166168]. incubation of either silk proteins in solution (e.g. B. mori
In vitro studies utilizing polystyrene substrates coated with broin [177] or of B. mori broin and collagen [102]) or
B. mori broin were washed in a calcium chloride solution, various water insoluble (i.e. -sheet rich) silkworm/spider
followed by incubation in a dessicator containing ammo- silk morphologies (lms [178,179], hydrogels [180] and
nium carbonate (which slowly releases carbon dioxide). foams [181,182]) in simulated body uids led to the depo-
The liberated carbon dioxide reacts with the calcium ions sition of hydroxyapatite nanoparticles on the surfaces of
complexed to the carboxylic acids displayed on the surface the silk scaffolds. Interestingly, incubation of -sheet rich
of the substrate, resulting in the formation of rhombohe- lms of a genetically engineered chimeric protein based
dral calcite crystals and a small number of vaterite crystals upon a major ampullate spidroin (from N. clavipes) and
[142]. The presence of magnesium ions in the solution used dentin matrix protein 1 in simulated body uid led to the
to wash the broin lm was observed to promote the for- growth of hydroxyapatite crystals on the surface of the lm,
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1107

whereas lms cast from the control protein without the A simple alternative to create silk/silica composites is
dentin matrix protein 1 domain did not induce biominer- to coat silk-based material templates with silica precursors
alization [183]. (such as tetraethylorthosilicate (TEOS)) and subsequently
Analogously it has proven possible to biomineralize heat them at 105 C for several hours, as was demonstrated
silkworm/spider silk scaffolds with hydroxyapatite by with major ampullate silk bers of N. madagascariensis spi-
repetitive cycles of: incubation in calcium rich solutions, ders [196] and cocoon bers of B. mori broin silkworms
washing with water, incubation in phosphate rich solu- [197]. Furthermore, the silk template can subsequently
tions, and washing with water (and minor variations of be removed by calcination at 600 C, yielding a porous
this procedure); as was demonstrated for bers [184,185], material in which the pore structure is determined by the
lms [186,187], particles [188,189] and foams composed silk-based material.
of silk alone [187] or mixtures of a silk protein and
poly(aspartic acid) [7880]. Interestingly, when naturally 3. Applications of composite materials based on silk
spun P. phalangoides spider silk bers were biomineralized proteins
with hydroxyapatite, the c-axis of the crystals of hydrox-
yapatite were aligned with the long axis of the ber due to Nature does not have a monopoly on the creative use of
interactions between the hydroxyapatite crystals and the composite materials, and in the nal section of the review
proteins aligned with the long axis of the ber by elonga- we will highlight some applications of man-made compos-
tional ow during the natural spinning process [190]. ite materials based on silk proteins.
An alternative approach is the generation of
silk/hydroxyapatite composites using preformed hydrox- 3.1. Textiles
yapatite nanoparticles. Examples include: hydrogels
formed from mixtures of B. mori broin and hydrox- B. mori silk bers have been used for centuries in the tex-
yapatite nanoparticles [191]; and bers composed of tiles industry due to their characteristic strength, moisture
mixtures of B. mori broin, poly(ethylene oxide) and absorbance and luster; yet the natural bers have short-
hydroxyapatite nanoparticles that were electrospun from comings as they are prone to photoyellowing, and have
solution in dilute aqueous phosphate buffer (in order to poor rub resistance and wrinkle recovery. These shortcom-
minimize aggregation of the nanoparticles), immersed ings have been addressed by chemical modication of the
in methanol (rendering the bers insoluble in water due naturally spun silk bers [32,33,198201] and by spinning
to -sheet assembly in the broin), then dried, yielding new bers composed of regenerated silk proteins (isolated
slightly rough bers with diameters of approximately from the natural bers) and other components.
510 nm. The polymers in the bers were phase sep-
arated, and there was evidence of both aggregates of 3.1.1. Composites based on silk proteins and synthetic
unaligned nanoparticles and well-dispersed nanopar- polymers
ticles that were aligned with the long axis of the ber Fibers composed of poly(acrylonitrile) and its copoly-
[118]. Other interesting examples utilize naturally spun mers are cheap, antibacterial and stable to photo-oxidation,
silk bers that were chemically modied with either and are commonly used for textiles despite their poor water
poly(-methacryloxypropyl trimethoxysilane) [192,193] absorption and tendency to collect static electricity. Mixing
or poly(4-methacryloyloxyethyl trimellitate anhydride) B. mori broin with poly(acrylonitrile) notably improved
[194] to which hydroxyapatite nanoparticles can subse- the bers water absorption and reduced their tendency
quently be grafted (covalently or ionically respectively). to collect static electricity [36,52]. The mechanical proper-
The resulting composite bers could be puried simply by ties of such bers were acceptable for textile applications
sonication, washing in water, and nally freeze-drying; [202]. The addition of 2 wt% of a copolymer compatibiliser
the hydroxyapatite nanoparticles were homogeneously bearing on average two short poly(acrylonitrile) chains (of
distributed on the bers and stable to extensive washing. ca. 65 kDa) improved the tensile strength by 50% (relative
to the ber in the absence of the compatibiliser) whilst
2.3.2.3. Silica. Silica is an amorphous biomineral found maintaining a broin-rich sheath that is important for good
in the cell walls of Diatoms. Silafns are low molecular luster and handle [53,54].
weight proteins involved in the formation of protein/silica
composites in nature. In vitro studies using a peptide 3.1.2. Composites based on silk proteins and biopolymers
derived from the repetitive motif found in silafn pro- Cellulose bers are widely used in the textile indus-
teins (known as the R5 peptide) demonstrated that this try (in the form of cotton, hemp or jute) as it is cheap,
peptide promotes and regulates silica formation at neutral biodegradable and moisture absorbent. Fibers composed of
pH. Genetically engineered chimeric proteins based upon 30 wt% B. mori broin and 70 wt% cellulose had improved
major ampullate spidroin (from N. clavipes) and R5 peptide elastic moduli at the expense of breaking strength and
are capable of both self-assembly and biomineralization. yield stress relative to equivalent bers produced solely
Fibers and lms (electrospun/cast from HFIP solutions from cellulose [131]. Chitin from crab and shrimp shells
respectively) were treated with methanol to induce - are readily available waste materials from food processing
sheet formation in the major ampullate domain of the companies. Wet-spun bers are relatively elastic yet weak,
chimeric protein, and their subsequent incubation with a their breaking strength could be slightly improved by addi-
water-soluble silicon species led to biomineralization of tion of 6 wt% B. mori broin allowing the manufacture of
silica [195]. socks [137].
1108 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

3.1.3. Composites based on silk proteins and metal 3.3.1. Composites based on silk proteins and other
nanoparticles proteins
Silver ions are well-known to be toxic to microorgan- Electrospun bers composed of B. mori broin and
isms, and B. mori broin coated with silver nanoparticles type I calf skin collagen that were cross-linked with glu-
was demonstrated to prevent the growth of the gram- taraldehyde, were demonstrated to support cell adhesion
positive bacterium Staphylococcus aureus on the surface of and proliferation of both human epidermal keratinocytes
the silk fabric for up to ve wash cycles, and inhibit the (NHEK) and broblasts (NHEF) cells in vitro. Interestingly,
growth even after 10 wash cycles [156]. electrospun bers composed of unmixed B. mori broin and
type I calf skin collagen (that were prepared by electrospin-
3.2. Sutures and dressings for wounds ning from two syringes simultaneously and subsequently
cross-linked with glutaraldehyde) showed much higher
Natural silk bers have been used as sutures for wounds levels of cell adhesion and proliferation for NHEK cells
for centuries due to their strength, biocompatibility and in vitro [95]. Foams composed of B. mori broin and
low immunogenicity. In analogy to textiles, the mechanical either type I bovine collagen or recombinantly produced
properties of bers determine if they can be used as sutures human-like collagen were demonstrated to support cell
for wounds. adhesion and proliferation of human hepatocellular carci-
noma (HepG2) cells in vitro, with signicant improvements
in both adhesion and proliferation in comparison to lms
3.2.1. Sutures
composed of B. mori broin alone [101,103,203].
Fibers composed solely of B. mori broin prepared by
Embedding collagen coated B. mori broin bers within
wet-spinning formic acid solutions into methanol had rea-
foams composed of mixtures of collagen, hyaluronic
sonable breaking strength, but were brittle, inexible and
acid and chondroitin-6-sulfate, yields materials that were
had low knot strength (a feature of importance for the
shown to support cell adhesion and proliferation of human
application of such bers as sutures for wounds). Adding
anterior cruciate ligament cells in vitro at levels markedly
up to 50 wt% of poly(vinyl alcohol) was shown to increase
improved from collagen coated B. mori broin bers alone.
the tenacity and elongation at break of the bers (at the
In vivo studies of these composites implanted in dogs
expense of the breaking strength) and the knot strength
showed the formation of new blood vessels and minimal
was observed to increase to ca. three times that of the bers
inammatory reaction after 6 weeks [155].
without poly(vinyl alcohol), which is sufciently high for
In vitro studies performed on foams composed of B. mori
application as sutures for wounds [71]. Furthermore, sil-
broin and hyaluronic acid showed improved adhesion and
ver nanoparticle coated B. mori broin bers may be useful
proliferation of human mesenchymal stem cells [154] and
antimicrobial sutures [156].
primary neural cells (cortical neurons from rat embryos)
[153] in comparison to either of the constituent polymers
3.2.2. Wound dressings alone.
Non-woven mats prepared via electrospinning are
currently being investigated for application as wound 3.3.2. Composites based on silk proteins and
dressings, and non-woven mats of B. mori broin bers polysaccharides
coated with silver nanoparticles may be useful as antibac- Composite materials (in which microspheres composed
terial wound dressings [159]. Interestingly, B. mori broin of alginate and A. mylitta broin were embedded in A.
lms containing titanium dioxide nanoparticles were mylitta broin foams) were shown to support the adhesion
demonstrated to inhibit the growth of Escherichia coli, P. and proliferation of AH927 broblast cells in vitro [130].
aeruginosa and S. aureus [165]. Interestingly, the differentiation of human mesenchymal
Hydrogels composed of B. mori broin and cross- stem cells in vitro was demonstrated to be determined
linked poly(ethylene oxide) were demonstrated to be by the growth factor released from B. mori broin micro-
non-cytotoxic in in vitro studies using mouse broblast spheres (containing either osteogenic bone morphogenic
(L929) cells, and were demonstrated to promote wound protein-2 or chondrogenic insulin-like growth factor) dis-
healing (relative to a Vaseline gauze control) in vivo in persed in alginate hydrogels [119].
rats [6466]. Similarly, B. mori broin hydrogels contain- Electrospun bers composed of B. mori broin and chitin
ing hydroxyapatite nanoparticles were demonstrated to were demonstrated to support cell adhesion and prolifer-
promote wound healing in vivo in pigs [191]. ation of both human epidermal keratinocytes (NHEK) and
broblasts (NHEF) cells in vitro [138]. Interestingly, electro-
3.3. Tissue scaffolds spun bers composed of unmixed B. mori broin and chitin
(that were prepared by electrospinning from two syringes
Natural extracellular matrices are predominantly com- simultaneously) showed much higher levels of cell adhe-
posed of nanobrous collagen and proteoglycans (such sion and proliferation for NHEK cells in vitro [139]. Foams
as chondroitin sulfates and hyaluronic acid), however the composed of B. mori broin and chitosan were shown to
widespread therapeutic use (e.g. for replacing/restoring support cell adhesion and proliferation of mouse brob-
malfunctioning tissues) of collagen and proteoglycans in last (L929) cells in vitro [144], and in vivo studies show
the clinic is limited due to their cost an attractive alter- that after 4 weeks of implantation in the abdominal wall of
native is the use of combinations of cheaper bio/synthetic guinea pigs, the implants had integrated seamlessly being
polymers. partially degraded and remodeled [150]. More recently,
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1109

such scaffolds have been implanted in the bones of sheep, foams composed of B. mori broin and poly(aspartic acid)
after up to 12 weeks in vivo the implants were shown to be have great potential for clinical application for bone regen-
partially degraded and remodeled, thereby demonstrating eration [79,80].
their potential for use in the clinic [204].
3.4. Biocompatible coatings
3.3.3. Composites based on silk proteins and synthetic
polymers Silk lms are promising candidates for biocompatible
Poly(l-lactide) is a popular polymer for biomedical coatings for biomedical implants owing to the facility with
applications due to its biodegradability. Electrospun bers which silk proteins can be processed into lms with differ-
composed of Agelena labyrinthica spidroin and poly(d,l- ing surface properties (morphology, hydrophilicity etc. . .),
lactide) were demonstrated to support cell adhesion and their biodegradability and low levels of immunogenicity in
proliferation of African green monkey kidney [CCL81 Vero] vitro/in vivo. Coating biomedical implants with silk lms
cells in vitro [93]. Electrospun bers with a core-shell potentially imbues their surfaces with anticoagulant prop-
morphology in which the core was composed of B. mori erties, or inhibits/promotes cell adhesion [208].
broin and gelatin and the surface was poly(l-lactide) were
demonstrated to support cell adhesion and proliferation of 3.4.1. Anticoagulant coatings
3T3 mouse broblasts and human umbilical vein endothe- Water-insoluble anticoagulants are important coatings
lial cells in vitro. Tubes formed of such bers are currently for stents, and water-insoluble lms composed of B. mori
being investigated for application as articial blood vessels broin and carboxymethyl keratin [122] or poly(acrylic
due to their biocompatibility, mechanical properties and acid) decorated with tetramethylpyrazine [209] were
porosity [205,206]. demonstrated to be more antithrombogenic than either of
Composite materials in which a B. mori broin foam the biopolymers alone in in vitro blood clotting tests.
lled the pores of a poly(-caprolactone) foam were shown Interestingly, it is also possible to control the release of
to support cell adhesion and proliferation of human brob- heparin (a natural anticoagulant) from lms composed of
last (ATCC HFL-1) cells in vitro better than foams composed pellethane and B. mori broin particles. The release pro-
solely of poly(-caprolactone) [85]. le of heparin could be controlled by variations in heparin
Foams composed of B. mori broin and poly(l-lactide) loading, lm thickness (thicker lms released heparin over
were demonstrated to support cell adhesion and prolifer- longer periods of time), and the ratio of pellethane to broin
ation of both mouse macrophage (RAW 264.7) and human (greater broin content leads to slower release, probably
hepatocellular carcinoma (HepG2) cells in vitro better than due to hydrogen bonding interactions between the broin
foams composed of poly(l-lactide) alone. Moreover, the and heparin) [94].
presence of B. mori broin reduced the inammatory reac-
tion of the macrophages [92]. 3.4.2. Antibacterial coatings
Foams composed of B. mori broin and poly(aspartic As discussed above in Section 3.1.3, antibacterial coat-
acid) supported the adhesion and proliferation of human ings are of great industrial importance, and coating both
mesenchymal stem cells in vitro, and osteogenic differ- poly(propylene) lms and poly(amide) foams with B. mori
entiation was induced by inclusion of bone morphogenic broin was demonstrated to inhibit the adhesion of Staphy-
protein-2 within the foam and culture medium; such foams lococcus epidermis gram-positive bacteria cells in vitro
may nd application for replacement of broken bones [78]. [210]. Additionally, composite materials based upon silk
proteins and inorganic nanoparticles (e.g. silver nanopar-
3.3.4. Composites based on silk proteins and inorganic ticles [159]) or titanium dioxide nanoparticles have been
particles demonstrated to inhibit the growth of E. coli, P. aeruginosa
Electrospun bers composed of B. mori broin and and S. aureus [165].
poly(ethylene oxide) supported the adhesion and prolifer-
ation of human mesenchymal stem cells in vitro, however 3.4.3. Improved cell adhesion
levels of differentiation towards osteogenic outcomes The biocompatibility of materials to be implanted in
were low despite the osteogenic media. Incorporation people (such as hip replacements) are sometimes rejected
of bone morphogenic protein-2 into the spinning dope from the body, and biocompatible coatings such as silks are
notably improved differentiation of the stem cells towards one method of improving their acceptance.
osteogenic outcomes. Furthermore, addition of hydroxya- Coating various poly(urethane) lms and foams [8789]
patite nanoparticles to the spinning dope yielded bers with B. mori broin was demonstrated to improve cell
with the nanoparticles embedded inside and was found adhesion and proliferation of human broblast cells
to improve bone formation [118,207]. Subsequent stud- in vitro relative to the uncoated materials. Similarly,
ies demonstrated that coating B. mori broin bers, lms poly(propylene) lms and poly(amide) foams coated with
or foams with hydroxyapatite nanoparticles improved B. mori broin were demonstrated to support cell adhesion
the adhesion and proliferation of mouse broblast (L929) and proliferation of mouse broblast (L929) cells in vitro
cells [193], mouse pre-osteoblastic (MC3T3-E1) cells [187], [210].
rat osteoblast cells [184], bone marrow stromal cells Poly(allylamine) is a polycationic polymer that is highly
[79,80], human mesenchymal stem cells [187] and human soluble in water that it utilized widely in biomedi-
osteoblast (MG-63) cells in vitro [182]. Recent in vivo stud- cal applications. Films composed of B. mori broin and
ies have also shown that hydroxyapatite biomineralized poly(allylamine) were found to be more stable in water
1110 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

than lms composed only of poly(allylamine), and such period of days in vitro and in vivo when implanted in rats
lms were demonstrated to support cell adhesion and pro- brains [215].
liferation of both insect (A. pernyi and B. mori silkworm)
and mammalian (mouse broblast L929) cells in vitro better
than either of the constituent polymers alone [56]. 3.5.2. High molecular weight drugs
The adhesion of mouse broblast L929 cells to lms Films composed of B. mori broin and various enzymes
composed of B. mori broin and poly(lactic acid) were [104108] have been shown to retain their catalytic activ-
shown to be better than either of the constituent polymers ity for many days with minimal leaching of the enzyme in
alone [91]. vitro. Microspheres composed of a model enzyme drug and
Mimicry of the extracellular matrix is a popular either alginate or poly(lactide-co-glycolide) were coated
approach to promote cell adhesion, and with this goal with B. mori broin, the silk coating slowed enzyme
in mind numerous materials have been prepared that release from the order of tens of hours to tens of days
incorporate components of the extracellular matrix. Films in vitro [90]. Growth factors or enzymes trapped within B.
composed of B. mori broin and rat-tail collagen were mori broin microspheres and subsequently dispersed in
demonstrated to support cell adhesion and proliferation either broin foams or alginate hydrogels were demon-
of rat liver cells in vitro [99]. Films composed of B. mori strated to be released over a period of days in vitro
broin and either type I bovine collagen or recombi- [119]. Likewise, high molecular weight drugs encapsu-
nantly produced human-like collagen were demonstrated lated in microspheres composed of alginate and A. mylitta
to support cell adhesion and proliferation of human hepa- broin suspended in B. mori broin foams were demon-
tocellular carcinoma (HepG2) cells in vitro, with signicant strated to be released without initial bursts for 35 days
improvements in both adhesion and proliferation in com- [130].
parison to lms composed of B. mori broin alone [97,98]. Films composed of B. mori broin and hyaluronic acid
Films composed of B. mori broin and recombinantly pro- were demonstrated to be responsive to both pH and elec-
duced proteins based upon N. clavipes spider dragline silk trical stimuli. At low pH values (between pH 2 and 3)
(incorporating the RGD integrin recognition sequence) was broin and hyaluronic acid form polyion complexes which
shown to support cell adhesion and proliferation of mouse dissociate at neutral pH, hence the -sheet cross-linked
osteoblast (MC3T3-E1) cells in vitro [111]. lms were observed to be more water permeable at neu-
tral pH (yet still mechanically stable); the complexes are
3.5. Drug delivery also responsive to electrical stimuli, and application of a
potential difference led to dissociation, and consequent
A reliable and controllable release prole is impor- swelling of the lms. In vitro studies showed that when
tant for a drug to have its optimal effect and minimize swollen the lms were permeable to Timolol maleate
side effects [211,212]. Composite materials based on silk which is well tolerated in humans upon transdermal
proteins are attractive drug carriers due to their biocom- application, and suggests that such lms may nd appli-
patibilility and highly tunable morphologies. cation in membrane-permeation controlled formulation
[216,217].
3.5.1. Low molecular weight drugs
Films composed of B. mori broin and chitosan (cross-
linked with glutaraldehyde) and various low molecular 3.6. Materials with novel electronic, magnetic and
weight drugs (ampicilline trihydrate, dichlofenac sodium, optical properties
salicylic acid and theophylline) released most of their pay-
load within 30 min of incubation in various pH buffered Coating naturally spun major ampullate silk bers with
solutions in vitro [213]. Films composed of B. mori broin poly(pyrrole) or gold nanoparticles (electrical conductors)
and between 12.5 and 25 wt% poly(ethylene oxide) were or cadmium sulde particles (a semiconductor) potentially
used to coat tablets of theophylline. Uncoated tablets dis- imparts novel electronic properties to the bers; likewise
solved completely within 210 min, whereas dissolution of bers coated with magnetite nanoparticles yields bers
the coated tablets was not complete until 300 min and dis- that respond to magnetic elds which have potential appli-
played zero order kinetics in vitro [214]. cation in audio reproduction [69,162]. Electrospun bers
Hydrogels composed of either a poly(ethylene oxide)- composed of B. mori broin that were coated with a layer
poly(propylene oxide) copolymer (Pluronic F-127) or of carbon nanotubes were demonstrated to have markedly
chitosan with B. mori broin were shown to release improved electrical conductivity (ca. 104 S cm1 ) in rela-
buprenorphine more slowly than hydrogels composed of tion to electrospun bers composed solely of B. mori broin
broin alone in vitro [67]. (ca. 1015 S cm1 ) [45]. B. mori broin microspheres coated
Adenosine is a potential therapeutic for epilepsy with a layer of carbon nanotubes were demonstrated to
which is a major health problem for many people. A have similar electrical conductivities (ca. 104 S cm1 ) [46].
complex B. mori broin composite material comprising: Such materials may nd application in textiles as they
broin/adenosine microspheres coated in silk, embedded would reduce the tendency of the fabrics to collect static
in a broin foam that is lled with broin/adenosine electricity. Moreover, it is also possible to prepare materials
solution, and subsequently coated in a multilayer lm (e.g. lms) with interesting nonlinear optical properties by
(composed of alternating layers of broin and adenosine) incorporation of green uorescent protein within the silk
was demonstrated to control the release of the drug over a matrix [117].
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1111

3.7. Enzymatically active biomaterials [8] Coyne KJ, Qin XX, Waite JH. Extensible collagen in mussel byssus:
a natural block copolymer. Science 1997;277:18302.
[9] Waite JH, Qin XX, Coyne KJ. The peculiar collagens of mussel byssus.
One method of preparing biosensors is the modica- Matrix Biol 1998;17:93106.
tion of the surface of electrodes with enzymes (via covalent [10] Hagenau A, Scheidt HA, Serpell L, Huster D, Scheibel T. Structural
immobilization). Yet a problem commonly encountered analysis of proteinaceous components in byssal threads of the mus-
sel mytilus galloprovincialis. Macromol Biosci 2009;9:1628.
with this approach is the denaturation of the enzyme due [11] Vepari C, Kaplan DL. Silk as a biomaterial. Prog Polym Sci
to non-covalent interactions with the electrode surface, 2007;32:9911007.
which renders them catalytically inactive. One solution to [12] Shao ZZ, Vollrath F. Materials: surprising strength of silkworm silk.
Nature 2002;418:7411741.
this problem is to embed the enzyme in a protein matrix [13] Putthanarat S, Eby RK, Adams WW, Liu GF. Aspects of the morphol-
which helps to maintain the enzymes native structure ogy of the silk of Bombyx mori. J Macromol Sci Pure Appl Chem
(thereby retaining its catalytic activity) [104108]. An ele- 1996;A33:899911.
[14] Putthanarat S, Stribeck N, Fossey SA, Eby RK, Adams WW. Investi-
gant example of this approach utilized lms composed
gation of the nanobrils of silk bers. Polymer 2000;41:773547.
of the enzyme horseradish peroxidase and B. mori broin [15] Inoue S, Tanaka K, Arisaka F, Kimura S, Ohtomo K, Mizuno S. Silk
stabilized gold nanoparticles (further cross-linked by inter- broin of Bombyx mori is secreted, assembling a high molecular
protein -sheet formation). The enzyme maintained its mass elementary unit consisting of h-chain, l-chain, and p25, with
a 6:6:1 molar ratio. J Biol Chem 2000;275:4051728.
catalytic activity and electrodes coated with such lms [16] Kundu SC, Dash BC, Dash R, Kaplan DL. Natural protective
responded swiftly to the reduction of hydrogen perox- glue protein, sericin bioengineered by silkworms: potential for
ide, demonstrating their applicability as biosensors [161]. biomedical and biotechnological applications. Progr Polym Sci
2008;33:9981012.
Interestingly, we have also demonstrated it to be possi- [17] Nirmala X, Kodrik D, Zurovec M, Sehnal F. Insect silk contains both
ble to covalently immobilize an enzyme (-galactosidase) a kunitz-type and a unique kazal-type proteinase inhibitor. Eur J
on a silk-based material with maintenance of the catalytic Biochem 2001;268:206473.
[18] Nirmala X, Mita K, Vanisree V, Zurovec M, Sehnal F. Identication
activity of the enzyme [218]. of four small molecular mass proteins in the silk of Bombyx mori.
Insect Mol Biol 2001;10:43745.
[19] Gatesy J, Hayashi C, Motriuk D, Woods J, Lewis R. Extreme diver-
4. Conclusions sity, conservation, and convergence of spider silk broin sequences.
Science 2001;291:26035.
Natural composite bers produced by silkworms and [20] Holland GP, Creager MS, Jenkins JE, Lewis RV, Yarger JL.
Determining secondary structure in spider dragline silk by
spiders have been used by humans for centuries. Human carboncarbon correlation solid-state nmr spectroscopy. J Am
creativity has produced a variety of composite materials Chem Soc 2008;130:98717.
(including bers, lms, gels, foams and particulates [3,34]) [21] Holland GP, Jenkins JE, Creager MS, Lewis RV, Yarger JL. Quanti-
fying the fraction of glycine and alanine in beta-sheet and helical
based on silk proteins with a number of exciting appli- conformations in spider dragline silk using solid-state nmr. Chem
cations. We envisage that in the future such composite Commun 2008:556870.
materials will be of economic importance not only in tex- [22] Holland GP, Jenkins JE, Creager MS, Lewis RV, Yarger JL. Solid-state
nmr investigation of major and minor ampullate spider silk in the
tile applications but in high-tech applications such as tissue native and hydrated states. Biomacromolecules 2008;9:6517.
scaffolds and drug delivery devices; and moreover, that de [23] Holland GP, Lewis RV, Yarger JL. Wise nmr characterization
novo designed silk-like proteins and polymers [28] will be of nanoscale heterogeneity and mobility in supercontracted
Nephila clavipes spider dragline silk. J Am Chem Soc 2004;126:
of great importance due to their highly tunable structures.
586772.
[24] Huemmerich D, Helsen CW, Quedzuweit S, Oschmann J, Rudolph
R, Scheibel T. Primary structure elements of spider dragline
Acknowledgements silks and their contribution to protein solubility. Biochemistry
2004;43:1360412.
JGH gratefully acknowledges nancial support from the [25] Huemmerich D, Scheibel T, Vollrath F, Cohen S, Gat U, Ittah S. Novel
assembly properties of recombinant spider dragline silk proteins.
Alexander von Humboldt Foundation, and TS acknowl- Curr Biol 2004;14:20704.
edges nancial support from Army Research Ofce [26] Annonymous. The 50 best inventions of 2009: spiderweb silk.
(W911NF-0810284). Time 2009; http://www.time.com/time/specials/packages/article/
0,28804,1934027 1934003 1933990,00.html.
[27] Vendrely C, Scheibel T. Biotechnological production of spider-silk
proteins enables new applications. Macromol Biosci 2007;7:4019.
References
[28] Hardy JG, Scheibel TR. Silk-inspired polymers and proteins.
Biochem Soc Trans 2009;37:67781.
[1] Sutherland TD, Young JH, Weisman S, Hayashi CY, Merritt DJ. Insect [29] Bajaj P. Finishing of textile materials. J Appl Polym Sci
silk: one name, many materials. Ann Rev Entomol 2010;55:17188. 2002;83:63159.
[2] Gerritsen VB. The tiptoe of an airbus. Protein Spotlight [30] Freddi G, Tsukada M, Kato H, Shiozaki H. Dyeability of silk fab-
2002;24:12. Online: http://www.expasy.org/spotlight/back rics modied with dibasic acid anhydrides. J Appl Polym Sci
issues/024/. 1994;52:76973.
[3] Hardy JG, Romer LM, Scheibel TR. Polymeric materials based on silk [31] Das S. The preparation and processing of tussah silk. J Soc Dyers
proteins. Polymer 2008;49:430927. Colourists 1992;108:4816.
[4] Scheibel T. Spider silks: recombinant synthesis, assembly, spin- [32] Arai T, Freddi G, Innocenti R, Kaplan DL, Tsukada M. Acylation of silk
ning, and engineering of synthetic proteins. Microbial Cell Factories and wool with acid anhydrides and preparation of water-repellent
2004;3(14):110. bers. J Appl Polym Sci 2001;82:283241.
[5] Jin HJ, Kaplan DL. Mechanism of silk processing in insects and spi- [33] Arai T, Freddi G, Innocenti R, Tsukada M. Preparation of water-
ders. Nature 2003;424:105761. repellent silks by a reaction with octadecenylsuccinic anhydride.
[6] Sponner A, Unger E, Grosse F, Klaus W. Differential polymerization J Appl Polym Sci 2003;89:32432.
of the two main protein components of dragline silk during bre [34] Hardy JG, Scheibel TR. Production and processing of spider silk pro-
spinning. Nature Mater 2005;4:7725. teins. J Polym Sci Part A Polym Chem 2009;47:395763.
[7] Sponner A, Vater W, Monajembashi S, Unger RE, Grosse F, Wiesshart [35] Fu C, Shao Z, Vollrath F. Animal silks: their structures, properties
K. Composition and hierarchical organisation of a spider silk. PLoS and articial production. Chem Commun 2009:651529.
ONE 2007;2:e998.
1112 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

[36] Cates DM, White HJ. Preparation and properties of bers contain- [64] Kweon HY, Park SH, Ye JH, Lee YW, Cho CS. Preparation
ing mixed polymers. 3. Polyacrylonitrile-silk bers. J Polym Sci of semi-interpenetrating polymer networks composed of silk
1956;21:12538. broin and poly(ethylene glycol) macromer. J Appl Polym Sci
[37] Moniruzzaman M, Winey KI. Polymer nanocomposites containing 2001;80:184853.
carbon nanotubes. Macromolecules 2006;39:5194205. [65] Yoo MK, Kweon HY, Lee KG, Lee HC, Cho CS. Preparation of semi-
[38] Khare R, Bose S. Carbon nanotube based composites a review. J interpenetrating polymer networks composed of silk broin and
Minerals Mater Charact Eng 2005;4:3146. poloxamer macromer. Int J Biol Macromol 2004;34:26370.
[39] Ayutsede J, Gandhi M, Sukigara S, Ye HH, Hsu CM, Gogotsi Y, [66] Kweon HY, Yeo JH, Lee KG, Lee HC, Na HS, Won YH, et al. Semi-
et al. Carbon nanotube reinforced Bombyx mori silk nanobers interpenetrating polymer networks composed of silk broin and
by the electrospinning process. Biomacromolecules 2006;7: poly(ethylene glycol) for wound dressing. Biomed Mater 2008;3,
20814. 034115/1-5.
[40] Kang M, Chen P, Jin HJ. Preparation of multiwalled carbon nan- [67] Fang HY, Chen JP, Leu YL, Wang HY. Characterization and evalua-
otubes incorporated silk broin nanobers by electrospinning. Curr tion of silk protein hydrogels for drug delivery. Chem Pharm Bull
Appl Phys 2009;9:S957. 2006;54:15662.
[41] Kim HS, Park WI, Kim Y, Jin HJ. Silk broin lms crystallized by [68] Guimard NK, Gomez N, Schmidt CE. Conducting polymers in
multiwalled carbon nanotubes. Int J Mod Phys B 2008;22:180712. biomedical engineering. Prog Polym Sci 2007;32:876921.
[42] Sashina ES, Novoselov NP, Vorbach D, Meister F. Conformational [69] Mayes EL, Vollrath F, Mann S. Fabrication of magnetic spider silk
changes in broin upon its dissolution in hexauoroisopropanol. and other silk-ber composites using inorganic nanoparticles. Adv
Polym Sci Ser A 2005;47:1096103. Mater 1998;10:8015.
[43] Blond D, McCarthy DN, Blau WJ, Coleman JN. Toughening of arti- [70] Swinerd VM, Collins AM, Skaer NJV, Gheysens T, Mann S. Silk
cial silk by incorporation of carbon nanotubes. Biomacromolecules inverse opals from template-directed beta-sheet transformation of
2007;8:39736. regenerated silk broin. Soft Matter 2007;3:137780.
[44] Cheung HY, Lau KT. Mechanical performance of silk-based struc- [71] Lee KH, Baek DH, Ki CS, Park YH. Preparation and characterization
tural composites. Key Eng Mater 2006;326328:45760. of wet spun silk broin/poly(vinyl alcohol) blend laments. Int J
[45] Kang M, Jin HJ. Electrically conducting electrospun silk membranes Biol Macromol 2007;41:16872.
fabricated by adsorption of carbon nanotubes. Colloid Polym Sci [72] Yamaura K, Kuranuki N, Suzuki M, Tanigami T, Matsuzawa S.
2007;285:11637. Properties of mixtures of silk broin syndiotactic-rich poly(vinyl
[46] Yoon SH, Kang M, Kim HS, Jin HJ. Electrically conducting silk broin alcohol). J Appl Polym Sci 1990;41:240925.
microspheres by incorporation of multiwalled carbon nanotubes [73] Tsukada M, Freddi G, Crighton JS. Structure and compatibility of
on their surfaces. Mater Sci Forum 2007;544545:97780. poly(vinyl alcohol)-silk broin (pva/sf) blend lms. J Polym Sci Part
[47] Kim HS, Yoon SH, Kwon SM, Jin HJ. Ph-sensitive multiwalled car- B Polym Phys 1994;32:2438.
bon nanotube dispersion with silk broins. Biomacromolecules [74] Sashina ES, Novoselov NP, Vnuchkin AV, Golubikhin AY. Prepa-
2009;10:826. ration and properties of lms of broin-polyvinyl alcohol blends
[48] Kim H, Kim HS, Lee HS, Chin IJ, Jin HJ. Dispersion of multiwalled from solutions in hexauoroisopropanol. Russ J Appl Chem
carbon nanotubes in aqueous silk broin solutions. J Nanosci Nan- 2007;80:46671.
otechnol 2008;8:55436. [75] Li MZ, Lu SZ, Wu ZY, Tan K, Minoura N, Kuga S. Structure and prop-
[49] Kwon SM, Kim HS, Jin HJ. Fabrication of organic silk erties of silk broin-poly(vinyl alcohol) gel. Int J Biol Macromol
broin/multiwalled carbon nanotube composite cryogels by 2002;30:8994.
freeze-drying method. Adv Mater Res 2008;4750:11058. [76] Li MZ, Minoura N, Dai LX, Zhang LS. Preparation of porous poly(vinyl
[50] Liu Y, Shao ZZ, Zhou P, Chen X. Thermal and crystalline behaviour alcohol)-silk broin (pva/sf) blend membranes. Macromol Mater
of silk borin/nylon 66 blend lms. Polymer 2004;45:770510. Eng 2001;286:52934.
[51] Freddi G, Tsukada M, Beretta S. Structure and physical proper- [77] Nair LS, Laurencin CT. Biodegradable polymers as biomaterials. Prog
ties of silk broin polyacrylamide blend lms. J Appl Polym Sci Polym Sci 2007;32:76298.
1999;71:156371. [78] Kim HJ, Kim UJ, Kim HS, Li CM, Wada M, Leisk GG, et al.
[52] Sun YY, Shao ZZ, Ma MH, Hu P, Liu YS, Yu TY. Acrylic polymer silk Bone tissue engineering with premineralized silk scaffolds. Bone
broin blend bers. J Appl Polym Sci 1997;65:95966. 2008;42:122634.
[53] Sun YY, Shao ZZ, Zhou J, Yu TY. Compatibilization of acrylic polymer [79] Zhao J, Zhang ZC, Wang S, Sun X, Zhang X, Chen J, et al. Apatite-
silk broin blend bers. I. Graft copolymerization of acrylonitrile coated silk broin scaffolds to healing mandibular border effects in
onto silk broin. J Appl Polym Sci 1998;69:108997. canines. Bone 2009;45:51727.
[54] Sun YY, Shao ZZ, Zhou J, Yu TY. Compatibilization of acrylic [80] Jiang X, Zhao J, Wang S, Sun X, Zhang X, Chen J, et al. Mandibular
polymer-silk broin blend bers: 2. Morphology and mechani- repair in rats with premineralized silk scaffolds and bmp-2-
cal properties of the compatilized blend bers. J Appl Polym Sci modied bmscs. Biomaterials 2009;30:452232.
1999;73:225564. [81] Qiao XY, Li W, Sun K, Xu S, Chen XD. Nonisothermal crys-
[55] Sun YY, Shao ZZ, Hu P, Yu TY. Hydrogen bonds in silk broin tallization behaviors of silk-broin-ber-reinforced poly(epsilon-
poly(acrylonitrile-co-methyl acrylate) blends: Ft-ir study. J Polym caprolactone) biocomposites. J Appl Polym Sci 2009;111:290816.
Sci Part B Polym Phys 1997;35:140514. [82] Qiao XY, Li W, Sun K, Xu S, Chen XD. Isothermal crystallization
[56] Arai T, Wilson DL, Kasai N, Freddi G, Hayasaka S, Tsukada M. Prepa- kinetics of silk broin ber-reinforced poly(epsilon-caprolactone)
ration of silk broin and polyallylamine composites. J Appl Polym biocomposites. Polym Int 2009;58:5307.
Sci 2002;84:196370. [83] Li W, Qiao XY, Sun K, Chen XD. Mechanical and viscoelastic
[57] Padma Priya S, Rai SK. Studies on the mechanical performance of properties of novel silk broin ber/poly(epsilon-caprolactone)
pmma toughened epoxy silk and pc toughened epoxy silk fabric biocomposites. J Appl Polym Sci 2008;110:1349.
composites. J Reinforced Plastics Compos 2006;25:3341. [84] Li W, Qiao XY, Sun K, Chen XD. Effect of electron beam irradiation on
[58] Jin HJ, Fridrikh SV, Rutledge GC, Kaplan DL. Electrospinning the silk broin ber/poly(epsilon-caprolactone) composite. J Appl
Bombyx mori silk with poly(ethylene oxide). Biomacromolecules Polym Sci 2009;113:10639.
2002;3:12339. [85] Chen G, Zhou P, Mei N, Chen X, Shao ZZ, Pan LF, et al. Silk broin
[59] Wang M, Jin HJ, Kaplan DL, Rutledge GC. Mechanical properties of modied porous poly(e-caprolactone) scaffold for human brob-
electrospun silk bers. Macromolecules 2004;37:685664. last culture in vitro. J Mater Sci Mater Med 2004;15:6717.
[60] Wang M, Yu JH, Kaplan DL, Rutledge GC. Production of submicron [86] Kesenci K, Motta A, Fambri L, Migliaresi C. Poly(epsilon-
diameter silk bers under benign processing conditions by two- caprolactone-co-d,l-lactide)/silk broin composite materials:
uid electrospinning. Macromolecules 2006;39:11027. preparation and characterization. J Biomater Sci Polym Ed
[61] Jin HJ, Park J, Valluzzi R, Cebe P, Kaplan DL. Biomaterial lms of Bom- 2001;12:33751.
byx mori silk broin with poly(ethylene oxide). Biomacromolecules [87] Chiarini A, Petrini P, Bozzini S, dal Pra I, Armato U. Silk
2004;5:7117. broin/poly(carbonate)-urethane as a substrate for cell growth:
[62] Hofmann S, Hagenmuller H, Koch AM, Muller R, Vunjak-Novakovic in vitro interactions with human cells. Biomaterials 2003;24:
G, Kaplan DL, et al. Control of in vitro tissue-engineered bone-like 78999.
structures using human mesenchymal stem cells and porous silk [88] Dal Pra I, Petrini P, Charini A, Bozzini S, Fare S, Armato U. Silk
scaffolds. Biomaterials 2007;28:115262. broin-coated three-dimensional polyurethane scaffolds for tissue
[63] Kim UJ, Park JY, Li CM, Jin HJ, Valluzzi R, Kaplan DL. Structure and engineering: interactions with normal human broblasts. Tissue
properties of silk hydrogels. Biomacromolecules 2004;5:78692. Eng 2003;9:111321.
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1113

[89] Petrini P, Parolari C, Tanzi MC. Silk broin-polyurethane scaffolds [111] Morgan AW, Roskov KE, Lin-Gibson S, Kaplan DL, Becker ML,
for tissue engineering. J Mater Sci Mater Med 2001;12:84953. Simon CG. Characterization and optimization of rgd-containing
[90] Wang X, Wenk E, Hu X, Castro GR, Meinel L, Wang X, et al. Silk silk blends to support osteoblastic differentiation. Biomaterials
coatings on plga and alginate microspheres for protein delivery. 2008;29:255663.
Biomaterials 2007;28:41619. [112] Gil ES, Spontak RJ, Hudson SM. Effect of beta-sheet crystals on
[91] Zhu HL, Feng XX, Zhang HP, Guo YH, Zhang JZ, Chen JY. Structural the thermal and rheological behavior of protein-based hydro-
characteristics and properties of silk broin/poly(lactic acid) blend gels derived from gelatin and silk broin. Macromol Biosci
lms. J Biomater Sci Polym Ed 2009;20:125974. 2005;5:7029.
[92] Hu K, Lv Q, Cui FZ, Xu L, Jiao YP, Wang Y, et al. A novel [113] Gil ES, Frankowski DJ, Spontak RJ, Hudson SM. Swelling behavior
poly(l-lactide) (plla)/broin hybrid scaffold to promote hepatocyte and morphological evolution of mixed gelatin/silk broin hydro-
viability and decrease macrophage responses. J Bioactive Compat gels. Biomacromolecules 2005;6:307987.
Polym 2007;22:395410. [114] Couet F, Mantovani D. Experimental validation of a new approach
[93] Zhou SB, Peng HS, Yu XJ, Zheng XT, Cui WG, Zhang ZR, et for the development of mechano-compatible composite scaf-
al. Preparation and characterization of a novel electrospun spi- folds for vascular tissue engineering. J Mater Sci Mater Med
der silk broin/poly(d,l-lactide) composite ber. J Phys Chem B 2008;19:25514.
2008;112:1120916. [115] Gil ES, Frankowski DJ, Bowman MK, Gozen AO, Hudson SM, Spon-
[94] Liu XY, Zhang CC, Xu WL, Ouyang CX. Controlled release of hep- tak RJ. Mixed protein blends composed of gelatin and Bombyx mori
arin from blended polyurethane and silk broin lm. Mater Lett silk broin: effects of so solvent-induced crystallization and com-
2009;63:2635. position. Biomacromolecules 2006;7:72835.
[95] Yeo IS, Oh JE, Jeong L, Lee TS, Lee SJ, Park WH, et al. Collagen-based [116] Gil ES, Frankowski DJ, Hudson SM, Spontak RJ. Silk broin mem-
biomimetic nanobrous scaffolds: Preparation and characteriza- branes from solvent-crystallized silk broin/gelatin blends: effects
tion of collagen/silk broin bicomponent nanobrous structures. of blend and solvent composition. Mater Sci Eng C 2007;27:
Biomacromolecules 2008;9:110616. 42631.
[96] Sell SA, McClure MJ, Garg K, Wolfe PS, Bowlin GL. Electro- [117] Putthanarat S, Eby RK, Naik RR, Juhl SB, Walker MA, Peterman E, et
spinning of collagen/biopolymers for regerneative medicine and al. Nonlinear optical transmission of silk/green uorescent protein
cardiovascular tissue engineering. Adv Drug Deliv Rev 2009;61: (gfp) lms. Polymer 2004;45:84517.
100719. [118] Li CM, Vepari C, Jin HJ, Kim HJ, Kaplan DL. Electrospun
[97] Hu K, Lv Q, Cui FZ, Feng QL, Kong XD, Wang HL, et al. Bio- silk-bmp-2 scaffolds for bone tissue engineering. Biomaterials
compatible broin blended lms with recombinant human-like 2006;27:311524.
collagen for hepatic tissue engineering. J Bioactive Compat Polym [119] Wang X, Wenk E, Zhang X, Meinel L, Vunjak-Novakovic G, Kaplan
2006;21:2337. DL. Growth factor gradients via microsphere delivery in biopolymer
[98] Lv Q, Hu K, Feng QL, Cui FZ. Preparation of insoluble broin/collagen scaffolds for osteochondral tissue engineering. J Control Release
lms without methanol treatment and the increase of its 2009;134:8090.
exibility and cytocompatibility. J Appl Polym Sci 2008;109: [120] Zoccola M, Aluigi A, Vineis C, Tonin C, Ferrero F, Piacentino MG.
157784. Study on cast membranes and electrospun nanobers made from
[99] Cirillo B, Morra M, Catapano G. Adhesion and function of rat liver keratin/broin blends. Biomacromolecules 2008;9:281925.
cells adherent to silk broin/collagen blend lms. Int J Artif Organs [121] Ki CS, Gang EH, Um NC, Park YH. Nanotibrous membrane of wool
2004;27:608. keratose/silk broin blend for heavy metal ion adsorption. J Membr
[100] Roh DH, Kang SY, Kim JY, Kwon YB, Kweon HY, Lee KG, et al. Wound Sci 2007;302:206.
healing effect of silk broin/alginate-blended sponge in full thick- [122] Lee KY, Kong SJ, Park WH, Ha WS, Kwon IC. Effect of surface prop-
ness skin defect of rat. J Mater Sci Mater Med 2006;17:54752. erties on the antithrombogenicity of silk broin/s-carboxymethyl
[101] Lv QA, Feng QL, Hu K, Cui FZ. Three-dimensional broin/collagen kerateine blend lms. J Biomater Sci Polym Ed 1998;9:90514.
scaffolds derived from aqueous solution and the use for hepg2 cul- [123] Lee KY, Ha WS. Dsc studies on bound water in silk broin/s-
ture. Polymer 2005;46:126629. carboxymethyl kerateine blend lms. Polymer 1999;40:41314.
[102] Wang J, Zhou W, Hu W, Zhou L, Wang S, Zhang S. Collagen/silk [124] Lee HK. Silk sericin retards the crystallization of silk broin. Macro-
broin bi-template-induced biomimetic bone-like structures. J mol Rapid Commun 2004;25:17926.
Biomed Mater Res Part A 2010;doi: 10.1002/jbm.a.32602. [125] Heim M, Keerl D, Scheibel T. Spider silk: from soluble protein to
[103] Hu K, Cui FZ, Lv Q, Ma J, Feng QL, Xu L, et al. Preparation extraordinary ber. Angew Chem Int Ed 2009;48:358496.
of broin/recombinant human-like collagen scaffold to promote [126] Rammensee S, Slotta U, Scheibel T, Bausch AR. Assembly mecha-
broblasts compatibility. J Biomed Mater Res Part A 2008;84: nism of recombinant spider silk proteins. Proc Natl Acad Sci USA
48390. 2008;105:65905.
[104] Liu HY, Qian JG, Liu YC, Yu TY, Deng JQ. Immobilization of glucose [127] Slotta U, Tammer M, Kremer F, Koelsch P, Scheibel T. Structural
oxidase in the composite membrane of regenerated silk broin and analysis of spider silk lms. Supramol Chem 2006;18:46571.
poly(vinyl alcohol): application to an amperometric glucose sensor. [128] Liang CX, Hirabayashi K. Improvements of the physical proper-
Bioelectrochem Bioenerg 1996;39:3038. ties of broin membranes with sodium alginate. J Appl Polym Sci
[105] Liu YC, Chen X, Qian JH, Liu HY, Shao ZZ, Deng JQ, et al. 1992;45:193743.
Immobilization of glucose oxidase with the blend of regener- [129] Lee KG, Kweon HY, Yeo JH, Woo SO, Lee JH, Park YH. Structural and
ated silk broin and poly(vinyl alcohol) and its application to a physical properties of silk broin/alginate blend sponges. J Appl
1,1 -dimethylferrocene-mediating glucose sensor. Appl Biochem Polym Sci 2004;93:21749.
Biotechnol 1997;62:10517. [130] Kundu SC, Mandal BB. Calcium alginate beads embedded in
[106] Asakura T, Kitaguchi M, Demura M, Sakai H, Komatsu K. silk broin as 3d dual drug releasing scaffolds. Biomaterials
Immobilization of glucose-oxidase on nonwoven fabrics with 2009;30:51707.
bombyx-mori silk broin gel. J Appl Polym Sci 1992;46: [131] Marsano E, Canetti M, Conio G, Corsini P, Freddi G. Fibers based on
4953. cellulose-silk broin blend. J Appl Polym Sci 2007;104:218796.
[107] Qian JH, Liu YC, Liu HY, Yu TY, Deng JQ. Immobilization of [132] Freddi G, Romano M, Massafra MR, Tsukada M. Silk broin/cellulose
horseradish peroxidase with a regenerated silk broin membrane blend lms preparation, structure and physical-properties. J Appl
and its application to a tetrathiafulvalene-mediating h2o2 sensor. Polym Sci 1995;56:153745.
Biosens Bioelectr 1997;12:12138. [133] Yang G, Zhang LN, Cao XD, Liu YG. Structure and microporous for-
[108] Tangkuaram T, Ponchio C, Kangkasomboon T, Katikawong P, Veera- mation of cellulose/silk broin blend membranes part II. Effect of
sai W. Design and development of a highly stable hydrogen post-treatment by alkali. J Membr Sci 2002;210:37987.
peroxide biosensor on screen printed carbon electrode based [134] Yang G, Zhang LN, Liu YG. Structure and microporous formation
on horseradish peroxidase bound with gold nanoparticles in the of cellulose/silk broin blend membranes I. Effect of coagulants. J
matrix of chitosan. Biosens Bioelectr 2007;22:20718. Membr Sci 2000;177:15361.
[109] Qiang L, Cao CB, Zhang Y, Man XL, Zhu HS. The preparation of [135] Sashina ES, Janowska G, Zaborski M, Vnuchkin AV. Compatibility
insoluble broin lms induced by degummed broin or broin of broin/chitosan and broin/cellulose blends studied by thermal
microspheres. J Mater Sci Mater Med 2004;15:11937. analysis. J Thermal Anal Calorimetry 2007;89:88791.
[110] Tsukada M, Freddi G, Kasai N. Physical-properties and phase- [136] Hirano S, Nakahira T, Zhang M, Nakagawa M, Yoshikawa M,
separation structure of antheraea-pernyi bombyx-mori silk broin Midorikawa T. Wet-spun blend biobers of cellulose-silk broin
blend lms. J Polym Sci Part B Polym Phys 1994;32:117582. and cellulose-chitin-silk broin. Carbohydr Polym 2002;47:1214.
1114 J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115

[137] Hirano S, Nakahira T, Nakagawa M, Kim SK. The preparation and [162] Singh A, Hede S, Sastry M. Spider silk as an active scaffold in the
applications of functional bres from crab shell chitin. J Biotechnol assembly of gold nanoparticles and application of the gold-silk
1999;70:3737. bioconjugate in vapor sensing. Small 2007;3:46673.
[138] Park KE, Jung SY, Lee SJ, Min BM, Park WH. Biomimetic nanobrous [163] Feng XX, Zhang LL, Chen JY, Guo YH, Zhang HP, Jia CI. Prepara-
scaffolds: preparation and characterization of chitin/silk broin tion and characterization of novel nanocomposite lms formed
blend nanobers. Int J Biol Macromol 2006;38:16573. from silk broin and nano-tio2. Int J Biol Macromol 2007;40:
[139] Yoo CR, Yeo IS, Park KE, Park JH, Lee SJ, Park WH, et al. Effect of 10511.
chitin/silk broin nanobrous bicomponent structures on inter- [164] Feng XX, Guo YH, Chen JY, Zhang JC. Nano-tio2 induced secondary
action with human epidermal keratinocytes. Int J Biol Macromol structural transition of silk broin studied by two-dimensional
2008;42:32434. fourier-transform infrared correlation spectroscopy and Raman
[140] Falini G, Weiner S, Addadi L. Chitin-silk broin interactions: rele- spectroscopy. J Biomater Sci Polym Ed 2007;18:144356.
vance to calcium carbonate formation in invertebrates. Calcif Tissue [165] Xia Y, Gao G, Li Y. Preparation and properties of nanometer titanium
Int 2003;72:54854. dioxide/silk broin blend membrane. J Biomed Mater Res Part B
[141] Wongpanit P, Tabata Y, Rujiravanit R. Miscibility and biodegrad- 2009;90:6538.
ability of silk broin/carboxymethyl chitin blend lms. Macromol [166] Kaplan DL. Mollusc shell structures: novel design strategies for syn-
Biosci 2007;7:125871. thetic materials. Curr Opin Solid State Mater Sci 1998;3:2326.
[142] Falini G, Albeck S, Weiner S, Addadi L. Control of aragonite or [167] Li CM, Kaplan DL. Biomimetic composites via molecular scale self-
calcite polymorphism by mollusk shell macromolecules. Science assembly and biomineralization. Curr Opin Solid State Mater Sci
1996;271:679. 2003;7:26571.
[143] Addadi L, Joester D, Nudelman F, Weiner S. Mollusk shell forma- [168] Furuhashi T, Schwarzinger C, Miksik I, Smrz M, Beran A. Molluscan
tion: a source of new concepts for understanding biomineralization shell evolution with review of shell caliccation hypothesis. Comp
processes. Chem Eur J 2006;12:9817. Biochem Physiol B 2009;154:35171.
[144] Wongpanit P, Sanchavanakit N, Pavasant P, Bunaprasert T, Tabata Y, [169] An XQ, Cao CB. Biomineralization of caco3 through the coopera-
Rujiravanit R. Preparation and characterization of chitin whisker- tive interactions between multiple additives and self-assembled
reinforced silk broin nanocomposite sponges. Eur Polym J monolayers. J Phys Chem C 2008;112:652630.
2007;43:412335. [170] An XQ, Cao CB. Coeffect of silk broin and self-assembly monolay-
[145] Park WH, Jeong L, Yoo DI, Hudson S. Effect of chitosan on mor- ers on the biomineralization of calcium carbonate. J Phys Chem C
phology and conformation of electrospun silk broin nanobers. 2008;112:158449.
Polymer 2004;45:71517. [171] Mehta N, Hede S. Spider silk composite. Hypothesis 2005;3:215.
[146] Chen X, Li WJ, Yu TY. Conformation transition of silk broin [172] Cheng C, Shao ZZ, Vollrath F. Silk broin-regulated crystallization
induced by blending chitosan. J Polym Sci Part B Polym Phys of calcium carbonate. Adv Funct Mater 2008;18:21729.
1997;35:22936. [173] Gebauer D, Verch A, Brner HG, Clfen H. Inuence of selected
[147] Park SJ, Lee KY, Ha WS, Park SY. Structural changes and their effect articial peptides on calcium carbonate. Crystal Growth Des
on mechanical properties of silk broin/chitosan blends. J Appl 2009;9:2398403.
Polym Sci 1999;74:25715. [174] Gebauer D, Colfen H, Verch A, Antonietti M. The multiple roles of
[148] Kweon HY, Um IC, Park YH. Structural and thermal characteris- additives in caco3 crystallization: a quantitative case study. Adv
tics of Antheraea pernyi silk bron/chitosan blend lm. Polymer Mater 2009;21:4359.
2001;42:66516. [175] Gebauer D, Volkel A, Colfen H. Stable prenucleation calcium car-
[149] Gobin AS, Froude VE, Mathur AB. Structural and mechanical char- bonate clusters. Science 2008;322:181922.
acteristics of silk broin and chitosan blend scaffolds for tissue [176] Cheng C, Yang YH, Chen X, Shao ZZ. Templating effect of silk
regeneration. J Biomed Mater Res Part A 2005;74:46573. bers in the oriented deposition of aragonite. Chem Commun
[150] Gobin AS, Butler CE, Mathur AB. Repair and regeneration of the 2008;(43):55113.
abdominal wall musculofascial defect using silk broin-chitosan [177] Kong XD, Cui FZ, Wang XM, Zhang M, Zhang W. Silk broin reg-
blend. Tissue Eng 2006;12:338394. ulated mineralization of hydroxyapatite nanocrystals. J Crystal
[151] Garcia-Fuentes M, Giger E, Meinel L, Merkle HP. The effect Growth 2004;270:197202.
of hyaluronic acid on silk broin conformation. Biomaterials [178] Kino R, Ikoma T, Monkawa A, Yunoki S, Munekata M, Tanaka J, et al.
2008;29:63342. Deposition of bone-like apatite on modied silk broin lms from
[152] Malay O, Bayraktar O, Batigun A. Complex coacervation of silk simulated body uid. J Appl Polym Sci 2006;99:282230.
broin and hyaluronic acid. Int J Biol Macromol 2007;40:38793. [179] Kino R, Ikoma T, Yunoki S, Monkawa A, Matsuda A, Kagata G, et al.
[153] Ren YJ, Zhou ZY, Liu BF, Xu QY, Cui FZ. Preparation and charac- Biodegradation of multilayer silk broin and hydroxyapatite com-
terization of broin/hyaluronic acid composite scaffold. Int J Biol posite material. Key Eng Mater 2006;309311:116972.
Macromol 2009;44:3728. [180] Nogueira GM, Aimoli CG, Weska RF, Nascimento LS, Beppu MM.
[154] Garcia-Fuentes M, Meinel AJ, Hilbe M, Meinel L, Merkle HP. Silk In vitro calcication of silk broin hydrogel. Key Eng Mater
broin/hyaluronan scaffolds for human mesenchymal stem cell 2008;361363:5036.
culture in tissue engineering. Biomaterials 2009;30:506876. [181] Weska RF, Nogueira GM, Vieira Jr WC, Beppu MM. Porous silk broin
[155] Seo YK, Yoon HH, Song KY, Kwon SY, Lee HS, Park YS, et al. Increase membrane as a potential scaffold for bone regeneration. Key Eng
in cell migration and angiogenesis in a composite silk scaffold for Mater 2009;396398:18790.
tissue-engineered ligaments. J Orthop Res 2008;27:495503. [182] Lin F, Li YC, Jin J, Cai YR, Wei KM, Yao JM. Deposition behavior and
[156] Gulrajani ML, Gupta D, Periyasamy S, Muthu SG. Preparation and properties of silk broin scaffolds soaked in simulated body uid.
application of silver nanoparticles on silk for imparting antimicro- Mater Chem Phys 2008;111:927.
bial properties. J Appl Polym Sci 2008;108:61423. [183] Huang J, Wong C, George A, Kaplan DL. The effect of genetically
[157] Dong Q, Su HL, Cao W, Han J, Zhang D, Guo QX. Biogenic synthe- engineered spider silk-dentin matrix protein 1 chimeric protein on
sis of hierarchical hybrid nanocomposites and patterning of silver hydroxyapatite nucleation. Biomaterials 2007;28:235867.
nanoparticles. Mater Chem Phys 2008;110:1605. [184] Zhao Y, Chou AHK, Li G, LeGeros RZ. Nonwoven silk broin
[158] Al SY, Gao JG, Zhu LS, Ma ZJ, Li XC. Preparation and spectroscopic net/nano-hydroxyapatite scaffold: preparation and characteriza-
studies of nanosilver/silk-broin composite. Spectros Spectral Anal tion. J Biomed Mater Res Part A 2009;91:11409.
2008;28:21269. [185] Wang J, Zuo Y, Yang W-H, Zhou GQ, Zhang L, Li Y-B. Study on prepa-
[159] Kang M, Jung R, Kim HS, Youk JH, Jin HJ. Silver nanoparti- ration of n-ha and silk broin bio-mineral material. J Inorg Mater
cles incorporated electrospun silk bers. J Nanosci Nanotechnol 2009;24:2648.
2007;7:388891. [186] Du CL, Jin J, Li YC, Kong XD, Wei KM, Yao JM. Novel silk
[160] Zhou Y, Chen WX, Itoh H, Naka K, Ni QQ, Yamane H, et al. Prepa- broin/hydroxyapatite composite lms: structure and properties.
ration of a novel core-shell nanostructured gold colloid-silk broin Mater Sci Eng C 2009;29:628.
bioconjugate by the protein in situ redox technique at room tem- [187] Vachiraroj N, Ratanavarapon J, Damrongsakkul S, Pichyangkura R,
perature. Chem Commun 2001;(23):25189. Banaprasert T, Kanokpanont S. A comparison of thai silk broin-
[161] Yin HS, Ai SY, Shi WJ, Zhu LS. A novel hydrogen peroxide based and chitosan-based materials on in vivo biocompatibility for
biosensor based on horseradish peroxidase immobilized on gold bone substitutes. Int J Biol Macromol 2009;45:4707.
nanoparticles-silk broin modied glassy carbon electrode and [188] Wang L, Nemoto R, Senna M. Effects of alkali pretreatment of silk
direct electrochemistry of horseradish peroxidase. Sens Actuators broin on microstructure and properties of hydroxyapatite-silk
B 2009;137:74753. broin nanocomposite. J Mater Sci Mater Med 2004;15:2615.
J.G. Hardy, T.R. Scheibel / Progress in Polymer Science 35 (2010) 10931115 1115

[189] Li L, Wei KM, Lin F, Kong XD, Yao JM. Effect of silicon on the forma- [203] Lv Q, Hu K, Feng Q, Cui F. Fibroin/collagen hybrid hydrogels with
tion of silk broin/calcium phospate composite. J Mater Sci Mater crosslinking method: Preparation, properties and cytocompatibil-
Med 2008;19:57782. ity. J Biomed Mater Res Part A 2008;84:198207.
[190] Cao B, Mao C. Oriented nucleation of hydroxylapatite crystals on [204] Rios CN, Skoracki RJ, Miller MJ, Sattereld WC, Mathur AB. In vivo
spider dragline silks. Langmuir 2007;23:107015. bone formation in silk broin and chitosan blend scaffolds via
[191] Okabyashi R, Nakamura M, Okabayashi T, Tanaka Y, Nagai ectopically grafted periosteum as a cell source: a pilot study. Tissue
A, Yamashita K. Efcacy of polarized hydroxyapatite and silk Eng Part A 2009;15:27157.
broin composite dressing gel on epidermal recovery from full- [205] Wang SD, Zhang YZ, Yin GB, Wang HW, Dong ZH. Electrospun
thickness skin wounds. J Biomed Mater Res Part B 2009;90: polylactide/silk broin-gelatin composite tubular scaffolds for
6416. small-diameter tissue engineering blood vessels. J Appl Polym Sci
[192] Furuzono T, Kishida A, Tanaka J. Nano-scaled hydroxyap- 2009;113:267582.
atite/polymer composite I. Coating of sintered hydroxyapatite [206] Wang SD, Zhang YZ, Wang HW, Yin GB, Dong ZH. Fabrication and
particles on poly(gamma-methacryloxypropyl trimethoxysilane)- properties of the electrospun polylactide/silk broin-gelatin com-
grafted silk broin bers through chemical bonding. J Mater Sci posite tubular scaffold. Biomacromolecules 2009;10:22404.
Mater Med 2004;15:1923. [207] Meinel L, Hofmann S, Betz O, Fajardo R, Merkle HP, Langer R, et al.
[193] Furuzono T, Yasuda S, Kimura T, Kyotani S, Tanaka J, Kishida Osteogenesis by human mesenchymal stem cells cultured on silk
A. Nano-scaled hydroxyapatite/polymer composite IV. Fabrication biomaterials: comparison of adenovirus mediated gene transfer
and cell adhesion properties of a three-dimensional scaffold made and protein delivery of bmp-2. Biomaterials 2006;27:49935002.
of composite material with a silk broin substrate to develop a [208] Kukreja N, Onuma Y, Daemen J, Serruys PW. The future of drug-
percutaneous device. J Artif Organs 2004;7:13744. eluting stents. Pharmacol Res 2008;57:17180.
[194] Korematsu A, Furuzono T, Yasuda S, Tanaka J, Kishida A. Nano- [209] Lian XJ, Wang S, Xu GL, Lin NN, Li Q, Zhu HS. The application with
scaled hydroxyapatite/polymer composite iii. Coating of sintered tetramethyl pyrazine for antithrombogenicity improvement on silk
hydroxyapatite particles on poly(4-methacryloyloxyethyl trimelli- broin surface. Appl Surf Sci 2008;255:4802.
tate anhydride)-grafted silk broin bers. J Mater Sci Mater Med [210] Cassinelli C, Cascardo G, Morra M, Draghi L, Motta A, Catapano G.
2005;16:6771. Physical-chemical and biological characterization of silk broin-
[195] Foo CWP, Patwardhan SV, Belton DJ, Kitchel B, Anastasiades coated porous membranes for medical applications. Int J Artif
D, Huang J, et al. Novel nanocomposites from spider silk-silica Organs 2006;29:88192.
fusion (chimeric) proteins. Proc Natl Acad Sci USA 2006;103: [211] Langer RS, Peppas NA. Present and future applications of bio-
942833. materials in controlled drug delivery systems. Biomaterials
[196] Huang LM, Wang HT, Hayashi CY, Tian BZ, Zhao DY, Yan YS. 1981;2:20114.
Single-strand spider silk templating for the formation of hierar- [212] Germeshausen KJ, Robert S. Langer Nature Rev Drug Disc
chically ordered hollow mesoporous silica bers. J Mater Chem 2004;3:5461546.
2003;13:6668. [213] Rujiravanit R, Kruaykitanon S, Jamieson AM, Tokura S. Preparation
[197] Xu Q, Li JB, Peng Q, Wu LL, Li SP. Novel and simple synthesis of of crosslinked chitosan/silk broin blend lms for drug delivery
hollow porous silica bers with hierarchical structure using silk as system. Macromol Biosci 2003;3:60411.
template. Mater Sci Eng B 2006;127:2127. [214] Bayraktar O, Malay O, Ozgarip Y, Batigun A. Silk broin as a novel
[198] Qing FL, Ji M, Lu RH, Yan KL, Mao ZP. Synthesis of peruoroalkyl- coating material for controlled release of theophylline. Eur J Pharm
containing multifunctional groups compounds for textile nishing. Biopharm 2005;60:37381.
J Fluorine Chem 2002;113:13941. [215] Wang X, Zhang X, Castellot J, Herman I, Iafrati M, Kaplan DL.
[199] Prachayawarakorn J, Boonsawat K. Physical, chemical.and dye- Controlled release from multilayer silk biomaterial coatings to
ing properties of Bombyx mori silks grafted by 2-hydroxyethyl modulate vascular cell responses. Biomaterials 2008;29:894903.
methacrylate and methyl methacrylate. J Appl Polym Sci [216] Malay O, Yalcin D, Batigun A, Bayraktar O. Characterization of silk
2007;106:152634. broin/hyaluronic acid polyelectrolyte complex (pec) lms. J Ther-
[200] Cai ZS, Jiang GC, Qiu YP. Chem modication of Bombyx mori silk mal Analy Calorimetry 2008;94:74955.
with epoxide epsib. J Appl Polym Sci 2004;91:357986. [217] Malay O, Batigun A, Bayraktar O. Ph- and electro-responsive char-
[201] Tsukada M, Imai T, Freddi G, Lenka S, Kasai N. Grafting of vinyl acteristics of silk broin-hyaluronic acid polyelectrolyte complex
monomers onto silk using redox systems. Yellowing of silk. J Appl membranes. Int J Pharm 2009;380:1206.
Polym Sci 1998;69:23946. [218] Huemmerich D, Slotta U, Scheibel T. Processing and modication
[202] Poole AJ, Church JS, Huson MG. Environmentally sustainable bers of lms made from recombinant spider silk proteins. Appl Phys A
from regenerated protein. Biomacromolecules 2009;10:110. 2006;82:21922.