123

Bioactive biomaterials
Jeffrey A Hubbell
The most important advances in the field of biomaterials over
the past few years have been in bioactive biomaterials.
Materials have been developed to incorporate bioactivity
through biological recognition, including incorporation of
adhesion factors, polyanionic sites that mimic the electrostatics
of biological regulatory polysaccharides, and cleavage sites for
enzymes involved in cell migration. Materials have also been
developed to be active in biological environments by
undergoing phase changes in situ, including transformations
from liquid precursors to solids and from soluble materials to
materials that are immobilised on tissue surfaces.
Addresses
Department of Materials and Institute for Biomedical Engineering,
Swiss Federal Institute of Technology and the University of Zurich,
Moussonstrasse 18, CH-8044 Zurich, Switzerland;
e-mail: [email protected]
Current Opinion in Biotechnology 1999, 10:123129
http://biomednet.com/elecref/0958166901000123
Elsevier Science Ltd ISSN 0958-1669

Introduction
Important advances have been made in the field of biomaterials over the past few years, and most of these have been
associated with rendering materials biologically active. It is
as logical to develop biomaterials that are bioactive as it is
to develop drugs that are bioactive. Pharmacological activity is based on the principles of biological recognition, for
example, to competitively inhibit receptors or enzymes, to
block binding sites, to regulate certain biological pathways,
and so on. One can, in most applications, consider the
function of a biomaterial-based device analogously to that
of a pharmaceutical. An example could be considered in
the context of the vascular graft. Certainly, the basic functions of the graft have nothing to do with biological
activity: it must carry blood flow, must resist the dilatory
pressures of the cardiovascular system, and must resist the
compressive and kinking forces of the tissues external to
the cardiovascular system. These goals can be met entirely in the absence of biological activity. Beyond these basic
performance goals, however, one may be well advised to
turn to bioactivity: to prevent coagulation, to encourage
endothelial cell attachment and retention, to promote capillary infiltration as a source of endothelial cells, to prevent
excessive smooth muscle cell proliferation and collagen
matrix expression. To turn to biological recognition to
accomplish these ends, for example, to incorporate biologically motivated, biomimetic adhesion-promoting sites or
to incorporate growth factors, is quite logically based on
the analogy of drug design. In addition to the highly
biospecific biological recognition phenomena described
above, some natural biological recognition proceeds by less
specific physicochemical interactions, such as binding of a

charged polysaccharide to a protein. One can mimic these

less specific interactions as well, in this case with polyelectrolytes, in biomaterials design.
This review considers biological recognition from one additional perspective, namely enzymatically modulated
material degradation. Most degradable biomaterials
degrade based on chemical clocks, such as ester hydrolysis
in the polymer backbone or sidechains. The rate of the
hydrolysis reaction is programmed via selection of the
detailed chemical environment around the ester bonds,
such as side groups, crystallinity and hydrophilicity, to set
the speed of the chemical clock. In this approach, the clock
is set in the laboratory, based on a prediction of a biological
response. An alternative approach, one involving the concept of bioactivity, is to let a degradation program proceed
along a coordinate of the healing process, rather than time,
by making the material sensitive to the feedback provided
by the cells involved in the healing response. Indeed, this
paradigm is employed naturally in tissue generation,
remodelling and regeneration, as cells enzymatically
degrade the extracellular matrix around them. By rendering
a biomaterial sensitive to these enzymatic activities, one
can pursue the goal of biomimetic material degradation.
This review also considers bioactivity from a perspective
other than biological recognition, namely active material
transformation from one state to another in the presence of
the biological system. Materials can be designed to transform
under some external stimulus, such as light, temperature or
chemical composition (e.g. from liquids to elastic solids, from
cell adhesive to cell non-adhesive, or from freely soluble to
bound to a tissue surface). By design of materials for bioactivity, it may be possible to develop materials that enable
new surgical procedures, such as closure of internal incisions
by bioactive adhesives, or organ culture methods, such as
culture of cell and cell aggregates within gels.

Bioactivity by incorporation of adhesion factors

One approach toward biological activity in biomaterials is
the incorporation of adhesion-promoting oligopeptides or
oligosaccharides. Cell adhesion to traditional biomaterials,
such as polyethylene, polytetrafluoroethylene or silicone
rubber, is based upon indirect recognition, that is, by proteins from the body fluids adsorbing nonspecifically to the
material surface and some subset of these adsorbed proteins, including fibronectin, fibrinogen, and vitronectin,
promoting the adhesion of cells by interacting with the corresponding adhesion receptors. As a more direct approach,
one that permits a greater degree of control by not depending upon a secondary mediator, several investigators have
explored the covalent or physicochemical incorporation of
adhesion-promoting oligopeptides and oligosaccharides.
The reader may refer to other reviews on the molecules

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Biochemical engineering

incorporated [1,2] and on methods that have been

employed to incorporate them, in the context of biomaterials for tissue engineering [24] and drug delivery [5].
Extensive research has been performed on the incorporation of adhesion-promoting oligopeptides into biomaterial
surfaces. These peptides are based on the primary structure of the receptor-binding domains of proteins such as
fibronectin and laminin, and often the corresponding linear or cyclised sequences can display similar receptor
specificity and binding affinity, as well as signalling of cellular responses, compared to the whole protein [1,2]. Early
work demonstrated an important possible advantage of
working with short adhesion peptides, rather than the
complete parent protein, that is, that the peptides could be
displayed in a manner such that nearly all of them were
available and active for binding to cell-surface receptors
[6]. When a bioactive tripeptide from fibronectin, the
sequence RGD (amino acid single letter code), was immobilised by its amino-terminal primary amine via a glycyl
spacer, approximately 105 copies per cell were required to
induce cell adhesion, spreading, focal contact formation
and cytoskeletal organisation, whereas many more copies
of the complete protein were required, presumably due to
unfolding of the protein associated with adsorption or
absorption of the protein in an orientation such that the
receptor-binding domain was not sterically available.
Likewise, early work also demonstrated, in the example of
the bioactive YIGSR pentapeptide from laminin, that peptides covalently immobilised in a single orientation could
have considerably different biological activity than the
same peptide adsorbed in a number of possible orientations, many of which were presumably sterically incapable
of binding to their receptors [7].
There are several important fundamental issues about how
such adhesion signals are displayed. For example, it has
been demonstrated that cell adhesion strength depends
upon the surface concentration of adhesion ligands, and
furthermore, that cell migration rates depend in a very sensitive manner on the strength of cell adhesion [8]. If cell
adhesion strength is very low, the cell is incapable of developing adequate traction to migrate, whereas if cell
adhesion strength is very high, this traction is also very
high and the rate of reassociation of dissociated adhesion
receptors is very fast, resulting in very slow migration.
Thus, in situations where one would incorporate adhesion
ligands to promote adhesion and migration (e.g. in the
pores of a vascular graft to promote capillary ingrowth or in
the pores of a wound healing membrane for promoting
healing of skin ulcers) one must search for an optimum in
cell migration with this phenomenon in mind.
Furthermore, there may be cases in which the spatial display of adhesion ligands is critical. For example, in the
immobilisation of galactose for promotion of hepatocyte
adhesion via the asialoglycoprotein receptor, the degree of
ligand mobility, as determined by the length of the flexible
tether used for immobilisation, strongly influenced cell

adhesion behaviour [9]. The extent of cell spreading

depended on both the overall amount of ligand immobilised as well as its ability to cluster into spatial
microdomains. This dependence is consistent with the
binding of the receptor to triads of galactose residues, in a
manner mimicking binding to the branched oligosaccharide in the natural ligand for the receptor.
What could be other advantages of employing short bioactive peptides or saccharides, rather than the complete
parent glycoprotein? One goal is selectivity for targeted
cell types. Cell-type selectivity is a common goal in drug
targeting [5], and it may also represent an important goal in
tissue engineering [2]. For example, in vascular graft
design, it would be beneficial to develop a material to support the adhesion and migration of vascular endothelial
cells, while at the same time rejecting the adhesion of
blood platelets [10]. The integrin receptor 41 represents
a target that is present on the endothelial cell but not the
blood platelet, and there exists an adhesion ligand specific
to this receptor, the tetrapeptide REDV, in the adhesion
protein fibronectin. Fibronectin does not provide a useful
ligand for this purpose, however, because the protein contains numerous adhesion-promoting domains, at least one
of which binds to most cells in the body. This lack of selectivity can be avoided by working with only the REDV
domain, and not the other, less specific domains. Thus,
one can accomplish a goal with a small domain of the protein that could not be accomplished with the complete
parent proteins, that is, cell-type selectivity. Other goals in
tissue engineering can also be obtained more easily with
small peptides than with the whole proteins, such as the
control of cell phenotype by exposure to certain adhesion
ligands, as has been demonstrated in culture with boneforming cells [11], and enhancement of cell adhesion
strength, which has been shown to be important in clinical
studies with endothelial cell seeded vascular grafts [12].
Several biomaterial systems have been explored and developed for display of bioactive adhesion-promoting ligands,
and a small number of these are discussed below to illustrate key points. More complete reviews on this topic can
be found elsewhere [2,13]. Most investigators have
approached the problem from the perspective of covalent
immobilisation of adhesion-promoting ligands upon a biomaterial surface, usually after some type of surface
modification [2,13], and this is usually adequate. For
degradable materials, which are particularly useful in tissue
engineering because they do not need to be removed from
the body, surface modification may be inadequate because
the surface can be rapidly degraded and removed. For such
situations, it may be necessary to develop new polymers
that display the ligand in the bulk of the material, and so
display new ligands on the surface continually as it is
degraded and remodelled. This has been addressed with
the practically important degradable biomaterial poly(lactic
acid) by the synthesis of the copolymer poly(lactic acid-colysine). The -amino groups on the lysyl comonomer

Bioactive biomaterials Hubbell

provide sites of ligand grafting in solution, thus presenting

the peptides on the transient surface as it moves though the
material during degradation [14]. Poly(lactic acid) has been
used extensively as sutures and surgical staples, and such
modification may make it more useful in tissue engineering
applications. Bulk incorporation is also useful in the case of
gels that are subject to cell infiltration. Bioactive migrationpromoting peptides, such as the YIGSR domain and the
SIKVAV domain from laminin, have been incorporated
within gels for use in nerve regeneration [15]. In this case,
the peptides were incorporated throughout the bulk of the
three-dimensional gels to permit the infiltrating neuronal
cells to contact the signals on all sides and at all times
during the infiltration process.
Other approaches to the presentation of bioactive adhesion-promoting peptides have been to include the
bioactive peptide sequences in the backbone of the polymer chains (e.g. in polypeptide biomaterials). For example,
elastin-like polypeptides have been developed to produce
elastic, protein-based biomaterials that contain adhesion
sites, such as the RGD tripeptide, engineered within the
polypeptide sequence [16].

125

Furthermore, specialised biomaterials have been developed to incorporate and modulate these important
biologically active molecules. For example, with the goal
of bone repair by delivery of the growth factor transforming growth factor , the affinity of the growth factor for
heparin has been exploited [20]. Many such growth factors bind heparin, as well as heparin sulphate
proteoglycans in the extracellular matrix. To exploit this
binding affinity, heparin was conjugated to collagen
matrices used in bone repair, and this immobilised
heparin served as an affinity site to bind and slowly
release the growth factor in the healing site. As an extension of this approach, the growth factor was chemically
conjugated to the collagen matrix via a poly(ethylene glycol) spacer. This growth factor was biologically active in
the immobilised state, presumably being able to bind to
the receptors on the surface of the cell and permit receptor dimerisation and signal transduction [21]. The
ability of immobilised growth factors to be biologically
active has also been demonstrated in the very well-characterised system of epidermal growth factor conjugated to
synthetic polymer surfaces, where it was shown to be
capable of directing hepatocytes to maintain their liverspecific morphology and function [22].

Bioactivity by incorporation of growth factors

Polypeptide growth factors are powerful regulators of a variety of cellular behaviours, including cell proliferation,
migration, differentiation, and protein expression, and
these molecules are being developed as important therapeutics in tissue regeneration (e.g. in closing bone defects
and in healing chronic ulcers in the skin). Growth factors
are also being explored as key components of biomaterials
and biomaterial systems, as discussed in the illustrative
examples below. Why is it useful to consider growth factors
as part of a system involving biomaterials, rather than on
their own? The biological activity of the growth factor
depends not only upon its identity, but also upon how it is
presented to the cells in space and over time. For example,
it has been demonstrated that some growth factors are more
effective when provided to cells through a controlled
release process, whereas others are more effective when
presented as a bolus [17]. This difference in behaviour may
be related to how the cells traffic and recycle their receptors
for these growth factors, and it may be possible to modulate
trafficking and recycling by altering either the growth factor or its interactions with a biomaterial that is releasing or
presenting it. Under the natural conditions of the body, the
extracellular matrix plays a role in storing, displaying and
releasing growth factors. Given that the natural biomaterial
of the body plays this important role, it seems reasonable to
explore this and related behaviour with biomaterials, to
mimic this function of the extracellular matrix.
Controlled release systems have been developed for
growth factors, for example, based on traditional biomaterials in delivering angiogenic growth factors in vascular
repair [18] or in delivering neuronal survival and differentiation factors in neurodegenerative diseases [19].

Exciting advances have been made with the display and

delivery of precursors for polypeptide growth factors, as well
as the growth factors themselves. It has been demonstrated
recently that plasmid DNA can be efficiently taken up and
the encoded gene expressed when the plasmid is presented
on the surface of a biomaterial. This has been shown in the
context of smooth and cardiac muscle cells by presentation
of plasmid DNA on sutures [23], as well as in the context
of bone repair, in which fibroblasts in the repair tissue took
up the plasmid DNA encoding the growth factor bone morphogenetic protein-4, and by expression of the growth factor
enhanced bone formation [24]. Cellular and DNA interactions with the biomaterial appear to be important in
determining the extent of uptake by the cells in the repair
site; as such, the DNA-presenting biomaterial surface
becomes a bioactive biomaterial that expresses its bioactivity for long periods of time by affecting the behaviour of the
cells that come in to contact with the biomaterial.

Bioactivity by physicochemically based

biological recognition
Some biological interactions are based on physicochemical
interactions that are less specific than those described in
the previous sections, such as adsorption due to electrostatic interactions, which can be readily mimicked and
incorporated into bioactive biomaterials. Heparins electrostatically dominated interactions in anticoagulation
present one example, and extensive work has been performed at immobilisation of heparin on biomaterials to
render them bioactive [25]. Extensive work has also been
performed to obtain other polymers that possess heparinlike activity, for example, by screening dextrans with
varying extents of suphonation [26,27] or by identification

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Biochemical engineering

of plant polysaccharides with a fortuitously appropriate

charge density and structure, as in the example of fucan
[28]. Such biological activity can be extended beyond the
anticoagulant activity of heparin (i.e. binding to the proteins antithrombin III and thrombin to catalyse complex
formation between these two proteins), to other activities
of heparin, such as binding to growth factors or interfering
with a growth factors binding to its receptor [29]. It is
interesting to note that similar biological activity can be
obtained completely outside the platform of a polysaccharide chain (as seen with heparin) for charge presentation.
For example, with appropriate degrees of suphonation,
polyurethanes can function as heparin-like water-soluble
polymers [30].
Novel biomaterials for biosensing have recently been
developed by combination of less specific activity based on
physicochemical interactions and highly specific activity
based on enzymatic activity. Synthetic polymer hydrogels
have been developed as three-dimensional conductors,
with redox active charge transfer centres chemically incorporated within the gel network [31,32]. Redox
enzymes, such as glucose oxidase, can be incorporated and
even chemically bonded throughout the network to provide
for efficient electron transfer from the enzymes active site,
through the gel by transfer from site to site, to a collection
electrode to form a concentration-dependent sensor for glucose. Thus, careful combination of individual biomaterial
properties, such as polymer chain and network dynamics
(which controls charge transfer through the gel by modulating transient close approaches to within electron tunnelling
distances between immobilised charge transfer sites) and
enzyme coupling (which controls charge transfer from the
enzyme to the gel), can lead to a biomaterial with a highly
specialised and selective biological activity.

Bioactivity by incorporation of enzymatic

recognition sites
The two sections above, dealing with incorporated adhesion and growth factors, addressed the transmission of
biological information from a biomaterial to the neighbouring cells. One can also consider the other direction, in
which the biomaterial is the recipient of information produced by cells. One such form of information is enzymatic
activity associated with the cell surface during cell migration. Cell migration through collagen [33] and fibrin [34]
gels, both natural biomaterials involved in the generation,
remodelling and regeneration of tissues, has been explored
and is known to depend mainly upon the sensitivity of the
material to proteases produced by the cells, upon the
amount of enzyme produced by the cells, and upon the
amount of material to be remodelled by the cells as they
migrate through the material.
Approaches have been developed to engineer biomaterials
that can be remodelled by cells through cell-associated
enzymatic activity. Cells naturally remodel the extracellular matrix in development, adaptation and healing, and

materials that are subject to the remodelling activities of

the cell may enable exploitation of these biological activities in tissue engineering. For example, a fascile route to
the chemical incorporation of bioactive signals has been
developed for fibrin, a natural biomaterial matrix that can
be remodelled proteolytically. In this modification scheme,
exogenous peptides bear in one domain a substrate for the
transglutaminase involved in coagulation, factor XIIIa, and
are thus covalently conjugated to the fibrin network as it
forms, incorporating the bioactive peptide within the gel.
Another domain of the peptide bears a bioactive peptide,
for example, with cell adhesion or growth factor binding
activity [35]. Through such a route, it is possible to incorporate the biological activity of a host of non-fibrin
proteins (e.g. laminin) as synthetic components added into
the platform of the biologically-derived fibrin gel.
Completely synthetic biomaterials have been designed
that are proteolytically degradable and that comprise
other bioactive components as well. Gels have been
formed based on poly(ethylene glycol) chains comprising
central oligopeptides, sites that are substrates for collagenase or plasmin, both of which are involved in cell
migration [36]. These water-soluble hybrid chains may
then be coupled at their termini to form three-dimensional elastic gels that are completely synthetic, but
which are also degradable by cell-associated enzymatic
activity. Additional biological activity can be conferred
upon the proteolytically remodable gels by copolymerisation of suitably reactive oligopeptides, such as
terminally reactive poly(ethylene glycol) grafted with
the adhesion peptide RGD [37]. It is thus possible with
these approaches to construct materials that possess a
large number of the characteristics of the natural extracellular matrix, but which are totally synthetic and
capable of fascile manufacture and customisation.

Bioactivity by material transformation

Biomaterials can possess biological activity (i.e. activity in a
biologically relevant context) via the ability to be transformed from one state to another. Several examples have
already appeared and some have already found clinical utility, and selected examples are presented below. The reader is
directed elsewhere for a review exclusively on this topic [38].
Materials that undergo phase transformations have a great
deal of potential for use in surgery, for example, as adhesives, sealants, and barriers to celltissue contact. A
water-soluble macromer has been developed based on
poly(ethylene glycol) central blocks with oligo(lactic acid)
flanking blocks and terminal acrylates; the large central
block provides water solubility, the flanking oligoester
degradability, and the terminal sites of unsaturation polymerisability [39]. These soluble materials can be rapidly
transformed into elastic hydrogels by exposure to light in
the presence of suitable photoinitiators, such as eosin yellowish. These materials have been employed to block
blood platelet adhesion to blood-vessel surfaces after

Bioactive biomaterials Hubbell

injury to thereby improve the postsurgical healing of the

vessel [37,40], and to serve as a barrier and depot for local
drug delivery to prevent the formation of scar tissue adhesions between organs after gynaecological surgery [41].
Photocurable, and photolysable, hydrogels have also been
developed based on cinnamylidene acetate dimerisation,
rather than acrylate polymerisation as in the examples
above [42,43]. Materials that can be converted from liquids
into elastic gels by reaction after mixing of two liquids
have also been developed, for example, by reaction of
N-hydroxysuccinimidyl activated esters on the end of a
difunctional poly(ethylene glycol) with amines on the end
of a tetrafunctional poly(ethylene glycol) [44]. This
scheme has also been employed to synthesise enzymatically degradable gels, by using an aminated hyaluronic acid
chain as one component of the two-liquid system
(D Aeschlimann, personal communication). All of the reactions described above (i.e. gel formation by photoinitiation
or by mixing of two liquids) can be sufficiently gentle to be
carried out in vivo. For example, photopolymerisation of
degradable poly(ethylene glycol) acrylates is currently
used to seal leaking tissue surfaces after surgery in man.
A second sort of material transformation that can be used
to modulate biological interactions is gel swelling or collapse in response to temperature changes. Material
chemistries that can be employed to lead to polymers displaying lower critical solution behaviour (giving gels on
warming of liquids) or upper critical solution behaviour
(giving gels on cooling of liquids) have been described
[45]. Such materials have been used as injectable depots
for drug delivery (after a liquid-to-solid transformation)
[46], as reversible cell culture substrates upon which cell
monolayers and multilayers can be cultured and then
released intact for transplantation (after a hydrophobic-tohydrophilic transformation) [47], and to block a
ligand-binding site on a protein in a thermally regulatable
manner (after an extended-chain-to-collapsed-chain transformation) [48,49]. Such transformations can also be
obtained in protein solutions, also based on changes in
temperature, as well as changes in salt concentration. For
example, self-assembly can be obtained in protein based
on silk and elastin to form gels [50], as well as by electrostatic interactions in other designed oligopeptides and
polypeptides employing leucine zipper domains [51] and
ionic self-complementary domains [52].
As a final example of approaches toward bioactivity through
material transformation, schemes to develop biomaterial
monolayers as therapeutics have been developed. In the
examples on photopolymerisable hydrophilic macromers
discussed above, a therapeutic goal was blockade of cell
interactions with a tissue surface after injury associated
with surgery. This might also be obtainable with polymer
monolayers, only nanometers thick, rather than gels tens of
microns thick. This goal has been approached by the design
of multifunctional polymers bearing domains that bind to
tissue surfaces based on electrostatic interactions via a

127

backbone of poly(lysine), in addition to other domains that

repel the binding of proteins and cells via pendant blocks of
poly(ethylene oxide) [53,54]. These graft co-polymers
were demonstrated to bind to complex biological surfaces,
through affinity for their net-negative charge, and sterically
stabilise them against subsequent adhesion, such as preventing lectin-induced agglutination of red blood cells
[53] or preventing the formation of postoperative adhesions [54]. Such approaches of assembly on biological
surfaces have been taken even further, to actually chemically graft amine reactive poly(ethylene glycol) derivatives
to lysine residues in proteins on tissue surfaces to inhibit
cell adhesive interactions after surgical tissue damage,
where such chemical grafting is performed under conditions sufficiently mild to permit grafting in vivo [55].

Conclusions
Biological activity has played an important role in modern
biomaterials development, employing the principles of
biological recognition that are used so frequently in pharmaceutical design in addition to other more
material-centric principles, for example, those that permit
material to respond to external stimuli. Only recently have
these novel bioactive biomaterials begun to make clinical
impact, but given the relatively long cycle from concept to
clinic this is to be expected. Indeed, it is probable that the
concepts of bioactivity reviewed here, as well as others,
will make much more direct clinical impact in the culture
laboratory and clinic in the next few years. It is clear that
the expenses associated with development and regulatory
approval of products based on bioactive biomaterials will
be higher than that of products based on traditional biomaterials, and as such these development activities must
be targeted at economically important applications with
largely unmet needs, where advantages in safety and efficacy associated with bioactivity compensate favourably for
the higher cost of development and regulatory approval of
the bioactive product.

References and recommended reading

Papers of particular interest, published within the annual period of review,
have been highlighted as:

of special interest
of outstanding interest
1.

Yamada KM: Adhesive redognition sequences. J Biol Chem 1991,

266:12809-12812.

2.

Hubbell JA: Biomaterials in tissue engineering. Biotechnology

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3.

Peppas NA, Langer R: New challenges in biomaterials. Science

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4.

Ratner BD: The engineering of biomaterials exhibiting recognition

and specificity. J Mol Recognit 1996, 9:617-625.

5.

Langer R: Drug delivery and targeting. Nature 1998, 392:5-10.

6.

Massia SP, Hubbell JA: An RGD spacing of 440 nm is sufficient for

integrin avb3-mediated fibroblast spreading and 140 nm for focal
contact and stress fiber formation. J Cell Biol 1991, 114:1089-1100.

7.

Massia SP, Rao SS, Hubbell JA: Covalently immobilized laminin

peptide Tyr-Ile-Gly-Ser-Arg (YIGSR) supports cell spreading and
co-localization of the 67-kilodalton laminin receptor with a-actinin
and vinculin. J Biol Chem 1993, 268:8053-8059.

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Biochemical engineering

8.

Palecek SP, Loftus JC, Ginsberg MH, Lauffenburger DA, Horwitz AF:
Integrinligand binding properties govern cell migration speed
through cellsubstratum adhesiveness. Nature 1997,
385:537-540. [Published erratum appears in Nature 1997, 388:210].
This paper provides very convincing evidence of a direct link between cell
adhesion strength and cell migration rates, providing for the biomaterials engineer important guidance that too much of a good thing (e.g. an adhesion ligand) can be bad (e.g. if cell migration was an important performance criterion).
9.

Griffith LG, Lopina S: Microdistribution of substratum-bound

ligands affects cell function: hepatocyte spreading on
PEO-tethered galactose. Biomaterials 1998, 19:979-986.

10. Massia SP, Hubbell JA: Vascular endothelial cell adhesion and
spreading promoted by the peptide REDV of the IIICS region of
plasma fibronectin is mediated by integrin a4b1. J Biol Chem
1992, 267:14019-14026.
11. Bearinger JP, Castner DG, Healy KE: Biomolecular modification
of p(AAm-co-EG/AA) IPNs supports osteoblast adhesion and
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9:629-652.
12. Meinhart J, Deutsch M, Zilla P: Eight years of clinical endothelial
cell transplantation. Closing the gap between prosthetic grafts
and vein grafts. ASAIO J 1997, 43:M515-521.
This clinical study with endothelial cell seeded vascular grafts demonstrated that graft performance in high flow regions (where stresses on the
attached endothelial cells were greatest) was better when the biomaterial
matrix included the adhesion protein fibronectin. Although the study was
performed with a complete protein, rather than an engineered peptide, it
provides direct clinical evidence of the need, and probability of success,
of such approaches.
13. Elbert DE, Hubbell JA: Surface treatments of polymers for
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14. Cook AD, Hrkach JS, Gao NN, Johnson IM, Pajvani UB,
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dimensional extracellular matrix engineering in the nervous

system. J Biomed Mater Res 1998, 40:392-400.
The investigators employed grafted adhesion peptides from laminin to
enhance the rate of neurite extension through a three-dimensional gel to
which the peptide was attached. This provides an example of peptide grafting in three dimensions and evidence that clinically relevant biomaterials are
likely to result.
16. Urry DW, Pattanaik A, Xu J, Woods TC, McPherson DT, Parker TM:
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Dinbergs ID, Brown L, Edelman ER: Cellular response to

transforming growth factor-beta1 and basic fibroblast growth
factor depends on release kinetics and extracellular matrix
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18. Zarge JI, Huang P, Husak V, Kim DU, Haudenschild CC, Nord RM,
Greisler HP: Fibrin glue containing fibroblast growth factor type 1
and heparin with autologous endothelial cells reduces intimal
hyperplasia in a canine carotid artery balloon injury model. J Vasc
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19. Haller MF, Saltzman WM: Nerve growth factor delivery systems.
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20. Schroeder-Tefft JA, Bentz H, Estridge TD: Collagen and heparin
matrices for growth factor delivery. J Controlled Release 1997,
49:291-298.
The investigators employed the affinity of the important growth factor
transforming growth factor for heparin to develop clinically relevant
bioactive biomaterials for bone regeneration. Heparin, grafted to the collagen substrate, served as a docking and release site to provide for
delayed release. When cells migrate into such matrices, they may also
release the growth factor by local enzymatic action, as occurs naturally in
the extracellular matrix.
21. Bentz H, Schroeder JA, Estridge TD: Improved local delivery of
TGF-b2 by binding to injectable fibrillar collagen via difunctional
polyethylene glycol. J Biomed Mater Res 1998, 39:539-548.
This report, along with [22], provides evidence that covalently immobilised
growth factor, in this case transforming growth factor , can be presented to
cells in a bioactive manner. This was not clear a priori, in that the ligandbound receptors must presumably dimerise in the plane of the membrane in
order to express biological activity.

22. Kuhl PR, Griffith-Cima LG: Tethered epidermal growth factor as a

paradigm for growth factor-induced stimulation from the solid
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biological activity. In addition to the points made to [21 ], this demonstrates that internalisation of the ligand-bound receptors is not critical in
order to express biological activity.
23. Labhasetwar V, Bonadio J, Goldstein S, Chen W, Levy RJ: A DNA
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skeletal and cardiac muscle. J Pharm Sci 1998, 87:1347-1350.
This pair of reports (with [24]) demonstrates that plasmid DNA, delivered
upon a biomaterial surface, can be efficiently taken up by cells and the
encoded gene expressed in a wound healing environment. This was demonstrated to have clinical relevance in repair of skeletal and cardiac muscle
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with the material can continue to express the gene encoded on the plasmid
DNA for an extended period of time.
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McCauley LK, Davidson BL, Roessler BJ: Stimulation of new bone
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Maaroufi RM, Jozefowicz M, Tapon-Bretaudiere J, Jozefonvicz J,

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therapeutic. It possesses only one set of biological signals, and thus obtains
generally only one sort of healing response, namely a scar. The investigators
present a simple approach by which to functionalise fibrin with exogenous

Bioactive biomaterials Hubbell

bioactive peptide signals, by using bi-domain peptides, one peptide containing a substrate site for the coagulation transglutaminase factor XIIIa and
the other containing a bioactive peptide of interest. This opens a fascile
route to make fibrin clots with customised biological activity.
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32:241-244.
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synthetic biomaterial gel that can be degraded by cell-associated proteases.
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this opens the door for synthetic materials that can be invaded and remodelled by cells in a healing response, in complete analogy with the way tissues
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Hern DL, Hubbell JA: Incorporation of adhesion peptides into

nonadhesive hydrogels useful for tissue resurfacing. J Biomed
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38. Hubbell JA: In situ material transformations in tissue engineering.

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40. West JL, Hubbell JA: Separation of the arterial wall from blood
contact using hydrogel barriers reduces intimal thickening after
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129

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48. Stayton PS, Shimoboji T, Long C, Chilkoti A, Chen G, Harris JM,

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53. Elbert DL, Hubbell JA: Self-assembly and steric stabilization at
heterogeneous, biological surfaces using adsorbing block
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Most drugs operate by highly specific biological recognition. In this case, a
soluble bioactive polymer was developed that recognised its target with relatively low specificity, via binding of a poly(cation) to net negatively charged
cell surfaces. Grafted poly(nonion) chains then sterically repelled high affinity biological interactions, providing an example of drug-like bioactivity via
physicochemical mechanisms.
54. Elbert DL, Hubbell JA: Reduction of fibrous adhesion formation by
a copolymer possessing an affinity for anionic surfaces. J Biomed
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55. Deible CR, Beckman EJ, Russell AJ, Wagner WR: Creating
molecular barriers to acute platelet deposition on damaged
arteries with reactive polyethylene glycol. J Biomed Mater Res
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