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As most cellmaterial interactions are mediated by the cell membrane, it is worth considering which molecules comprise the membrane and how they are arranged. There is enormous diversity among
cell types, especially when considering bacteria, which possess a cell
wall that is exterior to the cell membrane. In this Perspective, we will
focus only on cells from multicellular organisms, such as those that
Royal School of Mines, Prince Consort Road, Imperial College, London SW7 2AZ.
*e-mail: m.stevens@imperial.ac.uk
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PERSPECTIVE
Glycocalyx
1 m
Cell membrane
Endothelial cell
HO
O
OH
AcHN
O
HO
COO
O
O3S
O
HO
O3SHN
RBC
Normal
Diseased
HO
COO
O
O
HO
AcHN
60 nm
Desulfated
OSO3
O
HO
20 nm
30 nm
10 nm
Normal
OH
Figure 1 | Representation of cell surface properties. a, Diagram of a 3535nm section of the red blood cell surface showing the lipids (pink), major
proteins (blue) and carbohydrates (orange). Different proteins and carbohydrates are indicated by different shapes and shading. Although there is also
considerable heterogeneity in the lipid component of the cell membrane, this is omitted both for simplicity and because the nanoscale organization is still
a matter of debate. The relative dimensions, concentrations and chemistries of all membrane components can vary drastically depending on cell type,
disease state and life cycle, as shown in bd. b, Relative sizes of the glycocalyx for a red blood cell (RBC; <10nm) and a cell lining the blood vessel wall
(>200nm). c, Relative concentration of HER2 protein (also known as ERBB2) at the surfaces of a normal cell and a cancerous cell. In certain tumour lines,
this protein alone can cover more than 15% of the cell surface area57. d, The carbohydrate coat plays a chemical as well as physical role in cell regulation.
Negatively charged domains resulting from sulfation of disaccharide chains are capable of specific protein interactions. Disrupting these domains with
strong oxidizing agents can drastically alter cell behaviour, even preventing embryonic stem cells from undergoing differentiation. Ac, acetyl. Insert
shows carbohydrate chain based on N-acetylglucosamine (brown) and glucaronic acid (tan) monomers. Green indicates N- (circle), 6-O- (square) or
2-O-sulfation (star).
tens of nanometres, this layer can regulate the approach of the cell
to a solid support. Interference microscopy studies have shown that
for many cell types the membrane is initially held around 50 nm
away from a solid surface12,13. Over several minutes, this spacing
decreases12, allowing proteins in some regions of the cell membrane
to contact the support. Understanding how and why this process
occurs is of critical importance in determining which surfacegrafted ligands will be accessible to the cell, and when.
One way to study the effects of glycocalyx-mediated structuring
at the cell interface is by modifying ligand tethers. In one such investigation14, glycine spacers were systematically added to the end of
an adhesion-promoting peptide and it was found that, as expected,
adding a small spacer increased cell binding. Notably, spacers more
than nine amino acids in length actually decreased cell binding. It
was posited that this effect may be due to the peptide folding over,
underlining the need to consider the mechanical as well as dimensional properties of even a supposedly inert spacer. One study of this
effect looked at polymer brushes with a bimodal length distribution
(22- or 10-mer) where only one of the two chain populations bore
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PERSPECTIVE
Many cell types will not grow if floating freely, and must be attached
to a surface to function normally. However, some surfaces support cell growth much better than others. One important parameter is the materials wettability, which is determined mostly by the
charge, polarizability and polarity of the surface functional groups.
These factors do not, however, directly mediate cell response. The
effect of surface wettability is largely to alter the type (and state) of
proteins adsorbed from solution20, which in turn affects cell adhesion. Hydrophobic surfaces irreversibly adsorb large quantities of
albumin, an abundant serum protein that does not support cell
attachment. By contrast, moderately hydrophilic surfaces such as
glow-discharge-treated polystyrene (that is, tissue culture plastic)
a
f
b
Peptide chemistry
Polymer chemistry
Crystallography
Physical chemistry
c
Electrochemistry
Biochemistry
HO
O
HO
OH
Figure 2 | Interdisciplinary achievements and possibilities within chemistry for the development of bioactive surfaces. a, Grafting polymer scaffolds
(orange) with synthetic polypeptides (gray) allows for simultaneous control of structural and mechanical properties as well as incorporation of specific
cell-adhesion cues. b, Modifying the length and composition of a molecular tether can qualitatively alter functional presentation. A bimodal brush (shown
here) can increase ordering and promote ligand availability. c, With careful control of intermolecular spacing, it is possible to create a monolayer that
responds to electric fields with an altered conformation and consequently different surface properties. The example shown here undergoes a hydrophilic to
hydrophobic transition when the terminal carboxylic acid is electrostatically driven down onto the substrate. d, Through genetic engineering, it is possible
to introduce enzymes at the cell surface that will modify electroactive monolayers, enabling electronic transduction of biological activity, for instance
by conversion of 4-hydroxyphenyl valerate to quinones. e, The galactose-specific hepatic lectin has enabled carbohydrate-grafted tissue-engineering
scaffolds that promote liver cell growth. Many lectin-binding pockets are preformed, making them possible future targets for rational ligand design. f,
Cyclic RGD (shown here as part of a pentapeptide) is a more effective adhesion promoter than the linear sequence. Structural studies of native ligand
receptor pairs can reveal optimal conformations for synthetic peptides.
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Peptidepeptide
Integrin
Immunoglobulin
Lectin
Syndecan
10-mer NCAM-binding
peptide for neural
cell adhesion
Galactose-grafted
polymers for
hepatocyte culturing
HSBD-containing peptides
to simulate ECM protein
interactions better
Amino-acid sequence
Conformation (cyclization)
Modifications:
Methylation
Phosphorylation
Ligand
variables
Stereochemistry (epimers)
Chain length
Modifications:
Sulfation (domains)
Acetylation
Presentation variables
Material system
Polymer functionalization
Self-assembled monolayer
Tether properties
Co-functionalization
Density/clustering
Length
Flexibility
Synergystic binding
Protein-resistant background
Figure 3 | Molecular diversity of cell adhesion motifs available for surface engineering. The classic ligand-receptor pair of integrins and RGD has been
heavily optimized, for instance by cyclization (upper left) to increase affinity. More recently, alternative peptides have been explored that bind other
classes of molecules such as the immunoglobulin neural cell adhesion molecule (NCAM), potentially offering greater cell-type selectivity. In some cases
(such as syndecans, upper right), synthetic peptides (blue) can bind carbohydrates (orange) rather than the peptide portion of membrane components.
Conversely, some adhesion molecules such as lectins recognise carbohydrates, opening up a new class of surface functionalization possibilities. With this
potential comes a new set of molecular-level variables that can be tailored. However, many of the supramolecular considerations are the same. Regardless
of the type of molecule grafted on a surface, factors such as spacing and tether length will affect cellular response. Note that this is not a comprehensive
list of available classes of adhesion molecules; that subject has previously been covered in more detail58. Similarly, the applications given are intended only
as representative examples and the interested reader should consult more comprehensive reviews59. HSBD, heparan sulfate binding domain.
NATURE CHEMISTRY | VOL 3 | AUGUST 2011 | www.nature.com/naturechemistry
585
PERSPECTIVE
PERSPECTIVE
Natural role
Engineering challenge
Physical
reordering
Proteolytic
cleavage
Protein
secretion
T
Figure 4 | Some of the pitfalls and possibilities associated with cellular microenvironment remodelling. a, Cells naturally pull on the ECM, maintaining a
state of tension (indicated by arrows). On adsorbed protein surfaces, this tension can result in rearrangement, leading to variable properties. But grafting
adhesion molecules (blue) to tethers (red) allows controlled reordering. The ligand initially samples a volume determined by the conformation of the
tether and, on binding, can move a distance determined by the ultimate tether length. b, Most cells secrete proteases that, in vivo, remodel the ECM (grey)
but, in vitro, can cleave grafted adhesion molecules (red/blue). However, this process can be exploited through the creation of specifically degradable
gels (orange) that encourage cell penetration. By crosslinking polymers with synthetic peptides (chains of spheres, arrows indicate point of cleavage),
it is possible to control cell-mediated scaffold degradation. c, Cells naturally secrete proteins to build the ECM, but in vitro the resulting layer (grey) can
cover an engineered surface (orange). With the right surface, however, this protein layer can also be used as a weak link to allow the gentle removal of
adherent cells. By switching the material surface properties, cells can be released (vertical arrows) without the use of harsh chemical agents. One class of
such smart materials is thermoresponsive polymers, that is, polymers which transition between a collapsed state (top) and a hydrated, extended state
(bottom) in a physiologically relevant range of temperatures (T).
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PERSPECTIVE
will need to create interfaces that read and react to cellular outputs
rapidly and specifically. One approach to this challenge is measurement using surface-sensitive optical techniques such as surface
enhanced Raman scattering or sum frequency generation spectroscopy. Alternatively, electrochemical measurements may offer
higher spatial resolution or greater possibility for rapid feedback.
One example54 of this approach was the generation of monolayers of 4-hydroxyphenyl valerate on an electrode and the genetic
engineering of a population of cells to express a membrane-bound
enzyme capable of oxidizing those groups to quinones (Fig. 2d).
By quantifying the quinone density with cyclic voltammetry, it
was thus possible to measure biochemical cellular activity. More
recently, this concept was extended55 by cloning an entire transmembrane electron transport chain into bacteria, creating cells
that were capable of direct electron transfer to inorganic electrodes.
Future advances in functional surfaces will probably increase the
range of transduction possible, perhaps even obviating the need for
genetically modified cells. Regardless, it will remain absolutely necessary to consider dynamic changes initiated by both the cell and
the artificial material.
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Acknowledgements
We thank J. Weaver for constructive reading of this manuscript. M.M.S. thanks ERC
starting investigator grant Naturale for funding.
Additional information
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