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Beyond the lipid-bilayer: interaction of polymers and nanoparticles with
membranes
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Matthias Schulz,† Adekunle Olubummo† and Wolfgang H. Binder*

Received 19th October 2011, Accepted 22nd January 2012
DOI: 10.1039/c2sm06999g

Membranes can be fabricated either from lipid or polymer molecules, leading to the formation of
liposomes or polymersomes. In all types of liposomal membranes, the issue of phase separation plays
a central role not only in the membrane-formation itself, but also in the resulting structural features
taking place within or at the surface of such membranes. When nanoparticles or polymers interact with
lipid membranes, the final morphology is strongly determined by the charge, composition and size of
the interacting components, which in turn induce phase separation processes. The present review
assembles investigations on lipid/polymer/nanoparticle interaction with the main focus directed
towards model membrane systems published in the recent literature starting from
2005, providing
a deeper and more thorough understanding of these complex interactions and their effects on
membrane properties.

1. Introduction lipid-based liposomes have had a victorious path along the

Membranes can be fabricated either from lipids or polymers,
leading to the formation of liposomes or polymersomes. Starting
from the initial discovery of Kunitake1 and Ringsdorf et al.2
Martin-Luther University Halle-Wittenberg, Institut f€ ur Chemie,
Lehrstuhl Makromolekulare Chemie, Chair of Macromolecular
Chemistry, Faculty of Natural Sciences II (Chemistry, Physics and
Mathematics), Institute of Chemistry, Von Danckelmannplatz 4,
D-061120 Halle (Saale), Germany. E-mail: wolfgang.binder@chemie.
uni-halle.de; Fax: +49-(0)345/55-27392; Tel: +49-(0)345/55-25930
† MS and AO contributed equally to this article.
scientific world, using them most widely as delivery systems in
medicine for drug formulation or for gene delivery. However, the
significantly younger ‘‘polymersomes’’, whose membranes are
built from amphiphilic polymers, have gained increased atten-
tion, leading to a significant expansion of the structural vari-
ability of membranes. In contrast to lipid membranes, where the
overall physicochemistry (permeability, bending moduli, stiff-
ness or fluidity of components) is determined by
5 nm thick
lipid bilayer, the membranes of polymersomes offer significantly
more chemical-, spatial- and physicochemical variability. As
demonstrated by the early pioneers, such as Discher and

Matthias Schulz is currently Adekunle Olubummo is

a PhD candidate in Macromo-
lecular Chemistry at Martin
Luther-University Halle-Wit-
tenberg (since 2010). His
research interests include the
preparation and characterization
of amphiphilic block copolymers
and their interactions with bio-
logical membrane models. Being
born (1984) in Tangerm€ unde
(Germany), he studied chemistry
at the University of Halle and
Matthias Schulz conducted his MSc in Macro-
molecular Chemistry (2009)
under the supervision of Prof.
Binder.
Adekunle Olubummo
currently a PhD candidate in
Macromolecular Chemistry at
Martin Luther-University
Halle-Wittenberg (since 2010).
He studied Industrial chemistry
at the Federal University of
Technology (Akure, Nigeria)
and conducted his MSc in
Polymer Material Science at the
University of Halle under the
supervision of Prof. Binder. He
is interested in the synthesis and
characterization of polymer
functionalized nanoparticles and
their interactions with lipid
membranes.

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Eisenberg,3 Eisenberg and Shen,4 Meier et al.5–7 and many others
(for representative publications see e.g. ref. 8–11), the thickness
of a polymersomal membrane made via self-assembly of amphi-
philic di- and triblock copolymers can be adjusted ‘‘stepless’’ from

10 to
50 nm with a significant increase in its stability.12,13
Critical for this process is the relation of the (hydrophobic)
membrane thickness (d) scaling with the molecular weight (Mw) of
the polymer by dhydrophobic z Mw0.5,14 whereas the hydrophilic
part of the membrane often scales with dhydrophilic z Mw.15 Often,
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a significantly reduced lateral mobility of the polymers in
comparison to a conventional lipid bilayer membrane can result
as a consequence of specific conformational contraints.13,16 In all
types of liposomal membranes (see Fig. 1), the issue of phase
separation plays a central role not only in the membrane forma-
tion itself, but also in the resulting structural features taking place
within or at the surface of such membranes.17,18 Thus, lateral
phase separation in lipid membranes can lead to domain forma-
tion, which—in cellular membranes—are proposed to form
‘‘lipid-rafts’’ as an essential clustering phenomenon of biological
receptors.19,20 Interaction of polymers within a polymersomal
membrane can display features of lateral phase separation
between the inner/outer parts of a polymersome, further resulting
in macroscopic distortions by selective swelling of a specific
polymeric phase.21 When nanoparticles interact with lipid
membranes22,23 or polymersomes,24–28 the final morphology is
strongly determined by the charge and size of the interacting
components. Thus either adsorption or incorporation22 of nano-
particles can result, with a significantly higher load of nano-
particles being incorporated into polymersomal membranes due
to their higher stability.25 Fine-tuning of the polymer/surface
interaction can even result in a selective location of the nano-
particles in a specific part of the polymersomal membrane.29
The interaction between polymers and lipid membranes is
mainly governed by the hydrophobic/hydrophilic balance, as well
as by the polymeric architecture. Adsorption, followed by reor-
ganization of domains, can take place by inserting graft
Wolfgang H. Binder is currently
a full professor of Macromolec-
ular Chemistry at the Martin-
Luther-University Halle-Wit-
tenberg (since 2007). Being
born (1969) and raised in
Vienna (Austria), he studied
chemistry at the University of
Vienna and conducted a PhD in
organic chemistry (1995). After
postdoctoral stays (1995–1997)
with Prof. F. M. Menger
(Emory University, Atlanta,
Wolfgang H: Binder USA) and Prof. J. Mulzer
(University of Vienna), he
completed his habilitation at the
Vienna University of Technology (TU-Wien, 2004), where he was
an Associate Professor from 2004–2007. Subsequently he moved to
the MLU-Halle-Wittenberg as a Full Professor in 2007. His
research interests include polymer synthesis, supramolecular
chemistry and nanotechnology.
4850 | Soft Matter, 2012, 8, 4849–4864
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polymers.30–33 Hydrophilic polymers, especially those bearing
poly(ethylene oxides) (PEOs) or poly(oxazolines) (POZOs), can
insert into membranes, resulting in ‘‘mushroom’’- or ‘‘brush’’-type
decorations.34 Charged polymers (polyelectrolytes) such as poly-
carboxylates or polyamines can adsorb onto oppositely charged
lipid bilayers35 producing lateral phase separation via electro-
statically induced flip/flops.36–40 Finally, membrane distortions
can result if the interacting forces are larger than the bending
modulus, visible in increased curvature of the membranes.41,42
Pores are another structural feature which have generated
significant debate among membrane scientists,43 resulting in
different types of permeability, depending on the polymer’s
architecture.44 Polycarboxylate43 as well as di-, tri-,45–47 and
starblock copolymers48–51 composed of PEO–PPO–PEO and
PEO–PS-diblockcopolymers52–55 have been investigated proving
a strong impact of the molecular weight56,57 as well as displaying
sealing effects47,58,59 or membrane destruction in the case of
cationic polymers.60,61
When nanoparticles (up to
100 nm) interact either with lipid
or polymersomal membranes, a significantly different behavior is
observed, as stabilities of lipo- and polymersomes are different.
Thus hydrophilic nanoparticles are assembled onto the outside
or inside of lipid vesicles, whereas hydrophobic nanoparticles can
only be inserted into lipid membranes with much difficulty, as at
significantly higher loading immediate destruction of the
membranes follows.62,63 Macroscopically visible effects are
curving of the lipid membrane, budding or fission effects.64,65
Negatively charged nanoparticles (with sizes of 4–40 nm) can
lead to clustering effects thus reducing mobility and increasing
the gel-phase of the involved lipids.66 In contrast, membranes
composed of polymers can more easily be decorated with
nanoparticles, either by incorporation into the hydrophobic
interior25,67 or via outside attachment.26,67
A significant more complex picture is generated, when a direct
interaction between polymers, lipids and nanoparticles takes
place on the same liposomal surface.44 In these cases, a basic
understanding has not been achieved up to now.44,68,69 As both,
vesicles and polymersomes, are thermodynamically labile struc-
tures, an important aspect (often neglected) is the preparation
method of vesicles or polymersomes. Despite the fact that a large
variety of different methods is available, starting from the basic
methods such as film rehydration, dialysis, microfluidics70,71 and
spin-coating,72 the final morphology often depends on the
method of preparation. Thus, the electroformation method73 in
particular has found increased attention for the preparation of
large unilamellar vesicles, both for liposomes and for polymer-
somes. The present review assembles investigations on lipid/
polymer/nanoparticle interaction with the main stress directed
towards model membrane systems, putting a focus on recent
literature starting from
2005. The basic idea of this review is the
summation of recent publications on these basic interactions
with the aim to induce a deeper and more thorough under-
standing of these complex interactions.
2. Interaction between nanoparticles and lipid
membrane
The interaction of functional nanoparticles with membranes
can influence important aspects such as intercellular uptake,74
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Fig. 1 Selected structure-formation upon lipid/polymer and lipid/nanoparticle interaction in membranes. All drawings are schematic, focusing on the
dynamic as well as structural effects.

drug- and gene-delivery,75 localization of nanoparticles at
specific positions of the membrane or within cellular compart-
ment22 and their biological/biomedical application with minimal
cytotoxicity. Because of their flexibility and high elasticity, lipid
membranes can easily be deformed when nanoparticles interact
with them either by insertion or via adsorption onto their surface.
It has been found that the effective interaction between nano-
particles and membranes includes hydrophobic mismatch
effects,76 chain stretching of the lipids near the site of interac-
tion,77 nanoparticle induced spontaneous curvature of the
membrane,78 changes in lipid packing79 and macroscopic effects
such as pearling.80 The following sections will focus on how
nanoparticles interact with membranes with regards to nano-
particle surface hydrophobicity, effect of the nanoparticles
charge, size effect, and their shape (i.e. being round shaped or
tubular).
bilayer permeabilization, biocompatibility and liposomal release,
also useful for investigating biochemical reactions in vitro. In
order to embed NPs into the hydrophobic membrane interior,
the nanoparticle must fulfil two requirements. Firstly, the NP
must be small enough (<8 nm) to fit within the bilayer dimension
and secondly, it must possess a hydrophobic surface. In one of
the first successful examples of NP incorporation, Vogel et al.23
have described the fabrication of highly controllable lipid/
hydrophobic CdSe nanoparticle hybrid vesicles where nano-
metre-sized particles are confined within a 4 nm thick lipid
bilayer membrane (Fig. 2A) as proven by confocal microscopy
(see Fig. 2B). These hybrid vesicles were used as nanocontainers

2.1 Effect of nanoparticles on lipid membranes

2.1.1 Effect of hydrophobic nanoparticles. Hydrophobicity

plays a very important role when dealing with the interaction
between nanomaterials and lipid membranes, as nanoparticle
assembly and lipid stabilization are essentially driven by hydro-
phobic/hydrophilic (interfacial) effects.81 Upon interaction of
NPs with lipid membranes, numerous theoretical studies have
found that morphological reorganization and hole formation
strongly depend on the surface hydrophobicity or hydrophilicity
Fig. 2 (A) Hybrid lipid/QD vesicles showing the location of hydro-
of the interacting NPs.82,83
phobic nanoparticles in a hydrophobic lipid bilayer. (B1) Confocal
In principle, a nanoparticle can be encapsulated into a lipo- fluorescence cross-sectional image of hybrid lipid/QD vesicles. (B2) TEM
some, being either trapped within the aqueous vesicle core or micrograph of hydrophobic Au-NPs (2 nm) embedded in the membrane
embedded into the hydrophobic lipid bilayer part. Embedding of of lipid vesicles. (A) is taken from ref. 23, copyright 2006, Wiley-VCH
functional hydrophobic nanoparticles into lipid membranes Verlag GMbH & Co. KGaA, Weinheim. (B2) is taken from ref. 22—
provides a plethora of different applications for controlling reproduced by permission of the PCCP Owner Societies.

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for the release of small molecular weight compounds into living
cells. Bothun et al.84 have embedded hydrophobic 5 nm oleic acid
capped superparamagnetic iron oxide nanoparticles into a DPPC
bilayer resulting in magnetoliposomes. The embedded NPs
suppressed the DPPC pretransition (Tp ¼ 36  C) (describing the
transition from the ordered gel (Lb) to the rippled gel (Pb) phase)
by shifting it to higher temperature. This effect was more
pronounced with increasing nanoparticle loading. Furthermore,
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it was also demonstrated that the embedded nanoparticles lead to
controlled release of the liposomal contents.
We have demonstrated that modifying the surface of nano-
particles with a hydrophobic or hydrophilic moiety will direct the
location of the nanoparticles either onto the surface or within
a specific compartment of the membrane.22 It was found that
2 nm hydrophobic Au-NPs were selectively enriched into the
hydrophobic lipid bilayer of a lipid vesicle membrane (see
Fig. 2B).
The enrichment of hydrophobic nanoparticles within the lipid
bilayer interior has been explained by Korgel et al.85 as a result of
NP interaction with the hydrophobic tail of the lipid molecules,
as shown in Fig. 3A. Thus hydrophobic nanoparticles can cause
the bilayer to ‘‘unzip’’ when they are located at the center, leading
to changes in lipid packing and disruption of lipid–lipid inter-
actions between the lipid head groups and/or the lipid alkyl tails.
This unzipping creates void space around the nanoparticle
resulting in nanoparticle clustering and minimization of the free
energy of deformation (Fig. 3B).
Bothun86 has demonstrated that the incorporation of hydro-
phobic decanethiol-covered Au-NPs into a liposomal membrane
lowers the melting temperature at high lipid to nanoparticle
ratios (more than 15 : 1 w/w) with an additional increase in
bilayer fluidity of the gel phase by reduction of lipid ordering. As
it remains unknown how and to what extent NP surface hydro-
phobicity induces disruption and pore formation of the lipid
bilayers, Zhu and Jing87 have observed that the disruption of
supported lipid bilayer was noticed above a critical NP hydro-
phobicity and concentration (c* z 3 nM). Semi-hydrophobic
nanoparticles (cPS-NPs with hydrophobic polystyrenes
Fig. 3 (A) Cryo-EM image of hybrid/Au nanoparticle vesicles showing
hydrophobic nanoparticle embedding within the bilayer. (B) Schematic
illustration of the insertion of a hydrophobic nanoparticle into a lipid
bilayer inducing membrane deformation and ‘‘unzipping’’ of the bilayer.
When two nanoparticles are present in the bilayer, as in B2, the total
membrane curvature is reduced by lateral aggregation of the nano-
particles as shown in B3. The strained regions of the lipid bilayer are
circled in red in (2). Figures reprinted with permission from ref. 85,
copyright 2010, American Chemical Society.
4852 | Soft Matter, 2012, 8, 4849–4864
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displaying an ionizable end-group) were found to readily adsorb
on a-PC bilayers and drag lipids from the bilayer leading to the
formation and growth of lipid-poor regions referred to as pores
or holes (Fig. 4A). Hydrophobic attraction facilitated the
formation of patched lipid bilayers on large NPs which further
enhanced the formation and growth of lipid-poor regions on
SLBs. Proof of this observation was obtained by coarse-grained
lipid models (Fig. 4B)88 explaining that
10 nm hydrophobic
nanoparticles were included into the membrane leading to
a lateral distribution of lipids around the inclusion which is
generally referred to as hydrophobic ‘‘mismatch’’. In contrast
semi-hydrophilic nanoparticles were observed to adsorb on the
surface of the lipid bilayer because of the substantial energy
barrier needed to be subdued for the wrapping process.
2.1.2 Effect of surface charge. Besides adsorption of nano-
particles on lipid membranes, the exposure of membranes to
nanoparticles composed of iron oxide, CdSe-quantum dots or
gold nanoparticles bearing a significant surface charge can lead
to severe membrane disruption effects induced by such nano-
particles, which in turn can result in enhanced porosity of the
cellular membrane.89 Two general types of disruption namely (a)
nanoscale hole formation and (b) membrane thinning have been
explored by the use of oriented circular dichroism,90 molecular
modeling91 and atomic force microscopy.92 Differences in the
packing density of lipids in close proximity to regions where the
nanoparticles are adsorbed were one of the main described
disruption effects.
A significant number of theoretical simulations have addressed
the interaction of nanoparticles with lipid membranes. Li and
Gu93 showed by use of molecular dynamics simulation that in
contrast to uncharged particles (which prefer to remain in the
aqueous phase), charged nanoparticles induce membrane
deformation and influence the density distribution of the lipid
head groups driven by electrostatic interactions. Positively
charged nanoparticles interact with the phosphate terminus of
the lipids by increasing the tilt angle and thus enlarging the area
of the head groups. Other coarse-grained molecular dynamics
simulations94 showed that cationic gold nanoparticles (AuNPs)
are more disruptive to negatively charged lipid vesicles than their
anionic counterparts by altering the surface texture i.e. formation
of pores which induce defective areas and thus alter the bilayer
surface texture. This disruptive penetration depends on the
number of positive charges on the nanoparticles surface, which in
turn may generate a pore, with the ‘‘pore’’ region in the bilayer
being substantially hydrated.
Positively charged nanoparticles can also induce flipping of
membrane areas leading to particle inclusion and membrane
depolarization.95 Such positively charged nanoparticles may bind
to the plasma membrane as proven via TEM96 or even induce
aggregation. It has been shown97 that the presence of silanol
groups on the surface of cationic amino derived g-Fe2O3@SiO2
core–shell magnetite nanoparticles (CSMNs) can significantly
contribute to the driving force responsible for their strong
attraction with giant unilamellar vesicle membrane. The silanol/
membrane affinity could increase the tilt angle of the membrane
head groups (of mixed DOPC/DOPG 4 : 7) which in turn leads
to an enhancement of phospholipid density and subsequent
membrane stiffening. In addition to the mentioned effects,
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Fig. 4 (A) Fluorescence micrographs in panels (i–iv) show the morphological evolution of mixed a-PC and LR-PE SLBs after adding cPS-NPs of d ¼
28 nm at c ¼ 3.3 nM in PBS solutions, against the elapsed time. Panel (v) is the schematic illustration of the final morphology of lipid bilayers after the
adsorption of cPS-NPs of d ¼ 28 nm PBS solutions. (B) Trajectory snapshots of hydrophobic NP simulation in stereo views: (1) initial configuration of
the starting point (2) NP adsorption onto the surface of the bilayer, (3) embedding of NPs by inducing pore formation on the lipid bilayer, (4) final
equilibrium stage, with NPs located at the embedded pore in the lipid bilayer. (A) reprinted with permission from ref. 87, copyright 2011, American
Chemical Society. (B) reprinted with permission from ref. 88, copyright 2008, American Chemical Society.

membrane fusion is another effect caused by nanoparticles.

When liposomes are aggregated by oppositely charged NPs, the
interliposomal contact ensures that they can cling and fuse to
form larger vesicles, also inducing membrane fusion.98
In contrast to cationic nanoparticles, experiments93 have
shown that anionically charged nanoparticles induce the
formation of increased densities of the lipids around the nano-
particles by interacting with the positively charged head group of
the lipid, thereby causing a reduction in the tilt angle of the lipid’s
head group. Simulation studies as well as calorimetric and fluo-
rescence studies have demonstrated that membranes formed
from single component phospholipid bilayers can switch their
local phase state upon binding of charged nanoparticles.99

2.1.3 Effects of nanoparticle size. Despite the intense research

effort in the field of cellular uptake of nanoparticles, it remains
uncertain how the NP’s size relates to the extent of membrane
disruption or the structure and morphology of nanoparticle lipid
assemblies. Bothun86 reported that smaller dodecanethiol capped
gold nanoparticles (5.7 nm) in the proximity of a lipid bilayer
embed directly within the lipid bilayer, while larger Au-NPs were
capped and dispersed in the aqueous phase by a lipid monolayer
bringing the alkyl tails of the lipid in contact with the decanethiol
tails. In a similar experiment, Roiter et al.100 have studied the
interaction of DMPC with small (<22 nm) and large (>22 nm)
nanostructured silica nanoparticles on a silica surface with AFM
(Fig. 5A). It was found that small nanoparticles form a hole in
the lipid bilayer, whereas larger nanoparticles are mostly covered
with the lipid bilayers as a whole. A critical particle diameter
d0 z 22 nm was defined for a transition between the envelopment
and the free state of a particle (with d0 z 3l, where l is a specific
length of the membrane as defined by the bending modulus and
the adhesion constant). This size effect has been determined by
measuring the bilayer current in the presence of nanoparticles,101
using 50 nm and 500 nm aminated silica nanospheres in contact
with DOPC bilayers. The bilayer-disruption effect of these
Fig. 5 (A) Lipid bilayer formation in the presence of particles larger
than lipid bilayer thickness. (1 and 2) AFM images of lipid bilayer
formation over a silica nanoparticles surface with size smaller and bigger
than 22 nm. (B) Membrane curvature induced by noninteracting and
interacting nanoparticles. (A) reprinted with permission from ref. 100,
copyright 2008, American Chemical Society.
nanospheres was significantly more pronounced for the 500 nm
sized particles. As demonstrated by calculations,102 smaller,
interacting nanoparticles will cause a curving-away of the
membrane from the nanoparticles, whereas in the case of small,
non-interacting nanoparticles the membrane will curve towards
the nanoparticles (Fig. 5B). Non-anchored particles may be
repelled from or attracted towards the membrane surfaces. If
these particles are repelled from the membranes, depletion layers
are formed in front of these membranes which increase the
excess free energies of the membrane–water interfaces. In such

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a situation, the membranes bend towards the particle solution in
order to decrease the size of the depletion zone. On the other
hand, if relatively small particles were adsorbed onto the
membrane, the Gibbs adsorption equation implies that the
membrane tends to bend away from the particle solution in order
to increase its area.103
With larger nanoparticles, inclusion effects and the formation
of fission and budding structures104 could be derived. Computer
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simulations suggested that it is thermodynamically favorable for
2–8 nm sized NPs to embed within a lipid bilayer. Based on the
bilayer phase behavior, it was demonstrated that it may be
possible to embed nanoparticles that have a diameter in prox-
imity to, or even exceeding the thickness of the bilayer, which is
consistent with the numerical simulation.82
When dendrimers (which can be seen as small nanoparticles) in
noncytotoxic concentrations were interacting with lipids an
enhanced porosity with important implication for drug- and
gene-delivery was observed. Holl et al.105,106 showed with AFM
and conductance measurements that generation 5(G5) and
generation 7 (G7) amine terminated dendrimers induce a hole in
the membrane, while uncharged (hydrophobic) dendrimers can
only absorb onto the outer membrane surface. It was further
observed that such created holes led to dendrimer internalization
into cells and diffusion of cytosolic proteins out of cells.
2.1.4 Effects of fullerene (C60) and carbon nanotubes. Since
the discovery of fullerene (C60) in 1985 (ref. 107) lots of research
activities have been directed towards its use in biomedical tech-
nology, such as X-ray contrast agents,108 inhibitors to allergic
responses,109 transport of electrons across the host lipid
membranes,110 and as robust anti-HIV drugs.111 Therefore, it is
important to fully understand how fullerene interacts with
a bilayer membrane in order to habituate them for the above-
mentioned biomedical applications. Toxicology studies on
fullerene molecular aggregates suggest that they can enter cells,
alter their functions or cross the blood–brain barrier. However,
the mechanisms by which fullerenes penetrate and disrupt cell
membranes are still poorly understood.112 Coarse grained and
molecular dynamics simulation113,114 have been employed for
studying the interactions of both hydrophobic pristine C60 and
its various derivatives with a DPPC bilayer. It was shown that
the number of polar groups on the surface of C60 affects the
nature of fullerene/bilayer interactions which in turn can range
from partitioning into the bilayer to adsorption onto the bilayer
surface. Pure C60 enters the hydrophobic bilayer core and
remains inside. In contrast, more polar derivatives of fullerenes
drive rapidly into the bilayer from the aqueous phase, but were
found to be partitioned close to the head group/tail interface of
the bilayer. The amphiphilic C60-molecule is oriented such that
the hydrophobic surface interacts with lipid tails, while its polar
surface interacts with the lipid head groups.
Fullerene has been reported to suppress the phase transition of
DPPC bilayers, resulting in a ripple-like in-plane bilayer ordering
across the lipid bilayer with an enhanced correlation along the
bilayer plane.115 The bilayer ordering indicates that C60 can be
better accommodated into the host bilayer in the liquid crystal-
line state. Unlike small, hydrophobic solutes, it has been
shown116 that larger, non-hydrophobic fullerenes preferred to be
situated off the center plane of a hydrated DMPC membrane,
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which optimized the dispersion interactions between the atomi-
cally denser fullerene surface and the relatively dense atomic
environment. This tendency to be situated off the bilayer center
was mediated by the distortion of the bilayer structure by the
fullerene, which is induced as the fullerenes begin to aggregate.
2.2 Interaction between nanoparticles and polymersomes
Polymersomes as synthetic analogues to liposomes have attrac-
ted considerable attention due to their increased thickness,
rigidity and stability.28,117 In general, the interaction of nano-
particles and block copolymer vesicles has been shown to offer
a convenient way to control the arrangement of nanoparticles in
polymer films by segregating nanoparticles into a favorably
interacting polymer domain or at the interface between two
polymers.118,119 Morphological changes of block copolymer
assemblies can be induced by the incorporation of nanoparticles
which plays an active role in the self-assembly of the block
copolymer into membranes.120 This nanoparticle induced self-
assembly structure of the polymer is generally controlled by
solvent–nanoparticle and polymer–nanoparticle interactions.
Thus, either the internalization of NPs into the central hydro-
phobic position of the membrane is enabled, or its aggregation at
the interface (core–shell polymersomes) or the formation of
purely micellar aggregates.121 Maskos67 and Binder et al.122,123
showed that hydrophobic nanoparticles were successfully
enclosed inside the hydrophobic compartment of polymersomes
prepared from poly(butadiene)-b-poly(ethylene oxide) or poly-
isobutylene-b-polyethyleneoxide (PIB75-b-PEO52) block copoly-
mer. The quantum dots located between the hydrophobic
poly(butadiene) layers introduced curvature of the copolymer
layer around the guest particles (Fig. 6A). Loading of polymer
vesicles with ‘‘ultra-small superparamagnetic iron oxide’’
(USPIO) has been observed to increase the PDI and slightly
decrease the hydrodynamic radius which can be explained as
a result of larger hydrophobic effect when the copolymer is
combined with USPIOs coated with surfactant.28 Eisenberg and
Mai29 have shown that nanoparticles can selectively be incor-
porated into the central portion of block copolymer vesicle walls
by coating the particles with diblock copolymers of a structure
similar to that of the diblock copolymers used in the polymer-
some, which allowed the particles to be preferentially localized in
the central portion of the membrane’s wall (Fig. 6C).
To prepare stable nanoparticle-incorporated polymersomes, it
has been found124 that the mass ratio r ¼ mn/mp of nanoparticles,
mn, to polymer, mp, should be less than a critical ratio r* z 0.2 to
0.3, otherwise polymer/nanoparticle agglomerates will be
formed. This aggregation effect was observed by Lecomman-
doux125 where the incorporation of hydrophobic iron oxide
nanoparticles into vesicle-forming poly(butadiene)-b-poly(glu-
tamic acid) block copolymers was investigated. A rather aggre-
gated but still hollow, vesicle-like structure was observed, which
was deformable in external magnetic fields. Several factors such
as the nanoparticles surface functionality and the hydrophobic/
lipophilic balance determined their location in the polymersomal
membrane. Hydrophobic nanoparticles with a hydrophobic/
lipophilic balance (HLB) between the hydrophobic phase (PI)
and the hydrophilic phase (PEO/water) preferentially segregated
to the hydrophobic/hydrophilic interface with a penetration
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Fig. 6 (A) The inclusion of CdSe nanoparticles into the central portion of vesicles made from PIB-b-PEO. (C) Schematic illustration of the preparation
of diblock copolymer coated nanoparticles and TEM micrographs of vesicles with the incorporated nanoparticles prepared from the combined solution
of the PS235-b-PEO45 copolymer (0.5 wt%) and PEO45-b-PS155-b-P(APb)25 micelles (0.5 wt%). In (a) small vesicles; (b) a large vesicle; (c) an indented
vesicle with the magnified indentation of the vesicle membrane is shown. (B) Scheme of nanoparticle-induced bilayer pairing and bridging. The
nanoparticles are located in the hydrophobic/hydrophilic interface. (A) Reprinted from ref. 123, copyright 2008, Wiley-VCH Verlag GMbH & Co.
KGaA, Weinheim. (B) Reprinted with permission from ref. 124, copyright 2008, American Chemical Society. (C) Reprinted with permission from ref.
29, copyright 2010, American Chemical Society.

depth into the hydrophobic domain depending on its HLB
value.124 The nanoparticle decorated the interface of the poly-
mersomal membrane, which can induce the formation of bridges
from one bilayer to an adjacent bilayer, leads to bilayer pairing
(see Fig. 6B).
It has been reported126 that BCP vesicular bilayers composed
of nanoparticles can undergo a morphological transition into the
coexistence of spherical micelles and bilayers in water depending
on the NP volume fraction. This structural transformation
occurs through a budding process caused by the embedded
nanoparticles clustering. The driving process for this budding
process was attributed to the increased entropy of polymer
strands surrounding the nanoparticle aggregates due to the
excessive stretching of polymers at the edge of NP aggregates.
Hydrophobically functionalized nanoparticles have been repor-
ted to increase the membrane thickness (from 2.4 nm to 6.1  1.3
nm) of polymersomes prepared from PEO-b-PBO because they
interact with the hydrophobic domain of the polymer during the
self-assembly127 (see Fig. 7).
Increased functionality and enhanced mechanical strength of
polymersomes can be introduced by embedding of nanoparticles
into their curved membranes.128 Bon et al.129 have covered the
surface of positively charged polymer vesicles with negatively
Fig. 7 (1) Hydrophobic silver nano-particles entrapped in a polymer-
somal membrane; schematic representation of the nanoparticles thick-
ening the membrane and (2) hydrophilic gold nanoparticles encapsulated
in the core. Red and blue areas depict the hydrophobic and hydrophilic
domains of the block copolymer in the vesicles. Figures taken with
permission from ref. 127.
charged (24 nm) silica nanoparticles using electrostatic attrac-
tions as the driving force. The nanoparticles adhere to the outer
surface of polymersomes made of poly(N-butylmethacrylate)-b-
poly(ethylene oxide) without affecting the hollow bilayer,
thereby increasing the overall rigidity and altering their perme-
ability, useful for the uptake and release of drugs.
2.3 Biological impact of nanoparticle/membrane interactions
The interaction of nanoparticles with biological systems has
become one of the most significant areas of research in material
science, because the exposure of human beings to nanoparticles is
inevitable and therefore a deeper comprehension of the potential
risk and hazard is necessary. We will not go into a detailed
discussion about nanoparticle interactions with cell and their
eventual toxicological impact as this issue has been reviewed by
different authors, such as Stellacci,130 Brayner,131 Ren132 and
Drezek.133 However, it should be emphasized that smaller
nanoparticles with a diameter of tens of nanometres or less are
consistently more toxic than larger analogues with a diameter of
hundreds of nanometres. Thus various authors have shown that
the cytotoxicity effect of nanoparticles primarily depends on
their size.134,135 The entrance of nanoparticles into cellular
membranes has been described136 to follow the route of either
endocytosis or direct penetration. Research has suggested the
latter mechanism to have potential adverse effects to human
health because of its potential to induce membrane disruption
and lipid peroxidation.105,137 Systematic studies include the work
by Napierska et al.138 who investigated the viability of endothe-
lial cells in the presence of monodisperse silica nanospheres with
seven different diameters (ranging from 14 to 335 nm). LDH and
mitochondrial activity assays have demonstrated that a reduc-
tion in diameter leads to an increase in toxicity. It has been
shown139 that the presence of polymer-coated AuNPs caused
destruction and increased the permeability of mannitol across
epithelial Caco-2 monolayers by up to 4-fold. The increased
permeability was caused by loosening of tight junctions

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connecting the epithelial cells thereby allowing small molecules
such as mannitol to pass through the monolayer via the para-
cellular route. This result was also shown by Minchin et al.140
where they demonstrated that poly(acrylic acid) coated Au-NPs
can bind to and induce unfolding of the fibrinogen in plasma
leading to an inflammatory response.
3. Interaction of synthetic polymers with lipid
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membranes
Cell membranes mainly act as a selective barrier that regulates
the transport between the external and internal environment of
the cell protecting against foreign substances.141 The incorpora-
tion of molecules, in particular synthetic macromolecules
(such as polyelectrolytes,142,143 amphiphilic BCPs144–146 or den-
drimers147,148), can drastically change the physiochemical prop-
erties of a specific membrane. The advancing development in
designing macromolecules with well-defined architectures and
chemical composition, e.g. mimicking natural lipid structures,
enables the study of selective interactions between synthetic
polymers and model membrane systems.
Over the years up to 2008 numerous studies reporting on
interactions between synthetic polymers and model bio-
membranes were published.68,149 Control of non-covalent inter-
actions (e.g. hydrogen bonding, columbic association or
hydrophobic interaction) between macromolecules and lipid
membranes is affected primarily by the chemical structure and
charge properties of the interaction pairs. When macromolecules
interact with lipid membranes a large variety of effects could be
observed as illustrated in Fig. 8.
As illustrated in Fig. 8/1b, quite early150 an adsorption model
of cationic polymers was developed, which describes the ability
of a surface bound polymer (polylysine) to induce lateral reor-
ganization and segregation of single membrane components in
mixed lipid bilayers (DPPC/DPPA). Recently, detailed studies
on this topic have shown that polycation/liposome complexes
Fig. 8 Interaction pathways between synthetic polymers and lipid bilayers and their influence on membrane organization showing a large variety of
effects dependent on the polymer type.
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can lead to membrane fusion40 depending on the conformation of
the surface adsorbed polymer chains. Polymer anchoring onto
lipid bilayers (see Fig. 8/2b) via hydrophobical modifications of
their backbone as discussed earlier by Ringsdorf et al.31 is
nowadays an important strategy to reach an effective binding
between water-soluble polymers and lipid membranes.151 Tirrell
et al.152 were one of the first to study membrane solubilization by
a surface active polymer composed of (poly(2-ethylacrylic acid))
(PEAA).153 It could be shown that the polymer induced disrup-
tion of DMPC vesicular membranes increases with increasing the
acrylic acid fractions.
3.1 Effect of synthetic polymers on model lipid membranes
3.1.1 Effect of neutral polymers. Penetration of neutral
polymers into the hydrophobic membrane interior is primarily
driven by hydrophobic interactions. Recently, modern research
has been focused on the penetration ability of amphiphilic block
copolymers into lipid mono-154–157 and bilayer systems.146,158–160
Nonionic block copolymers (BCPs), such as pluronics or
poloxamers (PEO-b-PPO-b-PEO), have been studied extensively,
revealing the main parameters governing the interaction between
lipid membranes and such amphiphilic polymers.144 Qualitative
agreement between simulation and experimental results proved
that the hydrophilic/hydrophobic balance and the overall
hydrophobicity of the triblock copolymers drastically affect the
polymer incorporation into lipid monolayers. Thus, the incor-
poration activity of poloxamers is found to correlate with its
solubility in the subphase (water), as with increasing hydro-
phobic block lengths the solubility of the BCP decreases and
interactions with the lipid akyl-chains are stronger resulting in
higher squeeze-out pressures. An early experimental proof
showed the temperature dependent association between the
poloxamer F-68 (PEO76–PPO29–PEO76) and DPPC mono-
layer,154 resulting in a higher squeeze-out pressure of the triblock
copolymer chains and an increasing monolayer fluidity by
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decreasing the closed packing of the lipid molecules at higher
temperatures. Lee and Wu155 have shown that the lipid packing
density regulates the poloxamers (P188) insertion process leading
to a distinct effect on the lipid film morphology. If the hydro-
philic/hydrophobic balance of the poloxamers is kept constant,
the overall size of the polymer (molecular weight) determines the
squeeze-out behavior, whereas an increase in the total polymer
size leads to higher squeeze-out pressures.144 Thus the Langmuir
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effects were based on the fact that the incorporated hydrophobic
blocks (PPO) lower the van der Waals forces between the
hydrocarbon chains of the lipid molecules resulting in bilayer
expansion.159 Reports of several groups on the nonionic surfac-
tant properties of poloxamers have indicated that the used
polymer concentration (above or below their CMC) defines the
type of the formed lipid/polymer complex.146,160,162 When the
critical micellization concentration (CMC) of a specific polox-

balance technique coupled with spectroscopic methods such as
IRRAS (Infrared Reflection Absorption Spectroscopy) can give
a more detailed view on the molecular organization of the
interacting components in a mixed monolayer.156,157,161 Blume
et al.161 have studied the molecular arrangements in mixed
monolayers from perdeuterated lipids (DPPC-d62) and amphi-
philic triblock copolymers (PGMA14-b-PPO34-b-PGMA14),
which are comparable to poloxamers. Using IRRAS analysis it
was shown that the insertion of these amphiphilic polymers into
the spread lipid monolayer leads to mesoscopically to nano-
scopically dimensioned lipid cluster surrounded by a polymer
network (see Fig. 9A). Monitoring of the symmetric stretching
band of the perdeuterated methylene units (-CD2-) proved the
final conformational order of the lipid alkyl-chains showing
a large population of trans conformers.
It has been found that the interaction strength between
amphiphilic triblock copolymers and lipid membranes strongly
depends on the hydrophobicity of the polymer. Chen and
Chieng159 showed that the more hydrophobic pluronic L-61
(PEO2–PPO30–PEO2) has stronger interactions with DPPC
liposomes than F-127 (PEO100–PPO65–PEO100), which lead to
a complete disruption of the lipid membrane forming crew-cut
aggregates as illustrated in Fig. 9B. Such strong destabilization
amer is reached at a given temperature, the solubilization of the
lipid membranes occurred, forming mixed micelles (see
Fig. 9B).146,162 Based on isothermal titration calorimetry (ITC)
experiments it has been shown that pluronic P388 molecules are
incorporated into fluid-phase DMPC liposomes above their main
transition temperature (Tm ¼ 24  C), whereas gel-phase
membranes of DMPC (below Tm) prevent polymer incorpora-
tion.146 Starting from mixed complexes, decreasing the temper-
ature below the Tm of DMPC the spherically shaped lipid/
polymer vesicles change to disc-shaped architectures found by
phase separation between the polymer and the lipid gel-phase.
These matching results between the mono- and bilayer studies
describing the squeezing out of poloxamers chains from lipid
membranes can serve as a model system for membrane sealing. A
recent example between poloxamer (F127) and liposome asso-
ciation reported by Feitosa and Winnik160 suggested that the
formed types of lipid/polymer complexes depend not only on the
temperature and relative amounts of both components, but also
on the thermal pretreatment of the sample system with respect to
Tm of the lipid and the critical micellization temperature (CMT)
of the polymer component.
Non-covalent, hydrophobic interactions were shown to be
very effective in binding hydrophobically modified water-soluble

Fig. 9 (A) Schematic illustration of the morphology of the mixed lipid/polymer monolayer at the air/water interface. (1) Molecular structure of the
PGMA14-b-PPO34-b-PGMA14 block copolymer and (2) alkyl chain perdeuterated phospholipid structure of DMPC-d54 and DPPC-d62 (2). (B) Sche-
matic diagram of interaction and complexation between phospholipids and pluronic F-127 and L-61 depending on the polymer concentration. (A) is
taken from ref. 161—reproduced by permission of the Royal Society of Chemistry. (B) reprinted with permission from ref. 159, copyright 2009,
American Chemical Society.

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polymers to the surface of phospholipid vesicles. Zhang et al.151
investigated the relationship between the hydrophobic
chain structure with a series of hydrophobically end-capped
poly(ethylene glycols) and the insertion ability into (L-a-phos-
phatidylcholine) vesicles. It has been demonstrated that with
increasing hydrophobic chain length a reliable membrane
anchoring of the functionalized PEG molecules could be reached
modifying the liposomal surface with a polymer shell.
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In contrast to amphiphilic polymers, reports on the interaction
between dendritic polymers and model biomembranes describing
their nature and location are rather rare.148,163,164 The importance
of such interactions is related to the structure of dendrimers
exhibiting a well-defined highly branched architecture with
numerous functional groups on their surface suitable for phar-
maceutical applications. Recently, Demetzos et al.148 investigated
interaction between liposomal membranes and hyperbranched
polyesters, differing in their generation number and consequently
in the number of terminal hydroxyl groups. A deep penetration
of the dendritic molecules into the hydrophobic membrane
interior was strongly limited due to the polar structure of the
hyperbranched polyesters. Thus, it was shown that the dendritic
polyesters of 2nd and 4th generation displaying 16 and 64 terminal
hydroxyl groups exhibit the strongest interactions with lipid
membranes resulting in dehydration of the liposomal head group
region coupled to a decrease in membrane mobility. In
comparison to nonionic dendrimers, interaction studies of
cationic dendrimers with lipid membranes showed the electro-
static origin of their nature.147,165 Ionov et al.147 showed that
liposome/dendrimer interactions depend on the surface charge of
the lipid bilayer by studying the association of cationic phos-
phorus dendrimers with charged and neutral liposomes. Only in
the case of negatively charged liposomal surfaces strong den-
drimer/lipid interactions via electrostatic forces could be
observed, which resulted in changes of the main transition
behavior of the lipid bilayer indicating that the membrane
rigidity is reduced by polymer association.
3.1.2 Effect of charged polymers (polycations and poly-
electrolytes). A particular area of interest is columbic association
between charged polymers and lipid membrane. In comparison
to nonionic polymers, a deep incorporation of charged polymers
into the hydrophobic membrane interior is now strongly
restricted. Instead, the external binding of charged polymers
onto liposomal surfaces is driven by electrostatic interactions
between the charged polymer units and oppositely charged lipid
head groups, forming a polymeric corona on top of the liposome
surface.158 Most of the recent interaction studies have been per-
formed with positively charged polymers and negatively charged
liposomal surfaces.68,166–168 Few examples describe pH-depending
interactions between zwitterionic liposome surfaces and posi-
tively charged polymers143,169 or anionic polyelectrolytes.169,170
The adsorption of polymeric molecules onto the hydrophilic
corona of liposomes can lead to strong effects on the membrane
organization.
The binding of cationic polymers to lipid membranes has
shown irreversible rearrangements in the lipid organization up to
membrane disruption,92,171 or promote migration of lipid mole-
cules in one of the liposomal layers (segregation) or between the
inner and outer layers (‘‘flip-flop’’).158,172 It has been
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demonstrated that the ability of charged polymers to neutralize
the charge state of liposomal surfaces leads to effective polymer/
liposome complexation and could end in interliposomal polymer
migration173 or vesicular membrane fusion by reducing the
interliposomal repulsion.40 Menger et al.173 found that the
cationic polymer poly(N-ethyl-4-vinylpyridinium bromide)
(PEVP), when bound to lipid membranes, could undergo inter-
liposomal migration in a suspension of small unilamellar lipo-
somes. As an extension of this model, it was found that these
cationic PEVP polymers can reduce liposomal repulsion due to
their effective binding to the anionic liposome surface, and also
physically prevent interliposomal membrane fusion.40 In
contrast, surface adsorbed polylysine polymers were shown to
induce liposomal membrane fusion at low concentration.
The main reasons enabling polylysine molecules to induce
liposome fusion were the decrease in electrostatic rejection
between the negatively charged vesicles by the formed thin
polymer layer (see Fig. 10A/1 and 2), which ensured liposomal
contact and consequently the polymer decorated liposomes
become leaky to water, which promotes exchange of adjacent
membrane parts. As opposed to this external adsorbed PEVP
molecules form a physical barrier that prevents fusion between
vesicles due to the increased thickness of the polymer shell
compared to polylysine (see Fig. 10A/3 and 4).
Similar examples169 have shown the adsorption of poly-
electrolytes (hyaluronan or chitosan) onto liposomes observing
different degrees of surface coverage by polymers having the
opposite or same charge character as the membrane. As illus-
trated in Fig. 10B, it has been demonstrated that a low degree of
surface coverage leads to vesicle aggregation as described by
Yaroslavov.40 In case of polyelectrolyte excess, dissociation of
the polymer/liposome aggregates could be observed, followed by
their stabilization forming a homogeneous polymer layer, which
is caused by changes of their z-potential (see Fig. 10B). The effect
of salt concentration on the adsorption behavior of poly-
electrolytes onto lipid membranes was shown by several research
groups.143,170,173 Frisken et al.143 investigated the effect of the
cationic polyethylenimine (PEI) on zwitterionic DMPC vesicles
as a function of salt concentration (NaCl) demonstrating the
high sensitivity of the formed polymer/liposome complexes
against salt addition. At low NaCl concentration aggregation of
the polymer/liposome complexes was observed, whereas at high
salt concentrations vesicle aggregation was prevented, concom-
itant with drastic changes in the DMPC chain-melting transition
of the hydrocarbon tails, suggesting a strong penetration ability
of PEI molecules into the lipid bilayer at high salt concentrations.
A potential application of salt-sensitive polymer/liposome
complexes as light-harvesting devices was studied by Cosa and
Ngo,170 investigating interaction between the negatively charged
fluorescent polyelectrolyte poly[5-methoxy-2-(3-sulfopropoxy)-
1,4 phenylenevinylene] (MPS-PPV) and DOPC in the presence of
mono- and dications. The addition of Ca2+ ions showed
a significant increase in lipid/polymer interactions leading to
polymer uptake by the DOPC liposomes.
3.1.3 Pore-formation ability. An overview of various factors
important for the formation of channels and pores in lipid
membranes induced by synthetic polymers, including the chem-
ical composition and polymer architecture, is given in several
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Fig. 10 (A) Schematic illustration and the corresponding cryo-TEM images of the polylysine/liposome complexes before (1) and after fusion (2). (3)
Schematic illustration of looping in a PEVP/liposome complex, showing lateral phase separation (domain formation) induced by PEVP-molecules and
(4) cryo-TEM imaging demonstrates how PEVP physically prevents liposomal fusion. (B) Schematic illustration and microcopy observations of LUVs
suspension structure for three different cases of chitosan addition: (i) isolated bare vesicles when no chitosan is added, (ii) vesicle aggregation when
chitosan addition corresponds to the isoelectrical point of the LUVs (z-potential ¼ 0) and (iii) isolated chitosan-coated LUVs when chitosan addition
corresponds to coating saturation. (A) reprinted with permission from ref. 40, copyright 2011, American Chemical Society. (B) is taken from ref. 169—
reproduced by permission of the Royal Society of Chemistry.

reviews.68,149 A relationship between the polymer architecture 3.2 Hybrid lipid/polymer membrane models
and the pore formation pathway was extensively described by
Lipid/polymer hybrid systems and their mixing behavior offer
Binder et al.149 as illustrated in Fig. 11A.
new technical application in pharmaceutical or biomedical fields,
Recently, scientific interest has been focused on liposomal
where it is of specific interest to combine the biofunctionality of
release of encapsulated materials from the vesicle interior
liposomes with the enormous mechanical stability and func-
through the lipid bilayer by controlled breakup mechanisms,
tionality of polymeric membranes. Reports on such hybrid
which allow the control of membrane permeability. Accordingly,
membranes, which describe the mixing behavior of lipids and
Tribet et al.174 showed for the first time triggered permeabiliza-
synthetic polymers, are rather rare in literature.176–178 Vanderlick
tion of lipid membranes induced by external light irradiation.
et al.177 have opened this novel membrane platform by fabri-
They found that azobenzene modified poly(acrylic acid)
cating mixed lipid/polymer vesicles of various compositions.
copolymers embedded in DOPC GUVs can induce membrane
Confocal fluorescence microscopy studies proved that it is
perturbation by cis–trans photoconversion (see Fig. 11B). The
possible to incorporate high amounts of a non-biocompatible
biological relevance of this system was shown in cell culture
block copolymer (PBd46-b-PEO30) into a liposomal membrane
studies (mammal cells) stressing the remarkable influence on the
(POPC). The resulting mixed vesicles (GUVs) were prepared by
membrane properties under mild conditions. Previous investi-
an electroformation method starting from a dry mixed lipid/
gations proved nonspecific permeability of lipid bilayers induced
polymer film. Further investigations concerning the mixing
by hydrophobically modified poly(acrylic acids).175 These
behavior demonstrated that phase heterogeneities induced by
amphiphilic copolymers can undergo channel formation in
binding neutravidin to either biotinylated lipids or block
vesicular lipid membranes depending on the polymer concen-
copolymers lead to domain formations. Such strong clustering
tration. Furthermore, fluorescence microscopy studies of lipo-
effects are justified by the fact that one neutravidin molecule
somal release confirmed nanometre-sized polymer channels
binds to 4 biotin-labeled molecules as illustrated in Fig. 12A.
(channel radii of 1 to 5 nm) with the ability of transmembrane
Phase separation without supramolecular assemblies was
exchange of small molecules (albumin and dextran molecules).

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Fig. 11 Pore formation by full insertion of a polyelectrolyte (pore formation 1), by insertion of an amphiphilic graft copolymer (carpet model) and by
incorporation of amphiphilic triblock copolymers (pore formation 2). (B) Illustration of the light-induced membrane permeabilization by cis/trans
photoconversion of modified copolymer chains. The polymer-loaded GUVs remained impermeable when irradiated under UV light (436 nm) become
permeable and release their internal content—reprinted with permission from ref. 174, copyright 2010, American Chemical Society.

demonstrated by our group for the first time (Fig. 12B). We have
further shown the formation of truly biocompatible hybrid
vesicles178 composed of self-synthesized PIB–PEO block
copolymers and a natural lipid (DPPC). It was observed that the
incorporation of such biocompatible polymers into vesicular
DPPC membranes using different amphiphilic PIB–PEO BCPs
varying in their hydrophobicity leads to remarkable effects on
the lipid bilayer organization (e.g. flattening of holes and phase
borders or formation of domains).
At mixing ratios from 20 up to 28 mol% of the PIB87-b-PEO17
copolymer in the lipid membrane, the observed phase heteroge-
neities indicate a demixed system forming lipid- and polymer-
rich domains, which are obtained without the use of a strong
clustering effect via avidin/biotin interaction (compare Fig. 12A
and B). Recently, the control of membrane surfaces via selective
phase separation phenomena in mixed polymersomes containing
different vesicle-forming block copolymers was investigated by
Battaglia et al.18 They demonstrated that the domain dimensions
in demixed polymersome surfaces are linearly proportional to the
vesicle diameter and could be controlled by their polymer
compositions. Interestingly (see Fig. 12C), one of the block
copolymers (PMPC25–PDPA70) carries lipid functions (phos-
phorylcholine chains) in its backbone (polymerized phospholipid
block), the observed domain formation and their shapes on both
nano- and micrometre length scale can serve as a model system to
study the behavior of phase separated lipid/polymer vesicles.
Similar to bilayer membranes, Langmuir monolayers can serve
as a membrane model at the air/water interface investigating the
mixing behavior between synthetic polymers and lipids. Meier
et al.179 reported for the first time on mixed lipid/polymer
monolayers studying the morphological changes of binary mixed
monolayers from a lipid (DPPC or DOPC) and an amphiphilic
PMOXA-b-PDMS-b-PMOXA copolymer using the Langmuir
balance technique coupled with Brewster angle microscopy
(BAM). Depending on the lipid to polymer composition it was
found that saturated lipid monolayers (DPPC), forming rigid
films with a high lipid packing density, are much more sensitive
to the presence of the amphiphilic copolymer resulting in the
observation of phase separation phenomena (see Fig. 13A).
In particular the formation of pure lipid domains (DPPC) at
high compression states of the mixed film was observed (star-
shaped domains at p ¼ 25 mN m1, compare Fig. 13A/1 and B,
red square). Monolayers composed of DOPC (unsaturated lipid)
at specific polymer concentrations showed only in the low pres-
sure region a tendency to phase-separate, whereas at higher
surface pressure a homogeneous film was observed. The influence
of incorporated block copolymers on the phase transition
behavior of lipid monolayers was shown to depend on the lipid to
polymer mixing ratio, monitoring the lipid phase state by fluo-
rescence microscopy.178 We have observed that the increasing
polymer content in mixed lipid/polymer films leads to a shift of
the typical transition state (LE/LC) of DPPC films to higher
surface pressures, indicating that the PIB–PEO BCPs stabilize
the expanded state of the lipid monolayer. A complete suppres-
sion of the DPPC domain nucleation (LC-phase), forming
homogeneous films independent of the compression state, was
demonstrated by mixed monolayers containing 80 mol% of the
polymer component. Additional techniques monitoring mixed
lipid/polymer film morphologies were displayed by Romão
et al.180 They studied interactions and phase separation
phenomena between fluorescent thermo-responsive copolymers
(RhB-labeled PDMA207-b-PDEA177) and DPPG by BAM and
AFM combined with a laser scanning microscopy technique.
This allows a relationship between the observed film morphology
and the polymer induced changes by monitoring the fluorescently
labeled polymer molecules. It has been shown that the double

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Fig. 12 Examples of phase heterogeneities in hybrid membranes and their schematic illustration. (A) Confocal microscopy images of labeled hybrid
vesicles (BODIPY-DHPE, green) from POPC and biotinylated PBd46-b-PEO30 copolymers (30 mol%). (B) Confocal microscopy images (3D-recon-
struction) of hybrid vesicles (labeled with DiD-C18 dye) from PIB87-b-PEO17 copolymer and DPPC. (C) Hybrid PMPC25–PDPA70/PEO16–PBO22
polymersomal membrane obtained at different binary compositions. (A) reprinted with permission from ref. 177, copyright 2011, American Chemical
Society. (B) is taken from ref. 178—reproduced by permission of the Royal Society of Chemistry. (C) reprinted with permission from ref. 18, copyright
2011, American Chemical Society.

hydrophilic BCP molecules (below the LCST) were incorporated 3.3 Biological impact of polymer/lipid interactions
into the expanded phase of the lipid film, forming mixed
Depending on the chemical structure and composition of
domains, whereas an increase in surface pressure leads to poly-
synthetic polymers, different kinds of interactions and effects on
mer chain expulsion.

Fig. 13 (A) Brewster angle microscopy images of mixed monolayers from A65–B165–A65 triblock copolymer and DPPC at 25 mN m1 (image width
220 mm). (1) Mixed film obtained from a polymer to lipid ratio of 0.2 to 0.8 and (2) 0.1 to 0.9. (B) Schematic phase diagram (surface pressure versus
mixture composition) obtained for the binary system of PMOXA65–PDMS165–PMOXA65 and DPPC, illustrating the observations of phase hetero-
geneities in the mixed monolayers—reprinted with permission from ref. 179, copyright 2009, American Chemical Society.

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lipid membranes can be expected. For example, polymer
adsorption onto cell membranes can be optimized considering
surface modifications in cell transplants or lifetime of polymer/
liposome delivery systems. Herein, blood circulation times of
liposomal delivery systems in vivo are one of the most important
points in pharmaceutical and medical applications. For this
purpose synthetically prepared poly(ethylene glycols) (PEGs)
have shown to be an excellent polymeric shielding device for
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liposomes, forming a covalent linked hydrophilic corona, which
increases blood circulation times.181–183 Similar to the efficient
PEG long circulation properties in vivo, poly(2-oxazoline)-
grafted liposomes184 have shown comparable results suggesting
that this effect is species-independent. In view of cell surface
modifications, Iwata et al.145 demonstrated the behavior of
different synthetic polymers adsorbed or covalently bound to
mammalian cells over time. Thus, fluorescence microscopy
studies with fluorescently labeled polymers have shown that the
investigated polymers were gradually excluded from the cell
surface.
Cationic polymers are currently used as effective antimicrobial
agents due to the ability of disrupting cell membranes. In general
it has been found that the cationic group structure of polymers is
an important factor in the control of membrane activities (e.g.
binding, destabilization or membrane porosity), promising
applications in fields of gene-delivery185 or antimicrobial
agents.186 Thus, Kuroda et al.186 have proposed a potential
application of poly(methacrylamides) as surface coatings that
kill bacteria in the case of contact due to the fact that these
cationic copolymers selectively disrupt bilayer membranes.
Whitten et al.171 have studied interaction between poly-
electrolytes and model bacteria cell membranes to understand
the structural basis of the biocidal activity of these cationic
polymers. Such poly(phenylene ethynylene) (PPE)-based poly-
mers contain pendant quaternary ammonium groups, which are
promising candidates for antimicrobial action due to their
possibility to disrupt cell membranes. Their findings propose that
PPE-based polyelectrolytes associate with lipid membranes and
become incorporated into the hydrophobic membrane interior
leading to strong membrane disorganization.187 Herein, it was
shown that cationic copolymers with primary amines as pendant
groups exhibit stronger membrane interactions with high dis-
rupting abilities than their tertiary or quaternary ammonium
counterparts.142
4. Conclusions
To summarize, there is much room beyond the lipid bilayer,
which extends the principles of a bilayer-membrane into higher
aspects of supramolecular ordering and function. Thus both,
the lipid vesicle and the polymersome membrane order, can
be triggered significantly by interactions with nanosized
objects, such as macromolecules and nanoparticles. The
resulting effects now open a prospect for very subtle engi-
neering of membranes, their nano-porosity, (nano-) phased
structure and their mechanical properties and transporting
abilities. It is clear that applications in biomedicine, selective
drug-delivery or the sole deeper understanding of transport
phenomena will be the basis for future research and applica-
tion technologies.
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Acknowledgements
We thank the grants DFG BI 1337/6-1 (MS, AO) within the
Forschergruppe FOR-1145 and the grants DFG INST 271/249-
1; INST 271/247-1; and INST 271/248-1 for financial support.
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