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Current Opinion in Colloid & Interface Science 16 (2011) 491498

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Current Opinion in Colloid & Interface Science

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / c o c i s

Interactions of amphiphilic block copolymers with lipid model membranes

E. Amado, J. Kressler
Department of Chemistry, Martin Luther University Halle-Wittenberg, D-06099 Halle (Saale), Germany

a r t i c l e

i n f o

Available online 23 July 2011

Block copolymers
Planar phospholipid membranes
Biomedical applications

a b s t r a c t
Studies on interactions between amphiphilic block copolymers and lipid membranes have been focused
traditionally on ABA triblock copolymers of poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene
oxide), widely due to their commercial availability. However, new architectures of amphiphilic block
copolymer have been synthesized in recent years partially taking advantage of new polymerization
techniques. This review focuses on amphiphilic block copolymers with potential biological activity and on
model membrane systems used for studying interactions with such block copolymers. Experimental methods
to study block copolymerphospholipid interactions in Langmuir monolayers, liposomes, and planar bilayers
are summarized. This work is intended to convey a better understanding of amphiphilic block copolymers
used for in vitro and in vivo experiments in medicine and pharmacy. Recent developments and open questions
are addressed.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
The interactions between amphiphilic block copolymers and
phospholipid membranes have been attracting increasing research
interest in recent years partially stimulated by biochemical, pharmaceutical, and medical applications [1]. Amphiphilic block copolymers
are composed of covalently bonded hydrophilic and hydrophobic blocks
[2,3]. The hydrophobic block contains usually lipophilic segments but
in special cases it can be a block with peruorinated moeties instead.
Such uorinated blocks are not only hydrophobic but also lipophobic
[4,5]. Joining hydrophilic, lipophilic and uorophilic blocks together
yields so-called triphilic block copolymers [6,7]. Most commonly used
block copolymers are in the moment linear ABA triblock copolymers
where the middle block is composed of poly(propylene oxide) (PPO)
and the outer blocks are composed of poly(ethylene oxide) (PEO). They
are known under the generic name poloxamers (or their trademarks as
e.g. Pluronics or Synperonics) and are commercially available [8,9].
Although, the interactions of poloxamers with lipid membranes have
been studied for decades, several open questions remain. For instance,
the conformation of the PPO block once inserted into the non-polar
inner part of a lipid membrane is still under investigation [10]. Some
poloxamers are able to support the transport of drugs (including therapeutic proteins) across a cell membrane even across the blood brain
barrier [11,12]. But several poloxamers especially with large PEO blocks
lack the ability to be retained in mixed lms with phospholipids up to
surface pressures relevant for some biological membranes such as
membranes responsible for the bloodbrain barrier and erythrocyte

membranes [13]. To tackle these problems other block copolymer

architectures different to the linear ABA architecture might be more
suitable. The most relevant alternatives are outlined in the next section.
Despite the mentioned lack of comprehension, amphiphilic block
copolymers have been already applied for tumor therapy [14], as
chemo-sensitizing agent to ght multiple drug resistance (MDR) [15],
drug release systems [16,17], DNA transfection [18], and healing of
injured cell membranes [19,20] among others. A key prerequisite for
such medical effects are specic interactions with biological membranes.
In order to unveil the exact mechanism, the interaction with simpler
model membrane systems such as lipid Langmuir monolayers, liposomes, including giant unilamellar vesicles, and planar bilayers in form
of tethered lipid bilayers or black lipid membranes are studied rst. The
interactions are investigated by employing scattering methods, spectroscopic techniques, calorimetric measurements and several microscopy
techniques [21]. Typical model membranes and the most common
characterization techniques are reviewed in Sections 2 and 3, respectively. Model membranes allow frequently for a high lateral resolution or
for very sensitive measurements which are much more complicated to
achieve in studies with living cells or whole organisms. Nevertheless,
they are simple models neglecting many structural features of living cell
membranes as cytoskeleton, trans-membrane proteins, lipid domains of
varying chemical compositions etc. Thus, additional biological studies
encompass in vitro investigations using living cells as well as in vivo tests
in animals. Some typical examples are discussed with respect to the
pharmaceutical and medical applications of block copolymers.
1.1. Block copolymer architectures

Corresponding author. Tel.: + 49 3455525800.

E-mail address: joerg.kressler@chemie.uni-halle.de (J. Kressler).
1359-0294/$ see front matter 2011 Elsevier Ltd. All rights reserved.

Amphiphilic block copolymers are usually composed of hydrophilic and lipophilic blocks covalently joint together in several


E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498

different block copolymer architectures. Some of these architectures

are depicted in Fig. 1.
As already mentioned earlier partial or complete replacement of
the lipophilic blocks by uorophilic blocks leads also to amphiphilic
block copolymers since the uorophilic blocks are hydrophobic in
their nature. The simplest block copolymer architecture shown in Fig. 1a
consists of just two blocks A and B rstly synthesized by living anionic
polymerization [22]. In this review A is always considered to be the
hydrophilic block and B the hydrophobic block. Living ionic polymerization methods have been traditionally employed for the synthesis of
AB diblock copolymers. Though less important than anionic polymerization, also cationic polymerization is a suitable tool to prepare welldened block copolymers [23]. Recent developments in polymer
syntheses especially in the eld of controlled radical polymerization
(CRP), including nitroxide mediated radical polymerization (NMRP)
[24,25] transition-metal-catalyzed atom transfer radical polymerization (ATRP) [26 ]and reversible addition fragmentation chain transfer
(RAFT)/macromolecular design via the interchange of xanthates
(MADIX) [27,28 ,29], over the last decade have created enormous
opportunities to design block copolymers of almost any architecture
with unprecedented control. These advances are complemented by
polymer-analogous reactions with nearly complete conversion, socalled click chemistry [30 ,31]. It should be mentioned that for the
most popular click reaction between azides and alkynes a hydrophilic
junction point between the blocks consisting of a triazole is formed
[32]. The most important advantages of CRP techniques over conventional living ionic polymerizations are the requirements of less
stringent experimental conditions, tolerance to many functional groups,
and their ability to be carried out in aqueous medium.
The use of bifunctional initiators (or macroinitiators) allows
preparing ABA or BAB type triblock copolymers either by living ionic
polymerization or by CRP techniques (see Fig. 1a). Since the chain ends
are still functional after polymerization, the attachment of new blocks

can be continued for the synthesis of pentablock copolymers up to

multiblock copolymers. In the case that one of the end-blocks is rather
short or just a functional group, the amphiphilic polymer is referred to as
telechelic (BAB) or semi-telechelic (AB), see Fig. 1f. The most common
examples are telechelics formed by PEO with one or two hydrophobic
chain ends [33]. In this context, low molar mass PEOs are also frequently
called poly(ethylene glycol) (PEG). Their aggregation behaviour in
water and thus their biological activity depends strongly on the
character of the end group [34]. More complex architectures have
been obtained by using multifunctional initiators, as for the synthesis of
star block copolymers schematically depicted in Fig. 1c. The number of
arms is tailored by the number of functional groups carried by the
initiator molecule [35]. Of course, the A and B parts of an arm can easily
be exchanged strongly inuencing their aggregation behaviour in water.
A commercial example of four-arm star polymers are poloxamines
already mentioned earlier. Furthermore, it is possible to synthesize graft
block copolymers were the blocks are covalently attached as side chains
to a polymer backbone as shown in Fig. 1d. These polymers are called
graft block copolymers [36]. Furthermore, it has been possible in recent
years to synthesize cyclic amphiphilic block copolymers as demonstrated schematically in Fig. 1b [37,38]. Also block copolymers carrying
hyperbrached or dendritic blocks as depicted in Fig. 1e seem to have a
tremendous application potential [39].
For the block copolymer architectures mentioned earlier, the
hydrophobic blocks are typically composed of repeat units based on
n-alkyl acrylates (or methacrylates), styrenes, butadienes (isoprenes) or
its hydrogenated species, propylene oxide or tetrahydrofurane, among
others. Hydrophilic blocks are either neutral or charged species which
then belong to the polymer class of polyelectrolytes. Typical examples of
monomers used are ethylene oxide, acrylamide and their derivates,
hydroxy alkyl acrylates or methacrylates, acrylic or methacrylic acid,
ethyleneimine and glycidols. Furthermore, several stimuli responsive
water soluble polymers, i.e. whose solubility depends on temperature,

Fig. 1. Possible architectures for amphiphilic block copolymers. (a) Linear block copolymers with different numbers of A and B blocks. (b) Cyclic block copolymers. (c) Star block
copolymers. (d) Graft block copolymers (e) Block copolymers with dendritic or hyperbranched blocks. (f) Semitelechelic polymer (upper), telechelic polymer (middle),
asymmetrical telechelic polymer with different hydrophobic chain ends.

E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498

pH-value, presence of ions etc., have also been chosen as A block [40].
Another special class of block copolymers are so-called biomimetic
block copolymers. They contain biologically active units as phospholipids, cholesterol or proteins [41 ].


the different techniques available for characterizing the interaction of

block copolymers and lipid Langmuir lms and the kind of information
obtained can be found in Table 1.
2.2. Liposomes and giant unilamellar vesicles

2. Model membrane system

The main structural components of a cell membrane are phospholipids, which are amphiphilic in nature, and consequently organize
themselves in water into various aggregation states. In a cell membrane
they form a bilayer with their hydrophilic moieties exposed to water
and their hydrophobic tails hidden from water in the inner part of the
membrane. Other molecules incorporated in considerable amounts in
the phospholipid matrix are cholesterol, integral and peripheral membrane proteins, sphingolipids and glycolipids carrying a carbohydrate
moiety. Due to the complexity of a biological membrane simplied
model membranes are commonly used in order to study the interactions
with amphiphilic polymers. A scheme of such a bilayer together with the
general chemical structure of 1,2-diacylphosphocholines, one common
class of phospholipids occurring in cell membranes, is shown in Fig. 2.
The most relevant aggregation forms of lipids used as models of cell
membranes are shown in Fig. 3ad and are discussed subsequently.
2.1. Langmuir monolayers
Langmuir monolayers (Fig. 3a), spread at the airwater interface of a
Teon trough and compressed to a given surface pressure through
mobile barriers, are used as models of the outer leaet of a cell membrane. They allow for investigation of interactions with monolayers in
different physical states and packing order of the alkyl chains, from a
disordered gaseous-like up to a tightly packed solid analogue state. In
spite of its simplicity the results from such studies can be effectively
translated into more complex bilayer systems [42]. The large surface
areas that can be covered homogeneously by a Langmuir monolayer
enables to carry out X-ray or neutron reection measurements and
allows to follow the adsorption processes of block copolymers to
lipid monolayers by the time-resolved lm thickness variation. Special
spectroscopic techniques as IR or Raman can also be applied to Langmuir
monolayers. Particularly helpful for obtaining information at the
molecular level, such as molecular conformation and orientation, is
infrared reection absorption spectroscopy (IRRAS). An overview of

As already mentioned, Langmuir monolayers represent only half of a

cell membrane. Some model systems have also been developed that
mimic both leaets and allow studying interactions with a complete
membrane. Spherical liposomes (Fig. 3c) are formed from dried lipid
lms by hydration and subsequent mechanical treatment. Unilamellar
vesicles in the size range from 15 to 200 nm are easily obtained. They
are employed to study adsorption phenomena in conjunction with
thermal characterization techniques as ITC and DSC as well as dynamic
light scattering. Also vesicles with micrometer size, so called giant
unilamellar vesicles (GUVs), can be obtained with special techniques, as
electroformation. They are adequate for investigations with different
light microscopy techniques such as confocal laser scanning or
uorescence microscopy.
2.3. Black lipid bilayers, adsorbed and tethered lipid bilayers
Also planar phospholipid bilayers can be formed free-standing as
black lipid bilayers (BLMs) (Fig. 3e) or as tethered lipid bilayers (TLBs)
(Fig. 3d) on solid supports. Due to stability issues BLMs are normally
constrained to use phospholipids having a branched alkyl chain, such
as DPhDC (see structure in Fig. 2), and the obtainable free standing
lms have maximum diameter in the 100 m range. TLMs do not have
these disadvantages but the distance between the bilayer and the
solid support is limited to a few nanometers depending on the length
of the anchoring molecules.
2.4. Nanoscopic domains in model membranes
Recently, more complex ternary mixtures of cholesterol and two
other lipids (phospholipids or sphingolipids differing considerably in
their transition temperature (e.g. DPPC/DOPC/cholesterol, see Fig. 2) are
being increasingly used in model membranes in order to reect recent
discoveries about the compartmentalization of many types of cell
membranes into well-dened domains in different time and length
scales. The most widely investigated of such domains are known as lipid

Fig. 2. General organization of a phospholipid bilayer and chemical structure of common 1,2-diacylphosphocholines and cholesterol employed in model membranes.


E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498

Fig. 3. Typical model lipid membranes: a) Langmuir monolayer in expanded or condensed state. b) Lipid liposomes and GUVs. c) Supported lipid bilayers on mica or silicon and
tethered bilayer lipid membranes on a gold solid support. d) Chamber for studying electrical properties and ion transport of bilayers (black) lipid membranes.

rafts [43], which are enriched in sphingolipid/cholesterol, possess a

liquid-crystalline order, and are thought to play an important role in the
non-random distribution of some integral and peripheral proteins
within the plasma membrane and also in their biological function. Since
the domains are of nanoscale size (typically b 300 nm), analysis techniques with nanometer-level precision, well below the lateral resolution of conventional uorescence microscopy, are required. Only a
few techniques are available, among them atomic force microscopy
(AFM) allows for the visualization of the morphology of the different
nanoscopic domains [44]. Meanwhile, the most challenging determination of the spatially resolved chemical composition within the
domains has been reached by high-resolution secondary ion mass
spectrometry (SIMS) [45]. It requires the use of distinct isotopically
labelled lipids, though.
A summary of the characterization techniques commonly used for
investigating the interaction of block copolymers and lipid bilayers
can be found in Table 1 together with some key references.
3. Interactions of model membranes and block copolymers
Polyelectrolytes, i.e. polymers with charged moieties, and more
specically polycations, having positively charged groups, are able to
bind to the outer leaet of a cell membrane through electrostatic
interactions between the positively charged moieties in the polymer
and the negatively charged head group of anionic lipids within the
leaet. For example, the polycation quaternized poly(4-vinyl pyridine),
has been found to accelerate the lipid translocation from the inner to the
outer leaet in liposomes [46 ].
On the other hand, the interaction mechanism for non-ionic block
copolymers is totally different. They bind to the membrane by partial
or total insertion of their hydrophobic blocks into the inner, non-polar
region, of the lipid bilayer. Thus, a general requirement for a non-ionic
block copolymer to associate with a membrane is that the polymer
has to be amphiphilic, whereby the hydrophobic effect induces the

penetration of the polymer into the hydrophilic-hydrophobic interfacial layer of the membrane.
Depending on the particular molecular architecture and the size of
the hydrophobic block compared to the bilayer thickness there are
various possible modes of interaction. ABA triblock copolymers with a
hydrophobic middle block match closely the polar/non-polar/polar
structure of the bilayer. As the middle block shows a strong afnity
for the likewise hydrophobic inner part of the membrane it tends to
localize there, while the hydrophilic blocks extends into the water
phase [47 ]. ABA polymers whose hydrophobic block length is less
than the thickness of the bilayers insert only partially and weakly into
the membrane, leaving their hydrophilic blocks delocalized at the
membranewater interface; polymers whose hydrophobic block
length is enough to span the bilayer integrate tightly in the membrane,
projecting their hydrophilic blocks into the water phase on opposite
sides of the bilayer [47].
In this case, a further requirement is imposed on the chemical
nature of the hydrophobic middle block since an alkyl-based block
would be too hydrophobic and renders the whole molecule water
insoluble [48 ]. In contrast, polyether-based blocks, such as PPO,
exhibit a moderate hydrophobicity due to the presence of the polar
ether groups and are suitable for trans-membrane spanning insertion.
Another interaction type is found for amphiphilic copolymers bearing
a short hydrophobic block, in that case the binding to the membrane
through the hydrophobic anchor is facilitated. The anchor can be an
alkyl chain as in PEO-lipid conjugates [49] or a peruorinated moiety
as commonly used to enhance membrane binding of drugs [50 ].
As mentioned in the Introduction, most investigations on the interactions of polymers with lipid systems deal with ABA triblock
copolymers of the type PEO-b-PPO-b-PEO. This is due in part to the
intermediate hydrophobicity of PPO, but it is also partially the result
of their commercial availability. Poloxamers have already been the
subjects of several reviews, [51 ,52,53,54], and they will not be further
discussed here. In recent years novel amphiphilic block copolymers with

E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498


Table 1
Overview of the most common techniques for the characterization of interactions between block copolymers and lipid model membranes and the type of information obtained.
Experimental technique



Supported bilayers

Atomic force microscopy (AFM)

Secondary ion mass spectrometry (SIMS)


Langmuir monolayers

Surface pressure ()mean molecular

area (A) isotherms
Brewster angle microscopy (BAM)

Morphology of lipid domains in the tens of nanometer range

Chemical composition within lipid domains with spatial resolution
in the tens of nanometer range
Changes due to polymer adsorption: monolayer state, phase transitions,
monolayer compressibility and stability.
External probe-free visualization of changes in monolayer morphology,
formation of domains/aggregates, phase separation, reectivity. m-resolution
Visualization of domain formation with a uorescence label. m-resolution
Changes in lipid/block copolymer molecular conformation and orientation.
Monolayer composition.
Changes in structure of the lm in the surface normal direction from the
electron density prole, layer thickness.
Changes in structure of the lm in the surface normal direction, layer thickness.
Enhanced contrast compared to X-rays with deuterated blocks and
contrast matching in D2O.
Changes in type of molecular packaging within the monolayer

Fluorescence microscopy
Infrared reection absorption spectroscopy (IRRAS)
X-ray reectivity
Neutron reectivity


X-ray grazing incidence diffraction

Isothermal titration calorimetry (ITC)
Differential scanning calorimetry (DSC)
Fluorescence spectroscopy

Giant unillamelar vesicles

Bilayer lipid membranes

Cryo-transmission electron microscopy (c-TEM)

Quartz crystal microbalance (QCM)
Dynamic light scattering (DLS)
Fluorescence recovery after photobleaching (FRAP)
Confocal Laser Scanning Microscopy (CLSM)
Electrical/ion conductivity

Thermodynamic parameters for the binding of polymer to the membrane.

Partition coefcient, binding enthalpy.
Changes in the thermotropic phase behavior of the lipid. Enthalpy, phase transitions.
Inuence of polymer in the ip-op rate of membrane lipids,
membrane permeability towards uorescent molecules, pore formation.
Visualization of changes in liposome morphology and size.
Polymer binding isotherm
Changes in the hydrodynamic radius of liposomes, aggregation.
Intramembrane dynamics, lateral diffusivity
Visualization of domain formation on liposome surface with a uorescence label.
Variation in ion permeability of the membrane. Transient or permanent
pore formation. Membrane potentials.

hydrophobic blocks other than PPO and their interactions with lipid
systems have also been reported. Bio-inspired copolymers in the form of
conjugates of PEO with lipids have been studied for delivery of poorly
soluble drugs encapsulated in lipid-core micelles [55 ], and for tumortargeted quantum dots-containing micelles [56]. Closely related block
copolymers of PEO and short blocks of lipid-mimetic units [57] are
intended for the steric stabilization of liposomes in vivo. Also triblock
copolymers bearing two blocks of lipid-mimetic monomers and a
middle PEO block have been reported [58 ]. A triblock copolymer
intended for trans-membrane spanning insertion was synthesized
having a polystyrene middle block and two PEO outer blocks [59]. A
triblock copolymer with poly(p-dioxanone-co-L-lactide) as hydrophobic side blocks and a PEO middle block is studied to enhance DNA
uptake [60].
On the other hand, hydrophilic blocks other than PEO can also be
employed. In this case it has been found that not only the hydrophobicity of a copolymer determines its membrane disturbing ability, but
also its particular chemical structure plays eventually a role [61,62]. A
bulky hydrophilic block may induce a disturbance in the liquidcrystalline packing of lipid bilayers in addition to the effect brought
about by the hydrophobic block alone. In particular, block copolymers
having branched polyglycerol as hydrophilic block show a far more
pronounced effect on membrane structure as compared to copolymers
with linear PEO blocks [48]. ABA triblock copolymers having poly
(glycerol monomethacrylate) (PGMA) as hydrophobic block have evidenced a new type of molecular arrangement in mixed monolayers
resulting from the adsorption of PGMA-b-PPO-b-PGMA to DPPC monolayers [63 ]. An organization in ordered-lipid clusters with sizes in the
mesoscopic range surrounded by a network of block copolymers (lipid
corraling) was found. Such lipid corraling can be relevant for unveiling
the mechanism behind the healing effect of some poloxamers when
applied to cellular membranes injured by electroporation. Also polyoxazolines have been evaluated as hydrophilic blocks. In a recent
investigation poly(2-methyloxazoline)-b-poly(dimethylsiloxane)-bpoly(2-methyloxazoline) triblock copolymers showed a varying compatibility with DPPC or DOPC phospholipid monolayers depending on



the lipid uidity (stronger with more uid lipids), and on the polymer
size [64 ].
Several experimental techniques are commonly applied in these
kinds of studies of membraneblock copolymer interactions. The most
relevant experimental methods are summarized in Table 1.
4. Hybrid particles lipid-amphiphilic polymer
Recently, several self-assembled structures incorporating amphiphilic polymers and lipids, having sizes in the hundreds-of-nanometer
range are being assiduously investigated in order to exploit the advantages of natural and synthetic amphiphiles as nano-carriers.
4.1. Stealth liposomes
Among the various proposed nano-sized drug carriers, liposomes
are probably the highest developed, being already used in several
medical products mainly for cancer therapy and gene delivery [76 ].
Despite their numerous advantages including intrinsic excellent biocompatibility and the possibility for site-specic targeting, the low
stability, both in vitro and in vivo limits their widespread application.
Liposomes share also a drawback common for colloidal drug carriers in
general: they are rapidly removed from the blood stream by cells of the
reticulo-endothelial system. Therefore, there has been a continuous
search for strategies to prolong the circulation time in the body.
Among them, the steric stabilization through a corona from a lipid
anchored amphiphilic polymer surrounding the vesicle, producing socalled Stealth liposomes [77 ], has been the most successful strategy.
This effect is a result of the hydration of the adsorbed polymers that
slows downs the recognition by blood plasma opsonising proteins
mediating the clearance by the immune system, thus prolonging
the circulation time in the body [78]. The standard system for steric
stabilization of liposomes involves the incorporation of PEO-lipid
conjugates. However several alternative types of block copolymers
have also been evaluated for this purpose and some recent examples
incorporating a hyperbranched polyglycerol in the hydrophobic


E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498

corona [79 ], two or four lipid mimetic units anchors instead of normal
lipids [57], or a cholesterol anchor [79] are summarized in Fig. 4.
4.2. Lipopolymersomes
Vesicular structures made from the self-assembly of block copolymers are called polymersomes and can be regarded as synthetic
analogs of biological membranes [80]. Compared to liposomes they
are thicker (around 1020 nm compared to 5 nm), have a higher mechanical stability and elasticity. Therefore they have been used as model
for the study of various processes occurring also in cell membranes as

fusion and ssion, but with the advantage of a slower kinetics as

compared to liposomes [81 ]. On the other hand, polymersomes formed
from ABC block copolymers with an asymmetric composition of their
inner and outer hydrophilic leaet have been synthesized to mimic the
natural asymmetry of biological membranes. The water soluble blocks
are poly(2-methyloxazoline) and PEO surrounding an insoluble poly
(dimethylsiloxane) middle block [82]. Such polymersomes allow for
studies of the directed insertion of membrane proteins in a preferred
orientation that is not possible to achieve with liposomes of symmetric
Also hybrid vesicles of lipids and amphiphilic block copolymers,
named lipopolymersomes, have been recently prepared from POPC
and the block copolymer polybutadiene-b-poly(ethylene oxide) in an
attempt for combining the biocompatibility of lipids with the enhanced mechanical strength and stability of the copolymer matrix
[73]. A demixing process in polymer-rich and lipid-rich domains has
been observed in such hybrid membranes.
4.3. Solid lipid and cubic-phase lipid nanoparticles
Besides liposomes, other promising kind of self-assembled lipid
nanoparticles are cubic-phase liquid-crystalline nanoparticles (CPNPs),
also known as cubosomes, having an internal reverse bicontinuous cubic
structure [83], see Fig. 5. They are prepared by dispersing a glycerol
monooleate lipid in the presence of a non-ionic block copolymer, such
as poloxamer PEO98PPO57PEO98, as fragmentation aid and steric
stabilizer. CPNPs are cubes with sizes in the 100500 nm range [84].
The high internal interfacial area separating hydrophobic and hydrophilic domains makes such nonlamellar phase structures interesting
candidates as carriers of relatively large amounts of both hydrophilic
and hydrophobic molecules. Aqueous dispersions of analogous solid
lipid nanoparticles (SLNs) having a lipid crystalline matrix surrounded
by a emulsifying layer composed of an amphiphilic PEO based copolymer, such as polysorbates (PEO modied sorbitan esteried with fatty
acids) or poloxamers, are being tested as colloidal oral drug delivery
systems for both high and low water soluble drugs [85]. Their particular
advantages for oral drug delivery are the avoidance of organic solvents
and their increased stability in high concentrated suspensions under
physiological conditions typically found in the gastrointestinal tract as
compared to more traditional colloidal drug delivery systems based on
liposomes or polymersomes [85]. Also hybrid SLNs incorporating an
anionic polymer to facilitate the encapsulation of water-soluble cationic
drugs have been tested as delivery vehicles for the anticancer cytotoxic
drug doxorubicine with promising results [86].
5. In vitro investigations of the interactions of block copolymers
with living cells

Fig. 4. Scheme of stealth liposomes having a hydrophilic PEO-corona, (a) and (b), or a
PEO-hyperbranched polyglycerol-corona, (c) and (d). The hydrophobic part is formed
by lipid-like alkyl chains, (a), (b) and (c); or cholesterol (d). The leakage prole of a
uorescence dye through the bilayer (e) is normally used for evaluating liposomes'

Recently three different block copolymers were used to study the

reversal of multidrug resistance of tumor cells [87 ]. Pluronic L61 is in
the moment the most successful polymer in order to support the
transport of antitumor drugs across the cell membrane. It remains an
open question if this is the result of L61 phospholipid interactions or if
it is related to L61 P-glycoprotein (Pgp) interactions. Pgp upregulates
together with other multiple drug resistance proteins the cellular
drug efux system. In order to study the mechanism, the doxorubicin
uptake of cells with a high level of Pgp expression (human embryonic
kidney cells, HEK293) is measured after adding three different block
copolymers. Addition of L61 results in a strong uptake of doxorubicin.
When PFMA-b-PEO-b-PFMA, i.e. polymers consisting of a middle
hydrophilic PEO block embedded between two hydrophobic poly
(peruorohexylethyl methacrylate) (PFMA) blocks, is administered an
intermediate doxorubicin uptake is achieved. The third block copolymer
used, PEO-b-PHFPO-b-PEO; having poly(hexauoropropylene oxide)
(PHFPO) as substitute for the hydrophobic middle block of PPO, does not
induces drug uptake for cells with a high Pgp expression level, but it

E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498


Fig. 5. Scheme of a cubic-phase lipid nanoparticle having an internal reverse bicontinuous cubic structure and stabilized by an amphiphilic ABA triblock copolymer (a). It is able to
solubilize hydrophobic molecules within the bilayer (b) or to encapsulate hydrophilic molecules in the interior aqueous phase (c).

does for cell with lower Pgp expression. Since PEO-b-PHFPO-b-PEO

supports drug uptake but lacks any ionophoric activity, measured
separately on lipid bilayers of DPhPC through membrane potential
measurements, it is concluded that ion transport facilitation by these
synthetic block copolymers is not the determinative factor for MDR
reversal [87].
6. Conclusion
This review shows the enormous potential of new techniques of
polymer syntheses as controlled radical polymerization and click
chemistry to tailor polymer architectures for interacting with cell
membranes. This allows for the development of completely new
strategies in modern pharmacy and nanomedicine. Prior to in vivo and
in vitro testing, model membranes provide the possibility for detailed
studies of the interactions with block copolymers on a molecular level.
Modern microscopic, scattering, spectroscopic and calorimetric
methods have an unprecedented high spatial resolution or an extreme
sensitivity. This gives also a sound basis for comparison with data
obtained by computer simulations. These simulation techniques provide
more and more realistic pictures also of complicated systems as block
copolymers adsorbed on or incorporated in cell membranes. Nowadays,
simulations generate not only structural information, but simultaneously the dynamics of these complex systems can be studied. It can
be expected that the next years will show a continued interest in
the research on block copolymer cell membrane interactions triggered
by the fundamental scientic interest and simultaneously by the
large commercial interest in new pharmaceutical and biomedical
We acknowledge the nancial support by DFG (FOR 1145), and
T. Wieczorek for some of the graphic art work.
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of special interest.
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