a r t i c l e
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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
Amphiphilic block copolymers are usually composed of hydrophilic and lipophilic blocks covalently joint together in several
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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 ].
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Fig. 2. General organization of a phospholipid bilayer and chemical structure of common 1,2-diacylphosphocholines and cholesterol employed in model membranes.
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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.
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
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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
Information
Ref.
Supported bilayers
[44]
[45]
Langmuir monolayers
Fluorescence microscopy
Infrared reection absorption spectroscopy (IRRAS)
X-ray reectivity
Neutron reectivity
Liposomes
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
[65]
[63]
[66]
[63,65]
[67]
[68]
[67]
[69]
[70]
[48,51]
[71]
[72]
[70]
[73]
[74]
[48,75]
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
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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
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'
stablility.
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497
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).
Int Ed 2008;47:30925.
[2] Hamley I. Block copolymers in solution: fundamentals and applications. John
Wiley & Sons; 2005.
[3] Alexandridis P, Lindman B. Amphiphilic block copolymers: self-assembly and
applications. Elsevier; 2000.
[4] Busse K, Peetla C, Kressler J. Water surface covering of uorinated amphiphilic
triblock copolymers: surface pressure-area and X-ray reectivity investigations.
Langmuir 2010;23:697582.
of special interest.
of outstanding interest.
design, characterization and biological signicance. Adv Drug Del Rev 2001;47:
11331.
[19] Sharma V, Stebe K, Murphy JC, Tung L. Poloxamer 188 decreases susceptibility of
articial lipid membranes to electroporation. Biophys J 1996;71:322941.
[20] Maskarinec SA, Hannig J, Lee RC, Lee KYC. Direct observation of poloxamer 188
insertion into lipid monolayers. Biophys J 2002;82:14539.
[21]
Amado E, Blume A, Kressler J. Novel non-ionic block copolymers tailored for
2001;101:292190.
[27] Charmot D, Corpart P, Adam H, Zard SZ, Biadatti T, Bouhadir G. Controlled radical
polymerization in dispersed media. Macromol Symp 2000;150:2332.
[28]
Chiedari J, Chong YK, Ercole F, Krstina J, Jeffery J, Le TPT, et al. Living free-radical
498
E. Amado, J. Kressler / Current Opinion in Colloid & Interface Science 16 (2011) 491498
[30]
Binder WH, Sachsenhofer R. Click chemistry in polymer and materials science.
[42] Moghaddam B, Habib Ali M, Wilkhu J, Kirby DJ, Mohammed AR, Zhen Q, et al. The
application of monolayer studies in the understanding of liposomal formulations.
Int J Pharm 2011;417:23544.
[43] Brown DA, Rose JK. Sorting of GPI-anchored proteins to glycolipid-enriched membrane
subdomains during transport to the apical cell surface. Cell 1992;68:53344.
[44] Marqus JT, Viana AS, De Almeida RFM. Ethanol effects on binary and ternary
supported lipid bilayers with gel/uid domains and lipid rafts. Biochim Biophys
Acta 2011;1808:40514.
[45]
Anderton CR, Lou K, Weber PK, Hutcheon ID, Kraft ML. Correlated AFM and NanoSIMS
between the structure of amphiphilic copolymers and their ability to disturb lipid
bilayers. Biochemistry 2005;44:404254.
[49] Momekova D, Rangelov S, Lambov N, Karlsson G, Almgren M. Effects of
amphiphilic copolymers bearing short blocks of lipid-mimetic units on the
membrane properties and morphology of DSPC liposomes. J Dispersion Sci
Technol 2008;29:110613.
[50]
Gerebtzoff G, Li-Blatter X, Fischer H, Frentzel A, Seelig A. Halogenation of drugs
oxide and propylene oxide block copolymers on the permeability of bilayer lipid
membranes to small solutes including doxorubicin. Biochim Biophys Acta
2000;1468:7386.
[52]
Frey SL, Zhang D, Carignano MA, Szleifer I, Lee KYC. Effects of block copolymer's
architecture on its association with lipid membranes: experiments and simulations. J Chem Phys 2007;127:11490412.
[53] Firestone MA, Wolf AC, Seifert S. Small-angle X-ray scattering study of the interaction
of poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) triblock
copolymers with lipid bilayers. Biomacromolecules 2003;4:153949.
[54] Maskarinec SA, Wu G, Lee KYC. Membrane sealing by polymers. Ann NY Acad Sci
2005;1066:31020.
[55]
Torchilin VP. Lipid-core micelles for targeted drug delivery. Curr Drug Deliv
2005;2:31927.
[56] Papagiannaros A, Upponi J, Hartner W, Mongayt D, Levchenko T, Torchilin V.
Quantum dot loaded immunomicelles for tumor imaging. BMC Med Imaging
2010;10:2230.
[57] Momekova D, Momekov G, Rangelov S, Storm G, Lambov N. Physicochemical and
biopharmaceutical characterization of dipalmitoyl phosphatidylcholine liposomes
sterically stabilized by copolymers bearing short blocks of lipid-mimetic units.
Soft Matter 2010;6:591601.
[58]
Rangelov S, Almgren M, Edwards K, Tsvetanov C. Formation of normal and reverse
bilayer structures by self-assembly of nonionic block copolymers bearing lipidmimetic units. J Phys Chem B 2004;108:754252.
[59] Baekmark TR, Pedersen S, Jorgensen K, Mouritsen OG. The effects of ethylene
oxide containing lipopolymers and tri-block copolymers on lipid bilayers of
dipalmitoylphosphatidylcholine. Biophys J 1997;73:147991.
[60] Bhattarai SR, Yi HK, Bhattarai N, Hwang PH, Kim HY. Novel block copolymer
(PPDO/PLLA-b-PEG): enhancement of DNA uptake and cell transfection. Acta
Biomater 2006;2:20712.
[61]
Seelig A, Gerebtzoff G. Enhancement of drug absorption by noncharged detergents
through membrane and P-glycoprotein binding. Expert Opin Drug Metab Toxicol
2006;2:73352.
[62] Pavlov DN, Alexandrova NA, Krylova OO, Pohl P, Melik-Nubarov NS. Effect of block
architechture on the ability of polyalkylene oxides to overcome multidrug
resistance of tumor cells. J Drug Del Sci Technol 2006;16:25965.
[63]
Amado E, Kerth A, Blume A, Kressler J. Phospholipid crystalline clusters induced by
2004;43:3696703.
[76]
Torchilin VP. Recent advances with liposomes as pharmaceutical carriers. Nat Rev