Nano Today (2012) 7, 404—413

Available online at www.sciencedirect.com

journal homepage: www.elsevier.com/locate/nanotoday

REVIEW

Super-hydrophilic zwitterionic poly(carboxybetaine)

and amphiphilic non-ionic poly(ethylene glycol) for
stealth nanoparticles
Zhiqiang Cao, Shaoyi Jiang ∗

Department of Chemical Engineering, University of Washington, Seattle, WA 98195, USA

Received 7 May 2012; received in revised form 7 July 2012; accepted 6 August 2012
Available online 29 August 2012

KEYWORDS Summary This review compares two types of non-fouling polymers, the widely used non-
Polyethylene glycol; ionic poly(ethylene glycol) (PEG) and the recently established zwitterionic poly(carboxybetaine)
Zwitterions; (PCB), for their use in creating stealth nanoparticles (NPs) for drug delivery and protein pro-
Non-fouling; tection. While both types of polymers exhibit reasonable non-fouling properties, such as good
Protein conjugation; protein and colloidal stability and extended blood circulation time in vivo, amphiphilic PEG has
Liposome; negative effects on proteins and NPs due to its hydrophobic nature, including reduced protein
Drug delivery bioactivity, instability of assembled NPs, and lipid bilayer destabilization. These problems can
be overcome by super-hydrophilic PCB.
© 2012 Elsevier Ltd. All rights reserved.

Introduction their diagnostic or therapeutic functions. There are gener-

Avoiding non-specific interactions (or possessing non-fouling
properties) [1—7] is one of the most important character-
istics required by nanoparticles (NPs) in a wide range of
applications from drug delivery to diagnostics [8,9], espe-
cially for use in complex media. To achieve this stealth
property, non-fouling polymeric materials are used to mod-
ify the surface of particles. The treated particles exhibit
improved stability and blood circulation time, promoting
∗ Corresponding author. Tel.: +1 206 616 6509;
fax: +1 206 543 3778.
E-mail address: sjiang@u.washington.edu (S. Jiang).
URL: http://depts.washington.edu/jgroup/Index.htm
(S. Jiang).
ally two distinct types of non-fouling polymers; non-ionic
polymers, among which the most notable is polyethy-
lene glycol (PEG) [10], and zwitterionic polymers such as
poly(carboxybetaine) (PCB) [7,11—13] (Fig. 1).
Amphiphilic non-ionic poly(ethylene glycol)
PEG has been the most popular for modifying particles
because of its historical use since the 1970s and PEGyla-
tion has become a standard surface and particle modification
method for biological applications [14—23]. The non-fouling
property of PEG was attributed to its steric exclusion
effect [24,25]. More recently, it is generally agreed upon
that its hydration ability or hydrophilic nature plays a key
role in its non-fouling property [26,27]. Molecular simula-
tion studies of oligo(ethylene glycol) (OEG) self-assembled

1748-0132/$ — see front matter © 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.nantod.2012.08.001
Zwitterionic and non-ionic polymers for stealth nanoparticles 405

EG Headgroup (Amphiphilic) CB Headgroup (Superhydrophilic)

CH3 O

O CH2 CH2 CH2 N CH2 C O

CH3

Figure 1 Molecular structures of non-ionic EG and zwitterionic CB headgroups. PEG is amphiphilic with both hydrophilic and
hydrophobic properties. PCB is super-hydrophilic. The number of carbon groups (n) between the positive quaternary amine group
and the negative carboxylate group of each PCB unit can be varied. N = 1 is shown here, which is glycine betaine.

monolayers (SAMs) have shown a tightly bound water layer even humans [48], complicating clinical trials involving this
around OEG, generating large repulsive forces to repel material.
proteins when they are approaching the surface [28—30].
PEGylation on particles results in increased water solubil- Super-hydrophilic zwitterionic
ity and colloid stability, decreased interactions with blood
poly(carboxybetaine)
components (opsonization), and improved blood residence
time in vivo [15,16,31—33]. Because of these favorable
As alternatives to non-ionic PEG, zwitterionic non-
features, several PEGylated products have been launched
fouling polymers, namely poly(phosphobetaine),
to the market, such as PEGylated proteins and liposomes
poly(sulfobetaine), and PCB are promising [7,49—55].
[15,16,18,19,31—33].
While PEG and other non-ionic materials (e.g., tetraglyme
It should be pointed out that PEG is, in fact, amphiphilic.
[56], dextran [57], mannitol [58], polyamines function-
In addition to its hydrophilic property, PEG also has a
alized with acetyl chloride [59], PEG-mimetic peptoid
hydrophobic character [13,34,35] although PEG is often
[60], and serine-rich peptides [61]) bind water molecules
considered as a hydrophilic polymer. Because of this
via hydrogen-bonding hydration, zwitterionic polymers
amphiphilic feature, PEG is able to dissolve in many organic
achieve stronger hydration that is electrostatically induced
solvents in addition to water. This makes it convenient to
[62,63]. It has been shown that surfaces modified with
use PEG in both organic and aqueous solvents to modify
zwitterionic polymers have excellent non-fouling properties
both hydrophobic and hydrophilic particles. This accounts
against proteins, cells, and bacteria [7,64,65]. Take PCB
for its popularity as a modifying material. Unfortunately,
as an example, derived from naturally occurring betaines
due to its hydrophobic nature, the ease in its appli-
abundant in the animal kingdom including humans as well as
cation results in negative effects when it is attached
plants [66]. We have shown that PCB polymer brush coatings
to hydrophilic proteins or it is used with a hydropho-
resist non-specific protein adsorption from undiluted blood
bic polymer to form core—shell particles. For protein
plasma and serum to an undetectable level (<0.3 ng/cm2 )
protection, PEG is the standard material to be con-
using a surface plasmon resonance (SPR) sensor, and resist
jugated to a protein, increasing protein stability (e.g.
bacterial adhesion and delay biofilm formation for 10 days
against heat, salts, and proteolysis), water solubility, and
at room temperature [7,67—69]. Although both PEG and
circulation time in the body [13,15,36]. The resulting
PCB-coated gold NPs are stable in 10% blood plasma and
PEGylated protein, however, often significantly loses its
serum, PCB-coated NPs are stable in undiluted blood plasma
biological activity [31,37—40]. Taking PEG-interferon ␣2a
and serum for 3 days while PEG-coated NPs aggregate [70].
(PEGASYS® , to treat hepatitis C) as one example, PEGyla-
This review focuses on PCB as one example of zwitterionic
tion results in a 93% loss of its original bioactivity [31].
polymers and compares the pros and cons of PEG and PCB
This has been attributed to PEG chains interfering with
for creating stealth NPs.
the zwitterionic nature of protein surfaces and the func-
Unlike amphiphilic PEG, zwitterionic polymers are super-
tional pocket of the protein (mostly hydrophobic domains)
hydrophilic, and insoluble in most organic solvents. Such
through hydrophobic—hydrophobic interactions [12,13]. For
super-hydrophilic character circumvents the hydrophobic
carrier-based drug delivery systems, PEG is so far solely
character of PEG mentioned above. Three examples are
used to modify liposomes in commercial drug formula-
given below to illustrate the use of zwitterionic PCB and its
tions for long blood circulating purposes. For example,
advantage over the non-ionic PEG counterpart in construct-
LIPO-DOX® and DOXIL® , both PEGylated liposomes with
ing polymer—protein conjugates, self-assembled polymeric
doxorubicin encapsulated, have been clinically used to
NPs, and liposomes.
treat AIDS-related Kaposi’s sarcoma, breast cancer, ovar-
ian cancer, and other solid tumors [18,19,22,32,41,42].
Nevertheless, the inclusion of PEG tends to destabilize Three examples
the lipid bilayer because of its hydrophobicity, leading to
rapid drug leakage [43]. To compensate these destabiliz- Polymer—protein conjugates
ing issues arising from PEG, a large amount of cholesterol
(i.e., 39 mol%) is required in PEGylated liposomes to enhance PEGylation has been the benchmark for stabilizing proteins
drug retention [43]. In addition to these hydrophobic and increasing their blood circulation time [15,36]. How-
issues, PEG also suffers oxidation damage in biological ever, PEG reduces the bioactivity of conjugated therapeutic
media for long-term applications [5,26,34,44—46]. Further- proteins, antibodies, and enzymes [31,37—40]. Our recent
more, PEG antibodies occur in animals [15,34,47,48] and work has shown the stabilization of a protein by attaching
406 Z. Cao, S. Jiang

Figure 2 (a) Super-hydrophilic PCB increases enzyme—substrate hydrophobic—hydrophobic interactions, thereby increasing the
affinity of the substrate for the binding pocket [12]. (b) Amphiphilic PEG reduces enzyme—substrate hydrophobic—hydrophobic
interactions, and thus the bioactivity of the enzyme [12].

zwitterionic PCB to the protein without sacrificing its bioac-
tivity. Such a combination of high stability and bioactivity
has never been achieved before (Fig. 2) [12]. Specifically,
5 kDa PEG was attached to ␣-chymotrypsin, a proteolytic
digestive enzyme, to various degrees with different numbers
of polymer chains per protein. For fair comparison, PCB with
an equal molecular weight of 5 kDa (determined by NMR) and
PCB with an equal hydrodynamic size of 5 kDa PEG (deter-
mined by size exclusion chromatography) were synthesized
and conjugated to the same enzyme with similar degrees of
conjugation as PEG. Results show that enzymes can be bet-
ter stabilized by PCB than PEG, maintaining enzyme activity
at high urea concentrations and at elevated temperatures.
Fig. 3a shows the binding affinities between the substrate

Figure 3 (a) Binding affinities (Michaelis constant, Km ) of ␣-chymotrypsin (CT) conjugated with PEG (5 kDa molecular weight),
pCB Mn (the same molecular weight as PEG, 5 kDa), and pCB Rh (the same hydrodynamic size as 5 kDa PEG). Km increased as more
PEG conjugated to CT, showing the inhibitory effects of PEG. Unlike PEG, PCB showed either no significant effect on binding affinity
(i.e., pCB Mn conjugates) or even an increase in substrate affinity (i.e., pCB Rh conjugates) [12]. (b) Km of native CT in the presence
of 650 Mn PEG and ammonium acetate solutions. The effects of these small molecular weight solutes on Km are similar to their
respective high molecular weight versions (PEG 5 kDa and pCB Rh respectively). Error bars in (a) and (b) are values of standard error
(or standard error of the mean) [12].
Zwitterionic and non-ionic polymers for stealth nanoparticles
Figure 4 Structures of OEG (four EG segments), CB (1 carbon group between the positive quaternary amine group and the negative
carboxylate group), and chymotrypsin inhibitor 2 (CI2). The bar graph represents the SASAs of the hydrophobic domains of CI2 in
pure water, OEG solution, or CB solution, from molecular dynamics simulations [72].
and the polymer—protein conjugate as reflected by their
Michaelis constants (Km ) or the concentration of substrate
required to achieve 50% full activity. A lower Km indicates
higher binding affinity. It was shown that the attachment of
PEG chains decreases the enzyme binding affinity, while PCB
conjugates of similar hydrodynamic size, in contrast, have
no impact on or even increase binding affinity.
The reason why the binding affinity of a protein is influ-
enced differently by PEG and PCB has been attributed
to the hydrophobicity of these polymers [12]. Fig. 3b
illustrates the Km values of bare enzymes dissolved in solu-
tions of 650 Mn PEG and ammonium acetate (NH4 OAC). It
should be noted that NH4 OAC is composed of the most
protein-stabilizing and bioactive monovalent ions (kos-
motropic anion, OAC− , and chaotropic cation, NH4 + ) in
the Hofmeister series [71], and PCB can be considered
the polymeric version of these ions. The impact on Km or
binding affinity by 650 Mn PEG and free NH4 OAC molecules
was in a good agreement with that of the respective
high-molecular-weight versions of PEG and PCB polymers
attached to the proteins as shown in Fig. 3a. Results show
that polymers, whether chemically attached to the pro-
tein or not, will interfere with hydrophobic—hydrophobic
attractions between the substrate and the enzyme bind-
ing site, depending on the degree of their hydrophobicity.
As further exploration, the partitioning of the tested sub-
strate into free polymer solutions of PEG and PCB through
a semi-permeable membrane was also measured [12]. It
was found that the substrate had higher solubility in PEG
and lower solubility in PCB than that in buffer control.
Thus, the hydrophobic nature of PEG increases the local
hydrophobicity around the surface of a protein and near its
binding pocket, and results in higher solubility of the sub-
strate. As a result, the hydrophobic nature of PEG reduces
the substrate—enzyme hydrophobic—hydrophobic interac-
tion (Fig. 2). In contrast, the super-hydrophilic nature of PCB
facilitates the substrate—enzyme hydrophobic—hydrophobic
interaction in the reverse manner (Fig. 2).
Recently, further evidence was obtained, showing that
the amphiphilic PEG segment has more favorable inter-
actions with the hydrophobic domains of a protein than
407
super-hydrophilic PCB. The hydrophobic domains are gener-
ally recognized to play an essential role in the bioactivity of
a protein. We studied the effects of CB and ethylene glycol
(OEG) solutes [(EG)4 in this case] on chymotrypsin inhibitor
2 (CI2) using molecular dynamics simulations (Fig. 4) [72]. It
was found that CI2 has much less solvent access surface area
(SASA) for the hydrophobic domains in the OEG solution than
in the CB solution. With EG4 , nearly 1/3 of the hydrophobic
domains originally accessible in the water solvent become
inaccessible, whereas the corresponding loss for CB is only
around 1/10. For the hydrophilic domains, CI2 has identical
SASA in both OEG and CB solutions.
Self-assembled polymeric NPs
PEG has been widely used to modify a hydrophobic polymer
block to form amphiphilic block copolymers, which can be
further assembled into various nanostructures [17,73—78].
The assembled nanostructures have been explored for many
applications such as in drug delivery [17,73,78—81]. Some
of these NPs have been commercialized as chemo-drug
formulations such as Genexol-PM in a phase IIa trial, a
poly(D,L-lactide-b-ethylene glycol) (PLA-PEG) NP with pacli-
taxel [82] and BIND-014 in a preclinical trial, a poly(lactic
acid-co-glycolic acid-b-ethylene glycol) PLGA—PEG NP with
docetaxel that possesses cell targeting ability [83]. The
potential of these systems comes from the PEG block which
can be dissolved in aqueous media via hydrogen bonding
with water molecules, stabilizing the hydrophobic particle
cores of PLA or PLGA from aggregation in complete biological
media and improving their blood circulation time [78—81].
However, PEGylated NPs suffer limited stability under
harsh conditions such as freeze-drying, which is a common
way to prevent the degradation of polymers such as PLA
and PLGA, and to avoid drug leakage in aqueous storage
media. To the best of our knowledge, few polymer-based
NPs can survive lyophilization without cryoprotectant addi-
tives [17,84—86]. Even for PEGylated NPs, cryoprotectants
such as 10% sucrose are required to help stabilize these
NPs [17,86]. By replacing PEG with PCB, we synthesized
408 Z. Cao, S. Jiang

Figure 5 PCB—PLGA and PEG—PLGA assembled NPs. It is expected that all PCB chains migrate to the hydrated shell region of
the NP due to the super-hydrophilic nature. Amphiphilic PEG has dual solubility in both oil and water phases. PEG chains can exist
simultaneously in both the hydrated shell and hydrophobic core regions.

PLGA—PCB block copolymers (Fig. 5) and found that their
assembled NPs were remarkably stable after freeze-drying,
without any cryoprotectant additives (Fig. 6a) [11]. With
brief re-suspension with a pipette and without the need of
sonication, the freeze-dried PLGA—PCB NPs with or without
drug loaded have the same mean diameter and low polydis-
persity index (PDI) number as before freeze-drying (Fig. 6a).
Fig. 6b and c shows the morphology of PLGA—PCB NPs before
and after lyophilization in water under scanning electron
microscopy (SEM).
It should be noted that both PEG and PCB protected
polymeric NPs show reasonably good colloidal stability
in aqueous media (e.g., biological or physiological envi-
ronments) [11,78,80,81], since both polymers are fully
solvated. When NPs are dehydrated (e.g., freeze-dried),
the amphiphilic PEG crystallizes and loses its function to
prevent NP aggregation [85]. The super-hydrophilic PCB, in
contrast, has stronger hydration ability and binds a greater
number of water molecules more strongly to prevent crys-
tallization during lyophilization. Strong hydration ability
has also been used to explain few cases of NP systems
requiring no cryoprotectant through the lyophilization pro-
cesses, such as micelles protected by ionic polysialic acid
[87].

Figure 6 Stability of PLGA—PCB NPs throughout lyophilization. (a) NP sizes (mean ± SD, N = 3) for PLGA NPs, PLGA—PCB NPs, and
PLGA—PCB/Dtxl NPs with 1 wt% drug loading (shown as NP suspended), and those after lyophilization without any addition of a
cryoprotectant. Polydispersity indexes (PDIs, mean ± SD, N = 3) are indicated accompanying each size point. Dtxl = Docetaxel. (b)
SEM image for PLGA—PCB NPs before lyophilization. (c) SEM image for PLGA—PCB NPs after lyophilization and brief re-suspension
in water by pipettes without sonication. The scale bar for (b) and (c) is 1 ␮m [11].
Zwitterionic and non-ionic polymers for stealth nanoparticles 409

Figure 7 PEGylated and poly(zwitterionic) liposomes. For PEGylated liposome, amphiphilic PEG provides a stealth layer by itself
via hydration due to its hydrophilic nature, but dehydrates the polar head group region of the liposome due to its hydrophobic nature,
leading to negative consequences such as a rapid leakage of hydrophilic drugs. The addition of cholesterol is the most typical way
to neutralize these impacts from PEGylation by increasing the fluidity of the membranes. For poly(zwitterionic) liposome, super-
hydrophilic zwitterionic PCB provides a stealth layer via hydration, similar to PEG. Unlike PEG, PCB further hydrates the polar head
group region due to its super-hydrophilic nature, rearranging the membrane structure in a way opposite to PEG, leading to good
drug retention without the need for cholesterol [95].

Liposomes PEGylated liposomes [43]. Cholesterol plays the opposite

PEG is often conjugated to a lipid for liposome protec-
tion. The resulting PEGylated liposomes have achieved great
success in delivering chemo-drugs intravenously, such as
Doxil® (known as Caelyx® in Canada and Europe). The non-
fouling PEG reduces the interaction of plasma proteins with
liposomes and thus their uptake by the reticuloendothelial
system (RES) [22,88]. As a consequence, PEGylated lipo-
somes show prolonged blood circulation time, and target
tumor tissues through the enhanced permeability and reten-
tion (EPR) effect [18,19,22,32,88]. These positive effects all
come from the hydrophilic nature of PEG.
However, being also hydrophobic, PEG tends to desta-
bilize the lipid bilayer (Fig. 7). Specifically, PEG lowers
the polarity of the aqueous phase [89] and decreases the
hydration level of the lipid polar group region [90]. This fur-
ther enhances the lateral packing of the phospholipid acyl
chains, inhibits the diffusional motion of PEG, strengthens
PEG chain—chain interactions, and tends to induce bilayer
demixing [43]. This series of consequences cause the rapid
leakage of hydrophilic drugs [43], and are enhanced by PEG
with a higher molecular weight such as ≥5 kDa, and phos-
pholipids with a higher phase transition temperature (Tm )
such as 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC,
Tm = 55 ◦ C) [43,91]. To compensate for the destabilizing
effects of PEG, cholesterol is frequently required in these
role of PEG by increasing the hydration of the lipid polar
region and membrane fluidity, reducing PEG chain—chain
interactions and bilayer demixing, and improving drug
retention [43,92—94]. The necessity of cholesterol in PEGy-
lated liposomes has been shown in LIPO-DOX® and DOXIL® ,
both containing 39 mol% cholesterol of the total lipids. It
has also been shown in Fig. 8a, that PEGylation makes DSPC
liposomes leak more encapsulated carboxyfluorescein (CF, a
hydrophilic fluorescent dye) in the absence of cholesterol,
while CF retention was maintained with cholesterol present.
By replacing PEG with PCB, we synthesized DSPE-PCB
(DSPE: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine)
and formulated PCB-protected liposomes (Fig. 7) [95].
PCB-protected liposomes exhibit long blood circulating
characteristics in vivo, comparable to PEGylated liposomes
[95]. More importantly, they achieve drug retention without
cholesterol, which is otherwise required by PEGylated lipo-
somal formulations (shown in Fig. 8a). It was hypothesized
that unlike how the hydrophobic nature of PEG destabilizes
the lipid bilayer, super-hydrophilic PCB would behave
the opposite. This is supported in a differential scanning
calorimetry (DSC) study by measuring the major Tm of
DSPC in both PEG- and PCB-protected liposomes. As shown
in Fig. 8b, the addition of DSPE-PEG into DSPC liposomes
gradually increases the Tm from 55 ◦ C by less than 1 ◦ C. This
phenomenon has been related to the dehydration of the
410 Z. Cao, S. Jiang

(a) (b)
58
50 DSPC/DSPE-PEG 5K
DSPC/DSPE-PCB 5K
DSPC 57
CF Leakage in PBS (%) 40 DSPC/DSPE-PEG 5K 5%
DSPC/DSPE-PEG 2K 5% 56
DSPC/DSPE-PEG 2K 5%/Chol 39%
30 DSPC/DSPE-PCB 5K 10%

Tm ( C)
55

o
20
54

10
53

0 52
0 1 2 3 4 0% 5% 10% 15% 20%
Time (days) Molar Composition of DSPE-Polymer Conjugates

Figure 8 (a) Carboxyfluorescein (CF) was encapsulated into different liposome formulations. CF leakage profile was recorded
in PBS at 37 ◦ C (mean ± SD, N = 3). The percentage value in the formulation represents the molar composition of the specified
component. 2K or 5K represents the molecular weight of PCB or PEG. Chol, cholesterol; SD, standard deviation [95]. (b) Phase
transition temperature (Tm , mean ± SD, N = 3) of DSPC with the addition of DSPE-PCB 5K or DSPE-PEG 5K into the liposomes [95].

lipid polar region and a compact lateral packing of lipid
acyl chains [90]. The incorporation of DSPE-PCB, however,
decreases Tm , indicating an enhanced hydration around the
lipid polar region and a more fluidic membrane. It should be
noted that the addition of cholesterol will have the same
effect as PCB in lowing Tm [96,97].
How to connect super-hydrophilic PCB to a
hydrophobic segment
As discussed, super-hydrophilic PCB overcomes a number
of issues with amphiphilic PEG. However, linking PCB to a
hydrophobic segment, particularly a super-hydrophobic one
such as polydimethylsiloxane (PDMS), and a hydrolysable
one such as PLGA, is not as straightforward as PEG.
Amphiphilic PEG is soluble in a wide spectrum of solvents,
including aqueous and organic solvents, enabling the reac-
tion with both hydrophilic and hydrophobic moieties. The
PCB can dissolve mostly in aqueous solvents due to its super-
hydrophilicity. Thus, it is challenging to find a common
solvent in which a reaction of PCB with hydrophobic moieties
can occur. This solubility issue can be partially addressed by
using a mixed solvent system [70]. To fully resolve the solu-
bility issue, a hydrophobic precursor (i.e., CB—tBu in Fig. 9)
of the super-hydrophilic CB was introduced [11]. CB—tBu and

Figure 9 Synthesis of PLGA—PCB copolymers. DMF = N,N-dimethylformamide, HMTETA = 1,1,4,7,10,10-hexamethyltri-

ethylenetetramine, TFA = trifluoroacetic acid. (1) CB-tBu monomers are polymerized through ATRP initiated by an initiator,
2-aminoethyl 2-bromoisobutyrate. (2) TFA− is removed to generate PCB—tBu—NH2 . (3) PCB—tBu—NH2 is conjugated with PLGA—NHS
in anhydrous acetonitrile to form PLGA—PCB—tBu block copolymers. (4) PLGA—PCB is obtained by treating PLGA—PCB—tBu with
TFA to generate the zwitterionic CB structure while the PLGA remains intact [11].
Zwitterionic and non-ionic polymers for stealth nanoparticles
its polymer (PCB—tBu) have good solubility in many organic
solvents such as acetonitrile and dimethylformamide, mak-
ing it possible to incorporate PCB into hydrophobic blocks
such as PLGA (Fig. 9) [11] and lipids [95] in organic sol-
vents. After covalent binding of PCB—tBu to the hydrophobic
moieties, zwitterionic PCB can be readily regenerated by
hydrolysis of the tBu ester groups under mild acid condi-
tions such as trifluoroacetic acid (TFA). TFA does not degrade
typical ester bonds as observed in PLA [98—100], PLGA
[11,101], or DSPE [95]. In this way, amphiphilic diblocks with
a sharp polarity contrast between super-hydrophilic PCB and
hydrophobic PLGA or lipid can be prepared.
Conclusions and future directions
This review aims to compare non-ionic PEG with zwitterionic
PCB in creating stealth NPs for protein protection and drug
delivery. Although both polymers render non-fouling proper-
ties to NPs due to their respective hydrophilic natures, they
are quite different in their physical and chemical proper-
ties, leading to different effects from NP stability to protein
protection. PEG is amphiphilic in nature and is robust for
applications since it can dissolve in a wide range of polar
and non-polar solvents. However, the amphiphilic nature
of PEG will alter the bioactivity of PEGylated proteins and
the stability of assembled PEGylated polymeric particles
and liposomes. Some of these PEG issues may be over-
come, for example, by inserting cholesterol into PEGylated
liposomes to rescue their stability lost or using a cyropro-
tectant to maintain the stability of PLGA—PEG NPs during
lyophilization. PCB, in contrast, solves all these PEG issues
due to its super-hydrophilic nature. Although PCB and PEG
are used to illustrate the difference between zwitterionic
and non-ionic polymers, general conclusions are applicable
to other zwitterionic materials such as poly(sulfobetaine)
and poly(phosphobetaine), and other non-ionic polymers.
Among zwitterionic materials, PCB is unique in several
aspects desirable for medical applications. PCB is derived
from naturally occurring betaines abundant in the ani-
mal kingdom including humans as well as plants [66]. It
is generally considered to be biocompatible. For exam-
ple, results show that PCB-protected liposome or PLGA
NPs have no observable cytotoxicity even at high con-
centrations [11,95]. When PCB-protected liposome with
doxorubicin loaded was injected intravenously to mice at
the same recommended dose as DOXIL® , no severe tox-
icity was found such as >15% weight loss, symptom of
illness or death [95]. PCB is not only non-fouling, but
also functionalizable for the convenient immobilization
of bio-recognition elements for targeted drug delivery
or diagnostics via conventional coupling chemistry such
as 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and N-
hydroxysuccinimide (EDC/NHS) [11,67,68,102]. After func-
tionalization, unreacted moieties will be hydrolyzed back to
the zwitterionic CB structure, enabling the excellent non-
fouling properties of post-functionalized surfaces [67,70].
PCB can also be prepared as its hydrolysable cationic precur-
sor, which not only alters its solubility for use in the synthesis
of block copolymers with a sharp contrast in hydrophobicity,
but also to condense and release genes [103,104]. The non-
fouling, biocompatible, super-hydrophilic, functionalizable,
411
and hydrolysable properties make PCB and its derivatives a
unique class of materials for constructing next-generation
NPs for medical applications.
Acknowledgments
This work has been supported by the Office of Naval Research
(N000140910137, N000141010600, and N000141210441),
National Science Foundation (DMR-1005699 and CBET
0854298), Defense Threat Reduction Agency (HDTRA1-10-1-
0074), and Boeing-Roundhill Professorship.
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Zwitterionic and non-ionic polymers for stealth nanoparticles
Shaoyi Jiang is the Boeing-Roundhill Professor
of Chemical Engineering and Adjunct Profes-
sor of Bioengineering at the University of
Washington, Seattle. He received his Ph.D.
degree in chemical engineering from Cornell
University in 1993. He was a postdoctoral fel-
low at the University of California, Berkeley
between 1993 and 1994 and a research fellow
at California Institute of Technology between
1994 and 1996 both in chemistry. Currently, he
serves as a Senior Editor for Langmuir. He is a
Fellow of the American Institute of Chemical Engineering, a Fellow
of the American Institute of Medical and Biological Engineering and
a Member of the Washington State Academy of Sciences. His current
research focuses on the molecular understanding, design and devel-
opment of zwitterionic-based functional materials for biomedical
and engineering applications.
413
Zhiqiang Cao is currently a research fellow
in the David H. Koch Institute for Integrative
Cancer Research at Massachusetts Institute
of Technology, and the Department of Anes-
thesiology at Children’s Hospital Boston and
Harvard Medical School, under the direction
of Prof. Robert Langer. He received his Ph.D.
in Chemical Engineering from the University
of Washington in 2011 under the guidance
of Prof. Shaoyi Jiang. He received his B.Eng.
in polymer materials and engineering and
M.Eng. in biomedical engineering from Tianjin University, China
in 2004 and 2007, respectively. He will be an assistant profes-
sor in the Department of Chemical Engineering and Materials
Science at Wayne State University starting January 2013. His cur-
rent research is focused on zwitterionic materials and diabetes
research.