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Contents
1. An Introduction to Cubosomes: Self-Assembly of Lipids and Surfactants 2
2. Applications of Cubosomes in Nanotechnology 5
3. Preparation and Characterization of Cubosomes 14
3.1 Preparation of cubosomes 14
3.2 Characterization of cubosomes 15
4. Agents for the Stabilization of Cubosomes 15
4.1 Classes of steric stabilizers for cubosome dispersions 18
5. Future Developments in the Stabilization of Cubosomes 43
6. Conclusion 46
Acknowledgments 46
References 47
Abstract
Lyotropic liquid crystalline nanostructured particles, such as cubosomes, have grown in
popularity as drug delivery systems in the last few years. These systems require steric
stabilizers to maintain colloidal stability in an aqueous medium, with PluronicF127,
a block copolymer, being the most commonly employed stabilizer. However, in recent
years, alternative, more effective stabilizers, as well as rationally designed systems with
opportunities for further biofunctionalization have been reported. The purpose of this
chapter is to collate and collectively interpret studies in the field of steric stabilization of
this important emerging class of nanoparticles for drug and medical imaging agent
delivery.
Figure 1 Minimal surfaces of the Schoen gyroid (G), Schwartz diamond (D), and
Schwartz primitive (P). Images were generated using Mathematica V.9, based on the
equations obtained from [19].
Figure 2 An illustration of the main types of lyotropic liquid crystal phases depending
on the interface curvature (molecular shape or concentration in water). The mesophases
are denoted as I1, I2 (discrete micellar cubic phase), H1, H2 (hexagonal phase), V1, V2
(bicontinuous cubic phase), and L (lamellar phase).
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whereby CPP < 1. This conical geometry consequently results in the forma-
tion of spheres and cylindrical rods.
In contrast, type 2 mesophases, which include the reverse discontinuous
micellar cubic, reverse hexagonal, and reverse bicontinuous cubic phases,
are composed of surfactants or amphiphilic lipids with an inverse cone shape
space (Fig. 2), whereby CPP > 1. This geometry results in the formation of
inverse spheres and cylindrical rods (Fig. 2). Factors effecting v (volume of
the hydrophobic tail(s)), a (polar headgroup area), and l (length of the hydro-
phobic chain of the surfactant) are summarized by Malmsten [25].
In lyotropic liquid crystal phases, the solvent concentration is a variable
that will partly dictate the self-assembly behavior [26]. In contrast, thermo-
tropic liquid crystal phase transitions are only dependent on temperature and
pressure [26]. The order/sequence of self-assembly phases that are typically
observed as the surfactant/lipid concentration increases in an amphiphilic
lipid/water system is micelles, micellar cubic, hexagonal, bicontinuous
cubic, lamellar, and their respective reverse phases, as illustrated in Fig. 2
[2729].
Materials which have been reported to form cubic phase systems have
been listed by Fontell in 1990 [30]. These materials include anionic and cat-
ionic soaps, zwitterionic and nonionic surfactants, and amphiphilic lipids
of biological origin, such as monoglycerides, sphingolipids, and phospho-
lipids, and also galactolipids, glycolipids, and tetra ether lipids [30]. Since
then reviews have reported that inverse bicontinuous cubic phases have
been observed for many different types of lipids, including mono-
acylglycerides, glycolipids, urea, and urea-like amphiphiles and mono-
ethanolamides [3133]. The amphiphilic lipids most commonly used in
lipid lyotropic liquid crystal research have been glyceryl monooleate
(GMO), a food emulsifier, and phytantriol, a cosmetic ingredient (Fig. 3)
due to their low cost, ease of availability, and potential biocompatibility
based on their history of use in other fields.
2. APPLICATIONS OF CUBOSOMES IN
NANOTECHNOLOGY
The aforementioned lyotropic liquid crystal systems are thermody-
namically stable and for the case of the reversed phases or lamellar phase
can be dispersed into smaller particles that retain the complex internal nano-
structure in the presence of a stabilizer. Dispersions of these bulk parent
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Figure 3 The chemical structures of amphiphilic lipids: (A) phytantriol and (B) glycerol
monoleate.
phases have been given the suffix -osome. For example, dispersions from
the lamellar, hexagonal, and cubic phases are known as liposomes,
hexosomes, and cubosomes, respectively. Liposomes have been exten-
sively used as drug delivery vehicles with 12 clinically approved liposomal
drug formulations currently on the market and 22 liposomal drugs undergo-
ing clinical trials [34,35]. Particles based on other lyotropic liquid crystalline
structures such as cubosomes (inverse bicontinuous cubic phase, Fig. 4) and
hexosomes (inverse hexagonal phase) are also being developed as potential
drug delivery systems [37,38].
The key advantages of these nanostructured particles compared to lipo-
somes include their ordered 3D mesoporous internal structure with poten-
tial for controllable release and their increased lipid volume fraction per
particle, which provides a large lipophilic area for containing poorly
water-soluble lipophilic therapeutics [39,40]. The lyotropic liquid crystal-
line structure and dimensions of the phase, specifically the water channels,
determine the release rate of drugs from within the lyotropic liquid crystal
phase [41,42]. The phases can accommodate molecules of varying proper-
ties [43,44]. A recent study by Zabara and Mezzenga reported the controlled
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Herbal extracts (obtained from Poria cocos, Glyceryl monooleate PluronicF127 Skin (for hair [51]
Thuja orientalis, Espinosilla, Lycium chinense Mill, regrowth)
Coix lacryma-jobi, and Polygonum multiflorum
Thunberg)
Alpha lipoic acid Myverol 18-99 PluronicF127 Skin [52]
Saponin adjuvant Quil A and Phytantriol Pluronic F127 In vitro (skin) [53]
monophosphoryl lipid A
Triclosan Glyceryl monooleate PluronicF127 In vitro (skin) [54]
KIOM-MA-128 (water-soluble extract) Glyceryl monooleate Pluronic F127 In vitro (skin) [55]
Houttuynia cordata (water-soluble extract) Glyceryl monooleate Pluronic F127 In vitro (skin) [56]
Tacrolimus Glyceryl monooleate Pluronic F127 Intradermal [57]
Clotrimazole Glyceryl monooleate PluronicF127 Mucosal [58]
Dexamethasone Glyceryl monooleate PluronicF127 Ocular [59]
Flurbiprofen Glyceryl monooleate Pluronic F127 Ocular [60]
Cyclosporine A Glyceryl monooleate Pluronic F127 Ocular [61]
5-FC oleyl carbamate (pro-drug) 5-FC oleyl carbamate Pluronic F127 Oral [62]
Curcumin/piperine Phytantriol Pluronic F127 Oral [63]
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Amphotericin B Phytantriol Pluronic F127 Oral [64,65]
Cinnarizine Phytantriol or glyceryl Pluronic F127 Oral [66]
monooleate
Cyclosporine A Glyceryl monooleate PluronicF127 Oral [67]
Ibuprofen Phytantriol Pluronic F127 Oral [68]
Insulin Glyceryl monooleate Pluronic F127 Oral [69]
20(S)-protopanaxadiol/piperine Glyceryl monooleate Pluronic F127 Oral [70,71]
Simvastatin Glyceryl monooleate PluronicF127 Oral [72]
Omapatrilat Glyceryl monooleate Pluronic F127 Oral [73]
Omapatrilat Glyceryl monooleate Pluronic F68 Oral [73]
Coenzyme Q10 Glyceryl monooleate PluronicF127 Oral [74]
Coenzyme Q10 Glyceryl monooleate Pluronic F108 Oral [74]
Coenzyme Q10 Glyceryl monooleate Pluronic F68 Oral [74]
Continued
Table 1 A table listing examples of the therapeutics and lipid composition of drug-loaded cubosomes as drug nanocarrierscont'd
Administration
Bioactive molecule (peptide, drug) Matrix constituent Stabilizer route (if applicable) References
Coenzyme Q10 Phytantriol PluronicF127 Oral [74]
Coenzyme Q10 Phytantriol PluronicF108 Oral [74]
Coenzyme Q10 Phytantriol Pluronic F68 Oral [74]
Paclitaxel Soy Polysorbate 80 Oral [75]
phosphatidylcholine/
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glycerol dioleate
Baicalin/KiOM-C Glyceryl monooleate PluronicF127 In vitro (small [76]
intestine
adsorption)
S-164 (water-soluble extract) Glyceryl monooleate PluronicF127 In vitro (small [77]
intestine
adsorption)
Odorranalectin/streptavidin Glyceryl monooleate PluronicF127 Intranasal [78]
Indomethacin Glyceryl monooleate Pluronic F127 Percutaneous [79]
Bromocriptine Glyceryl monooleate Pluronic F127 Intraperitoneal [80]
Adjuvants imiquimod and monophosphoryl Phytantriol PluronicF127 Intravenous [81]
lipid A
Fluorescein isothiocyanate-ovalbumin/Quil Phytantriol PluronicF127 Intravenous [82]
A
Paclitaxel Glyceryl monooleate PluronicF127/ Intravenous [83]
mPEG2kDSPE
Propofol Soy Polysorbate 80 Intravenous [84]
phosphatidylcholine/
glycerol dioleate
Somatostatin Soy Polysorbate 80 Intravenous [85]
phosphatidylcholine/
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glycerol dioleate
Earthworm fibrinolytic enzyme (protein) Glyceryl monooleate/ PluronicF127 Intratympanic [86]
propylene glycol
Ovalbumin Phytantriol or glyceryl PluronicF127 [87]
monooleate
-Chymotrypsinogen A (protein) Glyceryl monooleate MO-PEG2000 [88]
[poly(ethylene
glycol) monooleate]
Carrier-free human recombinant brain- Glyceryl monooleate/ D--Tocopherol [89]
derived neurotrophic factor eicosapentaenoic acid poly(ethylene
glycol) 1000
succinate (V1000)
Annexin V (protein) Phytantriol PluronicF127 [90]
Curcumin Glyceryl monooleate Pluronic F127 [91]
Continued
Table 1 A table listing examples of the therapeutics and lipid composition of drug-loaded cubosomes as drug nanocarrierscont'd
Administration
Bioactive molecule (peptide, drug) Matrix constituent Stabilizer route (if applicable) References
Quercetin Glyceryl monooleate PluronicF108 [92]
Camptothecin Glyceryl monooleate PluronicF108 [93]
and folic acid
Dacarbazine Glyceryl monooleate PluronicF127 [9496]
Carbamazepine (CBZ), coenzyme Q10 Glyceryl monooleate Pluronic F127 [97]
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(CoQ10), cholesterol (Chl, sterol),
phytosterols (PSs, plant sterols)
Diazepam, griseofulvin, propofol, rifampicin Myverol 18-99 PluronicF127 [98]
50-Deoxy-5-fluoro-N4- 50- PluronicF127 [99]
(phytanyloxycarbonyl) cytidine (phytanyl pro- Deoxy-5-fluoro-N4-
drug analogue of capecitabine) (phytanyloxycarbonyl)
cytidine
Hydrocortisone Phytantriol PluronicF127 [39,100]
Atropine Phytantriol Pluronic F127 [39,100]
Transretinol Phytantriol Pluronic F127 [39,100]
Diazepam Phytantriol PluronicF127 [39,100]
Prednisolone Phytantriol Pluronic F127 [39,100]
Dexamethasone Phytantriol Pluronic F127 [39,100]
Progesterone Phytantriol PluronicF127 [39,100]
Haloperidol Phytantriol Pluronic F127 [39,100]
Levofloxacin Phytantriol Pluronic F127 [39,100]
Indometacin Phytantriol PluronicF127 [39,100]
Hydrocortisone Myverol 18-99K Pluronic F127 [39,100]
Atropine Myverol 18-99K Pluronic F127 [39,100]
PluronicF127
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Transretinol Myverol 18-99K [39,100]
Diazepam Myverol 18-99K Pluronic F127 [39,100]
Prednisolone Myverol 18-99K Pluronic F127 [39,100]
Dexamethasone Myverol 18-99K Pluronic F127 [39,100]
Progesterone Myverol 18-99K Pluronic F127 [39,100]
Haloperidol Myverol 18-99K PluronicF127 [39,100]
Levofloxacin Myverol 18-99K Pluronic F127 [39,100]
Indometacin Myverol 18-99K Pluronic F127 [39,100]
DOPURu (amphiphilic ruthenium-based 1,2-Dioleoyl-sn- [101]
molecule) glycero-3-
phosphocholine
(DOPC) and 1,2-
dioleoyl-sn-glycero-3-
phosphoethanolamine
(DOPE)
ARTICLE IN PRESS
Recent reviews by Rizwan et al. [104] and Conn and Drummond [33]
have collected examples where lyotropic liquid crystalline nanostructured
particles accommodate biologically active molecules such as vitamins,
enzymes, and other proteins, as well as crystallizing membrane proteins,
which have important application for membrane protein crystallization, bio-
sensors, biofuel applications, as well as in drug delivery. Owing to the high-
surface area of the internal mesophase structure (up to 400 m2/g) [105], the
cubic phase can be used to incorporate these biologically active molecules
(e.g., globular proteins), which have similar dimensions to the water chan-
nels in the bicontinuous cubic phases [89].
In other recent developments, cubosomes are also being investigated for
the containment of contrast agents for medical imaging applications
[106108], and capabilities as a cell-free bio-sensing platform [109]. Apart
from drug delivery and biomedical applications, the use and application
of lyotropic liquid crystalline nanostructured particles is also relevant within
the food industry (e.g., solubilization of food bioactives within lyotropic liq-
uid crystalline mesophases) [110,111] and agriculture industry (e.g., delivery
of plant agrochemicals) [112]. Therefore, any research into the colloidal
stability and retention of internal structure of nanostructured particles is
relevant to several research fields.
Figure 5 An illustration of some of the factors affecting steric hindrance between nano-
structured particles: (A) the concentration of steric stabilizer used and (B) PEG length in
steric stabilizer.
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PluronicF127 Phytantriol/dipalmitoyl phosphatidylserine (DPPS) Pn3m (Q224) or [154]
Im3m (Q229)
PluronicF127 1-O-(5,9,13,17-tetramethyloctadecanoyl)erythritol Pn3m (Q224) and [161]
(EROCO C22) Im3m (Q229)
PluronicF127 1-O-(5,9,13,17-tetramethyloctadecyl)--D- Pn3m (Q224) and [161163]
xylopyranoside (-XP) Im3m (Q229)
PluronicF127 glycolipid 1-O-phytanyl--D-xyloside (-XP) Pn3m (Q224) and [163]
Im3m (Q229)
PluronicF127 5-FC oleyl carbamate Pn3m (Q224) and [62]
Ia3d (Q230)
PluronicF127 50-Deoxy-5-fluoro-N4-(phytanyloxycarbonyl) Pn3m (Q224) and [99]
cytidine Ia3d (Q230)
PluronicF127 Monolinolein (MLO) Pn3m (Q224) at [164]
25 C
Continued
Table 2 Amphiphilic block copolymers used as steric stabilizers for cubosomes reported in the general literaturecont'd
Space group of inner
Stabilizer Lipid matrix constituent structure References
PluronicF127 Monolinolein (MLO)/oil Pn3m (Q224) [165168]
Fd3m (Q227) [165168]
224
Pluronic F127 Monolinolein (MLO)/diglycerol monooleate Pn3m (Q ) or [169]
(DGMO) or soybean PC/oil Im3m (Q229)
PluronicF127 Monoelaidin Im3m (Q229) [170]
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PluronicF127 Myverol 18-99K Pn3m (Q224) or [39,98,108,152,171]
Im3m (Q229)
PluronicF127 RYLO MG 19 Pn3m (Q224) and [96,171]
Im3m (Q229)
PluronicF127 Glyceryl monooleate Pn3m (Q224) or [48,49,51,54,55,
Im3m (Q229) 5761,66,67,
6973,7680,87,
91,9497,102,111,
119,152,155,159,
160,170,172188]
PluronicF127 Glyceryl monooleate/propylene glycol Pn3m (Q224) and [86]
Im3m (Q229)
PluronicF127 Glyceryl monooleate/soya phospholipids Im3m (Q229) [189]
229
Pluronic F127 Glyceryl monooleate/oil Im3m (Q ) [190]
Fd3m (Q227) [190]
PluronicF127 Glyceryl monooleate/1-glycerol monooleyl ether Pn3m (Q224) and [191]
(GME) Im3m (Q229)
PluronicF127 Dimodan U/J (96% monoglycerides: 62% linoleate Pn3m (Q224) and [192]
and 25% oleate)/tetradecane (oil) Im3m (Q229)
PluronicF127 Dimodan U/J (96% monoglycerides: 62% linoleate Pn3m (Q224) and [193195]
and 25% oleate) Im3m (Q229)
PluronicF127/mPEG2kDSPE Im3m (Q229)
ARTICLE IN PRESS
Glyceryl monooleate [83]
PluronicF127/mPEG350DSPE Phytantriol Pn3m (Q224), Im3m [196]
(Q229)
PluronicF127/mPEG750DSPE Phytantriol Pn3m (Q224) and [196]
Im3m (Q229)
PluronicF127/mPEG2kDSPE Phytantriol Pn3m (Q224) and [196]
Im3m (Q229)
PluronicF127/-casein Phytantriol Pn3m (Q224) [152]
mixture
PluronicF108 Phytantriol Pn3m (Q224) [118,160]
224
Pluronic F108 Glyceryl monooleate Pn3m (Q ) [92,93,160]
224
Pluronic F87 Phytantriol Pn3m (Q ) [160]
PluronicF87 Glyceryl monooleate Im3m (Q229) [160]
224
Pluronic F68 Phytantriol Pn3m (Q ) [160]
Continued
Table 2 Amphiphilic block copolymers used as steric stabilizers for cubosomes reported in the general literaturecont'd
Space group of inner
Stabilizer Lipid matrix constituent structure References
PluronicF68 Glyceryl monooleate Cubosome [73,160]
PluronicF68 Myverol 18-99K Cubosome [113]
229
Pluronic P123 Phytantriol Im3m (Q ) [160]
224
Pluronic P105 Phytantriol Pn3m (Q ) [160]
PluronicP105 Glyceryl monooleate Im3m (Q229) [160]
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229
Pluronic P104 Phytantriol Im3m (Q ) [160]
229
Pluronic P104 Glyceryl monooleate Im3m (Q ) [160]
224
Pluronic P84 Phytantriol Pn3m (Q ) and [160]
Im3m (Q229)
Polaxamine
Poloxamine 908 Myverol 18-99K Cubosome [113]
Poloxamine 908/ Myverol 18-99K Cubosome [113]
PluronicF127 combinations
Table 3 PEGylated lipid copolymers used as steric stabilizers for cubosomes reported in the general literature
PEG PEG Space group of inner
Stabilizer MW units Lipid matrix constituent structure References
PEGylated lipid
1,2-Dimyristoyl-sn- 550 12 Dielaidoylphosphatidylethanola Im3m (Q229) [197,198]
glycero-3-phosphoethanolamine-N- mine (DEPE)
PEG (DMPE-PEG550)
PEGylated monoolein (MO-PEG660) 660 15 1,2- Ia3d (Q230) and Pn3m [199]
ARTICLE IN PRESS
Dioleoylphosphatidylethanolami (Q224)
ne (DOPE)
1,2-Distearorylphosphatidylethanol 750 17 1,2- Cubosome [200]
amine-PEG (DSPE-PEG750) Dioleoylphosphatidylethanolami
ne (DOPE)
Polyoxyethylene (20) sorbitan 900 20 Myverol 18-99K Cubosome [113]
monopalmitate (Tween40)
Polyoxyethylene (20) sorbitan 900 20 Glyceryl monooleate Cubosome [102]
monooleate (Tween80)
Polyoxyethylene (20) sorbitan 900 20 Soy phosphatidylcholine/ Cubosome [75,84,85]
monooleate (Tween80) glycerol dioleate
Polyoxyethylene (20) sorbitan 900 20 Soy PE (L-- Cubosome [38]
monooleate (Tween80) phosphatidylethanolamine)
D-alpha-Tocopheryl PEO1000 succinate 1000 22 Phytantriol Im3m (Q229) [38]
(vitamin E TPGS)
Continued
Table 3 PEGylated lipid copolymers used as steric stabilizers for cubosomes reported in the general literaturecont'd
PEG PEG Space group of inner
Stabilizer MW units Lipid matrix constituent structure References
229
PEG1K20PHYT30 1000 22 Phytantriol Im3m (Q ) [201]
PEG1K25PHYT25 1000 22 Phytantriol Im3m (Q229) [201]
224
PEG1K30PHYT20 1000 22 Phytantriol Pn3m (Q ) and Im3m [201]
(Q229)
PEG1K40PHYT10 1000 22 Phytantriol Pn3m (Q224) and Im3m [201]
(Q229)
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1,3-Didodecyloxy-propane-2-ol-PEG 1300 30 GMO (RYLO MG 90) Ia3d (Q230) [202]
(DDP(EO)30)
PEG-40-stearate 1800 40 Phytantriol Im3m (Q229) [203]
1,2-Dioleoylphosphatidylethanolami 2000 45 Glyceryl monooleate and Cubosome [204]
ne-PEG (DOPE-PEG) cis-5,8,11,14,17-
eicosapentaenoic
acid (20:5, EPA)
PEGylated monoolein 2000 45 Glyceryl monooleate Pn3m (Q224) or Im3m [88]
(MO-PEG2000) (Q229)
1,2-Distearorylphosphatidylethanol 2000 45 1,2- Cubosome [200,205]
amine-PEG (DSPE-PEG2000) Dioleoylphosphatidylethanolami
ne (DOPE)
1,2-distearoylphosphatidylethanol 2000 45 Soy phosphatidyl choline (SPC) Cubosome [205]
amine-PEG (DSPE-PEG2000) and glycerol dioleate (GDO)
PEG-45-stearate 2000 45 Phytantriol Im3m (Q229) [203]
229
PEG2K10PHYT40 2000 45 Phytantriol Im3m (Q ) [201]
PEG2K20PHYT30 2000 45 Phytantriol Pn3m (Q224) and Im3m [201]
(Q229)
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PEG2K25PHYT25 2000 45 Phytantriol Im3m (Q229) [201]
229
PEG2K30PHYT20 2000 45 Phytantriol Im3m (Q ) [201]
PEG2K40PHYT10 2000 45 Phytantriol Im3m (Q229) [201]
229
PEG-50-stearate 2200 50 Phytantriol Im3m (Q ) [203]
229
1,3-Didodecyloxy-propane-2-ol-PEG 2300 52 GMO (RYLO MG 90) Im3m (Q ) and Ia3d [202,206]
(DDP(EO)52) (Q230) (coexisting with L3
phase)
PEG-55-stearate 2400 55 Phytantriol Im3m (Q229) [203]
PEG3K10PHYT40 3000 68 Phytantriol Pn3m (Q224) and Im3m [201]
(Q229)
PEG3K20PHYT30 3000 68 Phytantriol Pn3m (Q224) [201]
224
PEG3K25PHYT25 3000 68 Phytantriol Pn3m (Q ) [201]
PEG3K30PHYT20 3000 68 Phytantriol Pn3m (Q224) [201]
Continued
Table 3 PEGylated lipid copolymers used as steric stabilizers for cubosomes reported in the general literaturecont'd
PEG PEG Space group of inner
Stabilizer MW units Lipid matrix constituent structure References
224
PEG3K40PHYT10 3000 68 Phytantriol Pn3m (Q ) [201]
PEG4K10PHYT40 4000 90 Phytantriol Pn3m (Q224) [201]
224
PEG4K20PHYT30 4000 90 Phytantriol Pn3m (Q ) [201]
224
PEG4K25PHYT25 4000 90 Phytantriol Pn3m (Q ) [201]
PEG4K30PHYT20 4000 90 Phytantriol Pn3m (Q224) [201]
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224
PEG4K40PHYT10 4000 90 Phytantriol Pn3m (Q ) [201]
229
1,3-Didodecyloxy-propane-2-ol-PEG 4100 92 GMO (RYLO MG 90) Im3m (Q ) and Ia3d [202,206]
(DDP(EO)92) (Q230) (coexisting
with L3 phase)
PEG-100-stearate (Myrj59) 4400 100 Phytantriol Pn3m (Q224) [118,203]
229
1,3-Didodecyloxy-2-glycidyl-glycerol- 5000 114 GMO (RYLO MG 90) Im3m (Q ) and Ia3d [202,206]
PEG (Q230) (coexisting
(DDGG4-(EO)114) with L3 phase)
PEG6K10PHYT40 6000 136 Phytantriol Pn3m (Q224) [201]
224
PEG6K20PHYT30 6000 136 Phytantriol Pn3m (Q ) [201]
PEG6K25PHYT25 6000 136 Phytantriol Pn3m (Q224) [201]
224
PEG6K30PHYT20 6000 136 Phytantriol Pn3m (Q ) [201]
224
PEG6K40PHYT10 6000 136 Phytantriol Pn3m (Q ) [201]
1,3-Didodecyloxy-2-glycidyl-glycerol- 6000 136 GMO (RYLO MG 90) Im3m (Q229) and Ia3d [202,206]
PEG-1,3-didodecyloxy-2- (Q230) (coexisting
glycidyl-glycerol (DDGG2-(EO)136- with L3 phase)
DDGG2)
PEG-150-stearate 6600 150 Phytantriol Pn3m (Q224) [118]
224
PEG8K10PHYT40 8000 181 Phytantriol Pn3m (Q ) [201]
224
ARTICLE IN PRESS
PEG8K20PHYT30 8000 181 Phytantriol Pn3m (Q ) [201]
PEG8K25PHYT25 8000 181 Phytantriol Pn3m (Q224) [201]
224
PEG8K30PHYT20 8000 181 Phytantriol Pn3m (Q ) [201]
224
PEG8K40PHYT10 8000 181 Phytantriol Pn3m (Q ) [201]
224
PEG10K10PHYT40 10,000 227 Phytantriol Pn3m (Q ) [201]
224
PEG10K20PHYT30 10,000 227 Phytantriol Pn3m (Q ) [201]
PEG10K25PHYT25 10,000 227 Phytantriol Pn3m (Q224) [201]
224
PEG10K30PHYT20 10,000 227 Phytantriol Pn3m (Q ) [201]
224
PEG10K40PHYT10 10,000 227 Phytantriol Pn3m (Q ) [201]
PEG14K10PHYT40 14,000 317 Phytantriol Pn3m (Q224) [201]
224
PEG14K20PHYT30 14,000 317 Phytantriol Pn3m (Q ) [201]
Continued
Table 3 PEGylated lipid copolymers used as steric stabilizers for cubosomes reported in the general literaturecont'd
PEG PEG Space group of inner
Stabilizer MW units Lipid matrix constituent structure References
224
PEG14K25PHYT25 14,000 317 Phytantriol Pn3m (Q ) [201]
PEG14K30PHYT20 14,000 317 Phytantriol Pn3m (Q224) [201]
224
PEG14K40PHYT10 14,000 317 Phytantriol Pn3m (Q ) [201]
224
P(ODA)6-b-P(PEGA-OMe)27 Phytantriol Pn3m (Q ) [207]
Glyceryl monooleate Im3m (Q229) [207]
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224
P(ODA)6-b-P(PEGA-OMe)35 Phytantriol Pn3m (Q ) [207]
229
Glyceryl monooleate Im3m (Q ) [207]
224
P(ODA)6-b-P(PEGA-OMe)39 Phytantriol Pn3m (Q ) [207]
229
Glyceryl monooleate Im3m (Q ) [207]
P(ODA)10-b-P(PEGA-OMe)23 Phytantriol Pn3m (Q224) [207]
229
Glyceryl monooleate Im3m (Q ) [207]
224
P(ODA)10-b-P(PEGA-OMe)31 Phytantriol Pn3m (Q ) [207]
Glyceryl monooleate Im3m (Q229) [207]
224
P(ODA)10-b-P(PEGA-OMe)34 Phytantriol Pn3m (Q ) [207]
229
Glyceryl monooleate Im3m (Q ) [207]
PEG-based copolymers bearing lipid-mimetic anchors Glyceryl monooleate/sodium Cubosome (coexisting with [208]
cholate L3 phase)
Table 4 Additional miscellaneous steric stabilizers for cubosomes reported in the general literature
Space group of
Stabilizer Lipid matrix constituent inner structure References
Casein
-Casein Glyceryl monooleate Pn3m (Q224) [152]
Casein Myverol 18-99K Cubosome [113]
Albumin
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Albumin Myverol 18-99K Cubosome [113]
Lecithin
Partially hydrolyzed emulsifier lecithin Dimodan U/J (96% monoglycerides: 62% linoleate Im3m (Q229) [195]
(EmultopEP) and 25% oleate)
Modified cellulose
Hydroxypropyl methyl cellulose Glyceryl monooleate Pn3m (Q224) [161]
acetate succinate (HPMCAS)
Hydroxypropyl methyl cellulose 1-O-(5,9,13,17-tetramethyloctadecanoyl)erythritol Pn3m (Q224) [161]
acetate succinate (HPMCAS) (EROCO C22)
Hydroxypropyl methyl cellulose 1-O-(5,9,13,17-tetramethyloctadecyl)--D-xylopyranoside Pn3m (Q224) [161]
acetate succinate (HPMCAS) (-XP)
Modified starch
HI-CAP100 (hydrophobically Glyceryl monooleate Cubosome [209]
modified with octenyl succinate
groups)
Continued
Table 4 Additional miscellaneous steric stabilizers for cubosomes reported in the general literaturecont'd
Space group of
Stabilizer Lipid matrix constituent inner structure References
CAPSUL-E (hydrophobically Glyceryl monooleate Cubosome [209]
modified with octenyl succinate
groups)
Dextran Glyceryl monooleate Cubosome [209]
Laponite
Laponite XLG Phytantriol Pn3m (Q224) [151]
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224
Laponite XLG Dimodan U/J (96% monoglycerides: 62% linoleate Pn3m (Q ) [193]
and 25% oleate)
Laponite XLG Dimodan U/J (96% monoglycerides: 62% linoleate Pn3m (Q224) [192]
and 25% oleate)/tetradecane (oil)
Silica nanoparticles
4.1.1.1 Poloxamer
4.1.1.1.1 Poloxamer 407/PluronicF127 By far the most widely
and frequently used steric stabilizer for cubosomes is Poloxamer 407 (also
known as PluronicF127), a nonionic triblock copolymer composed of
PEG and polypropylene oxide (PPO): PEG100PPO65PEG100, with a molec-
ular weight of approximately 12,600 Da (Figs. 6 and 7). PluronicF127 is a
nonionic macromolecule that is used widely in pharmaceutical formulations
and personal care products. In lyotropic liquid crystalline dispersions,
PluronicF127 acts as a steric stabilizer through the incorporation or adsorp-
tion of its hydrophobic PPO block onto the surface of the nanostructured
particle. Whilst the PPO domain/block acts as an anchor to the particle,
the hydrophilic PEG chains extend to cover the surface, providing steric
shielding and stabilizing the colloidal particles in aqueous solutions [211].
PluronicF127 has been employed to stabilize cubosome dispersions in
various lipid systems, including GMO, glycerol monolinoleate, and
phytantriol. The GMO system has been the most extensively studied. At
low stabilizer concentrations (<4%, w/w, vs. GMO), PluronicF127 stabi-
lized GMO dispersions form Q224 cubosomes with Pn3m space group sym-
metry, whilst at higher stabilizer concentrations (i.e., 7.4 or 10%, w/w, vs.
GMO), Q229 cubosomes with Im3m space group symmetry are formed
[173]. Although using low stabilizer concentrations of PluronicF127 can
A B
4000 L121 P123 F127 4000 L121 P123 F127
L101 P103 P104 P105 F108 L101 P103 P104 P105 F108
Hydrophobe (MW of PPO)
L92 L92
L43 L43
Figure 6 Lyotropic liquid crystalline phases obtained from (A) phytantriol and
(B) monoolein dispersions using the Poloxamers/Pluronic surfactants. Image
reproduced from [160] with permission from The Royal Society of Chemistry.
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Figure 7 The chemical structures of stabilizers Pluronic and Tetronic. The blue (gray
in the print version) shading indicates the hydrophilic domain, whilst the yellow (light
gray in the print version) shading indicates the hydrophobic domain. A graphic illustra-
tion of the stabilizer structure is shown on the left hand side, with the dashed line rep-
resentative of the surface of a nanostructured particle.
4.1.1.2 Poloxamine
4.1.1.2.1 Poloxamine 908/Tetronic908 Poloxamine 908, also
known as Tetronic908, is a tetrafunctional PEGPPO ethylenediamine
block copolymer (Fig. 7). Boyd et al. reported stabilizing aqueous dispersions
of Myverol 18-99K in Poloxamine 908 solution [113]. Aqueous
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Figure 8 The chemical structures of some of the PEGylated lipid stabilizers used in the
literature for the preparation of cubosome dispersions. The blue (gray in the print ver-
sion) shading indicates the hydrophilic domain and the yellow (light gray in the print
version) shading indicates the hydrophobic domain. A graphic illustration of the stabi-
lizer structure is shown on the left hand side, with the dashed line representative of the
nanostructured particle's surface.
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[197,198]. Cubosomes were identified with cubic phase Q229, with Im3m
space group symmetry.
Figure 10 Accelerated stability assay results for (A) F127 (control stabilizer) at 0.3, 0.5,
0.7, 1, and 1.2 wt% stabilizer concentration, (B) PEG20PHYT30 copolymer series, where
PEG MW: 214K, (C) PEG4K-PHYT copolymer (from 20, 25, 30 and 40 PEG mol% copol-
ymer series), and (D) PEG6K-PHYT copolymer (from 20, 30 and 40 PEG mol% copolymer
series). ASA results after first spin 1800 rpm are represented in black columns, whilst ASA
results after second spin 2000 rpm are represented in blue (gray in the print version)
columns. Steric stabilizer concentration for ASA results presented in (B)(D) is 1 wt%,
with control standard steric stabilizer F127 at 1 wt% presented to the right. Image
reproduced with permission from [201].
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A B
600 2
500
Intensity (AU)
400
300 4 6
200 12
10 14
100
Figure 11 (A) SAXS diffraction pattern showing an Im3m space group symmetry and
(B) cryo-TEM image of monoolein dispersion stabilized using reduced P(ODA)10-b-P
(PEGA-OMe)34 copolymer at 25 C. Images reproduced from [207] with permission from
The Royal Society of Chemistry.
starch was mixed threefold higher than the weight of GMO, and the particle
size was 600 nm on average.
4.1.4.6 Nanoparticles
4.1.4.6.1 Laponite XLG (clay nanoparticles) Muller et al. and
Salonen et al. have reported cubic nanostructured particles from aqueous dis-
persions of two lipids: (i) phytantriol [151] and (ii) Dimodan U/J (consisting
of 96% monoglycerides, which contain 62% linoleate and 25% oleate), with
[192] and without [193] tetradecane (oil) in water, using Laponite XLG (clay
nanoparticles) as the steric stabilizer. The cubosomes formed were identified
with cubic phase Q224, with Pn3m space group symmetry.
6. CONCLUSION
We have discussed the stabilization of cubosomes. Due to their small
size and ordered 3D mesoporous internal structure, with a high surface area
for substance loading, cubosomes are being widely investigated as drug
delivery systems, to encapsulate hydrophilic, lipophilic, or amphiphilic ther-
apeutics, and/or imaging agents. However, these bicontinuous cubic
lyotropic liquid crystalline nanostructured particles require the presence
of a stabilizer for their colloidal stability. This review covers stabilizers which
come from amphiphilic block copolymers, PEGylated lipids, designer/cus-
tomized copolymer series, and alternative stabilizers. Although these few
groups of stabilizers have been identified and reported as suitable stabilizers
for these systems, researchers still rely quite heavily on commercially avail-
able stabilizers, such as PluronicF127.
However, advances in polymer synthesis techniques (e.g., RAFT poly-
merization) allow for new polymer structures (e.g., PEGylated brush copol-
ymers) to be investigated for their use as customized steric stabilizers for
cubosome systems. An important molecular design feature for creating an
effective customized stabilizer for cubosomes is having an asymmetric
amphiphilic polymer structure with a larger hydrophilic domain/moiety
(e.g., high HLB value). This can be achieved through the use of longer
PEG chains (e.g., high PEG MW) and/or multiple PEG chains (e.g., brush
or comb-like design). Designing and synthesizing new stabilizers for
lyotropic liquid crystalline nanostructured particles enable these stabilizers
to eventually be functionalized with a targeting moiety. This will enable
active targeting of these systems, which may be important for certain clinical
applications for different therapeutics. Future studies will focus on investi-
gating and screening for more effective stabilizers (e.g., both commercial
and customized) and/or solvent conditions for maintaining cubosome
dispersions.
ACKNOWLEDGMENTS
This work was supported by CSIRO and RMIT funding.
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REFERENCES
[1] V. Luzzati, et al., La structure des collodes dassociation. I. Les phases liquide
cristallines des syste`mes amphiphileeau, Acta Crystallogr. 13 (1960) 660667.
[2] V. Luzzati, P.A. Spegt, Polymorphism of lipids, Nature 215 (1967) 701704.
[3] V. Luzzati, et al., Structure of cubic phases of lipidwater systems, Nature 220 (1968)
485488.
[4] A. Tardieu, V. Luzzati, Polymorphism of lipids: a novel cubic phasea cage-like net-
work of rods with enclosed spherical micelles, Biochim. Biophys. Acta 219 (1970)
1117.
[5] W. Longley, T.J. McIntosh, A bicontinuous tetrahedral structure in a liquid-crystalline
lipid, Nature 303 (1983) 612614.
[6] M. Caffrey, Kinetics and mechanism of transitions involving the lamellar, cubic,
inverted hexagonal and fluid isotropic phases of hydrated monoacylglycerides moni-
tored by time-resolved X-ray diffraction, Biochemistry 26 (1987) 63496363.
[7] P. Mariani, V. Luzzati, H. Delacroix, Cubic phases of lipid-containing systems: struc-
ture analysis and biological implications, J. Mol. Biol. 204 (1988) 165189.
[8] P.J. Maddaford, C. Toprakcioglu, Structure of cubic phases in the ternary system
didodecyldimethylammonium bromide/water/hydrocarbon, Langmuir 9 (1993)
28682878.
[9] O. Diat, G. Porte, J.-F. Berret, Orientation and twins separation in a micellar cubic
crystal under oscillating shear, Phys. Rev. B 54 (1996) 1486914872.
[10] R.R. Balmbra, J.S. Clunie, J.F. Goodman, Cubic mesomorphic phases, Nature
222 (1969) 11591160.
[11] V. Luzzati, et al., Lipid polymorphism: a correction. The structure of the cubic phase of
extinction symbol Fdconsists of two types of disjointed reverse micelles embedded
in a three-dimensional hydrocarbon matrix, Biochemistry 31 (1992) 279285.
[12] P. Mariani, et al., Polymorphism of a lipid extract from Pseudomonas fluorescens:
structure analysis of a hexagonal phase and of a novel cubic phase of extinction symbol
Fd, Biochemistry 29 (1990) 67996810.
[13] J. Charvolin, J.F. Sadoc, Periodic-systems of frustrated fluid films and micellar cubic
structures in liquid crystals, J. Phys. 49 (1988) 521526.
[14] H. Delacroix, T. Gulik-Krzywicki, J.M. Seddon, Freeze fracture electron microscopy
of lyotropic lipid systems: quantitative analysis of the inverse micellar cubic phase of
space group Fd3m (Q227), J. Mol. Biol. 258 (1996) 88103.
[15] G.C. Shearman, et al., A 3-D hexagonal inverse micellar lyotropic phase, J. Am.
Chem. Soc. 131 (2009) 16781679.
[16] H.A. Schwarz, Gesammelte matematische Abhandlung, vol. 1, Springer-Verlag,
Berlin, 1890.
[17] A.H. Schoen, Infinite periodic minimal surfaces without self-intersections: NASA
Technical Note, TD-5541, Springfield, 1970.
[18] L.E. Scriven, Equilibrium bicontinuous structure, Nature 263 (1976) 123125.
[19] J.D. Enlow, Mathematical Modelling of Surfactant Liquid Crystal X-ray Diffraction,
Department of Mathematics and Statistics, University of Otago, New Zealand, 2001,
222.
[20] D. Yang, B. Armitage, S.R. Marder, Cubic liquid-crystalline nanoparticles, Angew.
Chem. Int. Ed. Engl. 43 (2004) 44024409.
[21] B. Angelov, Modulation of nanochannels hydration in lipid cubic phases studied by
SANS and SAXS, in: NSTI-Nanotech, 2007, pp. 58.
[22] B. Angelov, et al., Diamond-type lipid cubic phase with large water channels, J. Am.
Chem. Soc. 125 (2003) 71887189.
ARTICLE IN PRESS
[23] J.N. Israelachvili, D.J. Mitchell, B.W. Ninham, Theory of self-assembly of lipid bila-
yers and vesicles, Biochim. Biophys. Acta 470 (1977) 185201.
[24] V. Luzzati, A. Tardieu, Lipid phasesstructure and structural transitions, Annu. Rev.
Phys. Chem. 25 (1974) 7994.
[25] M. Malmsten, in: Surfactants and Polymers in Drug Delivery. Drugs and the Pharma-
ceutical Sciences, vol. 122, Marcel Dekker, New York, NY, 2002.
[26] A.M. Figueiredo Neto, S.R.A. Salinas, The Physics of Lyotropic Liquid Crystals:
Phase Transitions and Structural Properties, Monographs on the Physics & Chemistry
of Materials, Oxford University Press, New York, NY, 2005.
[27] P.A. Winsor, Binary and multicomponent solutions of amphiphilic compounds. Sol-
ubilization and the formation, structure, and theoretical significance of liquid crystal-
line solutions, Chem. Rev. 68 (1968) 140.
[28] G.J.T. Tiddy, Surfactant-water liquid crystal phases, Phys. Rep. 57 (1980) 146.
[29] J. Charvolin, Crystals of interfacesthe cubic phases of amphiphile water-systems,
J. Geophys. Res. 46 (1985) 173190.
[30] K. Fontell, Cubic phases in surfactant and surfactant-like lipid systems, Colloid Polym.
Sci. 268 (1990) 264285.
[31] C. Fong, T. Le, C.J. Drummond, Lyotropic liquid crystal engineering-ordered nano-
structured small molecule amphiphile self-assembly materials by design, Chem. Soc.
Rev. 41 (2012) 12971322.
[32] T. Kaasgaard, C.J. Drummond, Ordered 2-D and 3-D nanostructured amphiphile self-
assembly materials stable in excess solvent, Phys. Chem. Chem. Phys. 8 (2006)
49574975.
[33] C.E. Conn, C.J. Drummond, Nanostructured bicontinuous cubic lipid self-
assembly materials as matrices for protein encapsulation, Soft Matter 9 (2013)
34493464.
[34] H.-I. Chang, M.-K. Yeh, Clinical development of liposome-based drugs: formulation,
characterization, and therapeutic efficacy, Int. J. Nanomed. 7 (2012) 4960.
[35] J.-X. Zhang, et al., Application of liposomes in drug developmentfocus on gastro-
enterological targets, Int. J. Nanomed. 8 (2013) 13251334.
[36] S.B. Rizwan, et al., Characterisation of bicontinuous cubic liquid crystalline systems of
phytantriol and water using cryo field emission scanning electron microscopy (cryo
FESEM), Micron 38 (2007) 478485.
[37] A. Yaghmur, O. Glatter, Characterization and potential applications of nanostructured
aqueous dispersions, Adv. Colloid Interface Sci. 147148 (2009) 333342.
[38] J. Barauskas, M. Johnsson, F. Tiberg, Self-assembled lipid superstructures: beyond ves-
icles and liposomes, Nano Lett. 5 (2005) 16151619.
[39] X. Mulet, et al., High throughput preparation and characterisation of amphiphilic
nanostructured nanoparticulate drug delivery vehicles, Int. J. Pharm. 395 (2010)
290297.
[40] W.-K. Fong, et al., Controlling the nanostructure of gold nanorod-lyotropic liquid-
crystalline hybrid materials using near-infrared laser irradiation, Langmuir 28 (2012)
1445014460.
[41] K.W.Y. Lee, et al., Nanostructure of liquid crystalline matrix determines in vitro
sustained release and in vivo oral absorption kinetics for hydrophilic model drugs,
Int. J. Pharm. 365 (2009) 190199.
[42] R. Negrini, R. Mezzenga, Diffusion, molecular separation, and drug delivery from
lipid mesophases with tunable water channels, Langmuir 28 (2012) 1645516462.
[43] J. Clogston, et al., Controlling release from the lipidic cubic phase by selective alkyl-
ation, J. Control. Release 102 (2005) 441461.
[44] J. Clogston, M. Caffrey, Controlling release from the lipidic cubic phase. Amino acids,
peptides, proteins and nucleic acids, J. Control. Release 107 (2005) 97111.
ARTICLE IN PRESS
[45] A. Zabara, R. Mezzenga, Modulating the crystal size and morphology of in meso-
crystallized lysozyme by precisely controlling the water channel size of the hosting
mesophase, Soft Matter 9 (2013) 10101014.
[46] R. Mezzenga, et al., Polysaccharide-induced order-to-order transitions in lyotropic
liquid crystals, Langmuir 21 (2005) 61656169.
[47] N.M. Morsi, G.A. Abdelbary, M.A. Ahmed, Silver sulfadiazine based cubosome
hydrogels for topical treatment of burns: development and in vitro/in vivo character-
ization, Eur. J. Pharm. Biopharm. 86 (2014) 178189.
[48] T.K. Kwon, J.-C. Kim, Preparation and in vitro skin permeation of cubosomes con-
taining Hinokitiol, J. Dispers. Sci. Technol. 31 (2010) 10041009.
[49] T. Kwon, et al., In vitro skin permeation of cubosomes containing water soluble
extracts of Korean barberry, Colloid J. 72 (2010) 205210.
[50] T. Rattanapak, et al., Transcutaneous immunization using microneedles and cub-
osomes: mechanistic investigations using Optical Coherence Tomography and
Two-Photon Microscopy, J. Control. Release 172 (2013) 894903.
[51] S.R. Seo, et al., In vivo hair growth-promoting efficacies of herbal extracts and their
cubosomal suspensions, J. Ind. Eng. Chem. 19 (2013) 13311339.
[52] S. Sherif, E.R. Bendas, S. Badawy, The clinical efficacy of cosmeceutical application of
liquid crystalline nanostructured dispersions of alpha lipoic acid as anti-wrinkle, Eur. J.
Pharm. Biopharm. 86 (2014) 251259.
[53] T. Rattanapak, et al., Comparative study of liposomes, transfersomes, ethosomes and
cubosomes for transcutaneous immunisation: characterisation and in vitro skin pene-
tration, J. Pharm. Pharmacol. 64 (2012) 15601569.
[54] T.K. Kwon, S.K. Hong, J.C. Kim, In vitro skin permeation of cubosomes containing
triclosan, J. Ind. Eng. Chem. 18 (2012) 563567.
[55] S.K. Hong, J.Y. Ma, J.-C. Kim, In vitro skin permeation enhancement of KIOM-
MA-128 by monoolein cubosomes, J. Dispers. Sci. Technol. 33 (2012) 15031508.
[56] T.K. Kwon, J.-C. Kim, In vitro skin permeation and anti-atopic efficacy of lipid
nanocarriers containing water soluble extracts of Houttuynia cordata, Drug Dev.
Ind. Pharm. 40 (2014) 13501357.
[57] R.K. Thapa, et al., Preparation, characterization, and release study of Tacrolimus-
loaded liquid crystalline nanoparticles, J. Dispers. Sci. Technol. 34 (2013) 7277.
[58] E. Esposito, et al., Clotrimazole nanoparticle gel for mucosal administration, Mater.
Sci. Eng. C 33 (2013) 411418.
[59] L. Gan, et al., Self-assembled liquid crystalline nanoparticles as a novel ophthalmic
delivery system for dexamethasone: improving preocular retention and ocular bio-
availability, Int. J. Pharm. 396 (2010) 179187.
[60] S. Han, et al., Novel vehicle based on cubosomes for ophthalmic delivery of
flurbiprofen with low irritancy and high bioavailability, Acta Pharmacol. Sin.
31 (2010) 990998.
[61] Y. Chen, et al., Ocular delivery of cyclosporine A based on glyceryl monooleate/
poloxamer 407 liquid crystalline nanoparticles: preparation, characterization,
in vitro corneal penetration and ocular irritation, J. Drug Target. 20 (2012) 856863.
[62] X. Gong, et al., Nanostructured self-assembly materials from neat and aqueous solu-
tions of C18 lipid pro-drug analogues of capecitabinea chemotherapy agent. Focus
on nanoparticulate cubosomes (TM) of the oleyl analogue, Soft Matter 7 (2011)
57645776.
[63] Y.S. Tu, et al., Preparation, characterisation and evaluation of curcumin with piperine-
loaded cubosome nanoparticles, J. Microencapsul. 31 (2014) 551559.
[64] Z. Yang, et al., Optimization of the preparation process for an oral phytantriol-based
amphotericin B cubosomes. J. Nanomater. 2011 (2011), http://dx.doi.org/
10.1155/2011/308016.
ARTICLE IN PRESS
[84] M. Johnsson, et al., Physicochemical and drug delivery aspects of lipid-based liquid
crystalline nanoparticles: a case study of intravenously administered propofol,
J. Nanosci. Nanotechnol. 6 (2006) 30173024.
[85] C. Cervin, et al., A combined in vitro and in vivo study on the interactions between
somatostatin and lipid-based liquid crystalline drug carriers and bilayers, Eur. J. Pharm.
Sci. 36 (2009) 377385.
[86] H. Liu, et al., Protein-bearing cubosomes prepared by liquid precursor dilution: inner
ear delivery and pharmacokinetic study following intratympanic administration,
J. Biomed. Nanotechnol. 9 (2013) 17841793.
[87] S.B. Rizwan, et al., Preparation of phytantriol cubosomes by solvent precursor dilution
for the delivery of protein vaccines, Eur. J. Pharm. Biopharm. 79 (2011) 1522.
[88] B. Angelov, et al., Protein-containing PEGylated cubosomic particles: freeze-fracture
electron microscopy and synchrotron radiation circular dichroism study, J. Phys.
Chem. B 116 (2012) 76767686.
[89] B. Angelov, et al., Multicompartment lipid cubic nanoparticles with high protein
upload: millisecond dynamics of formation, ACS Nano 8 (2014) 52165226.
[90] H.-H. Shen, et al., Targeted detection of phosphatidylserine in biomimetic membranes
and in vitro cell systems using annexin V-containing cubosomes, Biomaterials
34 (2013) 83618369.
[91] E. Esposito, et al., Curcumin containing monoolein aqueous dispersions: a
preformulative study, Mater. Sci. Eng. C 33 (2013) 49234934.
[92] S. Murgia, et al., Drug-loaded fluorescent cubosomes: versatile nanoparticles for
potential theranostic applications, Langmuir 29 (2013) 66736679.
[93] C. Caltagirone, et al., Cancer-cell-targeted theranostic cubosomes, Langmuir
30 (2014) 62286236.
[94] D. Bei, J. Marszalek, B.B.C. Youan, Formulation of dacarbazine-loaded cubosomes
Part I: influence of formulation variables, AAPS PharmSciTech 10 (2009) 10321039.
[95] D. Bei, J. Marszalek, B.B.C. Youan, Formulation of dacarbazine-loaded cubosomes
Part II: influence of process parameters, AAPS PharmSciTech 10 (2009) 10401047.
[96] D. Bei, et al., Formulation of dacarbazine-loaded cubosomes. Part III. Physicochem-
ical characterization, AAPS PharmSciTech 11 (2010) 12431249.
[97] R. Efrat, et al., Solubilization of hydrophobic guest molecules in the monoolein dis-
continuous Q(L) cubic mesophase and its soft nanoparticles, Langmuir 25 (2009)
13161326.
[98] B.J. Boyd, Characterisation of drug release from cubosomes using the pressure ultra-
filtration method, Int. J. Pharm. 260 (2003) 239247.
[99] X. Gong, et al., Lyotropic liquid crystalline self-assembly material behavior and
nanoparticulate dispersions of a phytanyl pro-drug analogue of capecitabinea che-
motherapy agent, ACS Appl. Mater. Interfaces 3 (2011) 15521561.
[100] T.C. Le, et al., Predicting the complex phase behavior of self-assembling drug delivery
nanoparticles, Mol. Pharm. 10 (2013) 13681377.
[101] G. Mangiapia, et al., Cubosomes for ruthenium complex delivery: formulation and
characterization, Soft Matter 7 (2011) 1057710580.
[102] J. Barauskas, et al., Interactions of lipid-based liquid crystalline nanoparticles with
model and cell membranes, Int. J. Pharm. 391 (2010) 284291.
[103] J.C. Bode, et al., Interaction of dispersed cubic phases with blood components, Int. J.
Pharm. 448 (2013) 8795.
[104] S.B. Rizwan, et al., Bicontinuous cubic liquid crystals as sustained delivery systems for
peptides and proteins, Expert Opin. Drug Deliv. 7 (2010) 11331144.
[105] M.J. Lawrence, Surfactant systems: their use in drug delivery, Chem. Soc. Rev.
23 (1994) 417424.
ARTICLE IN PRESS
[106] X. Mulet, B.J. Boyd, C.J. Drummond, Advances in drug delivery and medical imaging
using colloidal lyotropic liquid crystalline dispersions, J. Colloid Interface Sci.
393 (2013) 120.
[107] G.Z. Liu, et al., Colloidal amphiphile self-assembly particles composed of gadolinium
oleate and myverol: evaluation as contrast agents for magnetic resonance imaging,
Langmuir 26 (2010) 23832391.
[108] B.W. Muir, et al., Metal-free and MRI visible theranostic lyotropic liquid crystal
nitroxide-based nanoparticles, Biomaterials 33 (2012) 27232733.
[109] S.J. Fraser, et al., Development of cubosomes as a cell-free biosensing platform, Aust. J.
Chem. 64 (2011) 4653.
[110] I. Amar-Yuli, et al., Solubilization of food bioactives within lyotropic liquid crystalline
mesophases, Curr. Opin. Colloid Interface Sci. 14 (2009) 2132.
[111] K. Larsson, Lyotropic liquid crystals and their dispersions relevant in foods, Curr.
Opin. Colloid Interface Sci. 14 (2009) 1620.
[112] P.P. Nadiminti, et al., Nanostructured liquid crystalline particles as an alternative deliv-
ery vehicle for plant agrochemicals, ACS Appl. Mater. Interfaces 5 (2013) 18181826.
[113] B.J. Boyd, Y.D. Dong, T. Rades, Nonlamellar liquid crystalline nanostructured par-
ticles: advances in materials and structure determination, J. Liposome Res. 19 (2009)
1228.
[114] C. Guo, et al., Lyotropic liquid crystal systems in drug delivery, Drug Discov. Today
15 (2010) 10321040.
[115] H. Ljusberg-Wahern, L. Nyberg, K. Larsson, Dispersion of the cubic liquid crystalline
phasestructure preparation and functionality aspects, Chim. Oggi 14 (1996) 4043.
[116] D. Libster, et al., Soft matter dispersions with ordered inner structures, stabilized by
ethoxylated phytosterols, Colloids Surf. B 74 (2009) 202215.
[117] I. Amar-Yuli, et al., Hexosome and hexagonal phases mediated by hydration and poly-
meric stabilizer, Langmuir 23 (2007) 36373645.
[118] J.Y.T. Chong, et al., Accelerated stability assay (ASA) for colloidal systems, ACS
Comb. Sci. 16 (2014) 205210.
[119] K. Larsson, Cubic lipidwater phasesstructures and biomembrane aspects, J. Phys.
Chem. 93 (1989) 73047314.
[120] A.J. Tilley, C.J. Drummond, B.J. Boyd, Disposition and association of the steric sta-
bilizer Pluronic (R) F127 in lyotropic liquid crystalline nanostructured particle disper-
sions, J. Colloid Interface Sci. 392 (2013) 288296.
[121] K. Lindell, et al., Influence of a charged phospholipid on the release pattern of timolol
maleate from cubic liquid crystalline phases, in: B. Lindman, B.W. Ninham (Eds.), The
Colloid Science of Lipids, Steinkopff, 1998, pp. 111118.
[122] K. Nishikawa, H. Arai, K. Inoue, Scavenger receptor-mediated uptake and metabo-
lism of lipid vesicles containing acidic phospholipids by mouse peritoneal macro-
phages, J. Biol. Chem. 265 (1990) 52265231.
[123] K. Funato, R. Yoda, H. Kiwada, Contribution of complement system on destabiliza-
tion of liposomes composed of hydrogenated egg phosphatidylcholine in rat fresh
plasma, Biochim. Biophys. Acta 1103 (1992) 198204.
[124] J. Senior, Fate and behavior of liposomes in vivo: a review of controlling factors, Crit.
Rev. Ther. Drug Carrier Syst. 3 (1987) 123193.
[125] A. Chonn, P. Cullis, D. Devine, The role of surface charge in the activation of the
classical and alternative pathways of complement by liposomes, J. Immunol.
146 (1991) 42344241.
[126] M.L. Immordino, F. Dosio, L. Cattel, Stealth liposomes: review of the basic science,
rationale, and clinical applications, existing and potential, Int. J. Nanomed. 1 (2006)
297315.
ARTICLE IN PRESS
[127] U. Wattendorf, H.P. Merkle, PEGylation as a tool for the biomedical engineering of
surface modified microparticles, J. Pharm. Sci. 97 (2008) 46554669.
[128] E. Ostuni, et al., A survey of structureproperty relationships of surfaces that resist the
adsorption of protein, Langmuir 17 (2001) 56055620.
[129] J.M. Harris, N. Martin, M. Modi, Pegylation, Clin. Pharmacokinet. 40 (2001)
539551.
[130] J.M. Harris, R.B. Chess, Effect of pegylation on pharmaceuticals, Nat. Rev. Drug
Discov. 2 (2003) 214221.
[131] V.P. Torchilin, Immunoliposomes and PEGylated immunoliposomes: possible use for
targeted delivery of imaging agents, Immunomethods 4 (1994) 244258.
[132] A.L. Klibanov, et al., Amphipathic polyethyleneglycols effectively prolong the circu-
lation time of liposomes, FEBS Lett. 268 (1990) 235237.
[133] R. Gref, et al., Stealth corona-core nanoparticles surface modified by polyethylene
glycol (PEG): influences of the corona (PEG chain length and surface density) and
of the core composition on phagocytic uptake and plasma protein adsorption, Colloids
Surf. B 18 (2000) 301313.
[134] M. Morra, On the molecular basis of fouling resistance, J. Biomater. Sci. Polym. Ed.
11 (2000) 547569.
[135] S.I. Jeon, J.D. Andrade, Proteinsurface interactions in the presence of polyethylene
oxide: II. Effect of protein size, J. Colloid Interface Sci. 142 (1991) 159166.
[136] S.I. Jeon, et al., Proteinsurface interactions in the presence of polyethylene oxide: I.
Simplified theory, J. Colloid Interface Sci. 142 (1991) 149158.
[137] A. Halperin, Polymer brushes that resist adsorption of model proteins: design param-
eters, Langmuir 15 (1999) 25252533.
[138] B. Zhu, et al., Chain-length dependence of the protein and cell resistance of
oligo(ethylene glycol)-terminated self-assembled monolayers on gold, J. Biomed.
Mater. Res. 56 (2001) 406416.
[139] K.L. Prime, G.M. Whitesides, Adsorption of proteins onto surfaces containing end-
attached oligo(ethylene oxide): a model system using self-assembled monolayers,
J. Am. Chem. Soc. 115 (1993) 1071410721.
[140] T. McPherson, et al., Prevention of protein adsorption by tethered poly(ethylene
oxide) layers: experiments and single-chain mean-field analysis, Langmuir 14 (1998)
176186.
[141] M. Malmsten, K. Emoto, J.M. Van Alstine, Effect of chain density on inhibition of
protein adsorption by poly(ethylene glycol) based coatings, J. Colloid Interface Sci.
202 (1998) 507517.
[142] P. Kingshott, H. Thissen, H.J. Griesser, Effects of cloud-point grafting, chain length,
and density of PEG layers on competitive adsorption of ocular proteins, Biomaterials
23 (2002) 20432056.
[143] S. Pasche, et al., Poly(L-lysine)-graft-poly(ethylene glycol) assembled monolayers on
niobium oxide surfaces: a quantitative study of the influence of polymer interfacial
architecture on resistance to protein adsorption by ToF-SIMS and in situ OWLS,
Langmuir 19 (2003) 92169225.
[144] J.V. Jokerst, et al., Nanoparticle PEGylation for imaging and therapy, Nanomedicine
6 (2011) 715728.
[145] C. Thies, Adsorption of styrene/butadiene copolymers and stabilization of silica dis-
persed in perchloroethylene, J. Colloid Interface Sci. 54 (1976) 1321.
[146] W. Heller, T.L. Pugh, Steric protection of hydrophobic colloidal particles by
adsorption of flexible macromolecules, J. Chem. Phys. 22 (1954) 1778.
[147] J. Lee, P.A. Martic, J.S. Tan, Protein adsorption on pluronic copolymer-coated poly-
styrene particles, J. Colloid Interface Sci. 131 (1989) 252266.
ARTICLE IN PRESS
[148] J.A. Neff, K.D. Caldwell, P.A. Tresco, A novel method for surface modification to
promote cell attachment to hydrophobic substrates, J. Biomed. Mater. Res.
40 (1998) 511519.
[149] Y.-D. Dong, et al., Bulk and dispersed aqueous phase behavior of phytantriol: effect of
vitamin E acetate and F127 polymer on liquid crystal nanostructure, Langmuir
22 (2006) 95129518.
[150] B.J. Boyd, et al., Self-assembled geometric liquid-crystalline nanoparticles imaged in
three dimensions: hexosomes are not necessarily flat hexagonal prisms, Langmuir
23 (2007) 1246112464.
[151] F. Muller, A. Salonen, O. Glatter, Phase behavior of phytantriol/water bicontinuous
cubic Pn3m cubosomes stabilized by laponite disc-like particles, J. Colloid Interface
Sci. 342 (2010) 392398.
[152] J. Zhai, et al., Revisiting -casein as a stabilizer for lipid liquid crystalline nanostruc-
tured particles, Langmuir 27 (2011) 1475714766.
[153] T. Kojarunchitt, et al., Development and characterisation of modified poloxamer 407
thermoresponsive depot systems containing cubosomes, Int. J. Pharm. 408 (2011)
2026.
[154] H.H. Shen, et al., The influence of dipalmitoyl phosphatidylserine on phase behaviour
of and cellular response to lyotropic liquid crystalline dispersions, Biomaterials
31 (2010) 94739481.
[155] C.E. Conn, et al., Enhanced uptake of an integral membrane protein, the dopamine
D2L receptor, by cubic nanostructured lipid nanoparticles doped with Ni(II) chelated
EDTA amphiphiles, Soft Matter 7 (2011) 567578.
[156] H.H. Shen, et al., The interaction of cubosomes with supported phospholipid bilayers
using neutron reflectometry and QCM-D, Soft Matter 7 (2011) 80418049.
[157] S.J. Fraser, et al., Surface immobilization of bio-functionalized cubosomes: sensing of
proteins by quartz crystal microbalance, Langmuir 28 (2012) 620627.
[158] T.E. Hartnett, et al., Size and phase control of cubic lyotropic liquid crystal
nanoparticles, J. Phys. Chem. B 118 (2014) 74307439.
[159] S.J. Fraser, et al., Controlling nanostructure and lattice parameter of the inverse
bicontinuous cubic phases in functionalised phytantriol dispersions, J. Colloid Interface
Sci. 408 (2013) 117124.
[160] J.Y.T. Chong, et al., Steric stabilisation of self-assembled cubic lyotropic liquid
crystalline nanoparticles: high throughput evaluation of triblock polyethylene oxide
polypropylene oxidepolyethylene oxide copolymers, Soft Matter 7 (2011) 47684777.
[161] M. Uyama, et al., Useful modified cellulose polymers as new emulsifiers of cubosomes,
Langmuir 25 (2009) 43364338.
[162] T. Abraham, M. Hato, M. Hirai, Glycolipid based cubic nanoparticles: preparation and
structural aspects, Colloids Surf. B 35 (2004) 107118.
[163] T. Abraham, M. Hato, M. Hirai, Polymer-dispersed bicontinuous cubic glycolipid
nanoparticles, Biotechnol. Prog. 21 (2005) 255262.
[164] L. de Campo, et al., Reversible phase transitions in emulsified nanostructured lipid
systems, Langmuir 20 (2004) 52545261.
[165] A. Yaghmur, et al., Oil-loaded monolinolein-based particles with confined inverse
discontinuous cubic structure (Fd3m), Langmuir 22 (2005) 517521.
[166] A. Yaghmur, et al., Emulsified microemulsions and oil-containing liquid crystalline
phases, Langmuir 21 (2004) 569577.
[167] A. Salonen, S. Guillot, O. Glatter, Determination of water content in internally self-
assembled monoglyceride-based dispersions from the bulk phase, Langmuir 23 (2007)
91519154.
[168] S. Guillot, et al., Direct and indirect thermal transitions from hexosomes to emulsified
micro-emulsions in oil-loaded monoglyceride-based particles, Colloids Surf. A
291 (2006) 7884.
ARTICLE IN PRESS
[169] A. Yaghmur, et al., Control of the internal structure of MLO-based isasomes by the
addition of diglycerol monooleate and soybean phosphatidylcholine, Langmuir
22 (2006) 99199927.
[170] A. Yaghmur, et al., Self-assembly in monoelaidin aqueous dispersions: direct vesicles to
cubosomes transition, PLoS One 3 (2008) e3747.
[171] E. Esposito, et al., Lipid-based supramolecular systems for topical application: a
preformulatory study, AAPS PharmSci 5 (2003) 6276.
[172] J. Gustafsson, et al., Cubic lipidwater phase dispersed into submicron particles,
Langmuir 12 (1996) 46114613.
[173] J. Gustafsson, et al., Submicron particles of reversed lipid phases in water stabilized by a
nonionic amphiphilic polymer, Langmuir 13 (1997) 69646971.
[174] K. Larsson, Colloidal dispersions of ordered lipidwater phases, J. Dispers. Sci.
Technol. 20 (1999) 2734.
[175] K. Larsson, Aqueous dispersions of cubic lipidwater phases, Curr. Opin. Colloid
Interface Sci. 5 (2000) 6469.
[176] K. Larsson, Bicontinuous cubic lipidwater particles and cubosomal dispersions,
in: T. Osamu (Ed.), Studies in Surface Science and Catalysis, Elsevier, 2004, pp. 4151.
[177] C. Neto, et al., Imaging soft matter with the atomic force microscope: cubosomes and
hexosomes, J. Phys. Chem. B 103 (1999) 38963899.
[178] P.T. Spicer, et al., Novel process for producing cubic liquid crystalline nanoparticles
(cubosomes), Langmuir 17 (2001) 57485756.
[179] J. Barauskas, et al., Cubic phase nanoparticles (cubosome): principles for controlling
size, structure, and stability, Langmuir 21 (2005) 25692577.
[180] M. Monduzzi, H. Ljusberg-Wahren, K. Larsson, A 13C NMR study of aqueous dis-
persions of reversed lipid phases, Langmuir 16 (2000) 73557358.
[181] M. Nakano, et al., Small-angle X-ray scattering and 13C NMR investigation on the
internal structure of cubosomes Langmuir 17 (2001) 39173922.
[182] O. Svensson, K. Thuresson, T. Arnebrant, Interactions between chitosan-modified
particles and mucin-coated surfaces, J. Colloid Interface Sci. 325 (2008) 346350.
[183] O. Svensson, K. Thuresson, T. Arnebrant, Interactions between drug delivery particles
and mucin in solution and at interfaces, Langmuir 24 (2008) 25732579.
[184] P. Vandoolaeghe, et al., Adsorption of cubic liquid crystalline nanoparticles on model
membranes, Soft Matter 4 (2008) 22672277.
[185] P. Vandoolaeghe, et al., Neutron reflectivity studies of the interaction of cubic-phase
nanoparticles with phospholipid bilayers of different coverage, Langmuir 25 (2009)
40094020.
[186] P. Vandoolaeghe, et al., Adsorption of intact cubic liquid crystalline nanoparticles on
hydrophilic surfaces: lateral organization, interfacial stability, layer structure, and inter-
action mechanism, J. Phys. Chem. C 113 (2009) 44834494.
[187] T. Landh, Phase-behavior in the system pine oil monoglycerides-poloxamer-407-
water at 20-degrees-C, J. Phys. Chem. 98 (1994) 84538467.
[188] A. Gupta, et al., Nanoassemblies of Gd-DTPA-monooleyl and glycerol monooleate
amphiphiles as potential MRI contrast agents, J. Mater. Chem. B 2 (2014) 12251233.
[189] B. Siekmann, et al., Preparation and structural investigations of colloidal dispers-
ions prepared from cubic monoglyceridewater phases, Int. J. Pharm. 244 (2002)
3343.
[190] M. Nakano, et al., Dispersions of liquid crystalline phases of the monoolein/oleic acid/
Pluronic F127 system, Langmuir 18 (2002) 92839288.
[191] G. Popescu, et al., Liquid crystalline phases and their dispersions in aqueous mixtures of
glycerol monooleate and glyceryl monooleyl ether, Langmuir 23 (2006) 496503.
[192] A. Salonen, F. Muller, O. Glatter, Dispersions of internally liquid crystalline systems
stabilized by charged disklike particles as pickering emulsions: basic properties and
time-resolved behavior, Langmuir 24 (2008) 53065314.
ARTICLE IN PRESS
[213] M.-S. Martina, et al., The in vitro kinetics of the interactions between PEG-ylated
magnetic-fluid-loaded liposomes and macrophages, Biomaterials 28 (2007)
41434153.
[214] M.-S. Martina, C. Wilhelm, S. Lesieur, The effect of magnetic targeting on the uptake
of magnetic-fluid-loaded liposomes by human prostatic adenocarcinoma cells,
Biomaterials 29 (2008) 41374145.
[215] J. Patton, M. Carey, Watching fat digestion, Science 204 (1979) 145148.
[216] M. Lindstrom, et al., Aqueous lipid phases of relevance to intestinal fat digestion and
absorption, Lipids 16 (1981) 749754.
[217] W. Buchheim, K. Larsson, Cubic lipidproteinwater phases, J. Colloid Interface Sci.
117 (1987) 582583.
[218] M. Almgren, et al., Dispersed lipid liquid crystalline phases stabilized by a
hydrophobically modified cellulose, Langmuir 23 (2007) 27682777.
[219] Y. Luo, et al., Dry coating, a novel coating technology for solid pharmaceutical dosage
forms, Int. J. Pharm. 358 (2008) 1622.
[220] C.D. Kablitz, M. Kappl, N.A. Urbanetz, Parameters influencing polymer particle lay-
ering of the dry coating process, Eur. J. Pharm. Biopharm. 69 (2008) 760768.
[221] H. Konno, et al., Effect of polymer type on the dissolution profile of amorphous solid
dispersions containing felodipine, Eur. J. Pharm. Biopharm. 70 (2008) 493499.
[222] C.D. Driever, et al., Layer-by-layer polymer coating on discrete particles of cubic
lyotropic liquid crystalline dispersions (cubosomes), Langmuir 29 (2013)
1289112900.
[223] B. Angelov, et al., DNA/fusogenic lipid nanocarrier assembly: millisecond structural
dynamics, J. Phys. Chem. Lett. 4 (2013) 19591964.
[224] J.D. Du, et al., A novel approach to enhance the mucoadhesion of lipid drug
nanocarriers for improved drug delivery to the buccal mucosa, Int. J. Pharm.
471 (2014) 358365.