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INTRODUCTION
Recently, there has been increased interest in novel drug
Doxorubicin (Dox) is an antineoplastic agent widely used in
the treatment of several types of cancer. The drug binds to
double-stranded DNA, which results in the inhibition of tumor
delivery systems (DDS) based on lipidic mesophases because of
cell replication.17,18 The drug has low selectivity; it aects both
their large drug loading capacity, stability, biocompatibility, and
healthy and diseased cells, causing a number of unwanted side
sustained release of drugs.13 In the case of anticancer drugs,
eects. One of the approaches to reduce these eects relies on
encapsulation of such drugs into the DDS would reduce the
the lower pH of the tumor cell environment compared with
side eects caused by the drug, especially reducing toxic eects that of healthy cells. Release proles of Dox from LCP were
on healthy cells. Lipidic cubic and hexagonal phases are monitored electrochemically by measuring the voltammetric
advantageous materials for drug delivery since they are stable in peak current of doxorubicin reduction. It was found that drug
excess water and can accommodate relatively large loads (drug) release from the phase is pH-dependent. The pKa of
due to a high surface area of ca. 400 m2/g.26 The highly doxorubicin is 8.2.19 Protonated doxorubicin, which carries a
structured reverse bicontinuous lipidic cubic phase (LCP), positive charge, is soluble in water, whereas the unprotonated
composed of highly curved lipid bilayers surrounded by two drug prefers a hydrophobic lipidic environment, where its
identical, nonintersecting aqueous channels, exhibits interesting diusion is much slower.15,16 Recently, we have voltammetri-
properties for applications as a drug carrier.1,3,5,6 Because of cally evaluated the release kinetics of Dox by analyzing a
their internal structure, LCPs can incorporate hydrophilic, decrease in its peak current over time. The results suggest that
amphiphilic, and hydrophobic drugs. Hydrophobic drugs tend the release mechanism is in agreement with the Higuchi model
to partition into the lipid bilayer, while hydrophilic drugs reside at low pH, while at higher pH values, non-Fickian transport
preferentially in the aqueous channels.79 LCPs have therefore dominates.16
been extensively investigated as vehicles for the sustained In the present study, we designed the following hybrid
release of bioactive molecules of various sizes and molecular materials based on monoolein cubic mesophases containing
weights. The location of the drug is an important parameter two types of magnetic nanoparticles (NPs) diering in their
aecting the diusion and release rate.1016 Partitioning
between the lipidic and aqueous compartments, which is Received: October 11, 2016
related to the size and polarity of the drug, largely determines Accepted: December 28, 2016
the release kinetics. Published: December 28, 2016
stabilizing coatings around the magnetic core: hydrophilic to molten MO. In the case of hydrophobic nanoparticles, the
nickelzinc ferrite nanoparticles stabilized with a citric acid appropriate amount of NP dispersion in hexane (NPoleic) was added to
adlayer (NPcitric) and lipophiliciron oxide core nanoparticles molten MO and sonicated. The mixture was left in a desiccator to
coated with an oleic acid adlayer (NPoleic). The addition of evaporate the solvent. Then, water or Dox solution was added. The
ratios of the hybrid systems were 59.8/0.2/40 or 58/2/40 and 59.3/
magnetic nanoparticles to the mesophase would open new 0.2/0.5/40 or 57.5/2/0.5/40 (% w/w) for MO/NP/H2O and MO/
possibilities for directing a liquid crystal drug carrier to the NP/Dox/H2O, respectively. To obtain homogeneous, transparent, and
desired target using a magnetic eld,20,21 as well as for magnetic viscous LCPs, all the samples were left for at least 24 h in tightly closed
hyperthermia.22 The mesophase can be dispersed into nano- vials at room temperature in the dark. Phase diagrams of the obtained
particles: cubosomes,23 which have the same properties as the hybrid phases were evaluated.
bulk phase but have lower density. Cubosomes doped with The obtained phases were used to prepare cubosomes.23 First, 0.1 g
nano-objects are of interest not only as the drug delivery of LCP phase with NPs (34.5 mg of NPs per gram of monoolein) was
systems3,2429 but also as contrast agents for magnetic placed in a glass vial, and 1.85 mL of Pluronic F-127 solution (1%) in
resonance imaging (MRI).30,31 deionized water was added. The cubic phases were sonicated for 20
min in a bath-type sonicator.
We studied the physical and chemical properties of the Synthesis of Hydrophilic Mixed Ferrite Nanoparticles
hybrid materials as a thin lm on the electrode surface and as (Ni0.5Zn0.5Fe2O4). Hydrophilic nanoparticles (NPcitric) were synthe-
cubic phase nanoparticles, i.e., cubosomes, both modied with sized according to ref 33 with minor modications. Briey, the
the magnetic nanoparticles, NPcitric and NPoleic. Due to their nanoferrites were prepared using the bottom-up technique by
dierent hydrophilic/hydrophobic properties, the two types of coprecipitation of nanoferrites from the solution of their precursors
NPs used are located in dierent phase domains of the LCPs. with a strong base according to the following equation:
All samples exhibited a double-diamond cubic Pn3m phase at Ni 2 + + Zn 2 + + 4Fe3 +16OH 2Ni 0.5Zn 0.5Fe2O4
+ 8H 2O
room and human body temperature, with some dierences in
the diameters of the water channels. For the mesophase doped First, the precursor solution was prepared by mixing the heated
with 2% w/w of hydrophilic NPcitric above 40 C, a switch from aqueous nitrate salts of 0.6667 M Fe3+, 0.1667 M Ni2+, and 0.1667 M
the cubic to the hexagonal phase takes place, which results in Zn2+ with a hot aqueous solution of sodium hydroxide. Then, in a
reaction vessel, 40 mL of this solution was mixed with 8 mL of 2 M
slower drug release from the mesophase. Drug release from the
HNO3 (to avoid hydrolysis), and 152 mL of H2O was added. The
bicontinuous cubic phase is signicantly faster than from the resulting solution was heated with continuous stirring to 95 C under
hexagonal phases.5 reux. An aqueous solution of NaOH (400 mL, 0.75 M) was
The release proles of Dox were evaluated using electro- preheated to 95 C in a separate container and then transferred to the
chemical methods, which monitor the changes in drug reaction vessel. A dark-brownish precipitate appeared, and the reaction
concentration directly in the cubic phase layer covering the was left heated for the next 12 h with stirring. Afterward, the reaction
electrode. A comparison of structures and drug elution proles was cooled to room temperature. Nanoparticles were precipitated with
reveals that the hybrid material with NPcitric changes the a magnet and washed three times with deionized water using
properties of the cubic phase. In the case of hydrophobic sedimentation and decantation in the eld of a permanent magnet.
The NP surfaces were treated with citrate ions by incubating for 1 h in
nanoparticles, the structure remains unaected. All results show
a 0.5 mg/mL citric acid solution with continuous stirring at 90 C.
that the hybrid material with hydrophobic magnetic NPs is a Then, the obtained suspension was cooled to room temperature and
promising matrix for drug delivery. Hybrid cubosome washed several times with an excess of deionized water under magnet-
dispersions were also prepared. The magnetocubosomes assisted sedimentation to remove the unbound citric acid. The
containing both types of magnetic nanoparticles have magnetic hydrophilic nanoparticles grafted with citric acid were located
properties similar to those of magnetic nanoparticles alone and primarily in the LCP aqueous domain.
can be considered for drug delivery directed by means of a The choice of these particular magnetic cores was intentional since
magnetic eld. it allowed control of the crystallite size and magnetic properties of the
a rotary evaporator and replaced with 100 mL of hexane. The lattice parameter of corresponding phase, and l is the lipid chain
hydrophobic nanoparticles stabilized with oleic acid were mainly length/monolayer thickness.
embedded in the lipidic part of the cubic phase; however, since they Finally, the water channel radius rw was obtained using equation:38
are also larger than the bilayer width, parts of the nanoparticle are
protruding into the aqueous channels (Scheme 1).36 rw = ( /2 )1/2 a l (4)
Polarized Microscopy. Polarized light microscopy (Olympus BX
Scheme 1. Characteristics over Length Scales of the 50) conducted with a heating/cooling stage was used for visual
Magnetic Nanoparticles, Cubic Mesophase, and inspection of the samples.
Magnetocubosomes Dynamic Light Scattering (DLS) and Zeta Potential. The
hydrodynamic diameter of the cubic phase particle dispersions was
determined using a Malvern Zetasizer instrument (Nano ZS, UK)
tted with a 4 mW HeNe laser ( = 632.8 nm) as the light source at
a scattering angle of 173. The solutions were equilibrated for 2 min
before measurement. The zeta potential was measured using the same
instrument. The values were reported as averages from 5 measure-
ments of each sample.
Thermogravimetric Analysis (TGA). Thermogravimetric analysis
of coated nanoparticles was performed at a temperature range of 25
600 C with a heating rate of 10 Cmin1 under oxygen using the
TGA Q-50 thermal analyzer (TA Instrument). From these measure-
ments, we evaluated that the amount of citric acid on NPcitric and oleic
acid on NPoleic was 25.0 and 24.6% w/w, respectively (Figure S1A).
Transmission Electron Microscopy (TEM). TEM measurements
(Libra 120 microscope, Zeiss) were used to assess the size of the
magnetic nanoparticles (Figure 1A,B). Before the measurement, a
METHODS
Small-Angle X-ray Scattering. The phase identity and structural
parameters of the lipidic samples were determined by small-angle X-
ray scattering (SAXS). The experiments were carried out with a
GADDS system (Bruker) using a 3-pinhole collimation system
working with Cu K radiation (CuK = 0.1542 nm), a Nanostar
system (Bruker) working with CuK radiation equipped with a Vantec
2000 area detector, a SAXSess mc2 instrument (Anton Paar) using a Figure 1. TEM images of the citrate-coated NPs (A) and oleic-
line collimation system and equipped with a sealed X-ray tube stabilized NPs (B); scale bars = 50 nm.
(PANalytical, CuK = 0.1542 nm), and a CCD detector (Princeton
Instruments). Samples were introduced into special thin glass
capillaries, which were immediately ame-sealed. SAXS spectra were drop of a sample suspension was placed on a Formvar-coated copper
recorded in the range of 1060 C. The scattering intensities I(q) grid and allowed to dry in the air. Hydrophilic nanoparticles NPcitric
were represented as a function of the magnitude of the scattering revealed an average diameter of 13 3 nm, while the average diameter
vector q = (4/) sin(), where 2 is the total scattering angle. The of hydrophobic NPoleic was ca. 7 2 nm (Figure S1B,C).
SAXS patterns obtained from the Bruker apparatus were analyzed with Cryo Scanning Electron Microscopy (Cryo-SEM). A cryo
the Bruker Topas 3 software. For the SAXS mc2 apparatus, all data scanning electron microscopy (cryo-SEM) image was taken using a
were corrected for background scattering from the empty capillary and Carl Zeiss Microscopy at a voltage 2 kV and a working distance of 2.6
for slit-smearing eects by a desmearing procedure from the suppliers mm. Sample was loaded into a holder and frozen in the liquid
software using the Lake method. nitrogen. The sample was transferred into the cryochamber, which was
The size of water channels was calculated based on the lattice held at 140 C. Then the sample was sublimed at 85 C for 23
parameter and the composition of cubic phases.37 Initially, the water min.
volume fraction was calculated by the equation: Magnetization Measurements. The magnetic properties of both
types of nanoparticles as a function of the magnetic eld at
w = c w /(c w + (1 c w)(w /l )) (1) temperatures ranging from 2 to 300 K were reported by us earlier.39,40
Those results were comparable, regardless of the surface modication
where w is the water volume fraction, cw is the water weight fraction,
(citric or oleic shell), and in general revealed typical ferromagnetic-like
w is the density of water =0.997 g/cm3, and l is the density of lipid =
behavior, with no hysteresis loop at temperatures close to room
0.942 g/cm3.
temperature. Here, we complemented our previous results with
The lipid volume fraction was determined from the equation:
magnetization measurements of NPs and hybrid materials performed
l = 1 w (2) with an automatic Faraday balance at a constant magnetic eld of 1 T.
The results are shown in Figure S2. Temperature was controlled with
The lipid chain length (l) was determined by solving the following an accuracy better than 0.5 K. The magnetization values for
equation:37 nanoparticles and hybrid materials were calculated using the weight
l = 2(l/a) + 4/3 (l/a)3 of nickelzinc ferrite and magnetite measured by TGA (Figure S1A).
(3)
The temperature variation in the mass magnetization of the
where is the ratio of the minimal surface in a unit cell to the quantity nanocrystalline NPcitric sample, containing nickelzinc ferrite, is
(unit cell volume)2/3, is the EulerPoincare characteristic, a is the plotted in Figure S2A and shows ferromagnetic ordering up to the
Curie temperature of TC 267 C. This Curie temperature is These results prompted us to use both types of magnetic nanoparticles
comparable with the TC = 264 C of the polycrystalline Ni0.5Zn0.5 as prospective vectors incorporated inside the cubosomes loaded
Fe2O4.41 At room temperature, the magnetic moment per nickelzinc with doxorubicin, forming a nanoparticulate hybrid drug delivery
ferrite equals 1.5 B (Table 1, vide inf ra) and is smaller compared to system as described further.
Electrochemical Measurements. Electrochemical measurements
Table 1. Magnetic Moment per Molecule of Citric Acid- were performed using the CH Instruments bipotentiostat model
Coated NP, Oleic-Coated NP, Hybrid Cubic Phases, and CHI750B. The three-electrode system, consisting of a glassy carbon
electrode (GCE, A = 0.07 cm2), an Ag/AgCl reference electrode, and a
Hybrid Cubosomes at Room Temperature platinum foil as a counter electrode, was employed. Prior to the
magnetic moment (B) measurements, the GCE electrodes were polished with an alumina
slurry (1.0, 0.3, and 0.05 m) on a polishing cloth. The electrodes
NPcitric 1.5
were then rinsed with a direct stream of ultrapure water, sonicated in
phase with NPcitric 1.4 an ultrasonic bath, and left to air-dry. After cleaning, the working
cubosomes with NPcitric 1.5 electrodes were modied with a thin lm of the cubic phase.
NPoleic 2.3 Measurements at elevated temperatures were performed using a BVT
phase with NPoleic 2.7 MT-1 minithermostat. Argon-saturated solutions were obtained by
cubosomes with NPoleic 1.9 bubbling high-purity argon gas for 15 min into the solution and
continuing with a ow of pure gas (Ar) over the solution during
experiments. Release of Dox from LCPs deposited on a GCE was
the 3.17 B of the polycrystalline Ni0.5Zn0.5Fe2O4. The reduced monitored using dierential pulse voltammetry (DPV). For each type
magnetic moment of nanocrystalline ferrite is typically attributed to of hybrid cubic phase, triplicate experiments were performed.
the structural and magnetic disorder within the crystallite surface layer.
The temperature variation in the mass magnetization of the
nanocrystalline NPoleic sample, containing magnetite Fe3O4, is plotted
RESULTS AND DISCUSSION
in Figure S2B and shows the ferromagnetic ordering up to the Curie Structural Characterization of the Cubic Phases. To
temperature of TC 490 C, which is lower than the 585 C reported evaluate the eect of the addition of two types of magnetic
for the polycrystalline magnetite42 and the 580 C reported for the nanoparticles on the cubic phase properties, SAXS and cross-
magnetite Fe3O4 coated with citric acid.43 polarized microscopy experiments were carried out.
The magnetic moment per Fe3O4 equals 2.3 B at room Polarized light microscopy and SAXS are the most widely
temperature (Table 1, vide inf ra) and is reduced compared to the used techniques for characterization of liquid crystalline phases.
3.77 B of the polycrystalline.36 The minute magnetization hump
observed between 220 and 320 C may reveal some change in the Cubic phases are isotropic; therefore, a dark image appears
interparticle interaction caused by removal of the surface-coating oleic when viewed under a microscope equipped with cross-
layer, which is also observed in the same temperature range for the polarizers. Other structures found in the MO/H2O system,
TGA spectra (Figure S1A). An analogous eect has been reported for lamellar and hexagonal phases, are anisotropic and have a
citrate-stabilized Fe3O4 aqueous colloidal magnetic nanoparticles.43 characteristic textures when observed by polarized light
Figure 2. Left panel: Monooleinwater partial phase diagrams with 2% w/w of hydrophilic nanoparticles (A) and 2% w/w of hydrophobic
nanoparticles (C). Right panel: Comparison of SAXS spectra at human body temperature obtained from the bulk cubic phases with 40% w/w of
water: MO/NPcitric/H2O (B) and MO/NPoleic/H2O (D) with MO/H2O system.
Figure 3. SAXS spectra of the various monoolein-based systems with 40 wt % of water at dierent temperatures: pure MO (A); with 2% w/w of
hydrophobic NPs (B); with 2% w/w of hydrophilic NPs (the intensities are represented on a logarithmic scale) (C). Polarized-light microscopy
images for phase with NPcitric at 25 C (Pn3m) (D), 40 C (Pn3m and H2 phases) (E), and 50 C (H2 phase) (F).
Table 2. Results of SAXS Measurements for Monoolein and Hybrid LCPs Depending on the Phase Composition and
Temperaturea
phase composition (% w/w) T (C) phase symmetry a (nm) l (nm) dw (nm)
25 Pn3m 9.5 1.6 4.2
35 Pn3m 9.3 1.6 4.1
MO/H2O 60/40
40 Pn3m+water 9.0 1.5 4.0
45 Pn3m+water 8.7 1.5 3.8
25 Pn3m 10.1 1.7 4.5
36 Pn3m 9.7 1.7 4.3
MO/NPcitric/H2O 59.8/0.2/40
40 Pn3m+water 9.3 1.6 4.1
44 Pn3m+water 8.9 1.5 4.0
25 Pn3m 8.9 1.5 3.9
36 Pn3m 8.5 1.5 3.8
8.3 1.4 3.7
MO/NPcitric/H2O 58/2/40 40 Pn3m and trace H2
nd nd nd
8.1 nd nd
44 Pn3m and H2
6.0 nd nd
25 Pn3m 9.4 1.6 4.2
35 Pn3m 9.3 1.6 4.1
MO/NPoleic/H2O 59.8/0.2/40
40 Pn3m+water 9.2 1.6 4.1
45 Pn3m+water 8.8 1.5 3.9
25 Pn3m 9.6 1.6 4.2
36 Pn3m 9.3 1.6 4.1
MO/NPoleic/H2O 58/2/40
40 Pn3m+water 9.0 1.5 4.0
44 Pn3m+water 8.7 1.5 3.8
a
Phase identity, lattice parameter a, lipid length l, and water channel diameters dw.
microscopy. The space group of the phases and the structural content varying between 20 and 40% w/w. By adding
parameters can then be determined by SAXS. hydrophilic nanoparticles that are located mainly in the water
A monooleinwater phase diagram has been described in the channels of the cubic phases, substantial changes in the phase
literature;32,4446 therefore, the inuence of the incorporation diagram in the studied region can be observed (Figure 2A).
of 2% w/w of NPcitric and NPoleic was investigated. The phase Below 40 C, the sequence of liquid crystals remains
diagrams were determined in the range of 1060 C for a water unchanged, except for the lamellar phase. Both the Ia3d and
2800 DOI: 10.1021/acsami.6b12889
ACS Appl. Mater. Interfaces 2017, 9, 27962805
ACS Applied Materials & Interfaces Research Article
Pn3m cubic phases are less temperature-stable. Indeed, Ia3d The incorporation of doxorubicin into MO does not
cubic phase melts at 45 C, and Pn3m phase disappears at signicantly aect the lattice parameter and size of water
approximately 60 C. However, for MO with hydrophobic NPs, channels since the drug molecule is small (ca. 15 nm) and is
these two phases are stable over 60 C. Interestingly, a phase easily accommodated in the structure (Table S3).15 However,
transition from Pn3m to H2 occurs at 40 C for a water content the incorporation of Dox into hybrid materials leads to an
of 40% w/w, as shown on the SAXS spectra and conrmed by increase in the lattice parameter and the width of the water
the texture of the samples observed with an optical microscope channels, which is more pronounced in the case of 2% w/w of
equipped with cross polarizers (Figure 3), whereas the hydrophilic nanoparticles.
transition appears at 90100 C for pure MO in a limited Properties of Cubosomes. Nanoparticles of the cubic
range of water content (1525% w/w of water). The decrease phasecubosomeswere prepared, and their characteristics
in the phase transition temperature (Pn3m cubic phaseH2) were evaluated (Figures S4 and S5). The diameter of cubosome
may be induced by the presence of the hydrophilic NPs that dispersions, in the absence/presence of NPs, was obtained from
swell the water channels. Therefore, the lattice axes are the DLS measurements. The cubic particle sizes are similar,
increased, and this results in a reduced curvature of the bilayer between 110 and 150 nm, which is suitable for a drug delivery
and a less pronounced wedge shape for the molecules. This system (Table 3 and Figure S4A). With the DLS technique, we
behavior favors the formation of a reverse hexagonal phase.45
By adding hydrophobic nanoparticles that are solubilized in the Table 3. Characteristic Values of Cubosomes
hydrophobic chains, no signicant modication of the phase
cubosomes with cubosomes with
behavior was observed (Figure 2C). Indeed, in comparison cubosomes NPcitric NPoleic
with pure MO, the same sequence of liquid crystals (lamellar zeta potential (mV) 18.9 4.4 17.3 1.3 19.6 2.3
phaseIa3d cubic phasePn3m cubic phase) was obtained as a size (nm) 111 10 137 8 150 13
function of the water composition, and the stable areas polydispersity index 0.35 0.26 0.33
associated with the temperature and concentration are similar. (PDI)
This is an important observation for future applications of
various forms of mesophases as drug delivery platforms with can perform measurements in the natural environment of the
NPs acting as delivery vectors. sampleswater solutions. Lipidic particles are rather fragile, and
Comparing the SAXS experiments of samples with 40% w/w preparation of samples for TEM measurements can change/
of water, the spectra of 2% w/w and 0.2% w/w NPs (Figures destroy the structure. Nevertheless, we used TEM imaging to
2B,D) at human body temperature are similar to that of pure conrm the presence of NPs inside the cubosomes. Figure S4
MO and present a cubic structure that corresponds to the presents TEM images of cubosomes with hydrophilic NPcitric
Pn3m space group, as conrmed by the relative positions of the (Figure S4C) and hydrophobic NPoleic (Figure S4D) nano-
reection lines 1:3/2:2:3:2:9/2. For the hybrid phase particles (please note the dierent scale bars on these gures).
with 0.2% w/w of citric NPs, the peaks are shifted to lower q The cubosomes are visible as gray discs with much darker dots
values (Figure 2B), while for the phase with 2% w/w NPcitric, of iron oxide nanoparticles because NPs have a much higher
the situation is opposite. For NPoleic, no shift of the peaks is electron density than lipids.
observed (Figure 2D), which indicates that the incorporation of Cryo-SEM image shows the spherical morphology of the
hydrophobic NPs does not aect the organization of lipidic cubosomes (Figure S4B). The cubic nature of the LCP
mesophases. Moreover, for the SAXS spectra (Figure 2, 3), it dispersion was characterized by SAXS. The SAXS pattern for
should be noted that for samples with NPs, diusion at small q MO cubosomes shows the local cubic structure with Im3m
values is observed, due to the presence of NPs in the system. space group (the relative positions of the diraction signals are
Increasing the temperature induces a shift of the peaks 2:4:6) (Figure S5). The crystallographic lattice param-
toward higher q values for hydrophobic NPs, and the Pn3m eter is 14.6 nm. It was reported earlier that sometimes during
cubic phase is maintained (Figure 3B), as observed for pure ultrasonic dispersion of material the Pn3m cubic phase
MO (Figure 3A). For the samples with 2% w/w of hydrophilic underwent transition to the cubic structure with Im3m
NPs, the reverse hexagonal phase H2 appears at 40 C, as symmetry.4750 In the case of cubosomes with NPoleic, the
conrmed by the relative positions of the reection lines scattering from nanoparticles masks completely the rst Bragg
1:3:2 on the SAXS spectra (Figure 3C), which is character- reection and also partly the second Bragg peak. However, the
istic of the hexagonal symmetry. For the samples with 0.2% w/ position of the third peak is clearly visible and at the same q
w of hydrophilic NPs, the peaks are shifted to higher q values, value as the corresponding peak for MO cubosomes, which
and no phase transition is observed in the temperature range conrms that incorporation of NPs does not aect the structure
from 25 to 60 C (data not shown). of cubosomes (Figure S5, inset), similar to the case of bulk
The calculated values of the lattice parameter, lipid length cubic phases (Figures 2 and 3).
and diameter of water channels are reported in Table 2. The Since the zeta potential for cubosomes with or without NPs
values conrm the observations of the SAXS spectra. At room is quite similar (see Table 3) and the zeta potential for NPs
temperature, the addition of 0.2% w/w of hydrophilic NPs alone is much higher (39 mV for NPcitric), we can conclude
leads to an increase in the water channel width by 0.3 nm. The that the nanoparticles remain inside the lipidic particle and not
addition of 2% w/w of NPcitric leads to a decrease in both the on the surface. The cubosomes in the TEM images were larger
lattice parameter and channel width by 0.6 and 0.3 nm, in size than those observed with DLS, which was likely caused
respectively. The addition of hydrophobic NPs has no inuence by attening during the drying process.
on the lattice parameter compared to pure MO. As expected, Doxorubicin Release Prole from Cubic Phases with
after increasing the temperature, the lattice parameter and the Nanoparticles. The drug release proles of the pure
diameter of water channels decrease for both NPs similar to the monoolein and hybrid phases were investigated based on the
pure MO phase.32 changes in the DPV voltammograms with time (Figures 4 and
2801 DOI: 10.1021/acsami.6b12889
ACS Appl. Mater. Interfaces 2017, 9, 27962805
ACS Applied Materials & Interfaces Research Article
Results obtained for Dox in systems containing 0.2% w/w of
both types of NPs embedded in the cubic mesophase suggest
that the release mechanism proceeds according to the Higuchi CONCLUSIONS
model. The same situation is found for the phase with 2% w/w The cubic phase is a promising material for the delivery of
of NPoleic at room temperature, but when increasing the hydrophilic and hydrophobic drugs. Insertion of magnetic
temperature, the inuence of the matrix is observed (n deviates nanoparticles can extend its applications in medicine. We
from 0.5). In the case of 2% w/w of NPcitric incorporated into presented and characterized hybrid materials based on a
the phase, clearly non-Fickian transport contributes to the monoolein cubic phase doped with two types of magnetic
transport, and the process cannot be described by the Higuchi nanoparticles: hydrophilic and hydrophobic. The system was
model even at room temperature (Table 4). tested as a cancer drug delivery platform; therefore, doxorubicin
Magnetic Properties of Hybrid Systems. Magnetic was chosen as an eective drug, which is, however, highly toxic
characterization was conducted for nanoparticles covered with to healthy cells. We showed that the Pn3m cubic phase
the citric/oleic layer, for the cubic phases containing 2% w/w of structure is retained upon addition of 0.22% w/w magnetic
nanoparticles, and for the cubosomes with nanoparticles. nanoparticles and 0.5% w/w Dox. The lipidic hybrid material
Initially, it was observed that both the cubic phase and the exhibited magnetic properties and thus could be moved by the
cubosome suspension reacted to the magnetic eld of a application of a magnetic eld. The drug release proles from
neodymium magnet, involving a movement of particles toward the phases were established using dierential pulse voltammetry
the magnetic eld. (DPV). Introduction of 0.2% w/w nanoparticles results in
Both nanocrystalline nickelzinc ferrite and magnetite removal of more drug by ca. 10% compared with the pure
covered with the citric/oleic layer were incorporated into the monoolein cubic phase. Adding 2% w/w hydrophilic magnetic
cubic phases. Figure 6 shows that mass magnetization decreases nanoparticles leads to a 3-fold slower release of the drug
with temperature. At room temperature, the magnetic moment because the negative charge of these nanoparticles makes them
per nanocrystallite contained in the bulk cubic phase with reside mainly in the aqueous channels, and attractive
NPcitric and NPoleic equals 1.4 B and 2.7 B (Table 1), interactions with the positively charged drug retain it in these
respectively. As expected, these results conrm that the channels. However, the presence of 2% w/w hydrophobic NPs
magnetic particles of the ferrite and magnetite still retain the has almost no eect on the channel transport of the drug since
magnetic moment in the cubic phases. The minute magnet- the NPs are located mainly in the lipidic part of the cubic phase.
ization enhancement observed between 70 and 110 C Temperature increases the rate of drug release: In case of
seems to be related to evaporation of water; however, the NPcitric, it is 53% at 37 C, whereas for NPoleic it is increased to
detailed mechanism is not understood at this time. 85%. This result is interesting from the viewpoint of
The temperature variation of magnetization was also magnetothermal applications of the system. Eective drug
measured for the cubosomes (Figure 6). Qualitatively, the delivery systems should exhibit a large internal surface where
temperature variation is similar to that of the cubic phase. The the drug can be accumulated and should release the drug only
absolute magnetization values are reduced approximately 3 and in the cancer cell environment. Interactions with the drug are
12 times compared to those of the corresponding bulk cubic not favorable, and this is observed in the case of LCP with 2%
phases. When taking into account the content of magnetic w/w hydrophilic magnetic nanoparticles. In the case of
nanoparticles in the cubosomes, the magnetic moments are hydrophobic nanoparticles, the T50 value is 50 min for the
found to be 1.5B and 1.9B for the ferrite (NPcitric) and 0.2% w/w NPoleic phase and 30 min for material with 2% w/w
magnetite (NPoleic) systems, respectively. These magnetic NPoleic. At human body temperature, there was no change in
moment values are comparable to the values obtained for the the elution rate of drug for the phase with 2% w/w NPoleic. All
2803 DOI: 10.1021/acsami.6b12889
ACS Appl. Mater. Interfaces 2017, 9, 27962805
ACS Applied Materials & Interfaces Research Article
these results favor the material with hydrophobic magnetic NPs (12) Clogston, J.; Craciun, G.; Hart, D. J.; Caffrey, M. Controlling
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