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Novel drug delivery system based on doxorubicin-

encapsulated magnetic nanoparticles modified
with PLGA-PEG1000 copolymer

Eommolbanin Ebrahimi, Abolfazl Akbarzadeh, Elham Abbasi, Amir Ahmad

Khandaghi, Farhad Abasalizadeh & Soodabeh Davaran

To cite this article: Eommolbanin Ebrahimi, Abolfazl Akbarzadeh, Elham Abbasi, Amir
Ahmad Khandaghi, Farhad Abasalizadeh & Soodabeh Davaran (2016) Novel drug delivery
system based on doxorubicin-encapsulated magnetic nanoparticles modified with PLGA-
PEG1000 copolymer, Artificial Cells, Nanomedicine, and Biotechnology, 44:1, 290-297, DOI:
10.3109/21691401.2014.944646

To link to this article: https://doi.org/10.3109/21691401.2014.944646

Published online: 11 Aug 2014.

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Artificial Cells, Nanomedicine, and Biotechnology, 2016; 44: 290–297
Copyright © 2014 Informa Healthcare USA, Inc.
ISSN: 2169-1401 print / 2169-141X online
DOI: 10.3109/21691401.2014.944646

Novel drug delivery system based on doxorubicin-encapsulated

magnetic nanoparticles modified with PLGA-PEG1000 copolymer
Eommolbanin Ebrahimi2, Abolfazl Akbarzadeh1,2, Elham Abbasi2, Amir Ahmad Khandaghi3,
Farhad Abasalizadeh2 & Soodabeh Davaran1,2
1Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran, 2Department of Medical Nanotechnology,

Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran, and 3Faculty of Medicine,
Tabriz University of Medical Sciences, Tabriz, Iran

eventually causes death. Most cancers that create in lung,

Abstract
New drug delivery systems delivered the active molecules
to the target site in a definite manner to produce the desired
effects without disturbing the delicate bio-environment. The
Fe3O4 magnetic nanoparticles were prepared by chemical
precipitation of Fe salts in the ratio of 1:2 under alkaline
and inert condition. PLGA-PEG1000 triblock copolymer was
synthesized by ring-opening polymerization. The properties
of this copolymer were characterized using Fourier transform
infrared spectroscopy. In addition, the resulting particles were
characterized by X-ray powder diffraction, scanning electron
microscopy, and vibrating sample magnetometry. The in
vitro doxorubicin (DOX) release profiles were obtained by
representing the percentage of DOX release. In this report, we
used this new method to fabricate PEGylated PLGA particles,
and examined the anticancer agent DOX.
Keywords: biomedical applications, doxorubicin, PLGA, polymeric
nanoparticles
Background
Cancer is a class of diseases described by uncontrolled cell
growth and this out-of-control cell growth starts with cells
in the large intestine. Recent developments in nanotech­
nology have allowed new research strategies to flourish
in the field of drug delivery. There has been considerable
interest in developing nanoparticles as effective drug deliv-
ery carriers. Lung cancer is the second most common can-
cer diagnosed in the UK after breast cancer. Around 43,500
people were diagnosed with lung cancer in the UK in 2011,
that is around 120 people every day (Cancer Research UK
2013). Lung cancer develops when normal lung cells sus-
tain genetic damage that eventually leads to uncontrolled
cell proliferation. Like all cancers, lung cancer cells have the
ability to invade neighboring tissues and spread or metasta-
size to distant parts of the body. Left untreated, lung cancer
known as primary lung cancers, are carcinomas that gain
from epithelial cells. The main types of lung cancer are small
cell lung carcinoma (SCLC), also called oat cell cancer, and
non-small cell lung carcinoma (NSCLC) (Quattrin 2004).
In most people, lung cancer is related to cigarette smoking.
Smoking causes nearly nine out of 10 cases (86%). A further
3% of cases of lung cancer are caused by exposure to second
hand smoke in non-smokers (passive smoking). Some other
causes of increasing lung cancer risk are: (Cancer Research
UK 2013)
•• Exposure to radon gas
•• Exposure to certain chemicals
•• Air pollution
•• Previous lung disease
•• A family history of lung cancer
•• Past cancer treatment
•• Previous smoking-related cancers
•• Lowered immunity
Appropriate primary treatment for NSCLC is determined
predominantly by stage of disease, but also by performance
status, co-morbidity and weight loss (Table I). Where per-
formance status is good with minimal co-morbidity and
weight loss, surgery, with intention to cure, is appropri-
ate for stage I, II and some IIIA patients. Radical radiation
therapy (RT) or combined chemo/RT should be considered
for locally advanced inoperable stage IIIA or IIIB disease.
Palliative chemotherapy should be considered for meta-
static stage IV disease (O’Connell 2010).
Magnetic nanoparticles are spherical nanocrystals of
10–20 nm of size with a Fe2 and Fe3 core surrounded by
dextran or poly(ethylene glycol) (PEG) molecules. Their
magnetic properties make them excellent agents to label
biomolecules in bioassays, as well as magnetic resonance
imaging contrast agents. They are also amenable to surface
functionalization for active targeting in vivo or for in vitro

Correspondence: Soodabeh Davaran, and Abolfazl Akbarzadeh, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz 5154853431,
Iran. Tel/Fax:984113355789. E-mail: akbarzadehab@tbzmed.ac.ir (A.A.), davaran@tbzmed.ac.ir (S.D.)
(Received 29 June 2014; revised 5 July 2014; accepted 10 July 2014)

290
 Novel Drug Delivery System 291

Table I. Approach to treatment of NSCLC. Table II. Some calculated physicochemical properties of
Performance doxorubicin–PLGA and doxorubicin (Bagheri et al. 2012).
Stage status Good Poor Physicochemical
Co-morbidity Min-mild Severe properties Doxorubicin–PLGA Doxorubicin
Weight loss Min-mild Severe Refractivity 214.17 135.50
Pulmonary Adequate Inadequate Polarizability 82.54 52.00
function Hydration energy  39.18  24.03
I Surgery Palliative care Surface area(A0) 987.26 729.45
Palliative RT where ∆G(solvation) (kcal/mol)  13.56  18.08
indicated Dipole moment (Debye) 7.736 7.767
II Surgery Palliative care BE (ev/mol)  4.034 –
Palliative RT where
indicated
III  Surgery (T3N1)
Radical RT Palliative care very suitable for the controlled delivery of medicaments in
 Chemo the body (Tamber et al. 2005).
IV  Chemo Palliative care
Palliative RT where The difficulty associated with surface-modifying PLGA
indicated particles has been the lack of functional chemical groups on
the aliphatic polyester backbone of the polymer. A diversity
of techniques have been developed for PEGylation of PLGA
diagnostics (Lu et al. 2007) .In the past few years, there has nanoparticles, for example adsorption (Faraasen et al. 2003).
been an increasing nanoparticles following intravenous Daunomycin (or daunorubicin) and adriamycin (or doxoru-
administration interest to develop stealth nanoparticles bicin or 14-hydroxydaunomycin) are renowned drugs used
as drug was determined over 24 h in rats carrier systems. in cancer chemotherapy (Di Macro et al. 1975).
One of the main methods for preparation of stealth nano- Fresh therapeutic approaches and unusual formulations of
particles or long-circulating nanoparticles is to modify their doxorubicin are directly needed to improve the value of treat-
surface with a hydrophilic, flexible and nonionic polymer, ment in lung cancer patients with a poor diagnosis (Allemann
PEG (Gref et al. 1994, Tobío et al. 1998, Quellec et al. 1998, et al. 1993). In this background, liposomes and polymer–drug
Peracchia et al. 1997). The biodegradable polyesters are all conjugates were developed in the 1960s and 1970s (Ma et al.
strongly hydrophobic, and this has caused some limita- 2011) (Figure 2). Doxorubicin is a highly potent antineoplastic
tions in practical drug formulations. To add hydrophilic and agent accepted for use against a large spectrum of tumors.
other physico-chemical properties, PEG has been incorpo- Regrettably, its long-term clinical use is compromised by tox-
rated into the biodegradable polyesters. PEG is a non-toxic, icities common to anthracycline drugs, the most serious being
water-soluble polymer with proven biocompatibility. Block irreversible cardiomyopathy and subsequent congestive heart
copolymers consisting of a hydrophobic polyester segment failure (CHF) (Yeh et al. 2004). There are a lot of procedures for
and a hydrophilic PEG segment have attracted large atten- drug delivery via drug–drug carrier combinations, for exam-
tion due to their biodegradability, biocompatibility, and ple encapsulation, hydrogel formation, nanoaggregation, and
tailor-made properties (Huh et al. 2003). Drug-loaded PLGA micellar delivery (Garg and Goyal 2014). Between these, dox-
nanoparticles (Figure 1a) are investigated for targeted and orubicin, encapsulation and micellar delivery have been the
non-targeted drug delivery. These nanoparticles have been focus of much attention because they can protect and carry
investigated especially in drug delivery systems for drug tar- the drug directed to its chosen target (Hassani and Bagheri
geting because of their particle size (ranging from 10 to 1000 2012). The geometric structure of doxorubicin–PLGA and
nm) and long circulation in blood. Thus, if the carrier size is daunorubicin–PLGA were optimized at B3LYP/6-311 g**
under 1 mm, an intravenous (i.v.) injection (the diameter of and HF/6-31g* level of theory and then the Gibbs free energy
the smallest blood capillaries is 4 mm) is enabled, minimizing of solvation (ΔG(salvation((was calculated at B3LY/6-31g*
any possible embolism (Mainardes and Evangelista 2005). level of theory using Gaussian 03 (Pourhassan-Moghaddam
Poly(d,l-lactide-co-glycolide) (PLGA or DLPLG) is et al. 2014) (Figure 3). Quantum mechanical molecular simu-
a very popular copolymer used for various medical, lation was also used to study drug delivery (Srinophakun and
pharmaceutical, industrial and other purposes (Verger Boonmee 2011) (Table II).
et al. 1998). The most attractive application of this copoly-
mer is, however, in medicine and pharmacy because it is
Experimental section
Materials
(a) (b) Ferric chloride hexahydrate (FeCl3∙6H2O), ferrous chlo-
ride tetrahydrate (FeCl2∙4H2O), and ammonium hydroxide
(32 wt%) were purchased from Fluka (Buchs, Switzerland).
d,l-lactide and glycolide were purchased from Sigma-
Aldrich (St Louis, MO) and recrystallized with ethyl acetate.
Drug Hydrophitic grup(PEG)
Stannous octoate (Sn(Oct)2:stannous 2-ethylhexanoate),
PEG (molecular weight 1000), and dimethyl sulfoxide were
Figure 1. Polymeric nanoparticles (a) prepared with poly(d,l-
lactide-co-glycolide) (PLGA) and (b) prepared with poly(d,l-lactide- purchased from Sigma-Aldrich. Doxorubicin hydrochlo-
co-glycolide) co-polymer. ride was purchased from Sigma-Aldrich. X-ray diffraction,
292 E. Ebrahimi et al.
Figure 2. Structures of adriamycin (R  OH) and daunomycin (R  H)
(Manchanda et al. 2010).
Rigaku D/MAX-2400 X-ray diffractometer with Ni-filtered
Cu Ka radiation, and scanning electron microscopy (SEM)
measurements were conducted using VEGA/TESCAN. The
drug-loading capacity and release behavior were determined
using an ultraviolet visible 2550 spectrometer (Shimadzu,
Tokyo, Japan). Infrared spectra were recorded in real-time
with a Perkin Elmer series FTIR. The magnetic property was
calculated on a vibrating sample magnetometer (Meghnatis
Daghigh Kavir, Iran) at room temperature. The samples
were homogenated using a homogenizer (SilentCrusher M,
Heidolph Instruments GmbH, Schwabach, Germany). The
organic phase was evaporated by a rotary (Rotary Evapora-
tors, Heidolph Instruments, Hei-VAP series).
Synthesis of superparamagnetic magnetic nanoparticles
Magnetic nanoparticles were synthesized using an improved
chemical coprecipitation method. According to this
method, 1.5868 g of FeCl2∙4H2O (0.008 mol) and 3.7842 g of
FeCl3∙6H2O (0.014 mol) were dissolved in 320 mL of deion-
ized water, such that Fe2/Fe3  1/2. The mixed solution
was stirred under nitrogen at 75°C for 1 h. Then, NH3∙H2O
40 mL was injected into the mixture quickly, stirred under
nitrogen for another hour, and then cooled to room tem-
perature. The precipitated particles were washed five times
with hot water and separated by magnetic decantation. At
last, the magnetic nanoparticles were dried under vacuum
at 70°C. (Yang et al. 2006).
Preparation of PLGA-PEG triblock copolymer
PLGA-PEG1000 copolymer as an initiator was synthesized
by a melt polymerization process under vacuum using
Figure 3. Structures of doxorubicin–PLGA (Bagheri et al. 2012).
stannous octoate [Sn(Oct)2:stannous 2-ethylhexanoate] as a
catalyst. d,l-lactide (7.2 g), glycolide (1.93 g), and PEG1000,
4 g (60% w/w), in a bottleneck flask were heated to 130°C
under a nitrogen atmosphere for complete melting. The
molar ratio of d,l-lactide and glycolide was 3:1. Then 0.05%
(w/w) stannous octoate was added and the temperature of
the reaction mixture was raised to 170°C. The temperature
was maintained for 3 h. The polymerization was carried
out under vacuum. The copolymer was recovered by dis-
solution in methylene chloride followed by precipitation in
ice-cold diethyl ether. A triblock copolymer of PLGA-PEG
was prepared by ring opening polymerization of d,l-lactide
and glycolide in the presence of PEG1000 (Jeong et al. 2000).
Preparation of doxorubicin-encapsulated Fe3O4 magnetic
nanoparticles modified with PLGA-PEG copolymer
Doxorubicin-encapsulated Fe3O4 magnetic nanoparticles
modified with PLGA-PEG copolymer were prepared using
the double emulsion method (w/o/w) employed by Song
et  al. with minor modifications. An aqueous solution of
doxorubicin 2.5 mg/2.5 mL was emulsified in 10 mL dichlo-
romethane, in which 120 mg of the copolymer and 1 mg
magnetic nanoparticles had been dissolved, using a probe
homogenizer or sonication at 20,000 rpm for 30 s. This w/o
emulsion was transferred to a 50 mL aqueous solution of poly-
vinyl alcohol 1% and the mixture was probe-homogenized
at 72,000  rpm for 1 minute. The w/o/w emulsion formed
was gently stirred at room temperature until evaporation
of the organic phase was completed or the organic phase
was evaporated (Heidolph Instruments). The nanopar-
ticles were purified by applying two cycles of centrifugation
(12,000 rpm for 1 h in a Biofuge 28 RS, Heraeus centrifuge)
and reconstituted with deionized and distilled water. The
nanoparticles were finally filtered through a 1.2 mm filter
(Millipore, Bedford, MA). In order to increase doxorubicin
entrapment in the nanoparticles, the external aqueous
phase used during the second emulsification step was satu-
rated with doxorubicin. Blank nanoparticles were also pre-
pared by the same method without adding doxorubicin at
any stage of the preparation. The content and encapsulation
efficiency of doxorubicin in Fe3O4 magnetic nanoparticles
modified with PLGA-PEG copolymer were determined by
the disintegration of nanoparticles in dichloromethane. The
doxorubicin concentrations were determined spectropho-
tometrically at 480 nm. Drug encapsulation efficiency was
calculated using the following equations:
Encapsulation Efficiency (%)  [amount of doxorubicin
drug in mg/amount of added drug in mg]  100%
In vitro drug release study
To study the drug release profile of the synthesized doxo-
rubicin-encapsulated Fe3O4 magnetic nanoparticles modi-
fied with PLGA-PEG copolymer, 3 mg of drug-encapsulated
nanoparticles were dispersed in 30 mL of phosphate-
buffered solution (pH 7.4). Samples were incubated at 37°C.
At designated time intervals, a 3 mL sample was removed
and same volume was reconstituted by adding 3 mL of fresh
phosphate-buffered solution and acetate buffer to each
sample. After the experiment, the samples were analyzed

using ultraviolet spectrofluorometry to find out the amount
of doxorubicin released (lex 470 nm and lem 585 nm for
doxorubicin measurement).
Results and discussion
Nanotechnology-based drug delivery is expected to dra-
matically change combination cancer therapy by controlling
accumulation and distribution patterns of multiple drugs
selective in disease sites. Drug targeting and mediators in
cancer magnetic hyperthermia (Chavanpatil et  al. 2007,
Nidhin et  al. 2008) as well as unique functional character-
istics as small size, high stability, low toxicity (Chouhan and
Bajpai 2009), easy purification and sterilization, etc. (Yadav
et  al. 2007). In addition, human cells’ diameter is nearly
between 10 and 20 mm and the size of cell organelles range
from nanometers to a few hundred nanometers (Yadav et al.
2007). Thus, nanoparticles can overcome physiological bar-
riers and readily interact with intracellular compartments
without any additional surgery (Chouhan and Bajpai 2009).
In cancer therapy, most proposed formulations present
certain drawbacks related to low drug loading, toxicity, and/
or an unsuitable release pattern (Immordino et  al. 2003).
An ideal formulation should supply biocompatible nano-
sized particles and high drug loading with sustained-release
characteristics, allowing drug release in the target site at a
therapeutic concentration, thereby minimizing drug inef-
ficiency and adverse effects. Various approaches have been
investigated in this line (Lee et al. 2011, Zhao et al. 2010, Gao
et al. 2008).
In this work, we reformulated doxorubicin in a better-
tolerated and less-toxic vehicle. To decrease or minimize
undesired interactions or undesired uptake into normal
sites, a biodegradable nanocarrier has been developed for
doxorubicin, wherein the amount and site of drug release is
controlled by the structure of copolymer-coated magnetic
nanoparticles and pH. This nanoparticle was designed and
prepared so that the carrier can be used for targeting an
extensive range of solid tumors. For this purpose, AB triblock
copolymer of PLGA-PEG was synthesized by ring open-
ing polymerization of lactide and glycolide in the presence
of PEG1000 (Valizadeh et  al. 2012, Akbarzadeh et  al. 2012c,
Akbarzadeh et al. 2013). FTIR spectra were consistent with the
structure of the PLGA-PEG copolymer. The molecular weight
was determined by gel permeation chromatography. In this
work, doxorubicin-encapsulated Fe3O4 magnetic nanopar-
ticles modified with PLGA-PEG copolymer were obtained
by encapsulation of doxorubicin in the nanoparticles. For
this purpose, the double emulsion (w/o/w) technique was
considered the most appropriate method. Nonetheless, the
influence of other factors on entrapment efficiency using
this technique is very complicated and includes copolymer
concentration in organic solution, volume of the inner aque-
ous phase, volume of the outer aqueous phase, doxorubicin
concentration in the inner aqueous phase, the first homog-
enized speed and time, the second homogenized speed
and time, and polyvinyl alcohol concentration. The loading
efficiency values achieved for doxorubicin were different
between the various Fe3O4-PLGA-PEG nanoparticles, which
Novel Drug Delivery System 293
could be attributable to the presence of different molecular
weights of PEG in the PLGA chains, but the mechanism is
indistinct.
The entrapment efficiency was 70%, and the particle size
was about 50–100 nm. The results demonstrated in vitro that
the doxorubicin-encapsulated Fe3O4-PLGA-PEG nanopar-
ticles show pH sensitivity and can be applied for targeting
extracellular pH and could be an effective carrier for antican-
cer drugs. It is expected that at tumor pH, the doxorubicin-
encapsulated nanoparticles made of Fe3O4-PLGA-PEG can
show enhanced cytotoxicity compared with that at normal
pH  7.4. In this paper, higher and faster doxorubicin release
was observed for Fe3O4-PLGA-PEG1000 nanoparticles at 12 h.
In conclusion, modification of the magnetic nanoparticles
could have potential benefit for drug delivery. Our results
show that magnetic Fe3O4-PLGA-PEG nanoparticles could
be an effective carrier for drug delivery.
Nanoparticle characterization
Power X-ray diffraction (Rigaku D/MAX-2400 X-ray dif-
fractometer with Ni-filtered Cu Ka radiation) was used to
investigate the crystal structure of the magnetic nanopar-
ticles. The size and shape of the nanoparticles were deter-
mined by SEM. The sample was dispersed in ethanol and
a small drop was spread onto a 400 mesh copper grid. The
magnetization curves of the samples were measured using
vibrating sample magnetometry at room temperature. The
infrared spectra were recorded by an FTIR spectrophotom-
eter (Perkin Elmer series), and the sample and KBr were
pressed to form a tablet. 1H NMR spectra were recorded in
real-time with a Brucker DRX 300 spectrometer operating
at 300.13 mHz.
X-ray diffraction patterns
The X-ray diffraction gives patterns for pure Fe3O4 and
doxorubicin-encapsulated Fe3O4 magnetic nanoparticles
modified with PLGA-PEG copolymer. The characteristic
diffraction peaks are marked, respectively, by their indices
(220), (311), (400), (422), (511), and (440), which could be
well indexed to the inverse cubic spinel structure of Fe3O4
(JCPDS card 85-1436). Characteristic diffraction peaks
were also observed for doxorubicin-encapsulated Fe3O4
magnetic nanoparticles modified with PLGA-PEG copo-
lymer. This demonstrates that modification of the Fe3O4
nanoparticles did not lead to any crystal phase change. The
average crystallite size D was about 10 nm and obtained
from the Sherrer equation D  Kl/(bcos q), where K is the
constant, l is the X-ray wavelength, and b is the peak width
of half-maximum.
Size and size distribution (SEM)
The surface morphology of the nanospheres during the
incubation period was observed by SEM. The nanographs
of pure Fe3O4 nanoparticles (Figure 4a) and doxorubicin-
encapsulated Fe3O4 magnetic nanoparticles modified with
PLGA-PEG copolymer (Figure 4b) are shown. Observing
the photograph, it can be seen that the nanoparticles
were well aggregated, which was due to the nanosize of
the Fe3O4 of about 10 nm. After encapsulation and modi-
294 E. Ebrahimi et al.
Figure 4. Scanning electron microscopy of (a) Fe3O4 magnetic nanoparticles and (b) doxorubicin-encapsulated Fe3O4 magnetic nanoparticles
modified with PLGA-PEG copolymer.
fication of the doxorubicin-encapsulated Fe3O4 magnetic
nanoparticles with PLGA-PEG copolymer, the size of the
particles changed to 50–100 nm and dispersion of the
particles was greatly improved (Figure 4b), which can be
explained by the electrostatic repulsion force and steric
hindrance between the copolymer chains on the encap-
sulated Fe3O4 nanoparticles. The samples were coated
with gold particles.
TEM
Figure 5 shows delegate TEM images of synthesized Fe3O4
nanoparticles. The TEM image indicates that magnetite
nanoparticles are nanocrystalline, though their shape is
primarily spherical with some hexagonal shaped nanopar-
ticles. The size of the spherical nanoparticles varies from 7 to
15 nm with average particle size of 12 nm and corresponds
to the size estimated using X-ray diffraction measurements.
Moreover, Fe3O4 nanoparticles are agglomerates due to
Figure 5. TEM images of magnetic Fe3O4.
high surface area and magnetic dipole–dipole interactions
between particles.
Magnetism test
The magnetic properties of the nanoparticles were analyzed
by vibrating sample magnetometry at room temperature.
Figure 6 shows the hysteresis loops of the samples. The
saturation magnetization was found to be 18 emu/g for
doxorubicin-encapsulated Fe3O4 magnetic nanoparticles
modified with PLGA-PEG copolymer, that is, less than for
the pure Fe3O4 nanoparticles (61 emu/g). This difference
suggests that a large amount of polymer encapsulated
the Fe3O4 nanoparticles and doxorubicin. With the large
saturation magnetization, the doxorubicin-encapsulated
Fe3O4 magnetic nanoparticles modified with PLGA-PEG
copolymer could be separated from the reaction medium
rapidly and easily in a magnetic field. In addition, there was
no hysteresis in the magnetization, with both remanence
and coercivity being zero, suggesting that these magnetic
nanoparticles are superparamagnetic. When the external
magnetic field was impassive, the magnetic nanoparticles
Figure 6. Magnetic behavior of magnetic nanoparticles.

Figure 7. Fourier transform infrared spectra of doxorubicin-
encapsulated magnetic nanoparticles modified with biocompatible
copolymer.
could be well dispersed by gentle shaking. These magnetic
properties are critical for application in the biomedical and
bioengineering fields.
FTIR spectroscopy
The FTIR spectrum is consistent with the structure of the
expected copolymer. FTIR spectroscopy was used to show
the structure of Fe3O4 and PLGA-PEG copolymer nano-
particles. From the infrared spectra shown in Figure 7, the
absorption peaks at 580 cm 1 belonged to the stretching
vibration mode of Fe–O bonds in Fe3O4. Also the absorp-
tion band at 3509.9 cm 1 is assigned to terminal hydroxyl
groups in the copolymer from which PEG homopolymer has
been removed. The bands at 3010 and 2955 cm 1 are due to
C–H stretch of CH and 2885 cm 1 due to C–H stretch of CH.
A strong band at 1762.6 cm 1 is assigned to C  O stretch.
Absorption at 1186–1089.6 cm 1 is due to C–O stretch.
In vitro release experiment
The in vitro doxorubicin release profiles were obtained by
representing the percentage of doxorubicin release with
respect to the amount of doxorubicin encapsulated. For three
nanoparticles, doxorubicin release occurred in two phases:
an initial burst release, with a significant amount of drug
released within 12 h, 33.1% for Fe3O4 magnetic nanoparticles
modified with PLGA-PEG1000 nanoparticles; and after 12 h,
the doxorubicin release profiles showed a sustained release
pattern (Figure 8). The collective amount of doxorubicin
release over 2 days was 81.4% from Fe3O4-PLGA-PEG1000.
The doxorubicin release rate from the Fe3O4-PLGA-PEG
nanoparticles was also pH-dependent and enhanced at pH
5.8. It is generally assumed that a drug is released by sev-
eral processes, including distribution through the polymer
matrix, release by polymer degradation, and solubilization
and diffusion through microchannels that exist in the poly-
mer matrix or are formed by erosion.
The magnetic-coated copolymer prepared in the present
work are AB triblock copolymer composed of hydrophobic A
blocks (lactide-co-glycolide) and hydrophilic B blocks (cen-
tral PEG). This copolymer is not soluble in water but exhibit
Novel Drug Delivery System 295
120
100
80
Total released Dox
60
40 pH=5.8
pH=7.4
20
pH=8
0
0 2 4 6 8 10 12 14
–20 Time(h)
Figure 8. Release profiles of doxorubicin from the hybrid nanoparticles
in different pHs (■: pH  7.4, : pH  8, : pH  5.8).
reverse thermal and pH-dependent gelation properties.
Hydrolysis of the ester linkage in these polymers will cause
the swelling to increase with time as hydrolysis proceeds.
The gel becomes increasingly pH-sensitive as hydrolysis
proceeds, and carboxylic acid groups are generated in the
structure. Within about 10 days, we can consider that drug
is released from the Fe3O4-PLGA-PEG nanoparticles by a
diffusion mechanism in vitro. The swelling of the particles
increases in acidic buffered solutions due to protonation
of central PEG groups and formation of positively charged
chains in the polymer structure.
Conclusions
Iron oxide nanoparticles were prepared using an improved
chemical coprecipitation method and then PLGA-PEG
copolymer was used to encapsulate Fe3O4 nanoparticles
by an emulsion method (w/o/w). The consequences indi-
cate that the copolymer chains effectively encapsulated
the Fe3O4 nanoparticles. Saturation magnetization was
found to be 18 emu/g. These particles were employed in
the encapsulation of doxorubicin under mild conditions
and could be used in drug delivery (Akbarzadeh et  al.
2012a, 2012b, 2012c, 2012d, Ebrahimnezhad et  al.
2013, Davaran et  al. 2014a, Rezaei-Sadabady et  al. 2013,
Nejati-Koshki et al. 2013, Ghasemali et al. 2013, Mollazade
et  al. 2013, Davaran et  al. 2014b, Davaran et  al. 2013,
Ahmadi et  al. 2014, Pourhassan-Moghaddam et  al. 2013,
Kouhi et  al. 2014, Sadat Tabatabaei Mirakabad et  al. 2014,
Alizadeh et al. 2014c).
Our results suggest that supercritical liquid technology is
a promising technique to produce drug–polymer magnetic
composite nanoparticles for the plan of controlled-release
drug systems.
Therefore, these nanoparticles could become a potent
chemopreventive and chemotherapeutic system for lung
cancer patients and constituents of these nanoparticles
could be appropriate candidates for drug development.
Future work will include an in vivo investigation of the tar-
geting capability and effectiveness of these nanoparticles in
the treatment of lung cancer (Eatemadi et al. 2014a, 2014b,
296 E. Ebrahimi et al.
Abbasi et al. 2014a, 2014b, 2014c, Alizadeh et al. 2014a, 2014b,
Nejati-Koshki et al. 2014a, 2014b, Gandomkar Ghalhar et al.
2014, Karnoosh-Yamchi et al. 2014, Zohre et al. 2014, Fekri
Aval et al. 2014, Ebrahimi et al. 2014).
Authors’ contributions
AA and SD conceived of the study and participated in its
design and coordination. EA and FA participated in the
sequence alignment and drafted the manuscript. All authors
read and approved the final manuscript.­­­
Acknowledgments
The authors thank Department of Medical Nanotechnology,
Faculty of Advanced Medical Science of Tabriz University
for all supports provided.
Declaration of interest
The authors report no declarations of interest. The authors
alone are responsible for the content and writing of the
paper.
This work is funded by 2013 Drug Applied Research
Center Tabriz University of Medical Sciences Grant.
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