Copyright © 2006 American Scientific Publishers Journal of

All rights reserved Nanoscience and Nanotechnology

Printed in the United States of America Vol. 6, 3095–3101, 2006

Characterization of Polymeric Micelles for Pulmonary

Delivery of Beclomethasone Dipropionate
Nazar Noureen Gaber, Yusrida Darwis, Kok-Khiang Peh, and Yvonne Tze-Fung Tan∗
School of Pharmaceutical Sciences, University of Science Malaysia, 11800 Penang, Malaysia

The potential of using poly-(ethylene oxide)-block-distearoyl phosphatidyl-ethanolamine (mPEG-

DSPE) polymer to prepare BDP-loaded micelles with high entrapment efficiency and mass median
aerodynamic diameter of less than 5
m demonstrating sustained release properties was evalu-
ated. The result showed that lyophilized BDP-loaded polymeric micelles with entrapment efficiency
of more than 96% could be achieved. Entrapment efficiency was affected by both the drug to poly-
mer molar ratio and the amount of drug used. Investigation using FTIR and DSC confirmed that
there was no chemical or physical interaction and the drug was molecularly dispersed within the
micelles. TEM images showed that the drug-loaded polymeric micelles were spherical in shape with
multivesicular morphology. Further analysis by photon correlation spectroscopy indicated that the
particle size of the BDP-loaded micelles was about 22 nm in size. In vitro drug release showed a
promising sustained release profile over six days following the Higuchi model. The mass median
aerodynamic diameter and fine particle fraction were suitable for pulmonary delivery. Moreover,
the small amount of deposited drug in the induction port (throat deposition) suggested possible
reduction in incidence of oropharyngeal candidiasis, a side effect normally associated with inhaled
corticosteroids therapy. IP:
The5.62.154.87
high encapsulation
On: Fri,efficiency, comparable
20 Dec 2019 inhalation properties, sus-
10:43:30
tained release behavior together
Copyright: American Scientific Publishers support the potential of
with biocompatibility nature of the polymer
BDP-loaded polymeric micelles as a versatile delivery system to be used in the treatment of asthma

RESEARCH ARTICLE
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and chronic obstructive pulmonary disease.
Keywords: Micelles, mPEG-DSPE, Beclomethasone Dipropionate, Pulmonary Delivery.

1. INTRODUCTION only release 15% of the drug over a period of eight

Inhaled glucocorticosteroid is considered the most effec-
tive treatment for asthmatic patients.1
2 However, the long
term use of inhaled glucocorticosteroid gives rise to sys-
temic side effects such as oropharyngeal candidiasis due
to the ingestion of the drug deposited in the patient’s
mouth and throat after inhalation. Recently, there has
been a growing interest in the formulation of beclometha-
sone dipropionate (BDP) pulmonary systems to achieve
therapeutic effect with added advantages of reduction in
frequency of application, decrease systemic absorption,
decrease side effects, and increase in patient compliance.
Nebulization of rehydrated freeze-dried BDP liposomal
preparations showed that the output of liposomes from
the nebulizer was dependent on the lipid used. Liposomes
formed from lipids with a high Tm exhibited a low out-
put of aerosols, due to aggregation during nebulization.3
BDP encapsulated in poly(L-lactic acid) yielded micro-
spheres in aerosolizable particle size range of less than
5 m. Nevertheless, degradation of the microspheres could
∗
Author to whom correspondence should be addressed.
days.4 As a result of nanoprecipitation, anhydrous BDP
in BDP-poly(L-lactic acid) nanoparticles was changed to
BDP monohydrate.5 Recently, non-phospholipid vesicles
loaded with BDP were fabricated with non-ionic surfac-
tant, polysorbate 20. Even though the fine particle fraction
(FPF) with aerodynamic diameter of less than 5 m was
95% and the permeation rate through model mucosal bar-
rier was greatly enhanced, the maximum entrapment effi-
ciency was only 20%.2
Polymeric micelles have gained importance in recent
years due to their promising colloidal carrier property
to deliver poorly water-soluble and amphiphilic drugs
to the site of action.6–10 The advantages of polymeric
micelles include better stability than surfactant micelles,
ability to solubilize an adequate amount of hydrophobic
drugs, prolonged circulation times in vivo, and capabil-
ity to accumulate in the target organs.7 Thus, deliver-
ing BDP-loaded polymeric micelles directly to the lung
in nanoparticle sizes in inhalation dosage form may be
deemed an effective means of treating asthma and chronic
pulmonary obstructive disease. This can be achieved since
the polymeric micelles are able to evade the mononuclear

J. Nanosci. Nanotechnol. 2006, Vol. 6, No. 9/10 1533-4880/2006/6/3095/007 doi:10.1166/jnn.2006.426 3095

 Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate Gaber et al.

phagocytic system due to their bulky hydrophilic outer
shell and sustain the release of the drug.9 In addition, drug-
loaded polymeric micelles were strongly suggested to pass
through the mucus layer associated with bronchial inflam-
matory diseases directly to their receptors in the epithelial
cells. Hence, the polymeric micelles will be very bene-
ficial in delivering hydrophobic corticosteroids, such as
BDP, for the treatment of chronic pulmonary obstructive
disease since the inability to penetrate through the mucus
layer to reach the target site was largely the result of fail-
ure in treatment in the past. In addition, it was reported
that a mammalian secreted phospholipase A2 (PLA2) was
able to degrade pegylated phosphatidyl ethanolamine. 11
12 and methanol was evaporated under vacuum at 40  C using
This finding supported the potential of pulmonary delivery
matrices containing pegylated lipid a prototype to target
glucocorticosteroids to the inflammed tissues of the lung.
The aim of this study was to evaluate the potential of
using poly-(ethylene oxide)-block-distearoyl phosphatidyl-
ethanolamine (mPEG-DSPE) polymer to prepare beclome-
thasone dipropionate loaded micelles with high entrapment
efficiency and mass median aerodynamic diameter of less
than 5 m, demonstrating sustained release properties. The
entrapment efficiency of beclomethasone dipropionate,
effects of drug-polymer molar ratio on particle size, phys-
icochemical properties, in vitro inhalation pattern, and
in vitro drug release profile of polymeric micelles were
also investigated.
IP: 5.62.154.87 On: Fri, 20
Copyright: American Scientific
RESEARCH ARTICLE
purchased from Merck, Germany. Acetonitrile of HPLC
grade was purchased from Labscan Asia, Thailand. All
other chemicals were of AR grade and used as received.
2.2. Preparation of Micelles by Solvent
Evaporation Method
2 mg/mL mPEG5000-DSPE stock solution and 0.5 mg/mL
BDP stock solution were prepared with methanol. Known
volumes of the two stock solutions were taken and the
final volume was adjusted with methanol. The content was
transferred to a round bottom flask, vortexed for 1 minute
a rotary evaporator (Eyela N-1000, Japan). 10 mL of dis-
tilled water was added to dissolve the drug/polymer film
in the round bottom flask. The aqueous solution was then
incubated at 40  C for 10 min and vortexed for 30 sec.
The micellar solution was filtered using 0.2 m PTFE
membrane filter to remove any non encapsulated BDP
and freeze-dried (Labconco 7753501, USA) overnight. The
various formulations are shown in Table I.
2.3. Determination of BDP Entrapment Efficiency
The lyophilized drug-loaded micelles were reconstituted in
a mixture of methanol and water (1:1, v/v) and the amount
of BDP was quantified using a UV spectrophotometer
(Hitachi U2000, Japan) at a wavelength of 241.5 nm. The
Dec 2019 10:43:30
entrapment
efficiency (EE) of the BDP-loaded micelles
Publishers

2. MATERIALS AND METHODS Delivered bywas determined.

Ingenta
amount of BDP in micelles
2.1. Materials EE % = × 100%
amount of BDP fed initially
Beclomethasone dipropionate (BDP) was purchased from The regression polynomial of entrapment efficiency was
Sigma, UK. Poly-(ethylene oxide)-block-distearoyl phos- calculated using the statistical software SPSS and was
phatidyl-ethanolamine (mPEG5000-DSPE) was obtained applied to approximate the response surface and contour
from NOF Corporation, Tokyo, Japan. Methanol was plot using the software Mathematica.
Table I. Effect of drug and mPEG5000-DSPE concentration on BDP entrapment efficiency and mean diameter by TEM.

Formula BDP mPEG5000-DSPE BDP:Polymer Entrapment efficiency Mean diameter

code (mol/mL) (mol/mL) (molar ratio) (% ± stdev) (nm ± stdev)

F1 0.1 0 1:0 17.22 ± 1.31 —

F2 0.1 005 1:0.5 24.80 ± 0.76 —
F3 0.1 01 1:1 66.69 ± 1.52 235.96 ± 48.99
F4 0.1 02 1:2 94.85 ± 1.75 71.47 ± 21.42
F5 0.1 03 1:3 96.83 ± 1.23 71.29 ± 15.34
F6 0.1 04 1:4 101.20 ± 3.77 52.62 ± 21.57
F7 0.1 05 1:5 102.91 ± 4.41 —
F8 0.2 0 2:0 6.84 ± 0.63 —
F9 0.2 005 2:0.5 18.47 ± 1.85 —
F10 0.2 01 2:1 31.06 ± 0.48 73.77 ± 20.27
F11 0.2 02 2:2 47.98 ± 3.63 150.50 ± 61.38
F12 0.2 03 2:3 30.35 ± 1.81 75.80 ± 33.65
F13 0.2 04 2:4 75.40 ± 3.39 65.72 ± 19.09
F14 0.2 05 2:5 52.36 ± 2.28 65.39 ± 13.15
F15 0.3 0 3:0 4.19 ± 0.71 —
F16 0.3 01 3:1 19.86 ± 1.56 45.60 ± 25.35
F17 0.3 03 3:3 19.42 ± 1.47 38.64 ± 19.57
F18 0.3 05 3:5 28.20 ± 0.83 37.13 ± 11.42

3096 J. Nanosci. Nanotechnol. 6, 3095–3101, 2006

Gaber et al. Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate
2.4. Characterization by FTIR and DSC
Approximately 1 to 2 mg of lyophilized drug-loaded
micelles and 20 mg of powdered potassium bromide was
triturated in an agate mortar with a pestle. The homoge-
nous mixture was then compressed at a compaction force
of 16 tons cm−2 and a holding time of 2 min to obtain
a potassium bromide disc. The spectrum was measured
using an IR spectrophotometer (Thermo Nicolet Corpora-
tion, Nexus model, USA equipped with Omnic 6.1 version
software). Samples of BDP, physical mixture of BDP and
polymer, lyophilized BDP-loaded polymeric micelles and
mPEG5000-DSPE were examined.
Differential scanning calorimetry (DSC) was carried out
using a thermal analysis controller (Perkin Elmer, Pyres 6,
UK). About 4 to 7 mg of the lyophilized drug-loaded micel-
les were placed into an aluminum pan and crimped.
Another pan was used to serve as the baseline. The ther-
mograms covered a temperature range of 0 to 250  C at a
heating rate of 10  C/min. The thermal properties of BDP,
physical mixture of BDP and polymer, lyophilized BDP-
loaded polymeric micelles, and mPEG5000-DSPE were
examined.
2.5. Particle Size Determination by TEM
The morphological examination of the drug-loaded micel-
les was carried out using a transmission electron micro-
IP: 5.62.154.87 On: Fri, 20 Dec of
Copyright: American
scope (TEM) (Phillips CM12, Netherlands) equipped with
Delivered by Ingenta
an image analysis system (Docu version 3.2, SIS, Ger-
many). The lyophilized micelles were first dispersed in
water and a drop of the sample was then placed onto a
400 mesh copper grid coated with a film of carbon. Excess
water was removed by blotting with filter paper. The sam-
ple was left to dry at room temperature before examination.
The TEM images were taken at different magnifica-
tions and labeled with the corresponding scale bars. Two
dimensional measurements (breadth and length) were car-
ried out manually for the particles. The particle size for
formulations containing different drug to polymer ratios
was determined. The mean and standard deviation were
calculated. No staining method was employed in the TEM
measurement.
2.6. Particle Size Determination Using Photon
Correlation Spectroscopy
The mean particle size and size distribution of BDP-loaded
micelles (F5) before freeze drying were determined by
photon correlation spectroscopy (PCS) using a Zetasizer
1000HS (Malvern Instrument, UK). The sample was mea-
sured for three cycles, with 10 measurements in each
cycle. The measurement was carried out at 25  C and
the angle of measurement was 90 . The mean Z average
particle diameter and polydispersity index (span) were
obtained using the Malvern Software System.
J. Nanosci. Nanotechnol. 6, 3095–3101, 2006
2.7. In Vitro Drug Release Study
Lyophilized BDP-loaded micelles (F5) (corresponding to
208 g of BDP) were dispersed in 30 mL of phosphate
buffer solution pH 6.8. 5 mL was transferred into a 5 cm
long dialysis tubing (Visking—dialysis tube size 4-22/32,
UK). The dialysis tubing was immersed in 70 mL of phos-
phate buffer solution pH 6.8 containing 0.1% of Tween
80 in a glass beaker. The latter were horizontally shaken
(30 rpm) in a thermostatic water bath maintained at 37 ±
05  C. The apparatus was protected from light throughout
the test. 0.1 mL samples were withdrawn from the beaker
at predetermined time intervals of 0, 2, 6, 12, 24, 48, 72,
and 144 hours. The amount of BDP released from the
micelles was determined using HPLC method. The release
studies were performed over a period of 6 days and each
experiment was conducted in triplicates.
The HPLC system was comprised of a pump (Shi-
madzu LC 6A), a UV detector (Shimadzu SPD-6A, Japan)
equipped with an integrator (Hitachi D2500, Japan).
A Rheodyne 7725 sample injector was fitted with a 20-L
sample loop. A Zorbax C18 (5 m, 250 mm × 4.6 mm ID)
column (Agilent, USA), fitted with a guard column was
used for the chromatography separation. The mobile phase
consisted of acetonitrile and distilled water (58:42, v/v).
Analysis was run at a flow rate of 1.0 mL/min. The sam-
ples were quantified using peak area at a detection wave-
length 2019
25410:43:30
nm. The HPLC method was validated.
Scientific Publishers
RESEARCH ARTICLE
2.8. Aerodynamic Characterization of
Nebulized Micelles
A known amount of lyophilized micelles F5 was dispersed
in distilled water and nebulized using a Pari LC Plus nebu-
lizer connected to a compressor (Pari Master Type 84G73,
Pari, Germany) through a 8-stage cascade impactor (Cop-
ley, NGI Model 170, UK) at a flow rate of 30 L/min using
a flow meter (Copley, Model DFM2, UK), at 25  C. BDP
was collected at different stages of the cascade impactor by
direct rupture of the polymeric micelles with acetonitrile.
The amount of BDP in each stage was determined using
HPLC. The determination was carried out in triplicates.
The mass median aerodynamic diameter (MMAD), geo-
metric standard deviation (GSD), and fine particle fraction
(FPF) were evaluated. MMAD and GSD were determined
after plotting the cumulative values of drug amount
deposited in each stage of the cascade impactor versus
their corresponding aerodynamic diameter as specified by
the cascade impactor using log-probability paper.13 The
fine particle fraction (FPF) was measured according to
Yamamoto et al. (2004).14
MMAD m = D50%


D841%
GSD =
D159%
3097
 Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate Gaber et al.

FPF < 66 m %

Total amount of drug from stage 2 and below
= × 100
amount of BDP loaded initially in the nebulizer
FPF < 39 m %
Total amount of drug from stage 3 and below
= × 100 100
amount of BDP loaded initially in the nebulizer
75 5
EE
50
2.9. Statistical Analysis 25
4

The results of the studies were treated statistically using
PC based software SPSS. General linear model univaraite
test, Post Hoc Tukey Honestly Significant Difference test,
and bivariate correlation test were applied, where appro-
priate. A statistically significant difference was considered
when p < 005.
3. RESULTS AND DISCUSSION
3.1. Effect of BDP and mPEG5000-DSPE
Concentrations on Entrapment Efficiency (EE)
The amount of BDP used were 0.1, 0.2, and 0.3 mol/mL,
while the amount of polymer used varied from 0 to
0.5 mol/mL. The drug to polymer ratios are shown in
Table I. The result shows that the entrapment efficiency
varied from 6.84 to 102.91% and can be expressed by the
following polynomial equation.
IP: 5.62.154.87 On: Fri, 20 Dec 2019
2
EE % = 2165 − 846x + 8549y −Copyright: American
3303xy − 1384y Scientific Publishers
RESEARCH ARTICLE
0 3
1
Polymer
1.5 2
2 1
BDP 2.5
3 0
Fig. 1. Response surface showing the entrapment efficieny (%) at var-
ious concentrations of BDP (×10−1 mol/ml) and mPEG5000-DSPE
(×10−1 mol/ml).
The importance of the initial amount of drug on entrap-
ment efficiency was also reported by other researchers.15–17
They found that when the amount of drug present excee-
ded the maximum loading capacity, precipitation of the
drug, and lower yield would be the outcome. In addi-
tion, EE for some hydrophobic drugs was reported to be
more than 90% when suitable formulation parameters were
employed. et al.18 studied the EE of hydrophobic
10:43:30
Wang
vitamin K3 in PEG3000-DSPE and achieved 980 ± 02%.

Delivered by Ingenta 19
2 2 2 3
+ 647xy − 134x y + 118x y + 023xy 3 Zhang et al. reported a high EE of approximately 100%
when PEG5000 conjugated with different phospholipids
+ 011x4 + 001y 4 including DSPE were used.
R2 = 0935
F = 6168
p < 001

Where 5

x = BDPmol/mL
y = mPEG5000-DSPEmol/mL 4
90 80 70 60 50 40 30
Figures 1 and 2 show the response surface and contour
mPEG5000-DSPE

plot of the entrapment efficiency of BDP, respectively. In 3

general, at any fixed concentration of polymer, an increase
in drug concentration showed a reduction in EE, while at
any fixed concentration of drug, an increase in polymer
2
concentration resulted in an increase in EE. In the absence
of polymer, EE was very low. At 0.1 mol/mL of BDP,
EE was about 17%, while at 0.3 mol/mL of BDP, EE was 20
only 4%. However, as the concentration of polymer was 1
increased to a range of 0.2 to 0.5 mol/mL, EE increased
drastically and reached more than 90% at BDP concen- 10
tration of 0.13 mol/mL or less. Nevertheless, when BDP 0
was increased to 0.3 mol/mL, maximum EE was between 1 1.5 2 2.5 3
20 to 30% even if the polymer concentration reached a BDP
high of 0.5 mol/mL. It is apparent that EE depended on Fig. 2. Contour plot showing the entrapment efficieny (%) at vari-
both the ratio of drug to polymer and also the concentra- ous concentrations of BDP (×10−1 mol/ml) and mPEG5000-DSPE
tion of drug present. (×10−1 mol/ml).

3098 J. Nanosci. Nanotechnol. 6, 3095–3101, 2006

Gaber et al. Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate

1662
bdp1 1

1617
1727
50 0

1184
%T

10

Heat flow
1755
–0
bdp-peg/mix 1662 20

1468
60
30
%T

1111
40
1615
1727

40
20
80 bdp-peg2 50
60 0 50 100 150 200 250 300
1662
1737

1111
%T

40
1468

20 Temperature
peg2
60
1468

40 0 2
%T

1110
1738

20 10

Heat flow
2000 1800 1600 1400 1200 1000 800 600 20
30
Wavenumbers (cm–1)
40
Fig. 3. FTIR spectra for BDP (1), physical mixture (2), lyophilized 50
BDP-loaded polymeric micelles (3), and mPEG5000-DSPE (4). 0 50 100 150 200 250 300
Temperature
3.2. FTIR
0 3
Figure 3 shows the FTIR spectra for BDP, physical mix-

Heat flow
10
ture, lyophilized BDP-loaded polymeric micelles, and 20
30
mPEG5000-DSPE. Both BDP and the physical mixture 40
showed the characteristic bands at 1662 cm−1 of car- 50
0 50 100 150 200 250 300
bonyl ketone and 1615 cm−1 of C=C stretching. How- Temperature
ever, for the lyophilized BDP-loaded polymeric micelles,
these peaks were diminished with the carbonyl ketone 0 4
band unchanged at 1662 cm−1 and the carbon double
Heat flow

10
bond shifted slightly to around 1630 cm−1 . The smaller 20
30
peak sizes were due to the relatively smaller amount of 40
BDP as compared to the pegylated lipid. In the spec- 50
trum of mPEG5000-DSPE, only theIP: 5.62.154.87
carbonyl ketoneOn:
bandFri, 20 Dec 20190 10:43:30
50 100 150 200 250 300
Copyright: American Scientific Publishers
was observed. Our result indicated that there wasDelivered
no clear by Ingenta Temperature

interaction between BDP and the polymer was detected.
Hyvönen et al.5 prepared BDP loaded poly(L-lactic acid)
nanoparticles and found clear shifts in both the carbonyl
and the carbon double bond peaks in the nanoparticles as
compared to the reference BDP.
3.3. DSC
Figure 4 shows the DSC thermograms of the various sam-
ples. It can be seen that BDP melted with decomposi-
 that the micelles were not homogenous in appearance and
tion at about 212.7 C. The peak for mPEG5000-DSPE
 resembled multivesicular liposome in structure21 but the
was at ca. 60 C, similar to the melting point provided
by the supplier of the polymer. The peaks for the physi-
 
cal mixture were at both 59 C and 205 C. Samples of
lyophilized BDP-loaded micelles showed only one peak
at 58  C, but no BDP peak, indicating that BDP was
molecular dispersed in the polymeric matrix. Nimodipine-
loaded poly(caprolactone)-poly(ethylene oxide)-polylac-
tide amphiphilic polymer nanoparticles were formulated by
Hu et al.20 They found the DSC of the drug loaded nanopar-
ticles showed no nimodipine peak and that the drug was
molecularly dispersed in the nanoparticles.
3.4. Morphology and Particle Size
Analysis by TEM
Most of the micelles were spherical in shape and majority
of the formulations yielding mean diameters of less than
RESEARCH ARTICLE
Fig. 4. DSC thermographs of BDP (1), physical mixture (2), lyophilized
BDP-loaded polymeric micelles (3), and mPEG5000-DSPE (4).
100 nm, except Formulations F3 and F11. In general, the
mean diameter was inversely correlated to both BDP con-
centration and polymer concentration (p < 001) (Table I).
Figure 5 shows the TEM of lyophilized BDP-loaded poly-
meric micelles, formulations having BDP to polymer ratios
of 2:1 and 2:3, respectively. The TEM micrographs showed
micelles obtained in the present study were below 200 nm
in size.
3.5. Particle Size Determination by Photon
Correlation Spectroscopy
The particle size of Formulation F5 before freeze-drying
determined by zetasizer was 218 ± 05 nm (polydispersity
of 0.14) smaller than the size obtained using TEM
(7129 ± 1534 nm). In both cases, the micellar samples
were dissolved in aqueous medium. The variation in par-
ticle size by TEM and PCS might be due to the difference
in the mechanism of the measurement and interpreta-
tion of results. TEM images involved a two-dimensional
measurement, while photon correlation spectroscopy was
based upon Mie Theory which allowed measured intensity

J. Nanosci. Nanotechnol. 6, 3095–3101, 2006 3099

 Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate Gaber et al.

y = 8.3092x–2.9772
100
R2 =0.99

% of BDP released
80

60

40

20

0
0 2 4 6 8 10 12 14
Square root of time (hr)

Fig. 7. Percent of BDP released from F5 versus square root of time

(Higuchi model).

100 nm
phosphate buffer solution pH 6.8 containing 0.1% of
Tween 80 was utilized as the dissolution medium in the
drug release study at 37  C. The release profile of F5 and
the reference BDP powder can be seen in Figure 6. Nearly
100% of BDP was released from the reference BDP pow-
der at the end of 24 hours. In contrast, about 12% of BDP
was released from F5 within the first 2 hours, followed
by a continuous release to reach 98.1% over six days.
The kinetic of BDP release from F5 was found to follow
Higuchi diffusion model with r 2 value of 0.99 (Fig. 7).

3.7. Aerodynamic Characterization

IP: 5.62.154.87 On: Fri, 20 Figure

Dec 82019
shows the distribution of the nebulized formula-
10:43:30
tion F5 Publishers
Copyright: American Scientific throughout the various stages of the cascade
RESEARCH ARTICLE

Delivered byimpactor.
Ingenta The concentration of BDP remaining in the neb-
200 nm
ulizer and mouth piece was approximately 56%. Three dif-
Fig. 5. TEM of polymeric micelles with BDP to polymer molar ratios ferent conventional jet nebulizers having small, medium,
2:1 (1) and 2:3 (2). and large droplet sizes were used to compare the aerody-
distributions to be converted to volume distributions for namic characterization of a commercial BDP preparation
particles of known optical properties. (ClenilR ).23 The amount of BDP retained in the nebulizer
and mouth piece was found to be directly correlated to the
3.6. In Vitro Drug Release relative droplet size produced by the nebulizers. This may
explain the relatively high deposition of BDP in the nebu-
BDP is highly hydrophobic and practically insoluble in lizer and mouth piece since Pari LC Plus, a conventional
water.22 BDP powder having a particle size of 44147 ± jet nebulizer having medium droplet size of 4 to 5 m
16972 nm was used as the reference powder, while was employed in this study. Similarly, high concentrations
of drugs retained in the nebulizers were also reported by
100
80.00
Deposition%
Drug released (%)

80 60.00

40.00 F5
60
20.00
40 F5
0.00
rt)
ct izer

7
1

r
po

te

BDP
e
e

fil
ag
ag

ag

ag

ag

ag

ag
l

20
in ebu

n
io

st
st

st

st

st

st

st
n
du
(

0
at
ro

0 24 48 72 96 120 144
th

Time (hr)
Fig. 8. Distribution of F5 in the nebulizer and throughout the cascade
Fig. 6. Drug release profile of BDP powder (reference) and lyophilized impactor following nebulization at a flow rate of 30 L/min for 15 min.
BDP-loaded polymeric micelles (F5). Mean ± SD, n = 3.

3100 J. Nanosci. Nanotechnol. 6, 3095–3101, 2006

Gaber et al. Characterization of Polymeric Micelles for Pulmonary Delivery of Beclomethasone Dipropionate

Table II. Aerodynamic parameters for BDP-loaded poly- The mass median aerodynamic diameter and fine particle
meric micelles (F5).
fraction were suitable for pulmonary delivery. Moreover,
Parameter Formulation F5 the small amount of deposited drug in the induction port
(throat deposition) may reduce the incidence of oropha-
FPF < 6.6 m (%) 31.6 ± 7.3
FPF < 3.9 m (%) 21.0 ± 4.3
ryngeal candidiasis, a side effect normally associated with
MMAD (m) 3.4 inhaled corticosteroids therapy.
GSD 2.1 The high encapsulation efficiency, comparable inhala-
FPF, MMAD, and GSD denote fine particle fraction, mass median aero- tion properties, sustained release behavior together with
dynamic diameter and geometric standard deviation, respectively. biocompatibility nature of the pegylated lipid support the
potential of BDP-loaded polymeric micelles as a versatile
Vaghi et al.23 and Darwis and Kellaway3 at 50 to 75% and delivery system to be used in the treatment of asthma and
22 to 74%, respectively. chronic obstructive pulmonary disease.
The deposition of the inhaled particles in the induction
port (which simulated the throat of the patient) was found Acknowledgments: The authors thank the University
to be as low as 3.6% of the nebulized fraction. The low of Science Malaysia, Penang, Malaysia, for providing the
amount of deposition suggests a possible reduction in the IRPA research grant and the Scholarship from Central Med-
incidence of oropharyngeal candidiasis, a side effect nor- ical Supplies, Khartoum, Sudan, in support of this work.
mally associated with inhaled corticosteroids therapy.
The mass median aerodynamic diameter (MMAD) and References and Notes
geometric standard deviation (GSD) of F5 were found to
1. S. Crowley, Pediatric Respiratory Reviews 4, 153 (2003).
be 3.4 m and 2.1, respectively. MMAD of 3.4 m was 2. C. Terzano, L. Allegra, F. Alhaique, C. Marianecci, and M. Carafa,
at the required level for asthma and chronic obstructive Eur. J. Pharmaceut. Biopharmaceut. 59, 57 (2005).
pulmonary disease treatment of less than 5 micron, while 3. Y. Darwis and I. W. Kellaway, Int. J. Pharm. 215, 113 (2001).
GSD of 2.1 indicated the polydisperse nature of the size of 4. M. M. El-Baseir, M. A. Phipps, and I. W. Kellaway, Int. J. Pharm.
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commercial BDP product (Clenil® )IP: 5.62.154.87
using On:®Fri,
Pari LC Plus . 207.Dec
M. C.2019
Jones10:43:30
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Copyright: American Scientific
48, 101Publishers
(1999).
The MMAD, FPF of <5 m (%) and FPF of Delivered <3 m, by Ingenta

were 7.5 m, 23%, and 7%, respectively. Furthermore,
MMAD of BDP micelles in this study was also compara-
ble to the MMAD of the liposomal formulations reported
by other authors.24
3 However, the liposomes are known
to be ruptured and lose their contents during nebulization.
The introduction of hydrophilic PEG segments in the poly-
meric micelles are most likely to prevent the direct contact
of the inner drug core to the shear forces generated by the
jet nebulizers and may be more effective for drug delivery.
4. CONCLUSION
In conclusion, BDP-loaded polymeric micelles with entrap-
ment efficiency of more than 96% were successfully for-
mulated. The entrapment efficiency was affected by both
the drug to polymer molar ratio and the amount of drug
used. The results obtained from FTIR and DSC confirmed
that there was no chemical or physical interaction and the
drug was molecularly dispersed within the micelles. TEM
images showed that the drug-loaded polymeric micelles
were spherical in shape. Analysis by photon correlation
spectroscopy indicated that the particle size of the BDP
loaded micelles before lyophilization was approximately
22 nm. In vitro drug release showed a promising sustained
release profile over 6 days following the Higuchi model.
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Received: 9 December 2005. Revised/Accepted: 27 January 2006.

J. Nanosci. Nanotechnol. 6, 3095–3101, 2006 3101