Pharmaceutical Development and Technology, 12:591601, 2007

Copyright Informa Healthcare USA, Inc.

ISSN: 1083-7450 print / 1097-9867 online
DOI: 10.1080/10837450701481181

LPDT

Development of Liposomal Systems of Finasteride for Topical Applications:

Design, Characterization, and In Vitro Evaluation
Rajiv Kumar, Bhupinder Singh, Gautam Bakshi, and Om Prakash Katare

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Development of Liposomal Systems of Finasteride for Topical Applications

University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, India

Finasteride (FNS) is a drug of choice for benign prostate

hypertrophy and prostate cancer. The drug has also been reported
to be useful orally in the treatment of some difficult-to-treat
androgen-dependent skin disorders, such as seborrhea, acne,
hirsutism, and androgenetic alopecia. However, the ideal route
for its administration (i.e., topical) remains unexplored. This has
logically suggested the search for strategic formulation
approaches to make the drug effective on topical applications,
hitherto unexplored. The present study targets the encasement of
drug molecules in the interiors of vesicular compartments (liposomes) made up of hydrogenated phospholipids, as an attempt
toward the development of a trans-epidermal therapeutic system
of FNS. Multilamellar drug-loaded liposomes were prepared by
thin-film hydration with sonication method and optimized with
respect to drug payload, entrapment efficiency, and size by formulating different vesicular compositions under different process
conditions. The vesicular systems consisting of saturated phospholipid (100 mg), cholesterol (50 mg), and FNS (5 mg) showed
highest drug payload (2.9 mg/100 mg of total lipids), and drug
entrapment efficiency (88.6%). Mean ( SD) vesicle size of the
prepared liposomes was found to be 3.66 1.6 m. Significantly
higher skin permeation of FNS through excised abdominal mice
skin of FNS was achieved from the liposomal formulations vis-vis corresponding solution and conventional gels. Liposomal
FNS formulations also showed more than fivefold higher deposition of drug in skin than the corresponding plain drug solution
and conventional gel. Stability studies indicated that the liposomal formulations were quite stable in the refrigerated conditions
for 2 months with negligible drug leakage or vesicle size alteration during the storage period. Results of the current studies
with FNS-loaded vesicular systems project the high plausibility

Received 22 November 2006, Accepted 25 April 2007.

Address correspondence to Professor Om Prakash Katare,
University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh-160 014, India; E-mail: drkatare@yahoo.com

of a topical liposomal formulation for effective localized delivery

of Finasteride.
Keywords phospholipids, drug delivery, transepidermal, skin,
permeation, vesicles, dermatological disorders

INTRODUCTION
Finasteride (FNS), a synthetic 4-azasteroid compound[1] (4-azaandrost-1-ene-17-carboxamide), is an antiandrogen used for benign prostate hyperplasia (BPH) in
low doses (1 mg/day), and for prostate cancer in higher
doses (5 mg/day).[27] Recently, oral administration of FNS
has also been found to be useful in the treatment of various
dermatological and follicular disorders, such as acne, seborrhea, and male pattern baldness (i.e., androgenetic alopecia).[813] It is well documented that some skin or hair
follicle disorders involve alteration in the rate of protein
synthesis in the nuclei of skin or hair follicle cells.[1,14]
Dihydrotestosterone (DHT), a derivative hormone (metabolite) of testosterone, has been shown to be critical in these
disorders, because alteration in the rate of protein synthesis
is caused by internalization of DHT-androgen receptor
complex in the nuclei of skin or of hair follicle cells.[15]
FNS blocks the production of DHT from testosterone by
competitively and specifically inhibiting type II 5- reductase
isozyme, thus decreasing many of its effects.[16] Hence,
FNS tends to plays a vital role in the treatment of clinical
problems related with skin and hair follicles.[8,17,18]
Although the importance and potential of FNS in the
treatment of dermatological problems has found a tacit
place, its oral route of administration has been a critical
issue.[19] The treatment of local skin problems involving
dermal layers and follicles would be more logical only
through the topical route, because oral administration of
FNS tends to cause several untoward effects in a vast
majority of patients. Common adverse effects following

591

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R. Kumar et al.

oral intake include impotence, impaired reproductive

function, erectile dysfunction, and gynecomastia.[2022]
Nevertheless, the oral route has hitherto been preferred on
account of unavailability of an effective topical formulation
to deliver FNS onto the desired site of action.
Lately, different drug delivery approaches have been
successfully explored to achieve effective drug transport
into and across the skin.[2325] Among these, phospholipidbased self-assembled supramolecular structures like
liposomes have shown promising results.[26,27] By virtue of
the unique physicochemical character and diversity in their
constructs, liposomes have proved to be a very effective
means of delivery, especially for topical administration.[2832]
Besides the characteristic merits of liposomes, their choice
for delivery of FNS to the mesodermal site of action
(including hair follicles) is also supported by the physicochemical nature of the drug, which is highly lipophillic (log
P = 4.277). It is supposed to get a more conducive milieu
within the unique amphiphillic interiors of vesicular compartments, favoring its partitioning and transport across the
skin barrier. Furthermore, the carrier-mediated delivery
would significantly augment the interaction of FNS with
androgen receptors of skin and follicles leading to the
enhanced drug action.[33] The latter has been recently
reported by Tabbakhian et al.[34] by preparing negatively
charged liquid state liposomes and niosomes of FNS and
evaluating them for follicular deposition of FNS in the
pilosebacious units of hamster flank skin and ears.
Thus liposomes were chosen in the current study to
exploit the potential of FNS in dermatological problems
through topical delivery. The current work relates to the
systematic development of multilamellar FNS-loaded vesicles considering various vesicle-specific as well as preparatory process-related parameters. Herein, the influence of
liposomal composition and process conditions was investigated on entrapment efficiency, drug payload, and vesicle
size. The effect of different lipid compositions, surface
charge, and vesicle size on the skin permeation behavior of
prepared FNS liposomes was also studied in vitro by using
excised abdominal mice skin, and the results obtained were
compared with the corresponding conventional FNS formulations. Finally, the stability studies on the optimized liposomal formulation were carried out to assess the influence of
storage conditions and time on drug retention characteristics, physical stability, and size of the prepared vesicles.

EXPERIMENTAL
Materials
FNS (Cipla Limited, India) and hydrogenated soy
lecithin (PC; Phosphatidylcholine content 9297%;

Phospholipon 90H; Phospholipids GmbH, Germany)

used in the current study were generous gifts from the
respective sources. Cholesterol (CHOL), Sephadex G-50
medium (bead size range 50150 m), and Hexadecyl
phosphate (Dicetyl phosphate; DCP) were procured from
Sigma Chemical Co. (St. Louis, MO, USA). Methylcellulose (viscosity grade: 30005000 mPas, 2%/20C) was
procured from LOBA-Chemie (India). All other materials
and solvents obtained from commercial sources were of
analytical grade. Double-distilled water was used throughout
the experimental studies.

Preparation of FNS-Loaded Multilamellar Liposomes

Different compositions used for preparation of FNS
containing liposomes are listed as Table 1. Multilamellar
vesicles were prepared by using film hydration with sonication technique.[35,36] For each formulation, a thin lipid
film was prepared by dissolving accurately weighed
amounts of the drug, PC, and/or CHOL in a minimum
volume of chloroform in a round-bottom flask. The solvent was then rotary evaporated at 40C (Buchi RE 121
Rotavapour, Buchi Laboratories, Switzerland) under nitrogen stream. The thin lipid film formed on the wall of the
flask was flushed with a stream of nitrogen for 1 min and
subsequently hydrated with distilled water for 20 min at
55 2C. The liposomal dispersion thus obtained was vortexed for 5 min and left undisturbed at room temperature
for 2 hr to allow annealing of the lipid bilayers. The
homogenous suspension of multilamellar vesicles was
subsequently sonicated (titanium microtip in.; Probe
sonicator-Misonix S-3000 USA; 4C; 36 Watts; 10 min-5
cycles of 2 min each) to reduce the size of MLVs. The
resulting liposomal suspensions were stored in nitrogenflushed vials at 4C until further studies. Liposomes, containing varied amounts of DCP, a surface charge inducer,
were also prepared analogously. The content of FNS with
respect to total lipids ranged between 20 and 50 ng per mg
of total lipids, and lipid concentrations used in different
liposomal suspensions ranged between and 10 and 17.5
mg mL1.

Estimation of Percent Drug Entrapment and Drug

Payload in Liposomes
Unentrapped drug from the prepared liposomes was
separated by mini-column centrifugation method.[37]
Liposomal suspension (0.2 mL) was placed in Sephadex
G-50 column, presaturated with empty liposomes, and
centrifuged at 2000 rpm (626 g) for 3 min. Elutes
containing drug-loaded liposomes were collected and

5
5
5
5
5
5
5
5
5
5
5

FNS (mg)
100
100
100
100
100
90
75
60
100
100
100

PL 90H (mg)
10
25
50
75
60
75
90
50
50
50

CHOL (mg)

Amount of FNS
entrapped*(mg)
3.17 0.13
3.49 0.23
3.93 0.08
4.43 0.18
4.24 0.12
4.11 0.19
3.60 0.12
3.50 0.03
4.37 0.12
3.90 0.09
2.50 0.12

DCP
(wt % of total lipids)

1
2
5

63.4
69.8
78.6
88.6
84.8
82.2
72.0
70.0
87.4
78.0
50

PDE

FNS: Finasteride; PL90H: Phospholipon 90H; CHOL: Cholesterol; DCP: Dicetylphosphate; PDE: Percent drug entrapment.
*Each value represents Mean SD (n = 3).
Highlighted rows indicate the chosen formulations for various studies.

FNS L1
FNS L2
FNS L3
FNS L4
FNS L5
FNS L6
FNS L7
FNS L8
FNS L9
FNS L10
FNS L11

Formulation
Code

Composition

31.7:1
31.7:1
31.4:1
29.5:1
24.2:1
27.4:1
24:1
23.3:1
29.1:1
26:1
16.6:1

FNS payload (g of drug:

mg of total lipids)

Table 1
Effect of composition of various FNS liposomal formulations on entrapment efficiency, drug payload, and vesicle size

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15.4 2.2
18.1 1.2
20.3 1.1
21.7 2.1
24.1 1.7
22.4 2.1
20.8 1.5
21.3 0.8
14.3 0.9
12.4 1.1
10.8. 0.8

Mean vesicle
diameter* (m)

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594

R. Kumar et al.

observed under optical microscope to confirm the

absence of unentrapped drug particles. Appropriate
amount of elute was digested with Triton-X (1% w/v),
and the clear solution was analyzed spectrophotometrically (UV-visible spectrophotometer, Shimadzu-1601,
Japan) for estimation of FNS content at a max of 212 nm
%
( E11cm
= 311). Liposomes prepared without drug, in a
similar manner, served as blank for the above studies
conducted in triplicate. Drug payload (DPL) in the
prepared liposomal formulations was calculated as
micrograms of FNS per mg of total lipids. Percent drug
entrapment (PDE) for the prepared liposomes was calculated as in Eq. (1).

under continuous gentle stirring. Subsequently, this

dispersion was gradually added to 80 mL of ice-cold
distilled water and stirred gently until complete gelling. Thereafter, appropriate amounts of FNS liposomal suspensions were gently levigated in the prepared
methylcellulose hydrogel to obtain corresponding liposomal gel formulations. For comparative studies, conventional hydrogel formulation of FNS was also
prepared analogously by replacing distilled water as
used in the above gel formulation with FNS-aqueous
solution (FNS solubilized in distilled water containing
10% v/v methanol).

In Vitro Skin Permeation Studies

Entrapped drug (mg)

PDE =
100
Total drug added (mg)

(1)

Vesicular Morphology and Size Analysis

Prepared liposomal dispersions were analyzed for
their morphological attributes viz. shape, surface characteristics, lamellarity, and size by using optical microscope
fitted with a CCD camera (Carton; Japan). Mean vesicle
size and size distribution profile of FNS-loaded liposomes
was determined by light-scattering particle size analyzer
(Malvern Mastersizer; Model-S, version 2.15, Malvern,
UK). The study was conducted in triplicate to assess the
batch to batch uniformity of vesicle size.

Transmission Electron Microscopy

Negative stain micrographs were prepared on copper
grids covered with a formvar/carbon film. The vesicle
dispersions were pipetted onto the grids and stained with
1% phosphotungstic acid. Subsequently, the stained
dispersions were viewed and photographed with a Philips
CM 10 transmission electron microscope at an accelerating
voltage of 80 kV.

Preparation of Methylcellulose Hydrogel as a

Secondary Vehicle
Methyl cellulose hydrogel (2% w/w) was prepared
as a secondary vehicle to make the prepared FNS liposomal formulations rheologically favorable for topical
application. Briefly, 2 g of methyl cellulose was dispersed in 18 mL of distilled water maintained at 50C

The experiments using animals were approved by the

university ethics committee. Trans-epidermal permeation
studies with FNS-containing liposomal (FNS-liposome
suspension and FNS-liposomes incorporated in methyl
cellulose gel 2% w/w) and nonliposomal formulations
(10% methanolic solution of FNS and FNS dispersed in
methylcellulose gel) were carried out by using excised
abdominal mice skin.
Briefly, following removal of the subcutaneous fat
from the full thickness abdominal mice skin, hair on the
dorsal side of the animal were removed with the help of a
0.1-mm animal hair clipper, in the direction of tail to
head. The dermis of the skin was wiped three to four
times with a lint-free adsorbent wipes soaked in isopropanol to remove any adhering fat material. The tissue
samples were stored frozen at a temperature of 20C for
a maximum of 2 weeks before use. Prior to permeation
experiments, skin tissue was thawed and clamped into
donor and the receptor compartment of the jacketed vertical Franz diffusion cell (cross-sectional area of 9.61 cm2;
capacity 30 mL; fabricated in-house). The receiver
medium consisting of phosphate buffer pH 6.4 USP containing 10% v/v methanol was thermostatted at 32 2C
under constant magnetic stirring up to 24 hr. Solubility of
FNS in the receptor medium was estimated prior to the
permeation experiments to ensure pseudo-sink conditions. Liposomal or non-liposomal FNS formulations
(equivalent to 5 mg of drug) were applied uniformly on
the dorsal side of mice skin. The donor chamber and the
sampling port were covered by parafilm to prevent evaporation during the study. Aliquots of 3 mL were withdrawn periodically and replaced with an identical volume
of receptor media (i.e., phosphate buffer pH 6.4 USP
containing 10% v/v methanol) to maintain the receptor
phase volume at a constant level. Samples were suitably
diluted and quantified spectrophotometerically at a max
%
of 210 nm ( E11cm
= 310).

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Development of Liposomal Systems of Finasteride for Topical Applications

Determination of FNS Deposition in Skin

RESULTS AND DISCUSSION

Following permeation studies, the skin tissue

mounted on the diffusion cell was removed and washed
thrice with saline solution, followed by blotting between
tissue paper to remove any adhering formulation from the
surface. Subsequently, the skin tissue was cut into small
pieces and homogenized with 10 mL of chloroform:methanol mixture (2:1 v/v) for extracting FNS. Homogenate
suspension thus obtained was centrifuged for 5 min at
5000 rpm (3913 g). The supernatant was filtered by
using a 0.45-, membrane filter (nylon; Millipore, MA,
USA) and quantified spectrophotometrically for drug con%
tent at a max of 245 nm ( E11cm
= 332). Fresh skin tissue
treated in the similar manner was taken as blank for the
above study. Each experiment was conducted in triplicate.

Preparation of FNS-Loaded Liposomes

Stability Studies
Stability of the FNS-vesicles was studied for 2
months. FNS-loaded liposomal suspensions (5 mL each)
were transferred into sealed ampoules of 10 mL capacity
after flushing with nitrogen prior to their storage in refrigerated condition (RF; 48C) and at room temperature
(RT; 25 2C). Ability of vesicles to retain the entrapped
drug (i.e., drug-retentive behavior) was assessed at the
fixed time intervals (i.e., once a week during the first
month, and every 15 days afterwards). Samples were withdrawn and analyzed for FNS content in the manner
described previously under drug entrapment studies.
The dispersions were transferred into clear test tubes,
at different time points, for visual and microscopic (optical
and TEM) observations such as vesicle fusion, aggregation disruption, and sedimentation. Physical stability using
these visual and microscopic characteristics was assessed
on an ordinal scale ranging between 0 and ++++ as below:
Score
0
+
++
+++
++++

Physical stability
:
:
:
:
:

Unstable
Poor
Average
Good
Excellent

Vesicle size was measured at all the time points with

0.1 mL of dispersion diluted 1000-fold with distilled water
on a Malvern Mastersizer. The pH of various liposomal
suspensions at different time points was measured by
using a high-sensitivity pH meter (Model: Metrohm 654,
Metrohm, Switzerland).

595

Multilamellar liposomes (MLVs) containing FNS

were prepared by thin film hydration technique, owing to
its stellar advantages, such as higher encapsulation efficiency for lipophillic drugs and ease of preparation.[38]
Preparation of FNS-loaded liposomes was affected by
studying the influence of drug-total lipid ratio on the drug
encapsulation efficiency of vesicles. Besides the significant
influence of process conditions, the drug-bearing capacity
(i.e., drug payload), and liposome size were found to be
invariably dependent on drug-lipid ratio and other vesicular
components viz. CHOL and DCP too.
Table 1 summarizes the influence of various formulation variables, such as drug-lipid ratio, CHOL and DCP on
percent drug entrapment (PDE), drug payload (DPL), and
mean size of prepared vesicles. As is evident from the
results, the cholesterol-free liposomes (FNS L1) could
entrap up to 63.4% of drug with DPL of 31.7 g/mg of
total lipids. Thereafter, considerable enhancement in PDE
was observed with the addition of cholesterol from 10 to
50 mg (formulation FNS L2 to FNS L4), at a fixed drug
level of 5 mg. This observation is in consonance with the
literature reports that incorporation of CHOL in liposomes
profoundly affects the membrane properties of the lipid
bilayers by reducing the rotational freedom of hydrocarbon chains.[39] In addition, the incorporation of cholesterol
also eliminates the gel-to-liquid phase transition of vesicle
bilayers and induces permanent transition of gel-state
bilayer to an ordered liquid crystalline state.[40] Both these
mechanisms make the lipid bilayers more stable and less
permeable to the encapsulated drug, leading to augmentation in the entrapment efficiency of vesicles. Although the
PDE of vesicles increased with CHOL addition in formulations FNS L1 to FNS L4, yet the effective drug payload
decreased due to net increase in the total amount of lipids,
with reference to the fixed amount (5 mg) of FNS. Maximum PDE was observed in formulation FNS L4, and the
same tended to decrease with further increase in the
amount of cholesterol or varying phospholipid-cholesterol
ratio (FNS L5 to FNS L8). Addition of cholesterol showed
a direct bearing on the vesicle size too as the mean vesicle
size increased from 15.4 m (FNS L1) to 21.3 m (FNS
L8) with increasing level of CHOL.
Based on the above results, the formulation FNS L4
with maximum PDE of 88.6% (drug payload 29.53 g of
drug:1 mg of total lipids) was selected for further study.
Addition of surface charge-imparting agent (i.e., DCP), up
to 1% of total lipids (FNS L9) did not influence the FNS
payload of liposomes. This observation is quite contrary to
the earlier studies[35,41] in which the improvement in drug

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R. Kumar et al.

payload in the vesicles containing surface charge inducer

was reported and ascribed to the improved structural integrity of the vesicles. However, the vesicle size in the current
study reduced further to a considerable extent (FNS L 9 to
FNS L11) vis--vis neutral FNS-liposomes, attributable to
the electrostatic repulsions in the lipid bilayers during the
vesiculation process.[38,41]
Regarding the process conditions, 5-min vortexing
was found to be adequate to obtain liposomal suspensions,
free from aggregates. The former did not affect drug
entrapment, as was confirmed by PDE determinations
both before and after vortexing. Because in liposomes
meant for topical applications, size also plays a key role in
the drug transport across the skin, sonication of formulation FNS L 9 was carried out in an attempt to obtain
smaller MLVs. The experimental conditions of the sonication process, especially the sonication time, was optimized
by looking into its considerable impact on drug encapsulation too.[38,42,43] As construed from Table 2, substantial
size reduction could be attained with 10 min of sonication,
as it yielded vesicles with mean size of 3.66 m (FNS
L13) with quite negligible impact on drug entrapment.
Figure 1 corroborates the same graphically. Further
increase in sonication affected PDE profoundly, probably
because of the disruption of drug-loaded vesicles leading
to fragmentation of lipid bilayers. Nevertheless, these
bilayers reassemble later to yield the aggregated liposomal
dispersions [41,42] as is evident from the increased vesicle
size (16.6 m) in formulation FNS L14. Based on the
results of this study, FNS L13 liposomes were further
characterized and subsequently investigated for skin
permeation behaviour in vitro.

Morphological and Micromeritic Characterization

Results of the optical and TEM characterization of FNS
L13 liposomes showed that the liposomes were homogenous
in shape and of lamellar structure. The optical photomicrograph (Figure 2A) of FNS L13 liposomes ratifies the multilamellear nature of the vesicles (magnification of 1500).

Further confirmation of vesicle formation and integrity of

bilayers was achieved by using TEM studies (Figure 2 B).
The vesicle size of all the liposomal formulations
tended to follow Gaussian (i.e., normal) size distribution. As
vividly apparent from Figure 3A and 3B, portraying vesicle
size distribution of unsonicated and sonicated liposomes,
respectively, the sonication reduced vesicle size and polydispersity significantly. Mean vesicle size in unsonicated
liposomes (FNS L4) was found to be 21.7 m with 90% of
the liposomal population equal to or below 54.17 m.
Smaller MLVs were, however, obtained with optimal sonication of FNS L4 liposomal dispersion with a mean vesicle
diameter of 3.66 m, having 90% of the population equal to
or below 5.77 m. Reproducibility of vesicle size of all the
FNS formulations was ratified from the low values of standard deviation observed for mean vesicle size (n = 3) of all
the liposomal-FNS formulations (Tables 1 and 2).

In Vitro Permeation and Skin Deposition Studies

The equilibrium solubility of FNS in phosphate buffer
pH 6.4 USP containing 10% v/v methanol was found to be
0.497 mg mL1. Table 3 reports the results obtained with
different formulations of FNS. Trans-epidermal permeation profile of FNS-containing liposomal formulations
[liposomal suspension (FNS L13) and liposomes incorporated in methyl cellulose gel (FNS L13-gel)] were studied.
The results obtained were compared with that of the nonliposomal formulations of FNS (i.e., 10% v/v methanolic
solution) and methylcellulose gel containing FNS in
equivalent amounts. To assess the effect of surface charge
and vesicle size on drug permeability, the other two liposomal formulations (i.e., FNS L4 and FNS L9) were also
included in the study.
After 24 hr the liposomal dispersion (FNS L13) and
liposomal gel (FNS L13-gel) showed higher FNS permeation across skin than that of the hydroalcohlic solution
(Control) and the conventional gel containing FNS (Figure 4).
As indicated in Table 3, the amount of FNS permeated in
24 hr was found to be 54% and 52.4% from the liposomal

Table 2
Optimization of sonication time: effect on PDE and vesicle size
Liposome composition
Formulation Code
FNS L9
FNS L12
FNS L13
FNS L14

FNS (mg)

PL 90H (mg)

CHOL (mg)

DCP (wt %)

Sonication
time (min)

PDE

Mean vesicle
diameter (m)#

5
5
5
5

100
100
100
100

50
50
50
50

1
1
1
1

5
10
15

87.4
88.0
85.8
71.2

14.3 0.9
6.1 1.8
3.66 1.6
16.6 3.4

Values represent Mean SD (n=3).

Development of Liposomal Systems of Finasteride for Topical Applications

PDE

(A)

Vesicle size

100

18

PDE

12
50
6
25
0

Vesicle size (m)

75

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597

0
0

10
15
Sonication time (min)

20

(B)

Figure 1. Effect of sonication on drug payload and size of

FNS-liposomes.

(A)

Figure 3. Particle-size distribution plots of FNS-liposomes (A)

unsonicated and (B) sonicated.

(B)

Figure 2. Photomicrogrpahs of FNS multilamellar liposomes

made up of saturated phospholipids (lipid concentration: 15 mg
mL1). (A) Optical photomicrogrpah. (B) Electron
photomicrograph.

suspension (FNS L13) and liposomal gel (FNS L13-gel),

respectively. Drug permeated in FNS-aqueous solution
(Control) and FNS dispersed in methylcellulose gel, however, was quite lower (i.e., 24% and 29%, respectively).
Higher flux obtained with FNS L13 (28.369 g/cm2/hr)
and FNS L13-gel (25.715 g/cm2/hr) than that obtained
with the aqueous solution (12.069 g/cm2/hr) and methylcellulose gel of FNS (10.377 g/cm2/hr), unequivocally,
substantiate the permeation-enhancing effect of drug
vesiculation. Nevertheless, the incorporation of liposomal
dispersion into a secondary vehicle (2% w/w methylcellulose hydrogel) did not affect the permeation characteristics
of the FNS-liposomal formulation. The observed permeation enhancement could be due to the increased fluidity
of the skin barrier on account of the interaction of phospholipid molecules of the membranous structures with that
of skin cells.[29,30,44,45] Moreover, the enrichment of the
skin layers with respect to hydrated lipids[46,47] is also supposed to enhanced the drug permeability.
The permeation behavior of FNS-bearing liposomes
has been shown to be influenced by the surface charge as
well as size.[48] Negatively charged smaller MLVs (FNS
L9 and FNS L13) showed higher permeation flux, as
indicated by the higher values of the enhancement ratio

598

R. Kumar et al.
Table 3
Cumulative amount, permeation flux, and skin deposition of Finasteride through abdominal
rat skin with different FNS formulations

Formulation

Qcum* (g/cm2)

Flux* (g/cm2h1)

ER$

Percent skin
deposition*

FNS L13-gel
FNS L13
FNS L9
FNS L4
FNS conventional gel
Control

280.95 5.89
286.16 8.97
218.52 5.51
197.71 6.87
130.07 7.71
156.08 4.33

28.369 1.321
25.715 1.917
21.954 2.301
18.862 3.101
10.377 1.091
12.069 2.342

2.35
2.13
1.82
1.56
0.85

31.56 2.9
30.87 2.4
21.76 3.4
16.75 2.9
4.99 0.9
2.56 1.1

*Values represent Mean SD (n=3).

ER: Relative flux enhancement ratio w.r.t. Control; Qcum: Cumulative amount of FNS permeated in 24 hr.
FNS L13-gel
: Sonicated liposomes incorporated in methylcellulose (2% w/w) gel.
FNS L13
: Sonicated liposomes.
FNS L9
: Negatively charged liposomes.
FNS L4
: Neutral liposomes.
FNS Conventional gel : Finasteride in methylcellulose (2.0% w/w) gel.
Control
: 10% methanolic solution of FNS.
$

FNS 4 L
FNS 13 L
Conventional gel

300
Cumulative drug amount per cm2

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Permeation

FNS 9 L
FNS 13 L gel
Control

200

100

0
0

12
Time (h)

18

24

Figure 4. Mean cumulative amount of Finasteride permeated

from various liposomal and non-liposomal formulations across
excised abdominal skin (n = 3). The error bars represent standard
deviation values.

(ER) [FNS L9: 1.81; FNS L13: 2.13] than that of the neutral liposomal formulations [FNS L4: 1.56] with higher
vesicle size. This observation is in contrast to the conventional understanding that the positively charged liposomes
are supposed to interact better with the negatively charged
skin cells.[49] However, in this report, the improved permeation of FNS with negatively charged liposomes may be
due to the other liposomal characteristics that tend to help
in skin penetration viz. size, skin hydration, and integra-

tion of liposomal phospholipids with the skin

lipids.[41,48,49]
The ability of vesicles in retaining drug within the
skin milieu (i.e., depot effect) was investigated by
determining the amount of drug deposited in the skin
samples used in drug permeation studies. The amount
of drug retained in the skin was considerably higher
(Table 3) in liposomal preparations than in the nonliposomal formulations. This observation may be
ascribed to the fact that the liposomal phospholipids
may mix with the intercellular lipids and thereby cause
the swelling of intercellular lipids. These swollen lipids
subsequently serve to provide local accumulation of the
drug and the consequent formation of intracutaneous
drug depots.[5052]

Stability Profile of FNS-Liposomes

Table 4 depicts the stability profile of the optimized
FNS-liposomal formulation at both the storage conditions.
The FNS-liposomal preparations showed remarkably better stability for 2 months when stored at RF than the formulation kept at RT. Substantial loss (nearly 13%) of drug
was evident from the samples stored at room temperatures
(i.e., 25 2C). At lower temperature (RF), on the other
hand, the liposomes could retain up to 97% of the incorporated drug. Higher drug leakage observed at elevated temperature suggests the need to store liposomal product
under refrigeration conditions. Loss of drug from the

Development of Liposomal Systems of Finasteride for Topical Applications

599

Table 4
Effect of storage conditions and time on drug leakage, physical stability, and vesicle size of optimized liposomal
preparation of Finasteride
Stability parameters
RF

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Time Points
Freshly prepared
(0 day)
1st week
2 weeks
3 weeks
4 weeks
6 weeks
8 weeks

Percent FNS
leakage

1.3 0.2
1.29 0.3
1.5 0.1
2.3 0.3
2.8 0.4

RT

Vesicle size

pH

Physical
stability

3.65 0.2

6.12 0.1

++++

3.55 0.1
3.75 0.2
3.49 0.1
3.30 0.2
3.77 0.3
3.89 0.2

5.85 0.2
6.07 0.4
5.98 0.2
6.32 0.03
6.11 0.2
5.93 0.3

++++
++++
++++
+++
+++
+++

vesicles stored at higher temperature may be assigned to

the effect of temperature on the gel to liquid transition of
lipid bilayers together with the possible chemical degradation of the phospholipids, leading eventually to defects in
the membrane packing.[30,31] Drug leakage of less than 3%
of the initial load at refrigeration conditions is well within
the permissible limits. No change in the pH values of vesicle suspensions, however, was seen at both the storage
conditions.
Microscopic (Optical and TEM) investigations
(Figures 5A and 5B) revealed that the liposomes stored
at RF were found to be quite stable in terms of aggregation and disruption tendencies over the studied storage
period. Drug-loaded vesicles also retained their multilamellar nature and shape uniformity to an appreciable
extent. Size determinations were performed, at different
time periods, as a part of stability protocol. Practically,
no change in the vesicle size was observed at RF

(A)

Percent FNS
leakage

3.7 0.2
5.5 0.1
6.3 0.3
9.8 0.4
13.3 0.5

Vesicle size

pH

Physical
stability

3.65 0.2

6.12 0.1

++++

3.34 0.3
4.11 0.2
5.89 0.5
6.4 0.1
8.9 0.2

5.91 0.1
6.13 0.3
5.98 0.2
6.16 0.3
6.36 0.1

+++
+++
++
+
0

vis--vis the significant increase observed at RT. The

latter may again be accredited to vesicle aggregation
and, hence, the poor physical stability of the vesicles
stored at higher temperatures. Considerable disruption
and drug leakage were observed from vesicles stored at
RT after 6 weeks, and the stability studies were thus
discontinued. The optimized FNS liposomes incorporated in the secondary vehicle (i.e., methylcellulose gel)
also exhibited similar stability profile (in terms of
microscopic characters, pH, and FNS content) at RF
conditions (data not shown). Moreover, the cellulosic
polymers are also known to stabilize the leakage of
lipid soluble drugs from the supersaturated systems,
besides improving their physical stability.[42,53] The stabilization effect of the polymer can be attributed to the
formation of a protective layer on the surface of the
vesicles, resulting in inhibition of drug leakage and
vesicle aggregation.

(B)

Figure 5. Photomicrogrpahs of FNS multilamellar liposomes after storage at RF for 2 months. (A) Optical photomicrogrpah. (B)
Electron photomicrograph.

600

R. Kumar et al.

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CONCLUSIONS
The present work on the preparation of topical liposomes is an attempt to exploit the immense potential of
multilamellar liposomal carriers to localize the drug onto
the desired target sites in the skin. For this, a steroidal
molecule, Finasteride, was identified owing to its lipophillic nature and delivery needs within the mesodermal layers
(including follicular sites). Following thorough investigations on its developed liposomal formulations, it can be
inferred that Finasteride-loaded liposomal constructs with
optimal characteristics viz. entrapment, drug payload, size,
lamellarity, and surface charge, are able to penetrate, partition, and permeate the skin barrier to access its delivery
destinations. Further, the nonchemical modification in the
drugs behavior by means of close supramolecular association of phospholipid molecules also promises to prolong
the drug action as revealed by drug deposition studies.
Conclusively, the experimental results and the supportive
theoretical analysis unambiguously indicate promising
avenues for Finasteride in dermatological problems while
exploiting the potential of liposomes through topical route
and call for further intensive investigations.

6.

7.

8.

9.

10.

11.

12.

ACKNOWLEDGMENTS
13.

The authors thank Cipla Limited, Ahmedabad, India,

and Phospholipids GmbH, Germany, for generously providing Gift samples of Finasteride and Phospholipon 90 H,
respectively. They also thank the University Grants Commission, New Delhi, India, for providing financial assistance for carrying out this research work.

14.

15.

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