Artificial Cells, Nanomedicine, and Biotechnology

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Oleic acid vesicles: a new approach for topical delivery of

antifungal agent

Shivani Verma, Ankur Bhardwaj, Mohit Vij, Pawan Bajpai, Nishant Goutam &
Lalit Kumar

To cite this article: Shivani Verma, Ankur Bhardwaj, Mohit Vij, Pawan Bajpai, Nishant
Goutam & Lalit Kumar (2014) Oleic acid vesicles: a new approach for topical delivery of
antifungal agent, Artificial Cells, Nanomedicine, and Biotechnology, 42:2, 95-101, DOI:
10.3109/21691401.2013.794351

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Artificial Cells, Nanomedicine, and Biotechnology, 2014; 42: 95–101
Copyright © 2014 Informa Healthcare USA, Inc.
ISSN: 2169-1401 print / 2169-141X online
DOI: 10.3109/21691401.2013.794351

Oleic acid vesicles: a new approach for topical delivery

of antifungal agent
Shivani Verma1, Ankur Bhardwaj2, Mohit Vij3, Pawan Bajpai4, Nishant Goutam5 & Lalit Kumar3
1Department of Pharmaceutics, Global College of Pharmacy, Punjab, India, 2Department of Pharmaceutics, ISF College

of Pharmacy, Punjab, India, 3Department of Pharmaceutics, Abhilashi College of Pharmacy, Himachal Pradesh, India,
4Department of Pharmaceutical Chemistry, Abhilashi College of Pharmacy, Himachal Pradesh, India, and 5Department of

Pharmacology, Abhilashi College of Pharmacy, Himachal Pradesh, India

perhaps the most demanding. Stratum corneum is the outer-

Abstract
most epidermal layer of skin acting as a principal regulatory
The occurrence of topical fungal infections is increasing nowadays.
barrier to the transcutaneous traffic of water and exogenous
Cutaneous fungal infections like cutaneous candidiasis are more
substances like bacteria or fungus (Downing 1992). There is
prominent in patients associated with AIDS. Current available
alteration in the structure of stratum corneum when skin is
strategies for the treatment of cutaneous fungal infections are
attacked by external agent like fungus leading to change in
creams or gels which show various adverse effects on skin along
permeability of skin (Golden et al. 1978). The incidences of
with systemic absorption. These drawbacks can be overcome
fungal infections are increasing nowadays especially in the
by using various novel drug delivery systems. So, the present
patients that are immunocompromised. This increase in
investigation aims at exploring the potential of fatty acid vesicles
the occurrence of topical/systemic fungal infection is due
(ufasomes) for the topical delivery of clotrimazole. Oleic acid
to coupling of causative fungus with the diseases like AIDS
was employed as a fatty material for the preparation of vesicles.
or due to excessive use of immunosuppressive drugs in this
Clotrimazole-loaded oleic acid vesicles were prepared using a
period of time when there is a technological advancement in
thin film hydration method. Prepared vesicles were characterized
the field of solid organ transplantation medicines, stem cell
for size, size distribution, shape, thermal behaviour (differential
transplantation and neonatology (Li et al. 2007). Topical fun-
scanning calorimetry), in vitro release, in vitro antifungal
gal infections are mainly found in the subcutaneous tissue.
activity, in vitro skin permeation and retention studies and for
They may be invasive in nature and can also reach deep into
in vivo antifungal activity. Transmission electron microscopic
epidermis (Kaur and Kakkar 2010). Oleic acid is generally
(TEM) images confirmed the formation of vesicular dispersion
employed as a penetration enhancer for delivery of differ-
(ufasomes) of clotrimazole. Oleic acid vesicles possessed high
ent bioactives into the skin. Oleic acid induces penetration
drug entrapment (49.5 ⴞ 1.0%) and optimum size (455 ⴞ 22
into skin due to subcutaneous lipid fluidization and phase
nm) along with good colloidal characteristics (polydispersity
separation (Naik et al. 1995). It has been reported that fatty
index ⴝ 0.210 ⴞ 0.035 & zeta potential ⴝ ⴚ 22.45 ⴞ 0.25 mV) at
acids like oleic acid and linoleic acid have tendency to form
4:6 drug-to-oleic acid ratio. In vitro drug release study showed
vesicular structures in the aqueous environment (Gebicki
sustained release of drug from the vesicular dispersion. Skin
and Hicks 1973). Methotrexate-entrapped deformable
permeation and skin retention studies suggested accumulation
liposomes prepared from phosphatidylcholine and oleic
of drug in the epidermal part of the skin. In vivo study confirmed
acid were compared with those of methotrexate-entrapped
prolonged release of drug from oleic acid vesicle up to five days
conventional liposomes prepared from phosphatidylcholine
indicating its usefulness for long-term therapy. So, it can be
and cholesterol for skin permeation. Liposomes containing
concluded from the present study that fatty acid vesicle may be
oleic acid showed high transdermal permeation as com-
a good approach to treat topical fungal infections.
pared with the other (Srisuk et al. 2012). Oleic acid vesicles
Keywords: colloidal, cutaneous, fatty acid, fungal infection, containing 5-flourouracil were developed for topical deliv-
oleic acid, sustained release ery. Results of the study showed that drug-loaded oleic acid
vesicles effectively penetrated stratum corneum and formed
drug depot in epidermal part of skin for localized delivery of
Introduction drug (Dhillon et al. 2011). Methotrexate was entrapped into
Skin is the outermost covering of human body performing oleic acid vesicles (ufasomes) for the effective topical deliv-
various important functions, but its protective role is ery against psoriasis. The methotrexate amount permeated

Correspondence: Lalit Kumar, Department of Pharmaceutics, Abhilashi College of Pharmacy, Mandi, Himachal Pradesh, India 175008. Tel: 91-8261876750.
Fax: 91-1905248539. E-mail: [email protected]
(Received 17 January 2013; revised 17 March 2013; accepted 6 April 2013)

95
96 S. Verma et al.

through rat skin was three- to four-fold higher using oleic at ambient temperature for 2 h with phosphate buffer
acid compared to that from plain drug solution or carbopol (pH 5.5). The prepared vesicular dispersion was sonicated
gel (Sharma and Arora 2012). Clotrimazole is a potent anti- to form the uniform size vesicular dispersion. Optimiza-
fungal agent used against topical fungal infections. One of the tion was performed by varying the ratios of oleic acid to
most common problems with this drug is that patients often clotrimazole. Unentrapped drug was separated from the
stop using it before the infection is completely removed, vesicle dispersion using gel chromatography (Sephadex
leading to re-infection in the body. It also shows absorption G-50 minicolumn) and borate buffer as an eluant.
into the systemic circulation on its frequent dosing (Souto
and Muller 2006). Entrapment efficiency
It has been shown that double layer membranes possess The entrapment efficiency was determined by disrupting
fusogenic characteristics because they reduce phase transi- the vesicles using sodium hydroxide solution (1 M) and
tion temperature of the lipids in the biological membrane sys- subsequence estimation of released clotrimazole, as ent-
tem. When this vesicular membrane comes into contact with rapped drug. Entrapment efficiency (%) was calculated using
skin, it shows its fusion with skin lipid bilayers and releases following formula:
its content. So, it is considered that fatty acid vesicles are
very effective carriers for enhancing the penetration of drug Entrapment (%)  Entrapped drug/Total drug  100 (1)
molecules through the stratum corneum with the reduction
of toxicity. Fatty acid vesicles are cheaper in cost and their
method of preparation is very easy. So in the present study
Vesicle size, zeta potential and morphology
we tried to develop an effective vesicular drug delivery sys-
of oleic acid vesicles
Vesicle sizes and zeta potentials of different formulations
tem (ufasomes) of clotrimazole for targeting fungus in deeper
were determined using particle size analyzer (DelsaTM
epidermal layer of skin and to provide a local effect of drug
Nano C, Beckman Coulter). Shape and surface morphol-
in the skin with reduction of dose. For achieving the goal,
ogy of oleic acid vesicles was examined using transmission
oleic acid vesicles containing clotrimazole were prepared
electron microscope (TEM) (HRTEM, H47500 Hitachi Ltd.,
using thin film hydration method. The effects of drug-to-
Tokyo, Japan).
fatty acid ratio on size and entrapment efficiency of vesicles
were studied. Vesicular dispersion of drug (Sample B) was
compared with marketed formulation (Sample A) for in-vitro Differential Scanning Calorimetry
skin permeation and in-vivo antifungal activity. Results of Thermal behaviour of drug, drug-loaded vesicles and blank
in-vivo studies revealed the prolonged effect of clotrima- vesicles were studied using differential scanning calorim-
zole ufasomes for 5 days in the treatment of experimentally eter (DSC) (Q-10, TA Instrument waters). Before adding
induced cutaneous candidiasis. Thus, vesicular dispersion sample for analysis, calibration of heat flow scale was done.
(ufasomes) of clotrimazole was found to be more effective as The samples were purged with dry nitrogen at a flow rate of
compared to marketed product 20 ml/min. The temperature was raised at a rate of 10°C/min
(Dhillon et al. 2011).

Materials and methods In vitro drug release study

Materials In vitro release study of oleic acid vesicles was carried out
Clotrimazole was provided as a gift sample by Edifice Labs using dialysis bag method (Dhillon et al. 2011). For this study,
(Ludhiana, India). Oleic acid was purchased from CDH 5 ml of vesicular dispersion was added to dialysis mem-
(New Delhi, India). SephadexG-50 and dialysis mem- brane-70 (Himedia, LA393-10MT). Dialysis membrane was
brane ⴚ 70 (LA393-10MT) was purchased from Himedia taken into 50 ml of phosphate buffer saline pH 5.5containing
(Mumbai, India). Fungal strain Candida albicans was pur- 0.01% SLS (Sodium lauryl sulphate), in a conical flask. The
chased from MTCC – Chandigarh, India. All other solvents flask was kept in an incubator shaker and the speed of the
used were of analytical grade and purchased from CDH shaker maintained was 60 rpm at 37°C. Samples (5 ml) were
(New Delhi, India). withdrawn and filtered. Samples were collected after dif-
ferent time intervals. Same volume (5 ml) of the phosphate
Preparation of oleic acid vesicles buffer, pH 5.5, containing 0.01% SLS was replaced after each
Oleic acid vesicles were prepared using film hydration sampling. The drug content in the sample was determined
method, as reported earlier (Sharma and Arora 2012, spectrophotometrically at 265 nm.
Murakami 1996). Briefly, oleic acid and clotrimazole were
dissolved in methanol in a round-bottomed flask fol- In vitro antifungal activity of vesicular dispersion
lowed by evaporation of solvent under vacuum using a In vitro antifungal activity of vesicular dispersion was deter-
rotary evaporator (Perfit equipments, Ambala, India) to mined using cup plate (cylinder plate) method. The over-
remove even the last traces of organic solvent. The com- night grown culture of Candida albicans was inoculated into
pletely dried film in rota-evaporator was left overnight for the sterilized Sabouraud dextrose agar (SDA) media plates.
the removal of any possible traces of methanol and also After solidification, in each plate, three wells, each 10 mm in
to prevent the formation of emulsion due to the residual diameter, were bored with a cork borer. In each plate, vesicu-
organic solvent. The dried film formed was then hydrated lar dispersion in phosphate buffer of pH 5.5, plain drug or
 Oleic acid vesicles for topical delivery of antifungal agent 97

control was placed in one of the wells in appropriate amount Prednisolone was administered four times (2 days and
and the plates were incubated at 37°C for 48 h. Phosphate immediately before, and 2 and 4 days after the inoculation of
buffer of pH 5.5 was used as a control. Zone of inhibitions fungal suspension) (Maebashi et al. 1995).
of drug, control and vesicular dispersion were measured at
different time intervals (El laithy and El-Shaboury 2002). Preparation of fungal inoculums. Candida albicans
(C. albicans MTCC Code-1637) fungus was used to induce
In vitro skin permeation studies of vesicular dispersion fungal infection in guinea pigs. A culture of fungal strain
In vitro skin permeation studies of vesicular dispersion (Sam- was grown on Sabouraud dextrose agar (SDA) for 48 h at
ple B) and marketed gel (Sample A) (Candid-V, Glenmark) 37°C. Grown cells from SDA plates were collected and re-
were carried out using Franz diffusion cell (Electrolab Ltd., suspended in sterile saline. Final concentration adjusted in
Mumbai, India) with a diffusion area of 3.3 cm2 and volume saline was 107 colony-forming unit/ml (cfu/ml) (Gupta and
of 60 ml. Penetration was studied using abdominal skin of Vyas 2012).
guinea pig. Hairs from the abdominal part of skin were care-
fully removed, and skin was excised from abdomen using Induction of fungal infection in skin. Each animal’s back was
surgical blade (Satturwar et al. 2005). Dermal side of skin shaved using an electric clipper, and two hairless patches
was carefully cleaned. Dermis part of the skin was washed of area 2.0 cm2 were developed there. Every patch was
with a cotton swab soaked in isopropanol for the removal of inoculated with 107 cfu/ml fungal dispersion by using a
fatty material (Bhatia et al. 2004). Later on skin samples were sterile cotton swab. Dispersion was rubbed into skin with
washed with normal saline and cut into appropriate sizes. a sterile cotton swab until no visible fluid appears on skin.
Skin was placed on donor side of Franz diffusion cell, main- Assessment of topical fungal infection was done by checking
tained at 37°C. Optimized vesicular formulation and mar- sign of more intense erythema at inoculation site on animal
keted gel (equivalent to 7.37 mg of clotrimazole) were placed body (Maebashi et al. 1995).
in the skin on donor side of Franz diffusion cell. A volume of
0.5 ml of sample was removed from the acceptor media at Treatment of fungal infection. Treatment began after 24 h of
regular time interval for a period of 24 h and replaced with assessment of infection. Animals were divided into three
the same amount of buffer to maintain sink condition. Sam- groups each containing four animals. First group was treated
ples were analysed at 265 nm using UV spectrophotometry with marketed formulation (Sample A) (Candid-V Gel,
(Pierre et al. 2001). Glenmark) (equivalent to 6.52 mg of clotrimazole). Second
group was treated with clotrimazole-loaded oleic acid
Skin retention of vesicular dispersion vesicles (Sample B) (equivalent to 6.52 mg of clotrimazole)
After performing skin permeation studies, skin was carefully and third group served as a control. Phosphate buffer of pH
removed from the Franz diffusion cell. Remaining formula- 5.5 was used as a control. One animal from each group was
tion was washed with cotton swab soaked in phosphate sacrificed just before applying formulation to determine
buffer of pH 5.5. The cleaned skin piece was mashed, and initial colony count before applying formulation (Day 0).
50 ml of methanolic PBS (4:6) of pH 5.5 was added to the Treatment was given once in a day for three consecutive
meshed mass and mechanically shaken in a water shaker days. After 24 h of last treatment, one animal from each
bath at 37°C for 1 h for the complete extraction of the drug. group was sacrificed for colony count (Day 1). After 72 h
After extraction, the resulting solution was filtered and the (Day 3) and 120 h (Day 5) of last treatment, similar process
amount of drug content in filtrate was determined using was carried out. For colony count, infected skin of animal
UV spectrophotometer at 265 nm (Agarwal and Katare was excised and homogenized in sterile saline solution. A
2002). Skin retention (in percentage) was calculated using portion of homogenate was cultured in Sabouraud dextrose
the following formula: agar (SDA) plates for 48 h at 37  1°C, and cfu values were
recorded using digital colony counter (Maebashi et al. 1995,
Skin retention (%)  Amount of drug in skin/Total drug Gupta and Vyas 2012).
 100 (2)
Statistical analysis
In vivo studies All the results are expressed as mean  standard devia-
All animal studies were performed according to the tion. The treated groups were compared with control using
guidelines compiled by the Committee for the Purpose analysis of variance (ANOVA) thanks to GraphPad PRISM
of Control and Supervision of Experiments on Animal software (GraphPad Software Corp., San Diego, CA). The
(CPCSEA, Ministry of Culture, Government of India). All p value  0.05 was considered as significant.
the study protocols were approved by the animal ethical
committee of the Abhilashi College of Pharmacy, Mandi
Results and discussions
(Himachal Pradesh), India.
Shape and morphology of clotrimazole-loaded
In vivo antifungal activity of gel oleic acid vesicles
Animals. Male guinea pigs weighing 400–450 g were used in Clotriamzole oleic acid vesicles were prepared effectively
the study. They were immunosuppressed by giving predni- used thin film hydration method. TEM image of clotrima-
solone injection subcutaneously (30 mg/kg of body weight). zole-loaded vesicles is shown in Figure 1. This image clearly
98 S. Verma et al.

Figure 1. Transmission electron microscopic (TEM) image of optimized clotrimazole-loaded oleic acid vesicles at 80 kV and 50,000 .

indicates that prepared fatty acid vesicles were spherical in vesicles and clotrimazole-loaded oleic acid vesicles are
shape and of size below 500 nm. shown in Figure 2 (A, B and C, respectively).
Sharp endothermic peak in thermogram A clearly indi-
cates the melting point of drug (146.05°C). A less sharp peak
Size, drug entrapment and colloidal behavior was observed at 285.54°C in case of blank vesicles (B). No
of vesicular dispersion peak related to drug was found in drug-loaded vesicles (C).
Prepared vesicular system was further characterized for size, These results clearly indicate that the drug was entrapped
drug entrapment, PDI and zeta potential. Table I represents inside the microspheres. These results were in accordance
optimization of oleic acid vesicles (Murakami 1996). It is with the previous findings (Dhillon et al. 2011).
clear from the table that maximum entrapment efficiency
(49.55  1.02) was found for formulation F4 having drug-
In vitro drug release study
to-oleic acid ratio of 4:6. It was seen that the drug-bearing
Drug release study was carried out using dialysis bag method.
capacity of the oleic acid vesicles depends upon the molar
Results of in vitro drug release studies are shown in Figure 3.
ratio of oleic acid to clotrimazole. The entrapment efficiency
It is clear from the graph that all the vesicular formulations
increased up to a drug/oleic acid molar ratio of 4:6, beyond
released 30–40% drug within 24 h. Kinetics of drug release of
this ratio further increase in the amount of drug reduced the
various formulations is shown in Table II. R2 value was found
degree of drug entrapment inside the vesicles. This may be
maximum in the case of zero-order graphs, which clearly
due to the drug saturation in the bilayer domain. Further
indicates that the formulations follow zero-order release.
addition of drug could have destabilized the vesicle mem-
The Korsmeyer–Peppas release exponent (n) was found in
brane leading to the leakage of drug . Formulation F4 was
the range of 0.354–0.408, which confirmed diffusion as the
having least particle size as compared to other formulations
principle mechanism of drug release.
which was considered good for the skin penetration (Sharma
and Arora 2012). Formulation F4 showed good colloidal
characteristics compared to other formulations so it was In vitro antifungal activity of vesicular dispersion
taken as optimized formulation for further studies. Results of in vitro antifungal activity are shown in Figure 4.
No zone of inhibition was observed in case of control at dif-
ferent time intervals, while initially there was a sharp increase
Differential Scanning Calorimetry in zone of inhibition in case of plain drug as compared to
Thermal behaviour of drug, blank oleic acid vesicles and vesicular dispersion. This is because oleic acid vesicles
clotrimazole-loaded oleic acid vesicles was checked using released drug in a controlled fashion and less in amount. But
DSC analysis. DSC thermograms for drug, blank oleic acid after 48 h, zone of inhibition was more in case of vesicular

Table I. Optimization of oleic acid vesicles. Values are expressed as mean  standard deviation (n  3).
Formulation Drug: Percentage of Zeta Potential
code Oleic acid Size (nm) entrapment PDI (mV)
F1 1:9 538  15 38.2  0.8 0.367  0.078 32.68  0.65
F2 2:8 495  12 43.4  1.3 0.390  0.056 28.67  0.34
F3 3:7 481  18 47.9  0.8 0.229  0.076 30.78  0.67
F4 4:6 455  22 49.5  1.0 0.210  0.035 22.45  0.25
F5 5:5 554  15 17.3  0.9 0.765  0.045 32.43  0.49
 Oleic acid vesicles for topical delivery of antifungal agent 99

Figure 2. DSC thermograms of pure drug (A), blank oleic acid vesicles (B) and drug-loaded oleic acid vesicles (C). Sharp endothermic peak in
thermogram A clearly indicates the melting point of the drug (146.05°C). A less sharp peak was observed at 285.54°C in the case of blank oleic acid
vesicles (B). No peak related to drug was found in the drug-loaded vesicles(C).

dispersion clearly indicating that there was a continuos con- and compared with marketed formulation. Amount of drug
trolled release of drug from dispersion up to 48 h. retained in skin after 24 h was 42.71  1.31% for vesicular
dispersion (Sample B) whereas for marketed gel (Sample A)
In vitro skin permeation studies of vesicular dispersion (Candid-V, Glenmark), it was found to be 10.90  1.54%.
In vitro skin permeation studies were performed on hairless High drug retention in case of microspheres gel may be due
guinea pig skin using Franz diffusion cell. Results of per- to formation of drug reservoir in epidermis due to deposi-
meation studies are shown in Figure 5. It is clear from the tion of clotrimazole oleic acid vesicles. Figure 6 shows results
graph that amount of drug permeated from marketed gel of skin retention studies.
was very high as compared to vesicular dispersion. Cumula-
tive amount of drug permeated in 24 h from marketed gel In vivo studies
(Sample A) was 53.67  1.98% while from oleic acid vesicles In vivo antifungal activity of microsphere gel was determined
(Sample B) it was near about 14.63  1.53%. Drug permeated by inducing fungal infection in animal with C. albicans. It is
from vesicular dispersion was low because clotrimazole- clear from the Figure 7 that vesicular dispersion possessed
loaded oleic acid vesicles increased the accumulation of drug high therapeutic effectiveness as compared to other formu-
in epidermal part of skin and thus decreased the permeation lation for prolonged period of time. On Day 0 (24 h prior to
of drug through skin. Drug permeated from marketed gel treatment), fungal colony count in all three group appeared
was very high because drug in this formulation was in free almost similar indicating that fungal infection was induced
form for penetration. effectively in dermal layer. After the assessment of this colony
count, treatment was started for three consecutive days.
On Day 1 (after 24 h of last treatment), control group
Skin retention of vesicular dispersion
showed a slight increase in fungal count. Animal group
Oleic acid vesicles showed drug reservoir effect in skin so
treated with marketed gel (Sample A) showed a significant
percent deposition of clotrimazole in skin was also calculated
reduction in fungal colony count on Day 1 which was due to
immediate release of drug from the marketed formulation.
Excess drug released from this formulation reduced fungal
colony to a very less amount as compared to control. In
case of vesicular dispersion (Sample B), reduction in colony
count was less as compared to marketed formulation which
may be due to less amount of drug released from it because
oleic acid vesicles retarded the drug release.

Table II. Kinetics of drug release of different formulations.

Korsmeyer–
Formulation Zero order First order Peppas release
code Higuchi (R2) (R2) (R2) exponent (n)
F1 0.302 0.946 0.941 0.358
F2 0.299 0.935 0.932 0.367
F3 0.289 0.928 0.923 0.354
F4 0.318 0.980 0.953 0.408
Figure 3. In vitro drug release profiles of different vesicular formulations. F5 0.303 0.959 0.944 0.396
Values are expressed as mean  standard deviation (n  3).
100 S. Verma et al.

Figure 4. In vitro antifungal activity of different formulations (plain Figure 7. In vivo antifungal activity of formulations (marketed
dug and drug loaded oleic acid vesicles) against C. albicans. Values are gel and drug-loaded oleic acid vesicles). Values are expressed as
expressed as mean  standard deviation (n  3). mean  standard deviation (n  3).

compared to oleic acid vesicular dispersion indicating that

the drug was still available in vesicular dispersion for further
action (P  0.01). So, there was a significant reduction in col-
ony count in the case of oleic acid vesicular dispersion even
after the fifth day of application. Furthermore, the results
may be due to reservoir effect of oleic acid vesicles in dermal
skin layer. So, the therapeutic efficacy of vesicular dispersion
was found to be better as compared to marketed gel.

Conclusions
From the present study, it is clear that oleic acid vesicles can be
used as an effective carrier for the delivery of antifungal agent
Figure 5. In vitro skin permeation of different formulations (marketed for the treatment of localized fungal infections. These deliv-
gel and drug-loaded oleic acid vesicles). Values are expressed as ery system has different advantages like cost-effectiveness,
mean  standard deviation (n  3). therapeutic viability and reduction in the dose of drug. These
delivery systems also showed good sustained release behav-
iour of drug and skin retention properties which proved
Further, after 72 h of last treatment (Day 3) colony count
their effectiveness for a long-term drug therapy. Oleic acid
was found to increase in case of marketed gel. This increase
vesicles can easily distributed in skin and may form depots in
in colony count may be due to lack of drug on skin after
the skin because of its better permeation properties in skin.
4 days because whole drug from marketed gel released on
All these studies proved the effectiveness of clotrimazole-
Day 1 and no drug was available for further action (P  0.05).
loaded oleic acid vesicle for treating cutaneous skin fungal
However, in case of vesicular dispersion, the colony count
infections, and thus, it can be taken as a preferred choice of
was low because of sustained release of drug. After 120 h
carrier system for drug delivery.
(Day 5), colony count was very high for marketed gel

Acknowledgement
The authors want to acknowledge the help of Dr. R. K.
Abhilashi, Chairman, Abhilashi College of Pharmacy,
Himachal Pradesh, for providing excellent research facilities.

Declaration of interest
The authors report no conflicts of interest. The authors alone
are responsible for the content and writing of the paper.

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