Drug Delivery, 2010; 17(6): 443–451

RESEARCH ARTICLE

Design and development of solid lipid nanoparticles for

topical delivery of an anti-fungal agent
Safal Jain1, Sanjay Jain2, Piush Khare1, Arvind Gulbake1, Divya Bansal1, and Sanjay K. Jain1
1
Pharmaceutics Research Projects Laboratory, Department of Pharmaceutical Sciences, Dr. Hari Singh Gour
Vishwavidyalaya, Sagar (M. P.) 470003, India, and 2Lipoxen plc, London Bioscience Innovation Centre, 2 Royal College
Street, London, UK
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Abstract
Topical application of the drugs at the pathological sites offers potential advantages of delivering the drug
directly to the site of action. The main aim of this work was to formulate and evaluate Miconazole nitrate (MN)
loaded solid lipid nanoparticles (SLN) for topical application. MN-loaded SLN were prepared by modified
solvent injection method and characterized for shape, surface morphology, particle size, and drug entrap-
ment. These solid lipid nanoparticles were spherical in shape with smooth surface and possessed mean
average size of 206.39 ± 9.37 nm. In vitro drug release of MN-loaded SLN-bearing hydrogel was compared
with MN suspension and MN hydrogel; MN-loaded SLN-bearing hydrogel depicted a sustained drug release
over a 24-h period. Tape stripping experiments demonstrated 10-fold greater retention with MN-loaded
For personal use only.

SLN-bearing hydrogel as compared to MN suspension and MN hydrogel. The in vivo studies were performed
by infecting the rats with candida species. It was observed that MN-loaded SLN-bearing hydrogel was more
efficient in the treatment of candidiasis. Results indicate that MN-loaded SLN-bearing hydrogel provides a
sustaining MN topical effect as well as quicker relief from fungal infection.
Keywords: Solid lipid nanoparticles; miconazole nitrate; topical application; hydrogel

Introduction Wissing & Muller 2003), maintain a sustained release

Solid lipid nanoparticles (SLN) have been established
as an alternative particulate carrier system by various
research groups (Siekmann & Westesen, 1992; Almeida
et al., 1997). Recently, increasing attention has been
focused on these SLN as colloidal drug carriers combin-
ing advantages of polymeric nanoparticles, fat emulsions,
and liposomes, but simultaneously avoiding some of
their disadvantages (Boltri et al., 1993). During the past
several years, SLN began to act as a topical carrier not
only for pharmaceutical molecules, but also for cos-
metic products. Compared with conventional carriers
such as cream, tincture, and emulsion, SLN combine
their advantages such as controlled release, in vivo good
toleration, and protection of active compounds (Muller
et al., 2000; Sylvia et al., 2003). Especially, SLN can favor
drug penetration into the skins (Jenning et al., 2000;
to avoid ­systemic absorption (Zur Muhlen et al., 1998),
act as a UV sunscreen system (Wissing & Muller, 2002a),
and reduce irritation (Maia et al., 2000; Sivaramakrishnan
et al., 2004). It was used for topical delivery of several
drugs including clotrimazole, flurbiprofen, prednicar-
bate, and betamethasone 17-valerate, and SLN also was
reported to have skin targeting potential (Maia et al.,
2000; Sivaramakrishnan et al., 2004; Jain et al., 2005; Song
& Liu, 2005). SLN was found to have skin targeting which
can result in a high accumulation of podophyllotoxin in
the skin (Chen et al., 2006).
The advantage of SLN is that the lipid matrix is made
from physiological lipids, which decrease the danger of
toxicity. Lipid and surfactants used in SLN have approved
status, e.g. GRAS status (Generally Recognized as Safe)
due to their low toxicity, or they are already used as
excipients in cosmetics or pharmaceuticals. The small

Address for Correspondence: Professor Sanjay K. Jain, Pharmaceutics Research Projects Laboratory, Department of Pharmaceutical Sciences, Dr. Hari Singh
Gour Vishwavidyalaya, Sagar (M.P.) 470 003, India. Tel: +91 7582 265457. Fax: +91 7582-264163. Email: drskjainin@yahoo.com

(Received 05 November 2009; revised 16 March 2010; accepted 01 April 2010)

ISSN 1071-7544 print/ISSN 1521-0464 online © 2010 Informa UK Ltd

DOI: 10.3109/10717544.2010.483252 http://www.informahealthcare.com/drd
 444   Safal Jain et al.

size of the lipid particles ensures close contact to the
stratum corneum and can increase the bioadhesive and
occlusive properties, which is a desired requirement
for topical application. Due to their solid lipid matrix,
controlled release from these carriers is possible. This
becomes an important tool when it is necessary to sup-
ply the drug over a prolonged period of time, to reduce
systemic absorption, and when drug produces irritation
in high concentrations. As a result of film formation after
topical application, occlusive properties have also been
reported for SLN (Wissing & Muller 2001; 2002b).
Miconazole nitrate (MN), one of the broad-spectrum
anti-fungal agents, is a weak base (pKa = 6.7) characterized
by relatively high molecular weight and melting point. The
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as a gift sample. Soya lecithin and Dialysis bag (mol.
­cut-off weight 12,000) were procured from Himedia Ltd.
(Mumbai, India). Carbopol 934 (hydrogelling agent)
was obtained from S.D. Fine Chem. Ltd. (Biosar, India).
Tristearin (solid lipid) was purchased from Colorcon Asia
Pvt. Ltd. (Mumbai, India). Methanol (HPLC grade) was
purchased from Sigma (St. Louis, MO). Other chemicals
were of analytical grade.
Fabrication of SLN
The solid lipid nanoparticles were prepared by solvent
injection method, as reported by Goymann and Schubert
(2003). Briefly, tristearin, soya lecithin, and drug (MN)

poor water solubility (1.03 µg/ml) of MN reduces its effi-
cacy for many therapeutic applications (Pedersen et al.,
1993). MCZ is usually employed at 2% (w/w) in topical
suspensions for the treatment of dermatophytoses, super-
ficial mycoses, and mixed infections, or as an oral gel for
the treatment of Candidal infections (Peira et al., 2008).
The main aim of our investigation was to develop and
evaluate an MN-loaded SLN-bearing hydrogel for topical
delivery of MN. The entrapped drug in lipid nanoparticles
can facilitate localized delivery of the drug and improve
For personal use only.
were dissolved in ethanol in a definite ratio and warmed
to ∼ 70 ± 2°C. Tween 80 in a definite amount was dissolved
in PBS solution (pH 7.4) to prepare aqueous phase. The
organic phase, i.e. alcoholic solution containing lipid
mixture, was added dropwise with stirring to pre-warmed
aqueous solution (70 ± 2°C) with the help of a hypodermic
needle. The mixture was then sonicated for varying time
periods to obtain nanoparticles with optimum size.
Optimization of different variables for MN-loaded SLN

local availability by means of a controlled release pat-

tern which can advance the anti-fungal activity and skin
­tolerability of miconazole nitrate.
Materials and methods
Materials
Medley Pharmaceutical Ltd. (Mumbai, India) gener-
ously supplied anti-fungal drug miconazole nitrate (MN)
Various formulation variables, i.e. lipid:soya lecithin
(1:1.25–1:0.25%w/w), drug:tristearin (2.5:97.5–10:90%w/w),
emulsifier tween 80 concentration (0.1–2.0%w/v), and
process variables, i.e. sonication time (2–6 min), which
could affect the properties of solid lipid nanoparticles were
optimized to get small SLN with maximum drug entrap-
ment. Various parameters were optimized by varying one
parameter while keeping others constant (Table 1) and
prepared nanoparticles were studied for their particle size,
polydispersity index, and percentage drug entrapment.

Table 1. Composition of various SLN formulations.

Composition Parameters
Tristearin:Soya Tween80 Sonication Particle size Polydispersity % Drug
Formulation code Lecithin (%w/v) Drug (mg) (%w/v) time (min) (nm) index entrapment
A 1:0.25 — 0.1 2 425.58 ± 10.5 0.38 ± 0.051 —
B 1:0.5 — 0.1 2 389.39 ± 8.74 0.34 ± 0.038 —
C 1:0.75 — 0.1 2 341.72 ± 5.48 0.29 ± 0.023 —
D 1:1 — 0.1 2 311.24 ± 7.20 0.24 ± 0.011 —
E 1:1.25 — 0.1 2 360.24 ± 8.29 0.44 ± 0.074 —
D1 1:1 5 0.1 2 322.20 ± 5.35 0.28 ± 0.020 87.62 ± 2.51
D2 1:1 10 0.1 2 326.63 ± 5.93 0.29 ± 0.031 89.35 ± 3.49
D3 1:1 15 0.1 2 328.84 ± 4.91 0.26 ± 0.019 92.74 ± 2.11
D4 1:1 20 0.1 2 352.79 ± 6.82 0.41 ± 0.081 84.95 ± 3.08
D31 1:1 15 0.5 2 305.57 ± 5.39 0.18 ± 0.011 92.81 ± 2.46
D32 1:1 15 1 2 292.53 ± 5.48 0.46 ± 0.051 88.47 ± 3.12
D33 1:1 15 2 2 255.27 ± 7.22 0.56 ± 0.072 83.68 ± 2.82
D311* 1:1 15 0.5 4 206.39 ± 9.37 0.21 ± 0.060 90.86 ± 5.20
D312 1:1 15 0.5 6 198.42 ± 11.48 0.45 ± 0.120 80.40 ± 3.40
* Optimized formulation, values represent mean ± SD (n = 3).
 Design and development of solid lipid nanoparticles   445
Particle size determination
The average particle size and polydispersity index of the
lipid particulate dispersions were determined by photon
correlation spectroscopy using a Zetasizer (DTS Ver. 4.10,
Malvern Instruments, UK). The sample of dispersion was
diluted to 1:9 v/v with double distilled water to ensure that
the light scattering intensity was within the instrument’s
sensitivity range. Double distilled water was filtered
through 0.45 µm membrane filters (Pall Life sciences,
Mumbai, India) prior to particle size determination.
Particle morphology (TEM)
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A transmission electron microscope was used as a visu-
alizing aid for particle morphology. The sample (10 µL)
was placed on the grids and allowed to stand at room
temperature for 90 s. Excess fluid was removed by touch-
ing the edge with filter paper. All samples were examined
under a TEM (Philips Morgagni 268, Eindhoven, The
Netherlands) at an acceleration voltage of 100 kV.
Surface morphology (SEM)
For personal use only.
A scanning electron microscope was used as a visualizing
aid for surface morphology. The samples for SEM were
prepared by lightly sprinkling the SLN powder on a dou-
ble adhesive tape, which was stuck on an aluminium stub.
All samples were examined under a scanning electron
microscope (LEO 435 VP, Eindhoven The Netherlands)
at an acceleration voltage of 30 kV.
Drug entrapment efficiency
SLN dispersion was ultracentrifuged for 1 h at 55,000 rpm
(ALC Centrifugette 4206). The supernatant was used for
MN analysis by HPLC and the quantity of free drug was
determined. The encapsulated amount of MN was calcu-
lated by subtracting the free amount of MN from the total
amount in the dispersion. Entrapment efficiency (%EE)
was calculated by the following equation, where Wi is the
amount of initial drug and Wf is the amount of free drug
(Evren et al., 2008).
%EE = (Wi − W f /WI ) × 100
Formulation of MN-loaded SLN-bearing hydrogels
First, Carbopol 934 was dispersed in minimum quan-
tity of double distilled water and stored overnight and
then neutralized by triethanolamine. After adding pro-
pylene glycol and glycerine the mixture was mixed well
with high-speed stirrer at 1000 rpm for 5 min. Finally,
the MN-loaded SLN dispersions were incorporated
into hydrogel and stirred continuous for the next 5 min
(Zhinan et al., 2005; Joshi & Patravale, 2006).
Determination of spreadability and pH
The spreadability of the hydrogel was determined using
the following technique: Hydrogel (500 mg) was placed
within a circle of 1 cm diameter pre-marked on a glass
plate over which a second glass plate was placed. A weight
of 500 g was allowed to rest on the upper glass plate for
5 min. The increase in the diameter due to spreading of
the hydrogels was noted. The pH of the MN-loaded SLN-
bearing hydrogel was determined using an Equip-tronic
Digital pH meter Model EQ 610, standardized using
pH 4.0 and 7.0 standard buffers before use (Shah et al.,
2007).
In vitro drug release studies
The in vitro drug release profile of MN suspension, MN
hydrogel, and MN-loaded SLN-bearing hydrogel were
studied using a dialysis bag. Different formulations
were taken into a dialysis bag (molecular weight cut-off,
12 KDa, Himedia, India) and placed in a beaker contain-
ing 20 ml of mixture of methanol:PBS (pH 6.4) (30:70).
Then, the beaker was placed over a magnetic stirrer
and the temperature of the assembly was maintained at
37 ± 1°C throughout the study. Samples (1 ml) were with-
drawn at definite time intervals and replaced with equal
amounts of fresh buffer. The samples were analyzed for
drug ­content using HPLC.
In vitro skin permeation, stripping, and retentivity
studies
In vitro skin permeation experiments for MN from dif-
ferent formulations were performed using excised full
thickness hairless abdominal skin of rats (Male albino
rats, Sprague Dawley; 100–150 g). The skin samples were
mounted on modified Franz diffusion cells (Crown
Glass Co., NJ) with a surface of 3.14 cm2 and a receptor
volume of 10 ml such that the dermal side of the skin
was exposed to the receptor fluid (methanol:PBS (pH
6.4), i.e. 30:70) ratio and the stratum corneum remained
in contact with the content of donor compartment.
Different formulations (MN-loaded SLN-bearing hydro-
gel, MN hydrogel, and MN suspension) were placed in
the donor compartment enabling one to to cover the
entire skin surface evenly. The temperature was main-
tained at 37 ± 1°C. Sampling was done at various inter-
vals up to 24 h and assayed for MN content using the
HPLC method.
Franz diffusion cells were then dismantled and skin
surface washed 10 times with a cotton swab and stripped
using a fixed diameter and length of Scotch Magic Tape.
 446   Safal Jain et al.

The 10 tape strips were mixed in 2-propanol and 0.1 M In vivo performance studies
HCl (90:10) mixture and shaken to dissolve adhered MN
Candida albicans (MTCC 183) was used to induce ­mycosis
in solvent mixture. Then, the solvent mixture was filtered
in experimental animals to study the in vivo perform-
through a 0.45 µm membrane filter and analyzed for
ance of the prepared drug delivery system. The study was
drug concentration using an HPLC method (Wissing &
approved by the Animal Ethical Committee of Dr. H.S.
Muller 2002a). Further, percentage drug skin deposition
Gour University Sagar, India. Cutaneous Candidiasis
after tape stripping was analyzed. The skin was washed
was induced on Sprague Dawley albino rats (100–150 g)
10 times with a cotton swab followed by weighing and
by slight modification of an earlier reported procedure
homogenizing in methanol. The resulting solution was
(Maebashi et al., 1995). Briefly, hairs on the back of the
centrifuged for 10 min at 7000 rpm and supernatant was
rats were removed using depilatory cream and an area
analyzed for drug using the HPLC method.
of 3 × 3 cm2 was marked. On the second day the marked
skin was slightly abraded with sandpaper and 500 mg
Skin-irritation testing of previously prepared Candida Albicans inoculum was
applied using a glass rod. Male albino rats (Sprague
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The skin irritation potential of the MN-loaded SLN-

Dawley; 100–150 g) selected for the study were weighed
bearing hydrogel in comparison to MN hydrogel and MN
and divided into four groups, including one control group,
suspension were evaluated by performing a Draize patch
with six animals in each group. The different formulations,
test on male albino rabbits (1.5–2 kg) (Draize et al., 1944;
i.e. MN-loaded SLN-bearing hydrogel, MN hydrogel, and
Joshi & Patravale, 2006). The experimental protocol was
MN Suspension, were applied topically separately once
approved by the Animal Ethical Committee of Dr. H.S.
daily for six consecutive days, starting on the day of post-
Gour University Sagar, India. Animals were divided into
infection to albino rats of respective groups, excluding the
four groups with three animals (rabbits 1.5–2 kg) in each
animals of the control group. The animals of the control
group as follows:
group were infected but did not receive any drug therapy.
The rats were continuously observed visually for the
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• Group I: Control (no drug treatment);

changes in texture of skin of the infected area after initia-
• Group II: MN suspension;
tion of treatment. The performance of various groups to
• Group III: MN Hydrogel; and
alleviate mycosis was compared with that of control after
• Group IV: MN-loaded SLN-bearing hydrogel.
6 days, skin texture was evaluated, and recovery time was
measured (Pershing et al., 1994).
The back of the rabbits were clipped free of hair, 24 h
Culture studies were done to assess the efficacy of treat-
prior to the application of the formulations. Different
ment. Six days after the respective treatment, each treated
formulations were applied on the hair-free skin of
site was wiped thoroughly with a cotton swab containing
rabbits by uniform spreading within the area of 4 cm2.
70% ethanol. The skin was excised from each treated site,
The skin was observed for any visible changes such as
minced with scissors, and homogenized in 4 ml of saline
erythema (redness) at 24, 48, and 72 h after the appli-
using the tissue homogenizer. A portion of homogenate
cation of various formulations. The mean erythemal
(each one) was streaked on the solidified growth medium.
scores were recorded (ranging from 0–4) depending
All plates were incubated at 25°C for 5 days in the incuba-
on the degree of erythema as follows: no erythema = 0,
tor. The numbers of CFUs in the agar plates were counted
slight erythema (barely perceptible-light pink) = 1,
and the logarithm of the number of CFUs per infected site
moderate erythema (dark pink) = 2, moderate-to-severe
was calculated. An animal was considered fungus positive
erythema (light red) = 3, and severe erythema (extreme
when more than one fungal colony was counted.
redness) = 4.

HPLC analysis of MN
MN content was determined using an HPLC apparatus
consisting of a pump (LC 10-AD), a UV detector (SPD-
10A), and a data station (Shimadzu, Kyoto, Japan). An
Ultra sphere® C-18 column (250 mm × 4.6 mm i.d.) was
used. After mixing, the mobile phase, which comprised
of methanol:phosphate buffer solution at pH 3.0 (90:10,
v/v), was degassed. The eluent was run at a rate of
1.0 ml/min and monitored at 230 nm following injected
volumes of 20 μL of sample. The relative retention time
was found to be 7.2 min (Peira et al., 2008).
Statistical analysis
Data are expressed as the mean ± standard devia-
tion (SD) of the mean and statistical analysis was car-
ried out employing the ANOVA test using the software
PRISM (Graph Pad). A value of p < 0.005 was considered
­statistically significant.
Results and discussion
Solid lipid nanoparticles-based hydrogel formulation
was prepared in two steps, first the drug-loaded SLN were
 Design and development of solid lipid nanoparticles   447

prepared using the solvent injection method reported by
Goymann and Schubert (2003) and then SLN dispersion
was dispersed in carbopol hydrogel. The various formu-
lation and process variables were optimized to obtain
nanosized particles with maximum drug entrapment
efficiency.
In the present investigation, ethanol was selected as
a miscible solvent for the preparation of SLN due to its
fairly good dermal acceptability as compared to other sol-
vents. Moreover, ethanol has good solubilizing potential
for tristearin at ∼ 70°C. No efforts were made to remove
ethanol and surfactants in the SLN dispersion, as all the
excipients used in this study, at the concentration present
in the dispersion, are acceptable for topical application.
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Moreover the emulsifier also helps to stabilize the newly
generated surfaces and particle aggregation (Goymann
& Schubert, 2003). However, the PDI values decreased
up to 0.5% w/v Tween 80 concentration and further addi-
tion of Tween 80 led to a sharp increase in PDI values
possibly due to irregular size reduction with high Tween
80 concentration (1 or 2%). Hence, the formulation D31
was taken further for optimization of sonication. In the
case of increasing sonication time from 4 to 6 min, no
significant difference in size was observed (p > 0.005),
whereas PDI and entrapment efficiency changed sig-
nificantly (p < 0.005). Hence, D311 was considered as an
­optimization formulation (Table 1).
A stable SLN formulation was achieved after stir-

ring ∼ 3000 rpm for 20 min and sonicating the formula-

tion for 4 min with a minimum average particle size
Optimization of different variables of 206.39 ± 9.37 nm and a maximum percentage drug
The different variables involved in the preparation of SLN entrapment of 90.86 ± 5.20% (Table 1).
were optimized, including lipid/lecithin ratio, drug/lipid
ratio, surfactant concentration, and sonication time to Characterization of SLN
obtain nanosized particles with maximum drug entrap-
Shape and surface morphology of the SLN prepared with
ment (Table 1).
optimized parameters was observed by transmission and
The lipid ratio (tristearin:soya lecithin) was first opti-
scanning electron microscopy. The study revealed that
mized by varying the ratio of tristearin:soya lecithin. It was
For personal use only.

most of the SLN were fairly spherical in shape, the sur-

found that on increasing the soya lecithin concentration in
face of the particle showed a characteristic smoothness,
the tristearin, the particle size decreased from 425.58 ± 10.5
and that the particle size was in the nanometric range
to 311.24 ± 7.2 nm. The decrease in particle size may be due
(Figures 1 and 2).
to the increased surfactant action of soya lecithin with
increase in its concentration for covering the fixed amount
of lipid concentration. Further increasing the lecithin con- Formulation of MN-loaded SLN-bearing hydrogel and
centration led to an increase in particle size, possibly due characterization
to the major effect of lecithin itself and probably due to Carbopol 934 was selected for the preparation of
formation of liposomal and other structures, as evident MN-loaded SLN-bearing hydrogel. The optimized SLN
from a sharp increase in PDI values (Table 1). dispersion was then incorporated into hydrogel. The
Drug:lipid ratio was optimized by varying the drug prepared formulation was first studied to establish the
concentration from of 2.5 to 7.5%. The drug entrapment release kinetics of the drug from the carrier systems.
efficiency increased from 87.62 ± 2.51% to 92.74 ± 2.11%.
On further increasing the concentration of drug to 10%,
the drug entrapment efficacy decreased. Further, there
was no significant increase in particle size on increas-
ing drug concentration up to 7.5% (p > 0.005). On further
increasing the concentration of drug to 10% a significant
increase in particles size was observed (p < 0.005). This
could be due to the saturation of lipid matrix with the
drug above 7.5% drug concentration and reduction of
surfactant effect of PC. Because of depletion of surfactant
effect of PC due to high concentration of drug, the particle
aggregation might occur, leading to an abrupt increase in
particle size of SLN (Table 1).
On increasing the concentration of Tween 80 from
0.1–2% w/v a decreased particle size was observed. This
may be due to the decrease of the surface tension between
organic and aqueous phases that possibly allows the for-
mation of initially smaller solvent droplets at the site of
solvent injection and causes the decreased particle size. Figure 1. SEM image of MN-loaded SLN.
 448   Safal Jain et al.

The pH was found to be 6.8 ± 0.1 which is acceptable for
­topical applications.
Spreadability is an important property for topical
­formulations. Application of the formulation on the
desired part would be more comfortable if the base
spreads easily, exhibiting maximum ‘slip’ and ‘drag’. The
diameter was found to be 5.8 ± 0.2 cm, which is indica-
tive of good spreadability of the MN-loaded SLN-bearing
hydrogel.
In vitro drug release studies
To evaluate the drug release pattern, the dialysis bag
method was used where a mixture of methanol:PBS (pH
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In vitro skin permeation, stripping, and retentivity
studies
A negligible amount of MN was obtained in the receptor
compartment from any of the applied formulations after
24 h in a rat skin permeation experiment (Table 2). On
conducting the tape stripping 10 times after 24 h, it was
observed that MN recovered in tape stripping was high-
est (145.5 ± 6.84 µg/cm2) with MN-loaded SLN-bearing
hydrogel (Table 2). In the case of MN suspension and MN
hydrogel it was 11.4 ± 2.95 µg/cm2 and 13.2 ± 3.52 µg/cm2,
respectively. The results confirm the presence of drug in
the upper layers of the skin which could further provide
a better treatment profile for pathological conditions like
superficial mycoses.

6.4) (30:70) was used as the diffusion medium. Different
release patterns were observed from the MN suspen-
sion, MN hydrogel, and MN-loaded SLN-bearing hydro-
gel. An initial rapid release was observed in the case of
MN suspension, whereas MN hydrogel and MN-loaded
SLN-bearing hydrogel depicted a slow initial release
with a lag time of 0.5 h and 1 h, respectively (Figure 3).
The direct exposure of MN hydrogel and MN suspension
to diffusion media may account for rapid initial release
in hydrogel and suspension. MN-loaded SLN-bearing
For personal use only.
Further, a skin retentivity study conducted after tape
stripping also suggested nearly 10-fold greater skin
retention in the case of MN-loaded SLN-bearing hydro-
gel as compared to the other two formulations (Table 2).
The retentivity profile could be attributed to occlusive
and bioadhesive properties of SLN, where occlusion
decreases the transepidermal water loss, which further
increases skin hydration and permeability. Further, it
may also lead to accumulation of bioactives into the
upper skin layers, reducing drug flux and creating a res-

hydrogel formulations depicted better controlled drug
release profile for a prolonged period, suggesting their
applicability in topical drug delivery. An earlier similar
release profile was obtained in the case of flurbiprofen
by Jain et al. (2005).
ervoir able to prolong skin residence time (Puglia et al.,
2006), and, hence, could provide better treatment to
upper skin infections and disorders. Similar results for
MN skin permeation and deposition were obtained by
100
Cummulative Drug Release (%)

90
80
70
60
50
40
30
20
10
0
0 5 10 15 20 25 30
Time (hrs.)

MN Suspension MN Hydrogel
MN loaded SLN bearing Hydrogel

500 nm Values represent mean ± SD (n=3)

Figure 2. TEM image of MN-loaded SLN. Figure 3. In vitro dug release studies of different formulations.

Table 2. Amount of MN permeated and retained in hairless rat skin from different formulations (tape stripping experiments).
Amount of MN permeated MN in tape strips % MN in skin after tape
S.No. Formulation (µg/cm2) (µg/cm2) stripping
1 MN suspension U.D. 11.4 ± 2.95 4.1 ± 1.18
2 MN hydrogel U.D. 13.2 ± 3.52 5.8 ± 2.73
3 MN-loaded SLN-bearing hydrogel U.D. 145.5 ± 6.84 65.3 ± 3.18
U.D. = Undetectable, values represent mean ± SD (n = 3).
 Design and development of solid lipid nanoparticles   449

Peira et al. 2008) using a microemulsion-based vehicle gel demonstrated a remarkable advantage over simple gel
for topical delivery. in improving the skin tolerability of MN, indicating their
potential in improving patient acceptance and topical
delivery of MN.
Primary skin irritation studies
One of the major disadvantages associated with the MN In vivo studies
therapy is skin irritation (erythema), which strongly limits
The in vivo efficacy of MN-loaded SLN-bearing hydro-
its utility and acceptability by the patients. Ideally, the
gel was assessed in a male albino rat model (Sprague
delivery system of MN should be able to diminish or abol-
Dawley; 100–150 g). Isolate of candida albicans was used
ish these erythematic episodes. It was hypothesized that
for production of cutaneous candidiasis in albino rats.
encapsulation of MN in SLN would reduce the direct con-
Table 4 depicts the efficacy of MN-loaded SLN-bearing
tact of the MN (the triggering factor for the erythematic
hydrogel formulation against cutaneous candidiasis in
events) with the stratum corneum, thus resulting in
rats as compared to that of MN hydrogel and MN sus-
reduced erythematic episodes. The results obtained from
pension. The isolates of viable candida albicans were
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the primary skin irritation studies are listed in Table 3.

removed from the lesions of all the treated animals. It
The skin-irritation studies indicated that MN-loaded SLN-
was observed that MN-loaded SLN-bearing hydrogel
bearing hydrogel resulted in considerably less irritation
depicts greater efficiency in the treatment of candidia-
as compared to MN hydrogel and MN suspension after
sis, as only one animal out of six exhibited a positive
24 h of application. The erythema continued to increase
culture test while in the case of MN hydrogel and MN
even after 24 h in the case of MN hydrogel and MN sus-
suspension treatment it was in three and four animals
pension, whereas no erythema was observed in the case
out of six, respectively. Fast recovery from fungal infec-
of MN-loaded SLN-bearing hydrogel. Thus, SLN-based
tion was found in the case of MN-loaded SLN-bearing
hydrogel. Such good performance of the SLN was pos-
Table 3. Mean erythemal scores observed for various formulations.
For personal use only.

sibly due to better occlusive and bioadhesive properties

Formulation Erythemal scores (n = 3)
(Figures 4 and 5).
24 hrs 48 hrs 72 hrs
Control (Group I) 0 0 0
MN suspension (Group II) 2 3 3
MN hydrogel (Group III) 1 2 2
Conclusion
MN-loaded SLN-bearing hydrogel 0 1 1
(Group IV) The MN-loaded SLN could be fabricated with the help
of a modified solvent injection method and successfully
incorporated into hydrogel for topical application. The
Table 4. Colony forming unit of Candidia albicans in skin of rats after
in vitro and in vivo data indicate that MN-loaded SLN-
treatment with different formulations.
bearing hydrogel provides sustained release of MN. The
No. of animals with
­positive culture/total Log CFU/ obtained results reflect the potential of SLN as a carrier
Treatment no. of animals infected sites for topical administration of MN which is demonstrat-
Control (Group I) 6/6 3.84 ± 0.47 ing greater drug deposition into skin and better in vivo
MN suspension (Group II) 4/6 3.11 ± 0.34 anti-fungal efficacy in a rat model with less erythemal
MN hydrogel (Group III) 3/6 2.46 ± 0.21 score. The study provides evidences that SLN could be
MN-loaded SLN-bearing 1/6 1.28 ± 0.09 a better module for prolonged release profile with wide
hydrogel (Group IV) applications in topical drug delivery.

A B

Figure 4. (a) Skin texture of rat skin for control at initiation of experiment, (b) Skin texture of rat skin for control after 6 days.
 450   Safal Jain et al.
A solid lipid nanoparticles for epidermal targeting. J Contr Rel.
B
Drug Delivery Downloaded from informahealthcare.com by University of Glasgow on 03/16/13
C tructured lipid carrier (NLC)-based hydrogel of valdecoxib. Drug
For personal use only.
Figure 5. Skin texture of rat skin after 6 days treatment with developed
formulations: (a) MN suspension, (b) MN hydrogel, (c) MN-loaded
SLN-bearing hydrogel.
Acknowledgements
The authors are grateful to All India Institute of Medical
Sciences, New Delhi for providing Scanning and Electron
Microscopy facility, and M/s Medley Pharmaceuticals
Mumbai, India for providing the gift sample of Miconazole
nitrate.
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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