122 Send Orders for Reprints to reprints@benthamscience.

ae
Current Drug Delivery, 2018, 15, 122-133

RESEARCH ARTICLE
ISSN: 1567-2018
eISSN: 1875-5704

Impact
Factor:
1.478

Formulation of Niosomal Gel for Enhanced Transdermal

Lornoxicam Delivery: In-Vitro and In-Vivo Evaluation
BENTHAM
SCIENCE

Mohamed Shafik El-Ridy1, Soad Aly Yehia2, Amira Mohamed Mohsen1,*, Sally A. El-Awdan3 and
Asmaa Badawy Darwish1

1
Pharmaceutical Technology Department, National Research Centre, Dokki, Cairo, Egypt; 2Pharmaceutics
Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 3Pharmacology Department, National Research
Centre, Dokki, Cairo, Egypt

ARTICLE HISTORY
Received: October 28, 2016
Revised: January 01, 2017
Accepted: February 13, 2017
DOI:
10.2174/1567201814666170224141548
Current Drug Delivery
Abstract: Background: The objective of this study was to investigate the potential of niosomal gels as
a transdermal delivery system to improve the permeation and anti-inflammatory activity of Lornoxicam
(LX).
Methods: LX niosomes were prepared by thin film hydration technique and were characterized using
Transmission Electron Microscopy (TEM), Differential Scanning Calorimetry (DSC), Particle Size
analysis and Zeta potential determination. LX niosomal gel/LX loaded gel were prepared using Car-
bopol 934 (2%) and were evaluated for their physical appearance, pH and rheological behaviour. Ex
vivo skin permeation test was performed on dorsal region of wistar rats. In vivo studies comprised skin
irritation test and anti-inflammatory activity study.
Results: The prepared LX niosomes exhibited an entrapment efficiency of more than 66% and a parti-
cle size diameter ranging from 295 nm to 1298 nm, with negatively charged zeta potential. TEM elec-
tron micrographs revealed spherical shaped vesicles. The release pattern of drug was analyzed and
found to follow Higuchi’s model. Rheology studies revealed the pseudoplastic behaviour of LX
niosomal gel. They exhibited a one and half fold increase in drug permeated through rat skin, when
compared to free drug. Skin irritation test proved the non-irritancy of LX niosomal gels, when applied
to dorsal region of Wistar rats. Percentage edema inhibition of LX niosomes was significantly higher
(P<0.05) than that of free LX group showing an enhanced anti-inflammatory activity of LX niosomes.
Conclusion: These findings revealed that LX loaded niosomal gels could be a potential transdermal

drug delivery system.

Keywords: Anti-inflammatory, lornoxicam, niosomal gel, niosomes, permeation, transdermal.

1. INTRODUCTION minor to moderate inflammation and pain in rheumatoid ar-

Controlling inflammatory disorders should comprise a
stepwise way to the usage of therapeutic agents. Critical
goals to reduce the severity of symptoms comprises relieving
of pain and reduction of inflammation [1]. Non-steroidal
anti-inflammatory drugs (NSAIDs) are among the most
widely used therapeutics, prescribed for the treatment of pain
and inflammation [2, 3].
Lornoxicam is one of the non-steroidal anti-inflammatory
drugs, belonging to the class oxicam, with analgesic, antipy-
retic and anti-inflammatory effect [4] .It is recommended for
*Address correspondence to this author at the Pharmaceutical Technology
Department, National Research Centre, 33 El-Buhouth Street, Dokki,12622,
Cairo, Egypt; Tel: +202 3335456; Fax: +202 33370931;
E-mail: amiramohsen80@hotmail.com
thritis and osteoarthritis [5]. Parenteral formulations that lead
to immediate release of LX are available for intramuscular
and intravenous (4 mg/ml) route of administration. The dose
that is recommended by oral route of administration is
8-16 mg/day and the total dose/day should not exceed 16 mg
[6, 7].
It was stated that its analgesic activity is analogous to the
analgesic effect of opioids. The oral bioavailability of LX is
90-100%. It has a short half-life (3-5 hrs) [8]. The most
common oral dosages of LX that was used in clinical trials
were 4 mg twice or 3 times daily or 8 mg twice daily for
controlling arthritic conditions [9]. In acidic pH of the stom-
ach LX has a very poor solubility [10]. Thus, it remains in
contact with the stomach wall for a long period leading to
gastric irritation and peptic ulcers [11]. In addition, the

1875-5704/18 $58.00+.00 © 2018 Bentham Science Publishers

Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery
mechanism of action of LX comprises inhibition of cy-
clooxygenase isoenzymes which causes inhibition of the
synthesis of prostaglandin from arachidonic acid. Prosta-
glandins have a significant role in protecting the mucosa of
the gastrointestinal tract (GIT). It acts by hindering the secre-
tion of gastric acid. Thus, inhibition of prostaglandin synthe-
sis usually leads to GIT complications such as ulceration,
bleeding and dyspepsia [12]. Although LX has better accept-
ability compared to other NSAIDs, different side effects,
revealed in the GIT, have been reported for LX. Upon oral
administration, renal and hematological side effects may also
occur. On the other hand, parenteral administration of LX is
not preferred due to chronic complaints [13].
An alternative route to overcome the GIT side effects of
NSAIDs, including LX, can be the transdermal route of ad-
ministration [14, 15]. Transdermal application of drugs has
many benefits over oral and intravenous routes of admini-
stration such as absence of first- pass metabolism and inci-
dence of systemic toxicity [16]. Inflammation can be treated
using transdermal patches that also exhibit systemic effects.
These systems show an improved patient compliance be-
cause of their easy application and ability to discontinue
treatment whenever being requested [17].
Nowadays, different nanotechnology based formulations
are available for topical application of drugs and cosmetics.
Proniosomes [18], solid lipid nanoparticles [12], nanoemul-
sion gels [19] and transdermal matrix patches [20] of LX
were previously studied and reported. Vesicular systems
such as liposomes and niosomes represent an important drug
delivery system [11]. The advantages of niosomes over
liposomes include enhanced chemical and physical stability
[21], lower cost and ease of use of surfactants [22].
Niosomes is a vesicular system which is composed of non-
ionic surfactants (NIS). In the aqueous media, these NIS self-
assemble to produce bilayer structures [23, 24]. Thus, it can
encapsulate both hydrophilic and hydrophobic drugs into this
bilayer structure [25]. Niosmes is a very beneficial drug de-
livery system with several applications [22, 25, 26].
The objective of the present study is to develop trans-
dermal niosomal gel of LX to improve the permeation and
anti-inflammatory activity of LX and to enhance its patient
compliance by providing sustained release medication. The
current research work comprises an in vitro evaluation in-
cluding preparation and characterization of LX niosomes,
in vitro release study as well as in vivo evaluation of the op-
timized LX niosomal gel.
2. MATERIALS AND METHODS
2.1. Materials
LX was a kind gift sample from Eva pharma Co., Cairo,
Egypt. Sorbitan monopalmitate (Span 40), Dihexadecylhy-
drogen-Phosphate (Dicetyl Phosphate) (DCP) and Choles-
terol (Ch) were obtained from Sigma-Aldrich Chemie GmbH,
Germany. Octadecylamine (Stearylamine) (SA) was pur-
chased from Sigma-Aldrich Co., St. Louis, Mo., USA. Chlo-
roform HPLC was obtained from Panreac Quimica SA, Bar-
celona, Spain. All other chemicals were of analytical grade.
Current Drug Delivery, 2018, Vol. 15, No. 1 123
2.2. Methods
An increasing number of NIS has been found to formu-
late niosomes among which sorbitan esters are much more
common. Hao et al. [27] suggested that the length of alkyl
chain is a crucial factor of permeability and that long chain
surfactants produce high drug entrapment into vesicles. Ac-
cordingly, Span 40 was selected as it is one of the surfactants
of choice for the formation of the non-ionic vesicles. Choles-
terol was used with Span40 as an enhancer of niosomal
membrane rigidity [28, 29]. Charge inducing agents (CIA)
such as DCP (negative charge inducing agent) and SA (posi-
tive charge inducing agent) were added to the niosomal bi-
layers for investigation of charge effect. The inclusion of
charge inducing agents in niosomes tends to stabilize them
[30]. Furthermore, it was reported that niosomes constructed
from cholesterol, NIS and dicetyl phosphate were effective
in increasing skin permeation of frusemide across mouse
skin as compared to conventional formulations [31].
2.2.1. Preparation of LX Niosomes
Niosomes were prepared by the thin film hydration
method [24]. In a 100 ml round-bottom flask, 10 mg LX and
accurately weighed quantities (100 mg) of Span40 and Ch
with or without CIA were dissolved in chloroform. The or-
ganic solvent was slowly evaporated under reduced pressure
using the rotary evaporator (Büchi rotavapor-M/HB-140,
Technik AG, Switzerland), at 56-58°C [27]. After chloro-
form evaporation, the thin film formed on the inner wall of
the rotating flask was then hydrated with 10 ml phosphate
buffered saline (PBS) pH=7.4 pre-warmed to 56°C-58°C.
Composition of different prepared LX loaded niosomes is
listed in Table 1.
2.2.2. Characterization of Prepared LX Niosomes
2.2.2.1. Entrapment Efficiency Percentage (E.E. %)
LX niosomes was separated from the free unentrapped
drug by cooling centrifugation at 5400 xg, at 4°C, for 30
minutes using the cooling centrifuge (Union 32R, Korea)
[32, 33]. The niosomal pellets were then washed with 10 ml
PBS and recentrifuged. The supernatant was filtered through
0.22
m Millipore filter (Millipore, USA). Then the free
drug content in supernatant was analyzed [11, 18] spectro-
photometrically at max 375 nm, after comparison to a pre-
constructed calibration curve (n=3, linearity range: 2-20

g/ml, R2=0.9999). Drug entrapment efficiency was calcu-
lated according to the following equation:
E.E. %= [Qt – Qs]/Qt * 100 Eq. (1)
Where E.E. is the entrapment efficiency, Qt is amount of LX
in suspension added, and Qs is amount of LX detected only
in the supernatant [17].
2.2.2.2. Transmission Electron Microscopy (TEM)
Transmission electron microscopy was performed to de-
termine the morphological characteristics of vesicles [34].
Samples were diluted five folds with bidistilled water and
mixed well prior to the examination. One drop of the sample
was placed onto 300-mesh carbon coated grid and allowed to
dry to a thin film [35]. Before complete drying of this film
124 Current Drug Delivery, 2018, Vol. 15, No. 1
Table 1. Composition of different LX niosomal formulations.
Formula Niosomes Type
Span 40
F1 Neutral 1 1
F2 Negative 1 1
F3 Positive 1 1
F4 Neutral 2 1
F5 Negative 2 1
F6 Positive 2 1
on the grid, it was negatively stained with 1% phosphotung-
estic acid. One drop of the freshly prepared stain was added
and allowed to air dry on a filter paper. After drying, the
samples were then examined by the TEM (JEOL, JEM-1230,
Tokyo, Japan.), with an accelerating voltage of 80KV [36].
The experiment was performed at room temperature.
2.2.2.3. Differential Scanning Calorimetry (DSC)
The thermal properties of selected LX niosomes were
analyzed using DSC calibrated with indium (DSC-60 with
TA-60 WS thermal analyzer, Shimadzu, Tokyo, Japan).
Thermal analysis was also performed on plain (drug-free)
niosomal formulations of the selected formulations F1 and
F4. Niosomal dispersions were lyophilized before DSC
analysis using freeze dryer (VirTis Freeze Drying Equip-
ment, Snijders Scientific). Thermograms were analyzed us-
ing Shimadzu TA-60 software. The niosomal individual
components of niosomes were also analyzed. A known
amount of the lyophilized niosomes was placed in a hermeti-
cally sealed aluminum pan and scanned at a rate of 5°C/min
[37] over a temperature range of 20-300°C with nitrogen
purging (25ml/min).
2.2.2.4. Vesicular Size Analysis
Vesicular size for the prepared niosomes was determined
by dynamic light scattering method based on laser diffraction
in a multimodal mode using the Malvern particle sizing sys-
tem (Malvern, Nano Series ZS90, Malvern Instruments, Ltd.,
UK) [38, 39] with He-Ne laser at 632.8 nm at a scattering
angle of 90.0° [40], which is capable of measuring vesicles
in the 1nm to 5
m size range. The niosomal preparation was
diluted with bidistilled water (1:100 vol/ vol). The sample
was then transferred to a quartz cuvette and examined at
room temperature. Size distributions were displayed in term
of number versus vesicle size. The polydispersity index (P.I.)
was determined as a measure of homogeneity [14].
2.2.2.5. Zeta Potential Determination
The zeta potential was measured on the bases of laser
diffraction analysis using Zeta sizer (Malvern, Nano Series
ZS90, Malvern Instruments, Ltd., UK). The niosomes were
El-Ridy et al.
Molar Ratio
Charge Inducing Agent
Ch
DCP SA
_ _
0.1 _
_ 0.1
_ _
0.2 _
_ 0.2
diluted tenfold with bidistilled water. The samples were then
transferred to a quartz cuvette and measured at room tem-
perature. Zeta potential studies were performed in triplicate.
2.2.3. Preparation of LX Niosomal gel/ LX Loaded Gel
Preparation of LX niosomal gel of selected niosomal
dispersion (F1) took place by adding 200 mg of Carbopol
934 in 10 ml niosomal dispersion with continuous stirring
using a glass rod. Triethanolamine (TEA) was added drop
wise for neutralization till reaching the desired skin pH (6.8-7)
[41].
A gel base was prepared by sprinkling the weighed
amount of Carbopol 934 (2% w/w) in appropriate amount of
distilled water to form an aqueous dispersion [42]. This dis-
persion was stirred, by magnetic stirrer, for 2 hrs. A known
amount of LX (equivalent to 1% w/w) was added to the gel
base and properly dispersed, with continuous stirring, until
homogenous gel was formed. Drops of TEA were added till
reaching the required pH. All the samples were stored in
refrigerator (4°C) for at least 24 hours prior to performing
rheological measurements [43].
2.2.4. Evaluation of Free Drug/LX Niosomal Gel
The prepared gels were evaluated via the following
studies.
2.2.4.1. Physical Appearance
The gels were observed for their clarity and homogeneity
by visual examination. They were also checked for the pres-
ence of any aggregates or clumps.
2.2.4.2. Measurement of pH
The pH of the gels was measured using digital pH meter
(Thermo Scientific Orion VERSA STARTM Advanced Elec-
trochemistry Meter, VSTAR 92, Thermo Fisher Scientific
Inc., USA) at room temperature. This study was performed
to assure that the pH of the developed gels are close to skin
pH, after being kept in refrigerator for 24 hrs. The determi-
nations were carried out in triplicate and the averages of
three readings were noted and S.D. was determined.
Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery
2.2.4.3. Rheological Behavior
Rheological behavior of prepared niosomal gel was
measured using parallel-plate rheometer (Anton Paar,
Physica MCR 301, Germany) at room temperature. The
shear stress and viscosity were determined while increasing
shear rate progressively (from 1 to 200 sec-1) and then
gradually decreasing (from 1 to 200 sec-1) [44]. Thus
rheogram curves were constructed. The flow behavior was
then analyzed by regression analysis of the log shear stress
vs. log shear rate through following equation:
Log G = n Log F – Log  Eq. (2)
-1
Where: G is shear rate (sec ), F is shear stress (Pa),  is vis-
cosity (Pa.s) and n (Farrow’s constant) is slope of log G
against log F plot, and it denotes deviation from Newtonian
flow behavior. It rises as the flow becomes increasingly non-
Newtonian. When n>1, it indicates pseudoplastic flow or
shear thinning and when n<1 this indicates dilatant flow or
shear thickening flow [45].
2.2.5. In Vitro Release Studies
2.2.5.1. In Vitro Release of LX Niosomes
This study was conducted on neutral and negatively
charged niosomal dispersions. Positively charged niosomes;
F3, and F6, were not included in this study and in further in
vivo studies due to their larger particle size revealed, low
stability indicated by zeta potential results, in addition to
their aggregation observed by TEM micrographs. The
amount of LX released from different niosomal formulations
was determined using the dialysis bag diffusion technique
[46]. The dialysis bag (molecular weight cut off 12000–
14000) was soaked in PBS (pH 7.4) for 12h before use [12].
The cellulose bag was filled with niosomal formulations
equivalent to 2 mg drug. Then, the cellophane bag was im-
mersed in a beaker containing 100 ml PBS (pH 7.4) [18, 46].
Sink conditions was maintained throughout the experiment
[47]. The beaker was placed in a water bath shaker to main-
tain the temperature at 32 ± 2ºC, with speed of 100 rpm.
At predetermined time intervals (1, 2, 3, 4, 5, 6, 7, 8 and
24 hrs); samples were withdrawn for analysis and replaced
with equal volume of fresh PBS (pH 7.4) at 32 ± 2ºC to
maintain a constant volume. The amount of drug in the col-
lected samples was detected spectrophotometrically. Each
result is the mean of at least four determinations. The per-
centage of drug release was plotted as a function of time.
2.2.5.2. In Vitro Release Study of LX Niosomal Gel
In vitro release study was conducted on selected LX
niosomes (F1) after being incorporated in gel, as well as free
LX loaded gel. The experiment was performed on the same
conditions as previously mentioned in 2.2.4.1.
2.2.6. Kinetic Study of Release Profiles of LX Niosomes
and LX Niosomal Gel
After completing in vitro release profiles of all the inves-
tigated batches, the data were treated with different mathe-
matical models, i.e., zero order kinetics, first order kinetics
[48] and Higuchi model [49], as described in Eqs. (2-4). The
release rate constant (K) values and the correlation coeffi-
Current Drug Delivery, 2018, Vol. 15, No. 1 125
cient (R2) were calculated from the linear curve obtained by
regression analysis of these plots.
Mt = M0 + k0t Eq. (2)
Mt = M0 (e –k1t) Eq. (3)
1/2
Mt = kH.t Eq. (4)
where Mt and M0 are amounts of drug released in the release
medium at time t and at t =0, respectively, k0, k1 and k H are
release constants of the zero-order, first-order and Higuchi
square root model, respectively.
2.2.7. Ex Vivo Skin Permeation Experiment
Skin permeation studies ware evaluated on hairless ab-
dominal Wistar rat skin using Franz diffusion cell [50]. The
receptor volume was 63 ml (phosphate buffer saline pH 7.4)
[11, 12] and the surface area of the release membrane was
3.14 cm2. The donor side was charged with 2 gm of the in-
vestigated gel (Free drug in gel or LX niosomal gel). The
solution in the receptor side was stirred with magnetic stirrer
bar adjusted at 500 rpm [51] and maintained at 37oC. Sam-
ples (2 ml) were withdrawn at different time intervals (1, 2,
4, 6 and 24 h) and replaced by fresh medium to maintain sink
condition [11]. Drug concentration was detected using a
validated HPLC method. HPLC Agilent 1260 infinity sys-
tem, equipped with MWD 1260 (Germany) was used. The
system employed consisting of a quaternary pump, 1260
ALS degasser and variable-wavelength detector. The sam-
ples were separated on a C18 column (Zorbax eclipse plus,
4.6 x 250 mm, Germany) with 0.1 M sodium dihydrogen-
phosphate buffer (pH 6)/ methanol (50:50, v/v) at a flow-rate
of 1 ml/min. The detection absorption was at 372 nm [52].
The linearity range of the calibration curve for LX was 10 -
2000 ng/ml. The standard curve of LX in plasma was linear
and the regression coefficients was R2=0.9992 (n=3), indi-
cating good linearity.
The cumulative amount of LX permeated per unit dialy-
sis membrane area (
g/cm2) was plotted versus time and the
permeation profiles were constructed. Linear regression
analysis was used to calculated steady state flux (Jss) of
drug. The permeability co-efficient (Kp) of LX, through the
stratum corneum, was calculated using the following equa-
tion [53]:
Kp = Jss / C Eq. (5)
Where, C is the initial concentration of the drug in the donor
compartment. The penetration enhancing effect was calculated
in terms of enhancement ratio (ER) by using the equation:
ER = Jss of formula/ Jss of control Eq. (6)
2.2.8. In Vivo Biological Evaluation of LX Niosomes
2.2.8.1. Animals
Albino Wistar rats weighting were used in all in vivo
studies. Animals were housed in polypropylene cages and
were given standard diet and water. They were kept at
25±1ºC with at 12 hr. light /dark cycle. The in vivo experi-
ments were conducted in accordance with ethical procedures
and policies approved by the Medical Research Ethical
126 Current Drug Delivery, 2018, Vol. 15, No. 1
Committee of the National Research Centre, Cairo, Egypt,
registration number 14078.
2.2.8.2. Skin Irritation Test
The skin irritation of LX niosomal gel formulation (F1)
was evaluated in terms of biological investigation, on white
albino Wistar rats (120-160 gm) based on the method de-
scribed by Draize et al. [54]. The rats were anesthetized with
thiopental (60 mg/kg) injection (i.p) then the dorsal side of
the rat was shaved 12 hr before the beginning of the experi-
ment. The animals were divided into five groups each group;
n=3 rats. Group I served as negative control while group II
acted as positive control and received 0.8% (v/v) aqueous
formalin solution as a standard irritant [55]. Group 3 re-
ceived niosomal gel free from LX. Groups 4 and 5 received
LX niosomal gel (F1) and free LX in gel, respectively, for 3
day. The sites of application was checked for edema and
erythema after 24 hours, and then graded from zero to 4 ac-
cording to a visual scoring scale, performed by the same
investigator. The scoring followed the following manner: 0,
none; 1, slight; 2, well defined; 3, moderate; and 4, scar or
ulcer formation. The edema scale was: 0, none; 1, slight; 2,
well defined; 3 moderate; and 4, severe. Photos were taken
for erythema visualized on some rats’ application sites. The
primary irritancy index (PII) was calculated for each group
according to the equation [54]:
Primary irritancy index (PII) = Mean of erythema + Mean of edema
Eq. (7)
The formulation were classified as non-irritant if (PII < 2),
irritant if (PII =2-5) and highly irritant if (PII=5-8).
2.2.8.3. In Vivo Anti-Inflammatory Activity
Anti-inflammatory activity of selected LX niosomal gel
was evaluated using carrageenan induced rat paw edema
model. Animals were divided into five groups; n=6. Group 1
acted as control. Group 2 received blank niosomal formula-
tions of (F1) in gel. Group 3 received free LX in gel. Groups
4 and 5 received LX niosomal (F1) gel and market product,
resoectively.
At the level of tibiotarsic articulation, a mark was made
on the right paw. The basal paw volume was measured by a
volume displacement method, previously described by
Swingle, et al. [56]; employing a water digital plethysmome-
ter 7500 (Ugo Basile, Comerio, Italy). Carrageenan suspen-
sion (0.1 ml of 1% w/v, in saline) was injected into the sub-
plantar surface of the right hind paw. Following this, the
formulations were applied on the shaved dorsal region of
animals and the paw volume was measured again at several
time intervals (1, 2, 3, 4, 5, 6 and 24 hrs) after carrageenan
injection. The increase in paw volume was calculated as per-
centage of edema compared to the basal paw volume accord-
ing to the following equation [57]:
Edema inhibition (%)
Vuntreated - Vtreated X 100/Vuntreated
Eq. (8)
Where, Vtreated is the volume of the paw after the topical
treatment of gel and Vuntreated is the volume of the paw with-
out treatment
El-Ridy et al.
2.2.9. Data Analysis and Statistics
Data are expressed as mean ± standard deviation (S.D.).
Statistical analysis was performed using SPSS software (ver-
sion 22.0; IBM Co., USA). Analysis of variance (ANOVA,
single factor) followed by Fisher LSD’s post-hoc test, was
employed in the statistical analysis of the determined pa-
rameters. A P-value < 0.05 was considered statistically
significant.
3. RESULTS AND DISCUSSION
3.1. Entrapment Efficiency Percentage (E.E. %)
The entrapment efficiencies (E.E. %) of all niosomal
formulations are revealed in Table 2. The effects of several
factors including the effect of different surface charges, dif-
ferent molar ratios and effect of non-ionic surfactant type on
the entrapment efficiency of LX were studied.
3.1.1. Effect of Surface Charge on LX E.E. %
Results, tabulated in Table 2, reveal that for the molar
ratio, Span 40: Ch: CIA (1:1:0.1), neutral LX niosomes ex-
hibited the highest E.E. % (66.11 ± 2.50%) showing a sig-
nificant difference (P< 0.05) when compared to negatively
(58.72 ± 1.09%) and positively (54.5 ± 2.31%) charged
niosomes. Same sequence was noticed for the molar ratio
Span 40: Ch: CIA (2:1:0.2). This can be attributed to the
presence of higher Ch content in neutral niosomes. Choles-
terol is known to be one variant in the niosomal structure
which could control retention of drugs by increasing the
packing of surfactants molecules, reducing bilayer perme-
ability [58]. The inclusion of DCP or SA tend to decrease the
amount of LX entrapped and that decrease may be due to the
repulsion between the niosomal bilayers which led to in-
crease of the leaking space in the bilayer membranes and
therefore permit the release of the encapsulated drug. These
results come in accordance with those reported for the incor-
poration of charge inducing agents into indomethacin
liposomes [59], acetazolamide liposomes [60], and gentamy-
cin niosomes [61].
3.1.2. Effect of Molar Ratio on E.E.%
The results revealed in Table 2 show that neutral, nega-
tively and positively charged LX niosomes of molar ratio
Span 40:Ch (1:1) and Span 40: Ch :CIA (1:1:0.1), F1, F2 and
F3, exhibited slightly higher entrapment efficiencies than
those prepared using the molar ratio Span 40:Ch (2:1) and
Span 40: Ch: CIA (2:1:0.2), respectively. These findings
might be attributed to the higher Ch content that reached up
to 50 mole % for the molar ratio Span 40: Ch (1:1) and Span
40: Ch: CIA (1:1:0.1) compared to the molar ratio Span: Ch
(2:1) and Span: Ch: CIA (2:1:0.2) with a Ch content of only
30 mole %. Recently, Confalonieri et al. [62] reported that
increasing the Ch content resulted in an improvement of the
total amount of drug encapsulated due to an increase in the
lipophilic behavior and crystallinity of the surfactant bilayer
of niosomes. Accordingly, we can conclude that LX, which
is a highly hydrophobic drug, intercalated in these lipophilic
portions.
Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery Current Drug Delivery, 2018, Vol. 15, No. 1 127

Table 2. Characterization of LX loaded Span 40 niosomes.

Formula Particle Size (nm) PDI Zeta Potential (mV) ± S.D. E.E. (%) ± S.D.

F1 295.3 0.391 -46.9 ± 9.83 66.11 ± 2.50

F2 531.2 0.410 -53.6 ± 6.78 58.72 ± 1.09

F3 1298 0.259 -21.2 ± 5.48 54.50 ± 2.31

F4 642.9 0.386 -56.9 ±11.6 65.68 ± 3.13

F5 740.9 0.325 -69 ± 7.4 57.62 ± 0.83

F6 955.4 0.492 -21 ± 5.31 42.83 ± 0.54

3.2. Niosomes Characterization
3.2.1. Transmission Electron Microscopy (TEM)
Fig. (1) reveals different vesicle shapes of LX niosomal
formulations, F1-F6, at different magnification powers. The
vesicles are well identified and exhibit a sphere like shape.
They also reveal well-stained niosomal vesicles, where the
outer lipophilic domain is black stained and the inner hydro-
philic domain is light stained.
3.2.2. Differential Scanning Calorimetery (DSC)
Fig. (2) reveal the themograms as well as the phase tran-
sition temperatures (Tc) of the free drug LX, LX loaded
niosomes as well as individual components of niosomes in-
cluding, Span 40, Ch, DCP and SA. Thermograms of span
40 showed sharp endothermic peak at 50.57°C, indicating its
crystallinity, while the thermogram of Lornoxicam showed
sharp exothermic peak at 221.54°C, which is in accordance
with its reported melting point [63].
DSC thermograms of plain niosomal formulations re-
vealed a change in the phase transition temperature of the
main constituent. A clear change in the phase transition tem-
perature of Span 40 was detected with clear changes in the
enthalpy in addition to peak broadening. The DSC thermo-
grams of LX niosomes, revealed the disappearance of the
melting exothermic peak of LX and shifting of the endo-
thermic peak of span 40. The disappearance of the character-
istic peak of drug and shifting and/or broadening of the en-
dotherms of surfactant recommends possible contact of drug
with bilayer components and may indicate the improved en-
trapment of LX into these vesicles [64].
3.2.3. Vesicular Size Analysis
3.2.3.1. Effect of CIA on Vesicle Diameter of LX Niosomes
The results, tabulated in Table 2, reveal that charged
niosomes (F2, F3/F5, F6) showed larger vesicular sizes
compared to their neutral counterparts (F1 and F4), respec-
tively. Incorporation of a CIA in niosomes usually leads to
an improvement in the spacing between adjacent bilayers,
leading to an increase in the size of the internal aqueous
compartment which causes the formation of larger niosomes
[65].

200 nm 500 nm 1 mm

F1 F2 F3

1 mm 1 mm 2 mm

F4 F5 F6

Fig. (1). Transmission electron microscopy micrographs of LX loaded niosomes.

128 Current Drug Delivery, 2018, Vol. 15, No. 1 El-Ridy et al.

0 50 100 150 200 250 300 respectively). The negative values of the zeta potential for
0.0 neutral niosomes might be attributed to the adsorption of
-0.3
-0.6 F1 LX Niosomes
counter ions or the preferential adsorption of hydroxyl ions
-0.9 at the vesicle surface [30].
0.00
-0.33 Negatively charged LX niosomes exhibited higher nega-
-0.66 F1 drug free niosomes
tive ZP values, i.e., -53.6 ± 6.78 mV and -69 ± 7.4mV for F2
-0.99
18.3
and F5, respectively, showing higher stability. Positively
12.2 Lornoxicam
charged LX niosomes showed lower ZP values (-21.2 ± 5.48
6.1 mV and -21 ± 5.31 mV) which indicates lower stability. Ac-
Endothermic

0.0
0.0
cordingly, positively charged niosomes were not included in
-6.3 further studies.
-12.6 Stearylamine
-18.9
0.0 3.3. Evaluation of Free Drug/LX Niosomal Gel
-4.1
-8.2 3.3.1. Physical Appearance
-12.3 Dicetyl Phosphate
0.0 The developed gels appeared clear and smooth. All gels
-2.9
examined were homogenous, showing no aggregates or
-5.8
-8.7 Cholesterol
clumps.
0.0
-5.6
3.3.2. pH
-11.2 S pa n 4 0
-16.8 The pH measured for the prepared LX niosomal gel/free
0 50 100 150 200 250 300 drug gel are listed in Table 3. Their pH ranged from 6.92-
Temperature (C) 6.96, showing similar pH as that of skin.

Fig. (2). Differential Scanning Calorimetry thermograms of drug,
niosomal components, LX loaded niosomes and drug free niosomes.
3.2.3.2. Effect of Molar Ratio on Vesicular Diameter of LX
Niosomes
From the results, it can be concluded that negative LX
niosomes of the molar ratio Span 40: Ch: CIA (1:1:0.1), ex-
hibited smaller vesicle sizes compared to their counterparts
of the molar ratio, Span 40: Ch: CIA (2:1:0.2). Incorporation
of DCP, which is an anionic surfactant in hydrophilic nature,
into the niosome membrane leads to water efflux into the
bilayer and promotes separation between bilayers [66]. Thus,
particle size and membrane thickness also increase.
On the contrary, in the case of positive niosomes of the
molar ratio Span 40: Ch: SA (1:1:0.1), containing higher
cholesterol content, exhibited larger vesicle size than posi-
tive niosomes of the molar ratio, Span 40: Ch: SA (2:1:0.2).
This can be attributed to the fact that cholesterol increases
the width of lipid bilayers and consequently increases the
vesicle size [67].
3.2.3.3. Polydispersity Index (PDI)
PDI values that are (< 0.3) indicate a homogenous popu-
lation, while higher PDI values moves towards heterogeneity
[68, 69] . PDI values, tabulated in Table 2 calculated for LX
niosomal dispersions ranged from 0.259 to 0.492, indicating
an intermediate homogeneity.
3.2.4. Zeta Potential (ZP)
The zeta potential is a measurement of stability of
niosomes. It was reported that if zeta potential values are
lower than (-30 mV) or higher than (+30 mV) the formula-
tion is considered to be stable [70]. Results, revealed in
Table 2, show that neutral LX niosomes showed negative ZP
values (-46.9 ± 9.83 mV and -56.9 ±11.6 mV, for F1 and F4,
3.3.3. Rheology Behavior
Fig. (3) illustrates the flow curve (shear stress vs. shear
rate) and viscosity curve (viscosity vs. shear rate) of LX
niosomal gel as well as that of LX loaded gel. The upward
and downward flow curves nearly overlapped, showing a
very small gap between each other, indicating that the devel-
oped gels exhibited shear thinning (pseudoplastic) behavior
[71]. This shear thinning behavior can be ascribed to the
instant arrangement of the colloidal network structure into
layers that can flow over each other more easily in the direc-
tion of shear, thus viscosity is decreased with the increase of
shear rate. Farrow’s constants (n), listed in Table 3, were
3.21 and 3.90 for LX niosomal gel and free drug loaded gel,
respectively, confirming the pseudoplastic or shear thinning
behavior (since n>1) [45]. These findings reveal that the de-
veloped systems acquired all the requirements needed for
topical application.
3.4. In Vitro Release Profile of LX Niosomes and LX
Niosomal Gel
The release profiles of drug from the investigated nioso-
mal dispersions are demonstrated in Fig. (4). The results
reveal that the release of LX from niosomes was biphasic.
There was an initial rapid drug release where about 11-33 %
of the encapsulated drug was released, from various nioso-
mal formulations as well as niosomes in gel, in the first hour.
This can be explained by the fact that the lipophilic drug LX
is mostly encapsulated between the fatty acid chains of the
lipid bilayers of niosomal vesicles. This caused rapid release
of drug on dispersing vesicles in buffer till reaching equilib-
rium [72]. During the following 5 h, further 15-29% of LX
was released from various niosomal preparations. After that,
the entrapped drug showed a sustained release profile. Dif-
ferences in the in vitro release patterns could be due other
factors such as lamellarity, vesicle size and membrane fluid-
ity which depends on chain length of surfactant [73].
Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery Current Drug Delivery, 2018, Vol. 15, No. 1 129

LX niosomal gel LX loaded gel

Up Up
600 1200 Down
Down

Shear stress (Pa)

500 1000

Shear stress (Pa)

400 800

300 600

200 400

100 200

0 0
0 50 100 150 200 0 50 100 150 200
Shear rate (1/s) Shear rate (1/s)

50 60

40 50
Viscocity (Pa.s)

Viscocity (Pa.s)
40
30
30
20
20
10
10

0 0
0 50 100 150 200 0 50 100 150 200
Shear rate (1/s) Shear rate (1/s)

Fig. (3). Rheograms of LX niosomal gels and LX loaded gel presenting shear stress vs. shear rate and viscosity vs. shear rate plots.

120 F1 niosomal
gel
100 Free LX gel

F1
80
Drug released (%)

F2
60
F4
40
F5
20
Free LX

0
0 5 10 15 20 25 30
Time (hrs)

Fig. (4). In vitro release profiles of different LX loaded niosomes and LX niosomal gel.

Table 3. pH and rheological behavior of LX niosomal gel and LX loaded gel.

Viscosity at Minimum Viscosity at Maximum

Formulation pH ( mean ± SD) Farrow's Constant
Shear Rate (Pa.s) Shear Rate (Pa.s)

LX niosomal gel 6.96 ± 0.11 47.3 2.86 3.21

LX loaded gel 6.92 ± 0.12 60 5.83 3.90

Fig. (4) also reveals the release pattern of the selected LX correlation coefficients (R2) values, showed that the release
niosomal gel (F1). It can be observed that the drug release of LX from niosomal formulations/niosomal gel is best-fitted
from niosomal gel occurred in two phases, a controlled re- to Higuchi’s model (diffusion mechanism), where it showed
lease phase, characterized by a relatively moderate drug re- the highest (R2) values. The results point to sustained release
lease rate, followed by a steady phase with a reduced and characteristics with a diffusion pattern of drug release, where
slow release rate which was maintained for 24 hours. niosomes act as a reservoir system for continuous delivery of
drug. These controlled release patterns of entrapped drug
3.5. Kinetics Studies of the Release Profiles reflect the high stability of these niosomal formulations [74].
Different mathematical models were employed to predict These findings confirmed that the drug was released from
the release mechanisms and compare the release profiles. niosomes by a diffusion-controlled manner, and they are in
Linear regression analysis of the mathematical models was agreement with results obtained and reported by previous
employed for the release data of LX from niosomes. The research studies [74-77].
130 Current Drug Delivery, 2018, Vol. 15, No. 1
35
30
Drug permeated (%)
25
20
15
10
5 Free LX gel
0
0 5
Fig. (5). Permeation profiles of LX loaded niosomal gel and LX gel.
(a)Erythema=0, Edema=0, PII=0.00 (b)Erythema=3, Edema=2, PII=5.0
(d)Erythema=0, Edema=0, PII=0.0
Fig. (6). In vivo skin irritation study showing photos of the dorsal region of wistar rats after application of (a) control; (b) positive control; (c)
drug free niosomal gel; (d, e) optimized LX niosomal gel (F1) (f) plain LX gel.
3.6. Ex Vivo Skin Permeation
Fig. (5) show the permeation profiles of drug from plain
gel as well as from F1 niosomal gel. Results show superior-
ity of permeation of LX encapsulated in niosomes (F1) over
plain LX gel. Table 4 reveals the permeability parameters;
steady-state flux (Jss), permeability coefficient (Kp), and en-
hancement ratio (ER). There was a significant (P<0.05) im-
provement in the permeation coefficient (Kp) of LX when
encapsulated in niosomes, 2.41x10-3 ± 9.52x10-5 cm/hr for
F1niosomal gel, compared to free LX gel (1.48x10-3 ±
12.72x10-5 cm/hr). There was about one and a half fold in-
crease in deposition of drug when incorporated into the
niosomal gel (F1), when compared to free drug in gel, as
revealed by Er (1.62 ± 0.23). Improved skin permeation of
LX from niosomal gel can be justified by the fact that non-
ionic surfactant (Span 40) in the niosomal formulation act as
permeation enhancers. The surfactants in vesicular form re-
duce the crystallinity of the intracellular lipid bilayers of the
skin and thus improve drug permeation [11]. Furthermore,
the niosomal vesicles in nanometer size range show higher
occlusive characteristics over plain aqueous gel. These char-
El-Ridy et al.
Niosomal
gel F1
10 15 20 25 30
Time (hr)
(c)Erythema=1, Edema=0, PII=1.0
(e)Erythema=0, Edema=0, PII=0.0 (f)Erythema=0, Edema=0, PII=0.0
acteristics enhance hydration of the skin and drug deposition
as well [61, 78].
3.7. In Vivo Evaluation of LX Niosomes
3.7.1. Skin Irritation Test
Fig. (6) reveals the photographs taken after application of
different formulations to Carrageenan injected wistar rats.
The results show that negative control group didn’t show any
erythema or edema. Positive control group (Fig. 5b) showed
redness, erythema and edema and scored 3 for erythrma and
2 for edema yielding a PII score of 5. Blank niosomal gel
showed some erythema, without any edema and PII was cal-
culated to be 1. Rats which received either LX niosomal gel
F1 (Fig. 5d,e) or free LX gel (Fig. 5f) were free of any irrita-
tion and there were no signs of erythema or edema
(PII=0).According to Draize et al. [54], the investigated
niosomal gels were considered to be non-irritant [PII< 2].
The primary irritation indices emphasized the non-irritancy
of the niosomal formulation components and showed that the
investigated formula is safe to be applied on the skin for the
intended period of time.
Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery Current Drug Delivery, 2018, Vol. 15, No. 1 131

Table 4. Comparison of LX loaded niosomal gel and free LX gel in terms of steady flux (ug/cm2/hr), permeation co-efficient (cm/hr)
and enhancement ratio from ex vivo skin permeation studies, n=3.

Formulation Jss ± S.D.* (ug/cm2 /hr) Kp ± S.D.* (cm/hr) Enhancement Ratio (Er)

F1 4.83 ± 0.19 2.41x10-3 ± 9.52x10-5 1.62 ± 0.23

Free LX 3.12 ± 0.31 1.48x10-3 ± 12.72x10-5 __
*: data are significantly different from one another at P<0.05.

Plain niosomal gel Sp40: Ch (1:1)

LX gel
Niosomal gel Sp40: Ch (1:1)
Market product
30

a, b
a, b
25

a, b
Edema inhibition (%)

a, b

a, b
a, b

a
a, b
20

a, c
a

a, c

a, c, d
15
a, c
a,c
a
a

b, c, d
b, c, d

b, c, d
b, c, d

10

b, c, d
c
c

0
0 1 2 3 4 5 6 7 8
Time (hr)
Fig. (7). Paw edema inhibition percentages after application of plain niosomal gel, LX gel, LX niosomal gel and market product to car-
rageenan induced rat paw. “a” significantly different from plain niosomal gel group, “b” significantly different from LX gel group, “c” sig-
nificantly different from LX loaded niosomal gel group, “d” significantly different from market product, at P<0.05.

3.7.2. In Vivo Anti-Inflammatory Activity revealed that the release of LX from niosomes is best-fitted to
Higuchi’s model. Skin permeation studies proved improved
The percentage edema inhibition is shown in Fig. (7).
skin permeation of LX from niosomal gel. Skin irritation test
The results reveal that the percentage edema inhibition for
proved the non-irritancy of the applied niosomal gel compo-
F1 LX niosomal gel was significantly higher (P<0.05) than nents. Paw Edema study revealed enhanced anti-inflammatory
free LX group, starting 3 hours and throughout the whole
activity of LX niosomal formulations. These findings reveal
study time till 24 hours (25.5±4.32% for F1 compared to
the potential for LX encapsulation in niosomes.
11.79 ± 6.97 for free LX group). Since LX niosomal gel F1
remained superior to the free LX in its ability to suppress ETHICS APPROVAL AND CONSENT TO PARTICI-
edema, this indicates improved anti-inflammatory activity of PATE
LX niosomal system. Statistical analysis, employing one
Animal procedures were conducted in accordance with
way ANOVA followed by LSD, revealed insignificant dif-
the Medical Research Ethics Committee of the National Re-
ference between LX niosomal systems and market product.
search Centre, Cairo, Egypt, registration number 14078.
CONCLUSION HUMAN AND ANIMAL RIGHTS
LX loaded niosomes were successfully prepared yielding No Humans were used for studies that are base of this
a high drug E.E.%. The vesicles prepared were in the research.
nanometric range and exhibited a sphere like shape. DSC The reported experiments in accordance with the stan-
thermogram indicated possible contact of drug with bilayer dards set forth in the 8th Edition of Guide for the Care and
components. Rheological studies revealed that LX niosomal Use of Laboratory Animals (http:// grants.nih.gov/grants/
gel acquired pseudoplastic behavior. The release pattern of olaw/Guide-for-the-care-and-use-of-laboratory-animals.pdf)
LX from the niosomal formulations as well as niosomal gel published by the National Academy of Sciences, The Na-
was biphasic, with an initial rapid drug release that was fol- tional Academies Press, Washington DC, United States of
lowed by a slower one. The correlation coefficients values America.
132 Current Drug Delivery, 2018, Vol. 15, No. 1
CONSENT FOR PUBLICATION
Not applicable.
CONFLICT OF INTEREST
The authors declare no conflict of interest, financial or
otherwise.
ACKNOWLEDGEMENTS
The authors express their gratitude to the Project’s Sector
at the National Research Centre, Cairo, Egypt, for funding
this work through the research group project fund number
10070115.
REFERENCES
[1] Foye, W.O.; Lemke, T.L.; Williams, D.A. Foye's Principles of
Medicinal Chemistry, Lippincott Williams & Wilkins, 2008.
[2] Lim, H.W.; Thorbecke, G.J.; Baer, R.L.; Gigli, I. Effect of indo-
methacin on alteration of ATPase-positive Langerhans cell density
and cutaneous sunburn reaction induced by ultraviolet-B radiation.
J. Invest. Dermatol., 1983, 81(5), 455-458.
[3] Bissett, D.; Chatterjee, R.; Hannon, D. Photoprotective effect of
topical anti-inflammatory agents against ultraviolet radiation-
induced chronic skin damage in the hairless mouse. Photodermatol.
Photoimmunol. Photomed., 1990, 7(4), 153-158.
[4] Homdrum, E.-M.; Likar, R.; Nell, G. Xefo® rapid, A novel effec-
tive tool for pain treatment. Eur. Surg., 2006, 38(5), 342-352.
[5] Staunstrup, H.O.J.; Larsen, U.T.; Elbaek, K.; Larsen, U.; Kroner,
K. Efficacy and tolerability of lornoxicam versus tramadol in post-
operative pain. J. Clin. Pharmacol., 1999, 39(8), 834-841.
[6] Hamza, Y.E.S.; Aburahma, M.H. Innovation of novel sustained
release compression-coated tablets for lornoxicam, formulation and
in vitro investigations. Drug Dev. Ind. Pharm., 2010, 36(3), 337-
349.
[7] Skjodt, N.M.; Davies, N.M. Clinical pharmacokinetics of Iornoxi-
cam. A short half-life oxicam. Clin. Pharmacokinet., 1998, 34,
421-428.
[8] Karna, N. Design, development and evaluation of novel sustained
release bi-layer tablets of lornoxicam based on the combination of
hydrophilic matrix formers and basic pH modifiers. Int. J. Pharm.
Biol. Sci., 2012, 3, 392-402.
[9] Godara, S.; Srivastava, R.; Godara, R.; Bhutani, G. Lornoxicam, a
review of its therapeutic potential in different clinical studies. J.
Drug Deliv. Therap., 2013, 3(2), 145-148.
[10] Skjodt, N.M.; Davies, N.M. Clinical pharmacokinetics of lornoxi-
cam. Clin. Pharmacokinet., 1998, 34(6), 421-448.
[11] Deepak, P.W.K.; Vavia, P. Niosomal gel of lornoxicam for topical
delivery: In vitro assessment and pharmacodynamic activity. AAPS
Pharm. Sci. Tech., 2013, 14(3), 1072-1082.
[12] Gönüllü, Ü.; Üner, M.; Yener, G.; Karaman, E.F.; Aydomu, Z.
Formulation and characterization of solid lipid nanoparticles,
nanostructured lipid carriers and nanoemulsion of lornoxicam for
transdermal delivery. Acta Pharmaceutica., 2015, 65(1), 1-13.
[13] Lin, S.-Z.; Wouessidjewe, D.; Poelman, M.-C.; Duchene, D. In
vivo evaluation of indomethacin/cyclodextrin complexes gastroin-
testinal tolerance and dermal anti-inflammatory activity. Int. J.
Pharm., 1994, 106(1), 63-67.
[14] Yener, G.; Üner, M.; Gönüllü, Ü.; Yildirim, S.; Kiliç, P, Aslan,
S.S.; Barla, A. Design of meloxicam and lornoxicam transdermal
patches, Preparation, physical characterization, ex vivo and in vivo
studies. Chem. Pharm. Bull., 2010, 58(11), 1466-1473.
[15] Kavitha, K.; Rajendra, M.M. Design and evaluation of transdermal
films of lornoxicam. Int. J. Pharma Biosci., 2011, 2(2), 54-62.
[16] Khadka, D.; Ahmed, M.G.; Acharya, A. Formulation and evalua-
tion of transdermal gel of lornoxicam and comparative diffusion
study by iontophoresis and chemical enhancers. IJAPR., 2015, 6(2),
40-49.
[17] Williams, A. Transdermal and topical drug delivery, from theory to
clinical practice, Pharmaceutical Press London, 2003.
El-Ridy et al.
[18] Chavan, P.; Jain, B.; Jain, P. Proniosomes as drug carrier system
for transdermal delivery of lornoxicam. World J. Pharm. Pharm.
Sci., 2012, 1(1), 393-404.
[19] Dasgupta, S.; Ghosh, S.K.; Ray, S.; Kaurav, S.S.; Mazumder, B. In
vitro & in vivo studies on lornoxicam loaded nanoemulsion gels for
topical application. Curr. Drug Deliv., 2014, 11(1), 132-138.
[20] Patel, B.B.; Shah, C.N. Formulation and in vitro characterization of
lornoxicam transdermal matrix patch. Int. J. Adv. Pharm., 2016,
5(2), 30-38.
[21] Vora, B.; Khopade, A.J.; Jain, N. Proniosome based transdermal
delivery of levonorgestrel for effective contraception. J. Control.
Rel., 1998, 54(2), 149-165.
[22] Manconi, M.; Sinico, C.; Valenti, D.; Lai, F.; Fadda, A.M.
Niosomes as carriers for tretinoin, III. A study into the in vitro cu-
taneous delivery of vesicle-incorporated tretinoin. Internat. J.
Pharm., 2006, 311(1), 11-19.
[23] Baillie, A.F.A.; Hume, L.; Muirhead, G.; Rogerson, A. The prepa-
ration and properties of niosomes-non-ionic surfactant vesicles. J.
Pharm. Pharmacol., 1985, 37(12), 863-868.
[24] Azmin, M.N.F.A.; Handjani-Vila, R.M.; Stuart, J.F.B.; Vanlerber-
ghe, G.; Whittaker, J.S. The effect of non-ionic surfactant vesicle
(niosome) entrapment on the absorption and distribution of
methotrexate in mice. J. Pharm. Pharmacol., 1985, 37(4), 237-242.
[25] Junginger, H.; Hofland, H.; Bouwstra, J. Liposomes and niosomes,
interactions with human skin. Cosmetics Toiletries, 1991, 106(8),
45-50.
[26] Handjani
Vila, R.; Ribier, A.; Rondot, B.; Vanlerberghie, G. Dis-
persions of lamellar phases of non
ionic lipids in cosmetic prod-
ucts. Int. J. Cosmetic Sci., 1979, 1(5), 303-314.
[27] Hao, Y.; Zhao, F.; Li, N.; Yang, T.; Li, K. Studies on a high encap-
sulation of colchicine by a noisome system. Int. J. Pharm., 2002,
244, 73-80.
[28] Manosroi, A.; Wongtrakul, P.; Manosroi, J.; Sakai, H.; Sugawara,
F.; Yuasa, M.; Abe, M. Characterization of vesicles prepared with
various non-ionic surfactants mixed with cholesterol. Colloids Sur-
faces B, Biointerf., 2003, 30(1), 129-138.
[29] Foye, W.O.; Thomas, L.L.; David, A.W. Principles of Medicinal
Chemistry. Williams and Wilkins, USA. 1995, 4th edn., 335.
[30] Junyaprasert, V.B.; Teeranachaideekul, V.; Supaperm, T. Effect of
charged and non-ionic membrane additives on physicochemical
properties and stability of niosomes. AAPS Pharm. Sci. Tech.,
2008, 9(3), 851-859.
[31] Azeem, A.; Ahmad, F.J.; Khan, Z.I.; Talegaonkar, S. Nonionic
surfactant vesicles as a carrier for transdermal delivery of
frusemide. J. Disp. Sci. Technol., 2008, 29(5), 723-730.
[32] El-Ridy, M.S.B.A.A.; Safar, M.M.; Mohsen, A.M. Niosomes as a
novel pharmaceutical formulation encapsulating the hepatoprotec-
tive drug silymarin. Int. J. Pharmacy Pharm. Sci., 2012, 4(1), 549-
559.
[33] Kuntsche, J.; Freisleben, I.; Steiniger, F.; Fahr, A. Temoporfin-
loaded liposomes, physicochemical characterization. Eur. J. Phar-
maceut. Sci., 2010, 40(4), 305-315.
[34] Omr,i A.A.M.R. Comparison of the bactericidal action of ami-
kacin,netilmicin and tobramycin in free and liposomal formulation
against Pseudomonas aeruginosa. Chemotherapy, 1996 b, 42, 170-
176.
[35] Scherphof, G.L.; Velinova, M.; Kamps, J.; Donga, J.; van der
Want, H.; Kuipers, F. Modulation of pharmacokinetic behavior of
liposomes. Adv. Drug Deliv. Rev., 1997, 24(2), 179-191.
[36] Omri, A.A.M.R. Preparation, properties and the effects of ami-
kacin, netilmicin and tobramycin in free and liposomal formula-
tions on Gram-negative and Gram-positive bacteria. Int. J. Antimi-
crob. Agents, 1996, 7, 9-14.
[37] Beaulac, C.; Clement-Major, S.; Hawari, J.; Lagace, J. In vitro
kinetics of drug release and pulmonary retention of microencapsu-
lated antibiotic in liposomal formulations in relation to the lipid
composition. J. Microencapsulation, 1997, 14, 335-348.
[38] Halwani, M.; Mugabe, C.; Azghani, A.O.; Lafrenie, R.M.; Kumar,
A.; Omri, A. Bactericidal efficacy of liposomal aminoglycosides
against Burkholderia cenocepacia. J. Antimicrob. Chemothera.,
2007, 60(4), 760-769.
[39] Mugabe, C.; Azghani, A.O.; Omri, A. Preparation and characteriza-
tion of dehydration–rehydration vesicles loaded with aminoglyco-
side and macrolide antibiotics. Int. J. Pharmaceut., 2006, 307(2),
244-250.
Formulation of Niosomal Gel for Enhanced Transdermal Lornoxicam Delivery
[40] Shekunov, B.Y.; Chattopadhyay, P.; Tong, H.H.; Chow, A.H.
Particle size analysis in pharmaceutics, principles, methods and ap-
plications. Pharmaceut. Res., 2007, 24(2), 203-227.
[41] Das, S.; Haldar, P.K.; Pramanik, G. Formulation and evaluation of
herbal gel containing Clerodendron infortunatum leaves extract.
Int. J. Pharm. Tech. Res., 2011, 1(3), 140-143.
[42] Fathalla, D.; Abdel-Mageed, A.; Abdel-Hamid, F.; Ahmed, M. In
vitro and in vivo evaluation of niosomal gel containing aceclofenac
for sustained drug delivery. Int. J. Pharm. Sci. Res., 2014.
http://dx.doi.org/10.15344/2394-1502/2014/105
[43] Islam, M.T.; Rodriguez-Hornedo, N.; Ciotti, S.; Ackermann, C.
Rheological characterization of topical carbomer gels neutralized to
different pH. Pharmaceut. Res., 2004, 21(7), 1192-1199.
[44] Basha, M.; Abd El-Alim, S.H.; Kassem, A.A.; El Awdan, S.;
Awad, G. Benzocaine loaded solid lipid nanoparticles, Formulation
design, in vitro and in vivo evaluation of local anesthetic effect.
Curr. Drug Deliv., 2015, 12(6), 680-692.
[45] Dhawan, B.; Aggarwal, G.; Harikumar, S. Enhanced transdermal
permeability of piroxicam through novel nanoemulgel formulation.
Int. J. Pharm. Invest., 2014, 4(2), 65-76.
[46] Dubey, A.; Prabhu, P. Development and evaluation of lornoxicam
loaded maltodextrin based proniosomes. Development, 2013, 5(3),
865-872.
[47] Bhaskaran, S.; Lakshmi, P. Comparative evaluation of niosome
formulations prepared by different techniques. Acta. Pharm. Sci.,
2009, 51(1), 27-32.
[48] Najib, N.; Suleiman, M. The kinetics of drug release from ethylcel-
lulose solid dispersions. Drug Dev. Ind. Pharm., 1985, 11(12),
2169-2181.
[49] Higuchi, T. Mechanism of sustained
action medication. Theoretical
analysis of rate of release of solid drugs dispersed in solid matrices.
J. Pharmaceut. Sci., 1963, 52(12), 1145-1149.
[50] Jain, S.; Chourasia, M.; Masuriha, R.; Soni, V.; Jain, A.; Jain, N.K.;
Gupta, Y. Solid lipid nanoparticles bearing flurbiprofen for trans-
dermal delivery. Drug Deliv., 2005, 12(4), 207-215.
[51] Yousuf, M.; Ahmad, M.; Usman, M.; Ali, I. Ketotifen fumarate and
salbutamol sulphate combined transdermal patch formulations, in
vitro release and ex vivo permeation studies. Ind. J. Pharm. Sci.,
2013, 75(5), 569.
[52] Radhofer-Welte, S.; Dittrich, P. Determination of the novel non-
steroidal anti-inflammatory drug lornoxicam and its main metabo-
lite in plasma and synovial fluid. J. Chromatography B, Biomed.
Sci. Appl., 1998, 707(1), 151-159.
[53] Edwards, D.A.; Langer, R. A linear theory of transdermal transport
phenomena. J. Pharm. Sci., 1994, 83(9), 1315-1334.
[54] Draize, J.H.; Woodard, G.; Calvery, H.O. Methods for the study of
irritation and toxicity of substances applied topically to the skin and
mucous membranes. J. Pharmacol. Exper. Therap., 1944, 82(3),
377-390.
[55] Mutalik, S.; Udupa, N. Pharmacological evaluation of membrane-
moderated transdermal system of glipizide. Clin. Exp. Pharmacol.
Physiol., 2006, 33(1
2), 17-26.
[56] Swingle, K.F.; Grant, T.J.; Jaques, L.W.; Kvam, D.C. Interactions
of anti-inflammatory drugs in carrageenan-induced foot edema of
the rat. J. Pharmacol. Exp. Ther., 1970, 172(2), 423-425.
[57] Charoo, N.A.; Shamsher, A.A.A.; Kohli, K.; Pillai, K.; Rahman, Z.
Improvement in bioavailability of transdermally applied flurbipro-
fen using tulsi (Ocimum sanctum) and turpentine oil. Colloids Surf.
B, Biointerf., 2008, 65(2), 300-307.
[58] Abd-Elbary, A.; El-Laithy, H.; Tadros, M. Sucrose stearate-based
proniosome-derived niosomes for the nebulisable delivery of cro-
molyn sodium. Int. J. Pharm., 2008, 357(1), 189-198.
[59] Srinath, P.; Vyas, S.; Diwan, P.V. Preparation and pharmacody-
namic evaluation of liposomes of indomethacin. Drug Dev. Ind.
Pharm., 2000, 26(3), 313-321.
Current Drug Delivery, 2018, Vol. 15, No. 1 133
[60] Hathout, R.M.; Mansour, S.; Mortada, N.D.; Guinedi, A.S.
Liposomes as an ocular delivery system for acetazolamide, in vitro
and in vivo studies. AAPS Pharm. Sci. Tech., 2007, 8(1), E1-E12.
[61] Abdelbary, G.; El-gendy, N. Niosome-encapsulated gentamicin for
ophthalmic controlled delivery. AAPS Pharm. Sci. Tech., 2008,
9(3), 740-747.
[62] Confalonieri, E.O.; Soraci, A.L.; Becaluba, M.; Denzoin, L.; Rod-
riguez, E.; Riccio, B.; Tapia, O. The disposition of free and
niosomally encapsulated Rac-flurbiprofen in dairy bovines. J. Vet-
erin. Pharmacol. Therap., 2010, 33(1), 9-14.
[63] Ahmed, M.O.; Al-Badr, A.A. Lornoxicam. Profiles Drug Subst.,
Excip. Relat. Methodol., 2011, 36, 205-239.
[64] ElMeshad, A.N.; Mohsen, A.M. Enhanced corneal permeation and
antimycotic activity of itraconazole against Candida albicans via a
novel nanosystem vesicle. Drug Deliv., 2016, 23(7), 2115-2123.
[65] Vyas, S.; Singh, R.; Asati, R. Liposomally encapsulated diclofenac
for sonophoresis induced systemic delivery. J. Microencapsulation,
1995, 12(2), 149-154.
[66] Carafa, M.; Santucci, E.; Alhaique, F.; Coviello, T.; Murtas, E.;
Riccieri, F.; Lucaniab, G.; Torrisib, M.R. Preparation and proper-
ties of new unilamellar non-ionic/ionic surfactant vesicles. Int. J.
Pharm., 1998, 160(1), 51-59.
[67] McIntosh, T. The effect of cholesterol content on the structure of
phosphatidylcholine bilayers. Biochim. Biophys. Acta, 1978, 51,
43-58.
[68] entjurc, M.; Vrhovnik, K.; Kristl, J. Liposomes as a topical deliv-
ery system, the role of size on transport studied by the EPR imag-
ing method. J. Control. Release, 1999, 59(1), 87-97.
[69] Centis, V.; Vermette, P. Physico-chemical properties and cytotox-
icity assessment of PEG-modified liposomes containing human
hemoglobin. Colloids Surf. B, Biointerf., 2008, 65(2), 239-246.
[70] Hanaor, D.A.H.; Michelazzi, M.; Leonelli, C.; Sorrell, C.C. The
effects of carboxylic acids on the aqueous dispersion and electro-
phoretic deposition of ZrO2 . J. Eur. Ceramic Soc., 2012, 32(1),
235-244.
[71] El-Say, K.M.; Abd-Allah, F.I.; Lila, A.E.; Hassan, A.E.-S.A.;
Kassem, A.E.A. Diacerein niosomal gel for topical delivery, devel-
opment, in vitro and in vivo assessment. J. Liposome Res., 2016,
26(1), 57-68.
[72] Mokhtar, M.; Sammour, O.A.; Hammad, M.A.; Megrab, N.A.
Effect of some formulation parameters on flurbiprofen encapsula-
tion and release rates of niosomes prepared from proniosomes. Int.
J. Pharm., 2008, 361(1), 104-111.
[73] Weiner, N.; Williams, N.; Birch, G.; Ramachandran, C.; Shipman,
C.; Flynn, G. Topical delivery of liposomally encapsulated inter-
feron evaluated in a cutaneous herpes guinea pig model. Antimi-
crobial Agents Chemother., 1989, 33(8), 1217-1221.
[74] Attia, I.A.; El-Gizawy, S.A.; Fouda, M.A.; Donia, A.M. Influence
of a niosomal formulation on the oral bioavailability of acyclovir in
rabbits. AAPS Pharm. Sci. Tech., 2007, 8(4), E106.
[75] Hashim, F.; El-Ridy, M.; Nasr, M.; Abdallah, Y. Preparation and
characterization of niosomes containing ribavirin for liver target-
ing. Drug Deliv., 2010, 17(5), 282-287.
[76] Bayindir, Z.S.; Yuksel, N. Characterization of niosomes prepared
with various nonionic surfactants for paclitaxel oral delivery. J.
Pharm. Sci., 2010, 99(4), 2049-2060.
[77] Guinedi, A.S.; Mortada, N.D.; Mansour, S.; Hathout, R.M. Prepa-
ration and evaluation of reverse-phase evaporation and multilamel-
lar niosomes as ophthalmic carriers of acetazolamide. Int. J.
Pharm., 2005, 306(1-2), 71-82.
[78] Ruckmani, K.; Sankar, V. Formulation and optimization of zi-
dovudine niosomes. AAPS Pharm. Sci. Tech., 2010, 11(3), 1119-
1127.