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Pharmaceutical nanotechnology
A R T I C L E I N F O A B S T R A C T
Article history: The objective of this study was to evaluate the transdermal delivery potential of a new curcumin-
Received 5 November 2014 containing microemulsion system. Three series of experiments were carried out to comprehend the
Accepted 1 February 2015 system characteristics: (a) examining the influence of water content on curcumin permeation, (b)
Available online 2 February 2015
studying the effect of curcumin loading on its permeability, and (c) assessing the contribution of the
vesicular nature of the microemulsion on permeability. The skin permeability of curcumin from
Keywords: microemulsions, which contained 5%, 10%, and 20% of water content (1% curcumin), was measured in vitro
Microemulsion
using excised rat skin. It has been shown that the permeability coefficient of CUR in a formulation
Curcumin
Transdermal drug delivery
containing 10% aqueous phase (ME-10) was twofold higher than the values obtained for formulations
Percutaneous penetration with 5% and 20% water (Papp = 0.116 103 0.052 103 vs. 0.043 103 0.022 103 and
0.047 103 0.025 103 cm/h, respectively. A reasonable explanation for this phenomenon may be
the reduction of both droplet size and droplets’ concentration in the microemulsion as the aqueous phase
decreased from 20% to 5%. It has also been shown that a linear correlation exists between the decrease in
droplet size and the increase of curcumin loading in the microemulsion. In addition, it has been
demonstrated that a micellar system, S/O-mix, and a plain solution of curcumin resulted in a significantly
lower curcumin permeation relative to that presented by the microemulsion, Papp = 0.018 103 0.011
103, 0.005 103 0.002 103, and 0.002 103 0.000 103 cm/h, respectively, vs. 0.110 103
0.021 103 cm/h for the microemulsion. The enhancement ratio (ER = Jss-ME/Jss-solution) of CUR
permeated via 1% loaded microemulsion was 55.
ã 2015 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ijpharm.2015.02.005
0378-5173/ ã 2015 Elsevier B.V. All rights reserved.
98 A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103
delivery systems while transdermal delivery of curcumin has 1:3 glyceryl oleate/Labrasol1. The microemulsion formed sponta-
received less attention so far. Patel et al. (2009) have developed a neously at room temperature as a clear monophasic liquid. The
dried polymeric film-type matrix containing curcumin, which its loaded concentration of CUR in the microemulsions was by default
in-vitro permeation followed Higuchi kinetics. In a series of three 1.0 wt% but can be increased to 4.0 wt%. As a convenient method to
publications dealing with microemulsion vehicles, the percutane- represent an increase of water content while decreasing CoS–S
ous delivery of 0.4% curcumin has been studied in terpene levels, ‘water dilution lines’ were drawn from the water apex to the
microemulsions, such as eucalyptol and limonene combined with opposite oil side of the phase diagram. The line was arbitrarily
polysorbate 80, ethanol and water (Liu and Chang, 2011; Liu et al., denoted as the value of the line intersection with the oil scale (e.g.,
2011), and in microemulsions composed of Pluronic F127 DL11 means line representing a surfactant-to-oil ratio of 89:11).
(surfactant), isopropanol (co-surfactant), myristic acid (oil), and Formulations along a dilution line DL11 was prepared to contain
water (Liu and Huang, 2012). Although microemulsions have not 5%, 10%, and 20% (w/w) of aqueous phase. The stability of CUR in
been widely explored for transdermal curcumin they are 10% water-containing microemulsion was tested at 4 C and at
extensively utilized for a variety of pharmaceutical purposes ambient temperature, resulting in decrease of 5.9% and 8.9%,
including transdermal and dermal drug delivery (Kreilgaard et al., respectively, after 7 days. However, no significant decrease in CUR
2000; Alvarez-Figueroa and Blanco-Mendez, 2001; Kreilgaard, level has been noted when including 0.01% a-tocopherol in the
2001; Kreilgaard et al., 2001; Rhee et al., 2001; Lee et al., 2003; formulation.
Sintov and Shapiro, 2004; Hua et al., 2004; Sintov and Botner,
2006; Sintov and Brandys-Sitton, 2006; Liu et al., 2007; Shevach- 2.3. Light scattering
man et al., 2008; Heuschkel et al., 2008; Gannu et al., 2010; Zhao
et al., 2011; Zhang et al., 2011; Zhang and Michniak-Kohn, 2011; The hydrodynamic diameter spectrum of microemulsion nano-
Sintov and Greenberg, 2014). Microemulsions, which are versatile droplets was collected using CGS-3Compact Goniometer System
systems (oil-in-water, water-in-oil, or bi-continuous systems as (ALV GmbH, Langen, Germany). The laser power was 20 mW at the
well as various structures and components), possess high He–Ne laser line (633 nm). Correlograms were calculated by ALV/
solubilizing capacity of diverse compounds and, in particular, is LSE 5003 correlator during 10 s for 20 times at 25 C. Measure-
capable of solubilizing poorly water-soluble drugs. The increase in ments were performed at permanent angle of 60 . The droplet size
solute concentration and the tendency of the vehicle to favor was calculated using the Stokes–Einstein relationship, and the
partitioning into the stratum corneum make microemulsions a analysis was based on regularization method as described by
useful vehicle to enhance transdermal drug permeability. Provencher (1982).
In the present report, we describe an alcohol-free micro-
emulsion system for transdermal drug delivery with respect to its 2.4. Viscosity
contribution to percutaneous permeability of CUR. The micro-
emulsion system, which contains no chemical enhancers, is The viscosity was measured by using a control stress rheometer
composed of water, isopropyl palmitate, propylene carbonate, CSL 50 (TA Instruments Ltd., UK) at 25 C. Cone and plate geometry
and the commonly-used topical nonionic surfactants, PEG- was used.
8 caprylic/capric glycerides (Labrasol1) and glyceryl oleate. We
studied the in vitro skin permeation of the microemulsions after we 2.5. In-vitro skin penetration study
had surmounted the technical difficulties of experimentation
regarding the high-rate of CUR degradability in aqueous solutions. The permeability of CUR through animal skin was determined
By using microemulsions as a vehicle, we have demonstrated an in vitro with a Franz diffusion cell system (Crown Bioscientific, Inc.,
increased solubilization as well as a significantly enhanced Clinton, NJ). The diffusion area was 1.767 cm2 (15 mm diameter
permeability of the water insoluble and poorly absorbable orifice), and the receptor compartment volumes was from 12 ml.
curcumin. The solutions in the water-jacketed cells were thermostated at
37 C and stirred by externally driven, Teflon-coated magnetic
2. Materials and methods bars. Each set of experiments was performed with at least four
diffusion cells (n 4), each containing skin from a different animal.
2.1. Materials All animal procedures were performed in accordance with
protocols reviewed and approved by the Institutional & Use
Curcumin was obtained from Sigma, Rehovot, Israel. Labrasol1 Committee, Ben Gurion University of the Negev, which complies
(PEG-8 caprylic/capric glycerides) was purchased from Gattefosse with the Israeli Law of Human Care and Use of Laboratory Animals.
(Saint-Priest Cedex, France), and glyceryl oleate was obtained from Sprague-Dawley rats (males, 300–350 g, Harlan Laboratories Ltd.,
Croda Europe Ltd. (East Yorkshire, England). Isopropyl palmitate Jerusalem, Israel) were euthanized by aspiration of CO2. The
and propylene carbonate were purchased from Sigma, Rehovot, abdominal hair was clipped carefully and sections of full-thickness
Israel. a-Tocopherol 1000 IU was obtained from BTSA Biotecno- skin were excised from the fresh carcasses of animals and used
logias Aplicadas (Madrid, Spain). High-performance liquid chro- immediately. All skin sections were measured for transepidermal
matography (HPLC) grade water and solvents were obtained from J. water loss (TEWL) before mounted in the diffusion cells or stored
T. Baker (Mallinckrodt Baker, Inc., Phillipsburg, NJ). at lower temperatures until used. TEWL examinations were
performed on skin pieces using Dermalab1 Cortex Technology
2.2. Preparation of microemulsions instrument, (Hadsund, Denemark) and only those pieces that the
TEWL levels were less than 10 g/m2/h were introduced for testing.
The formulation was prepared as previously published (Sintov The skin was placed on the receiver chambers with the stratum
and Botner, 2006; Sintov et al., 2010; Sintov and Greenberg, 2014). corneum facing upwards, and the donor chambers were then
Shortly, water-in-oil (W/O) liquid microemulsions were prepared clamped in place. The excess skin was trimmed off, and the
by mixing Labrasol1 and glyceryl oleate (surfactants), propylene receiver chamber, defined as the side facing the dermis, was filled
carbonate (co-surfactant) and isopropyl palmitate (oil), followed with 70:30 phosphate buffer (pH 7.4) – ethyl alcohol containing
by gently titration of distilled water. The co-surfactant/surfactants a-tocopherol (0.01%). After 15 min of skin washing at 37 C, the
(CoS/S) weight ratio was 2:5, while the surfactants’ ratio was buffer was removed from the cells and the receiver chambers were
A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103 99
Fig. 1. In vitro percutaneous permeation of curcumin through fresh rat skin after application of curcumin-loaded microemulsions containing various water contents. A
comparison was also made between 5% (diamonds), 10% (squares), and 20% aqueous phase (triangles) (n = 4).
100 A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103
Table 1
Skin permeability parameters of various curcumin-loaded microemulsion (ME)
formulations as measured in vitro by using rat skin (n 4).
Fig. 2. Nanodroplet size (average diameter, nm) and their prevalence (percent of total droplets) in microemulsions containing 5%, 10%, and 20% aqueous phase.
A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103 101
Fig. 5. In vitro percutaneous permeation of curcumin using fresh rat skin after its application in a W/O microemulsion (10% aqueous phase), in micellar system, and in a water-
free surfactants and oil mixture (all formulations contained 1% curcumin, n = 4). Data were compared with 1% curcumin solution in propylene carbonate.
102 A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103
Fig. 6. In vitro percutaneous permeation of curcumin through fresh rat skin after application of curcumin-loaded microemulsions at various curcumin concentrations. A
comparison was also made between 1% (diamonds), 2% (squares), and 3% aqueous phase (triangles) (n = 10).
Average droplet size (nm in diameter)
4 3.65 nm 100
3.5 90
80
1.5 40
1.26 nm
0.80 nm 30
1
20
0.5 10
0 0
0.0 1.0 2.0 3.0
Curcumin concentraon (% wt/wt)
Fig. 7. Nanodroplet size (average diameter, nm) and their prevalence (percent of total droplets) in microemulsions containing 0–3% concentrations of curcumin.
its permeability coefficient increased from 0.116 103 0.052 decrease was accompanied by increase in aggregation, the
103 cm/h in 1% CUR-loaded ME-10 to 0.233 103 0.051 diminution in size of more than 50% of the droplets may explain
103 cm/h and 0.295 103 0.050 103 cm/h in 2% CUR and the improvement in CUR permeability.
3% CUR-loaded ME-10, respectively (Table 1). It should be noted
that the Papp values for CUR permeation obtained in this study 4. Conclusion
are comparable with those previously published by Liu et al.
(2011), who used transdermal microemulsions composed of 20% From the results presented in this paper, it has been concluded
limonene (oil), 10% water, 35% polysorbate 80 (surfactant) and that the W/O microemulsion vehicle for transdermal CUR
35% ethanol (cosurfactant). There is no statistically significant permeation is advantageous over a micellar system and a
difference between the 2% and 3% CUR-loaded ME-10, however, surfactant-oil mixture, composed of the same proportions of
there is a significant difference between CUR permeability of ingredient. This indicates that the mechanism of CUR delivery is
these high loaded formulations and 1% CUR-loaded micro- based on the nanostructural characteristics of the microemulsion.
emulsion (p < 0.05, ANOVA). It seems that although micro- In addition, it has been demonstrated that one of the mediating
emulsion of the ME-10 type can be loaded up to 4% CUR, the factors of skin permeation and retention is the droplet size of the
effect of loading on CUR permeation through the skin is microemulsion. One more important factor is apparently the
practically expressed from 1% to 2% CUR concentration. Fig. 7 density of the dispersed droplets in the microemulsion, which
presents the effect of CUR loading on droplet size, showing a depends on both droplet size and the volume fraction of the inner
linear correlation between the decrease in droplet size and the phase. Finally, this report joins to a series of studies demonstrating
increase of CUR loading in the microemulsion. Although this the advantage of the microemulsion system in enhancement of
A.C. Sintov / International Journal of Pharmaceutics 481 (2015) 97–103 103
percutaneous and cutaneous permeability of pharmacologically- Li, L., Ahmed, B., Mehta, K., Kurzrock, R., 2007. Liposomal curcumin with and
active agents. without oxaliplatin: effects on cell growth, apoptosis, and angiogenesis in
colorectal cancer. Mol. Cancer Ther. 6 (4), 1276–1782.
Lim, K.J., Bisht, S., Bar, E.E., Maitra, A., Eberhart, C.G., 2011. A polymeric nanoparticle
Acknowledgment formulation of curcumin inhibits growth, clonogenicity and stem-like fraction
in malignant brain tumors. Cancer Biol. Ther. 11 (5), 464–473.
Liu, A., Lou, H., Zhao, L., Fan, P., 2006. Validated LC/MS/MS assay for curcumin and
The author is grateful for the professional assistance of Mr. Igor tetrahydrocurcumin in rat plasma and application to pharmacokinetic study of
Greenberg and Ms. Lillia Shapiro at the Laboratory for Biophar- phospholipid complex of curcumi. J. Pharm. Biomed. Anal. 40 (3), 720–727.
maceutics. Liu, C.H., Chang, F.Y., 2011. Development and characterization of eucalyptol
microemulsions for topic delivery of curcumin. Chem. Pharm. Bull. (Tokyo) 59
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