Scientia

Pharmaceutica

Review
Nanoemulsion: A Review on Mechanisms for the
Transdermal Delivery of Hydrophobic and
Hydrophilic Drugs
Dalia S. Shaker 1 , Rania A. H. Ishak 2 , Amira Ghoneim 1, * and Muaeid A. Elhuoni 3
1 Department of Pharmaceutics &Pharmaceutical Technology, Faculty of Pharmaceutical Sciences and
Pharmaceutical industries, Future University in Egypt (FUE), 11835 Cairo, Egypt
2 Department of Pharmaceutics& Industrial Pharmacy, Ain Shams University, 11591 Cairo, Egypt
3 Quality Control Department, Elnajah Medical Services, Benghazi, Libya
* Correspondence: amiraghoneim@gmail.com; Tel.: +(202)-2414-2927; Fax: +(202)-2618-6111




Received: 1 June 2019; Accepted: 9 July 2019; Published: 12 July 2019


Abstract: Nanoemulsions (NEs) are colloidal dispersions of two immiscible liquids, oil and water,
in which one is dispersed in the other with the aid of a surfactant/co-surfactant mixture, either
forming oil-in-water (o/w) or water-in-oil (w/o) nanodroplets systems, with droplets 20–200 nm
in size. NEs are easy to prepare and upscale, and they show high variability in their components.
They have proven to be very viable, non-invasive, and cost-effective nanocarriers for the enhanced
transdermal delivery of a wide range of active compounds that tend to metabolize heavily or
suffer from undesirable side effects when taken orally. In addition, the anti-microbial and anti-viral
properties of NE components, leading to preservative-free formulations, make NE a very attractive
approach for transdermal drug delivery. This review focuses on how NEs mechanistically deliver
both lipophilic and hydrophilic drugs through skin layers to reach the blood stream, exerting the
desired therapeutic effect. It highlights the mechanisms and strategies executed to effectively deliver
drugs, both with o/w and w/o NE types, through the transdermal way. However, the mechanisms
reported in the literature are highly diverse, to the extent that a definite mechanism is not conclusive.

Keywords: nanoemulsion; transdermal; permeability; water-in-oil; oil-in-water; mechanisms

1. Introduction
Nanoemulsions (NEs) are submicron sized and have gained great interest as drug carriers for
improving the delivery of therapeutic agents. They are considered an advanced nanodroplet systems
for systemic, controlled, and targeted drug delivery. NEs are colloidal dispersions consisting of
two immiscible liquids (water and oil), in which one liquid is dispersed in the other by means of
an appropriate surfactant mixture, forming a system where the size of droplets falls in the range of
20 to 200 nm, as reported by Takur et al. [1]. Nanoemulsion (NE) in particular is a very cost-effective
technique, with high storage stability, as it is easily prepared and does not always require high energy
or complicated procedures [2,3]. Despite the similarities between NEs and microemulsions (MEs) in
terms of their physical appearance, components, and preparation techniques, NEs are kinetically stable
and thermodynamically metastable, while MEs are thermodynamically stable [4].
Transdermal drug delivery is a very established route of drug administration that is known to
have enhanced therapeutic efficacy, ease of administration, and an allow an immediate withdrawal
of treatment if necessary [5]. This administration route bypasses the hepatic metabolism to reach
systemic circulation [6], leading to an increase in drug bioavailability. Moreover, it reduces the adverse
effects of some medicines, e.g., non-steroidal anti-inflammatory drugs (NSAIDs), which are usually

Sci. Pharm. 2019, 87, 17; doi:10.3390/scipharm87030017 www.mdpi.com/journal/scipharm

Sci. Pharm. 2019, 87, 17 2 of 34

associated with gastrointestinal tract (GIT) drug administration [7,8]. Drug delivery via skin for
systemic circulation is suitable for a number of clinical conditions, such as hypertension, arthritis,
diabetes, cancer, and high blood cholesterol level [9]. Because the dose is given transdermally and is
diabetes, cancer, and high blood cholesterol level [9]. Because the dose is given transdermally and
sustained over a long period via tailored delivery systems, particularly in the case of NE, it is of
is sustained
utmostover a longtoperiod
importance improvevia tailored
patient delivery
compliance [10].systems, particularly in the case of NE, it is of
utmost importance to improve patient compliance [10].
2. Skin Permeation Pathways
2. Skin Permeation Pathways
The outermost layer of the epidermis, the stratum corneum (SC), forms a hydrophobic barrier
Thethat impedes the
outermost transport
layer of theofepidermis,
exogenous thechemicals,
stratum including
corneum drugs [11].
(SC), An important
forms requirement
a hydrophobic barrier that
for transdermal delivery is that the drug carried by a vehicle is able to reach the skin surface at an
impedes the transport of exogenous chemicals, including drugs [11]. An important requirement for
adequate rate and in sufficient amounts [12]. The SC is the hardest layer of skin due to its flattered
transdermal delivery
corneocytes whichis that the drug carried
are surrounded by lipidby a vehicle
bilayers is ableoftoceramides
consisting reach the[9]. skin
It issurface
arrangedatinan adequate
10–
rate and15inrows
sufficient amounts [12]. The SC is the hardest layer of skin due
that are 10 μm in thickness, composed of highly keratinized cells, known as corneocytes. to its flattered corneocytes
which are surrounded
These by lipidby
cells are surrounded bilayers consisting
a continuous of ceramides
lipid phase, known as the [9].intercellular
It is arranged in 10–15and
lipid lamellae, rows that
are 10 µmhaveinbeen said to resemble
thickness, composed a “brick and mortar”
of highly model [13].
keratinized Finally,
cells, known diffusion through the SC
as corneocytes. is thecells are
These
sum of
surrounded bya series of segments
a continuous of lateral
lipid phase,diffusion
knownand intramembrane
as the intercellular trans-bilayer and[14].
transport
lipid lamellae, have been said
A permeant applied to the skin has three possible routes across the epidermis, as illustrated in
to resemble a “brick and mortar” model [13]. Finally, diffusion through the SC is the sum of a series of
Figure 1: The transcellular route, a lipid domain associated with the proteins inside corneocytes, the
segments of lateralroute,
intercellular diffusion
and the and intramembrane
appendageal trans-bilayer
route, through transport
hair follicles, [14]. sebaceous glands
via associated
A permeant
and sweat ducts applied to the
[15]. The skin has
fractional three possible
appendage area availableroutes across the
for transport epidermis,
is only asThis
about 0.1%. illustrated
in Figure 1: The
route transcellular
usually contributes route, a lipid
negligibly domain
to the steady associated withThis
state drug flux. the proteins
pore pathwayinsidemaycorneocytes,
be
important for
the intercellular ions and
route, andlarge
the polar molecules that
appendageal struggle
route, throughto cross an follicles,
hair intact SC [15].
via The transcellular
associated sebaceous
route is a sequence of partitioning and diffusion into alternating hydrophilic and lipophilic domains
glands and sweat ducts [15]. The fractional appendage area available for transport is only about 0.1%.
of cells, respectively, in the extracellular matrix [16,17]. In the intercellular route, the permeant crosses
This route usually contributes negligibly to the steady state drug flux. This pore pathway may be
the hard path within the extracellular matrix, without traversing the cells. Small hydrophilic
important for ions and
molecules generally large polar
prefer molecules route
the transcellular that struggle to cross anroute,
over the intercellular intactandSCthe[15]. Theistranscellular
reverse true
route is for
a sequence
lipophilic of partitioning
molecules. and diffusion
The transcellular andinto alternating
intercellular hydrophilic
routes constituteand the lipophilic domains of
transepidermal
pathway [18].
cells, respectively, in the extracellular matrix [16,17]. In the intercellular route, the permeant crosses the
The combined
hard path within flux of thesematrix,
the extracellular two pathways,
without lipid and pore, determines
traversing the cells. the Smalloverall observed flux
hydrophilic molecules
across the skin. It is widely accepted that the trans epidermal pathway is usually the predominant
generally prefer the transcellular route over the intercellular route, and the reverse is true for lipophilic
pathway of skin permeation and that under sink conditions, diffusion across the SC constitutes the
molecules. The transcellular
rate-limiting and intercellular
step that determines the overall routes constitute
flux of the permeant.the transepidermal pathway [18].

Figure
Figure 1. 1. Permeation
Permeation pathways
pathways in the in the(the
skin skin (the stratum
stratum corneum,
corneum, SC, is illustrated).
SC, is illustrated). (A)transcellular
(A) The The
transcellular route associated with the proteins inside corneocytes. (B) The intercellular route and the
route associated with the proteins inside corneocytes. (B) The intercellular route and the appendageal
appendageal route through (C) hair follicles with associated sebaceous glands (D) via sweat ducts.
route through (C) hair follicles with associated sebaceous glands (D) via sweat ducts.

The combined flux of these two pathways, lipid 2 and pore, determines the overall observed flux
across the skin. It is widely accepted that the trans epidermal pathway is usually the predominant
pathway of skin permeation and that under sink conditions, diffusion across the SC constitutes the
rate-limiting step that determines the overall flux of the permeant.
Sci. Pharm. 2019, 87, 17 3 of 34

3. Determination of Transdermal Permeability Coefficient

The total experimental permeability coefficient (P) for full-thickness skin may be expressed as [19]:
1
P= 1 1
(1)
PSC + PD/E

where PSC is the permeability coefficient of the SC and PD/E is the permeability coefficient for
the epidermis-dermis combination, which can be estimated from permeation experiments with
tape-stripped skin. PSC can be further divided into parallel lipoidal and pore pathway components in
the SC via the following equation:
PSC = PL + PP (2)

where PL and PP are the permeability coefficients for the lipoidal and pore pathways, respectively.
By substituting Equation (2) into Equation (1), the following expression is obtained:
1
P= 1 1
(3)
PL + PP + PD/E

Highly lipophilic drugs may be retained in the lipophilic SC and resist partitioning into the more
hydrophilic viable epidermis [20]. Thus, clearance from, rather than diffusion across, the SC may then
become the rate-limiting step for highly lipophilic drugs. Similarly, the appendageal pathway may be
more important for highly hydrophilic molecules, electrolytes, and large molecules with low diffusion
coefficients [21,22]. For lipophilic permeants, Equation (2) may be well approximated as follows:

PSC ≈ PL (4)

In an analysis of experimental data obtained with hairless mice skin in phosphate buffer saline,
Equation (4) has been found to be true for lipophilic permeant corticosterone in a PBS, as PP « PL and
PD/E » PL [23,24].

4. NE in Transdermal Delivery
Nanosized drug delivery systems significantly enhance the bioavailability and solubility of
active constituents by penetrating vital cellular reservoirs [25,26]. Nanoemulsions (NEs) have been
reported to improve the transdermal permeation of many drugs when compared to conventional
topical formulations [27]. Many formulators have investigated the skin permeation mechanism of
many drugs using NEs and microemulsions (MEs) as delivery systems [20,28,29]. NEs and MEs share
similar components in different ratios and the same penetration mechanisms. Their main difference
lies in their droplet shape, size distribution, and kinetic stability [30]. To the best of our knowledge,
although different mechanisms have been suggested to explain the effect of NE on skin permeation in
the literature, an answer to the following question is not well understood: Do the NE properties or
NE components provide the transdermal permeation enhancement? Thus, this review focuses on the
different reported mechanisms by which NE enhanced the transdermal permeation of hydrophobic
and hydrophilic drugs.

4.1. Physical Properties of Transdermal NE

NEs are a transparent (translucent) liquid in a liquid colloidal dispersion system which is kinetically
stable, with a droplet size <100 nm, as reported by McClements [30]. However, he stated that other NE
particle size upper limits, such as 200 and 500 nm, were proposed in recent literature. The long-term
physical stability of NEs, with no flocculation or coalescence, provides unique properties and makes
them approach thermodynamic stability [31]. NEs consist of oil and aqueous phases, stabilized by
a surfactant and co-surfactant in defined ratios [32]. NE, when compared to conventional emulsion,
shows superior properties, namely sturdiness against gravitational separation, a droplet size in the
Sci. Pharm. 2019, 87, 17 4 of 34

nanometer range, improved stability, and larger capacity for drug encapsulation [33]. The dispersibility
of NEs is very high compared to MEs, owing to the small droplet size that prevents flocculation and
allows dispersion without separation [34]. NEs can be used to deliver either hydrophilic or lipophilic
drugs in water-in-oil (w/o) or oil-in-water (o/w) formulations, respectively [8].

4.1.1. Oil-in-Water (o/w) NE

Most of the drugs available in the market today that are developed by the pharmaceutical industry
are poorly soluble in water and lipophilic in nature [20]. Recently, scientists have shifted their interest
toward nanotechnology-based lipid systems, such as solid dispersions, solid lipid nanoparticles, liposomes,
MEs, and NEs, due to their enhanced ability to solubilize drugs, increase bioavailability, and increase
drug carrying capacity, which are also the most advanced commercial approaches. NE is a leading
trend in transdermal drug delivery systems since it enhances drug skin permeation and improves the
bioavailability of poorly soluble drugs when compared to other transdermal dosage forms [35,36].

4.1.2. Water-in-Oil (w/o) NE

NEs that are of the type w/o, issued for hydrophilic compounds, are less common than o/w NEs
for transdermal delivery. In w/o NEs, the drug resides in the water phase rather than the oil phase,
although its partitioning in the oil phase is inevitable, based on its oil-water partition coefficient.
Since the drugs used in this category are water-soluble, the selection of surfactants in these preparations
is based on a proper hydrophile-lipophile balance (HLB) value, in order to reduce the tension between
the water phase and the oil phase and produce a stable formula [2].

4.2. Components of Transdermal NE

4.2.1. Oil Phase

Oleic acid (OA) is a common oil phase that is employed in the composition of NEs. OA has
an inherent penetration enhancing ability, as it causes the SC to absorb more water and swell, and it
also compromises some of the structural components of the SC, thus increasing penetration through
this limiting and protective barrier [37]. Other oils that possess permeability enhancing abilities,
such as capryol 90 [5] and isopropyl myristate [8], have been reported in the literature. The viscous oil
α-tocopherol gives a very small droplet size, followed by hexyl laurate [31].

4.2.2. Surfactants
Surfactants are compounds that have the ability to enhance permeation through the skin, which
is assumed to be related to their ability to reversibly bind to keratin filaments, which in turn leads to
the disruption of corneocytes, thus altering the diffusion coefficient of the SC [38,39]. A study using
a NE containing medium-chain monoglycerides and diglycerides as surfactants demonstrated that the
concentration of surfactant blend can differentially affect the permeation of the studied hydrophilic
and lipophilic drugs across skin [36]. As reported, the transdermal delivery of hydrophilic drugs but
not lipophilic drugs was significantly enhanced when the concentration of the surfactant blend was
increased [40]. An explanation for this is that the very high concentration of surfactants modified the skin
barrier [41].
Surfactants are generally classified into non-ionic, anionic, cationic, or zwitterionic types [42].
Non-ionic surfactants are the most commonly used type of surfactants in transdermal NEs, due to
their low toxicity and minimum interference in NEs [43]. These surfactants have the ability to fluidize
SC lipids and hence enhance drug absorption [44]. Their rate of enhanced penetration is influenced by
two possible mechanisms. At the beginning, the surfactant enters the intercellular regions of the SC,
fluidizing, solubilizing, and extracting the lipid components [45]. Following this process, the surfactant
penetrates the intercellular matrix, interacting and binding with keratin filaments, hypothetically
resulting in the disruption of corneocytes [46,47].
Sci. Pharm. 2019, 87, 17 5 of 34

Anionic surfactants, for instance, better enhance the skin penetration ability of target molecules,
exerting a more powerful interaction with keratin and lipids [43,48–51]. Additionally, sodium lauryl
sulfate (SLS) alkyl chains were reported to be involved in the hydrophobic interaction with skin
structures, exposing the end sulphate group of the surfactant, creating additional bonding sites in the
membrane, thus leading to increased skin hydration [52,53].
Cationic surfactants also affect the cornified cells by interacting with the keratin fibrils and
disrupting the cell-lipid matrix. They may also change the electronic properties of the SC by interacting
with the anionic components there, enhancing the transfer of anionic drugs into the skin [43].
There is no perfect combination of surfactants that can be used in all formulations, since many
possibilities need to be explored. For the selection of a surfactant, the greater its degree of ethoxylation,
the lower the viscosity conferred to the formulation. This is because when the degree of ethoxylation of
the surfactant is higher, its solubility in water is higher, thus decreasing the viscosity of the medium [54].

4.2.3. Co-Surfactants
The addition of a co-surfactant reduces the interfacial tension and increases the fluidity of the
liquid-liquid interface by decreasing its bending stress. The interfacial tension continuously decreases
with an increasing co-surfactant concentration, until a minimum, beyond which the interfacial tension
increases again. The concentration of alcohol necessary to reach this minimum becomes higher
as the alkyl chain of the alcohol becomes shorter [55]. NEs prepared with n-butanol, n-hexanol,
and n-pentanol, considered as medium-chain alcohols, have reduced interfacial tension between the
surfactant and water phase. This ability is reduced with longer alcohol chains [56].
The size and region of NEs can be strongly influenced by the presence of the surfactant and
co-surfactant in the system [57], by altering the surfactant film rigidity, increasing its flexibility,
and taking up different curvatures required to form NEs over a wide composition range [58]. Short-chain
co-surfactants, like ethanol, tend to extend the phase diagram NE area, acting as a hydrotrope that
dramatically decreases the surfactant film interfacial tension between the oil and water phases [59].
The effect of the co-surfactant on the NE stability is dependent on its chain length. Accordingly,
when long-chain alcohols such as heptanol and hexanol are used as co-surfactants they result in the
separation of a closed water domain inside a continuum of a hydrocarbon layer. This leads to a less
uniform and less organized micelle system. On the other hand, medium-chain alcohols form stable oil
droplets within the system, with less sign of phase separation [60]. The selection of the co-surfactant
can control the release of the drug by tailoring the viscosity of the NE to either low or high viscosity,
without compromising the stability of the NE [61]. The inclusion of alcohol in the formulation exerts
a strong influence on the density and viscosity of the NE. Ethanol, for example, tends to lower the
viscosity of the overall preparation [59].
Furthermore, co-surfactants can improve the solubility of the drug loaded into the system [62].
Ethanol also has a low topical toxicity and can be mixed with a wide range of surfactants, increasing
the dissolving power of active compounds [63]. Multiple co-surfactants, when optimally selected,
can act in conjunction to improve the overall flux of the drug without the need to add a permeation
enhancer (PE) [64].

4.2.4. Permeation Enhancers (Accelerants)

Permeation enhancers (PEs) have been employed to temporarily alter the physicochemical nature
of the SC, thus reversibly facilitating the passage of the drug. PEs exert reversible modifying action
on the connections between corneocytes [16]. A different mode of action has been proposed, through
affecting the metabolic activity in the skin [65,66]. Many studies indicate the importance and influence
of the chemical structure of PEs (i.e., terpenes, azones, fatty alcohols, and fatty acids) on the permeation
enhancement of drugs [42].
Amongst the most commonly used PEs are fatty acids, e.g., OA (which is usually used as an oil
phase in NE formulations), esters of fatty acids such as isopropyl palmitate and ethyl oleate, terpenes,
Sci. Pharm. 2019, 87, 17 6 of 34

for instance limonene, menthol, and cineole, and short-chain alcohols such as ethanol. Many have
shown an effective transdermal permeation enhancing ability with low skin irritation [67].
In the past, OA was used as a PE before the emergence of nanoscale systems. A formulation
containing 6% OA showed an increase in penetration enhancement of up to 208-fold when compared
to a formulation containing no OA [68–71].
The most commonly used terpene is limonene. It was used to enhance the transdermal permeation
of both lipophilic and amphiphilic compounds while found ineffective with hydrophilic compounds [72].
One study compared transdermal drug delivery through rabbit skin using 5% menthol as a PE, emphasizing
the impact of the addition of PE to formulations to enhance systemic drug delivery [73].
Other PEs have been employed in enhancing drug transdermal permeation, for example azone,
dimethyl sulfoxide (DMSO), and N-methyl pyrrolidone (NMP). Azone (laurocapran) was the first
specifically designed as a skin PE, being effective at low concentrations ranging between 0.1 to 5%.
It has been reported to enhance the skin transport of a wide variety of drugs, including steroids,
antibiotics, and antiviral agents, through partitioning into the skin bilayer lipid and disrupting their
packing arrangement [65].
DMSO has been incorporated along with ethanol as a PE for the transdermal delivery of
acyclovir [74]. Comparisons of the enhancer potencies based on the free concentration of the enhancers
revealed a nearly semi-logarithmic linear relationship between enhancer potency and the carbon
number of the alkyl chain length [75]. DMSO interacts with keratin and works as an enhancer by
changing the protein conformation and opening up water channels within the corneocyte [76].
N-methyl pyrrolidone (NMP) was found to be a more effective enhancer for the aqueous phase
of MEs than the organic phase. Owing to its low isopropyl myristate/water partition, NMP resides
exclusively in the water phase of the system, where its enhancing effect from that phase should
dominate. In w/o MEs, NMP is sequestered in the encapsulated phase and unable to interact with skin.
This might explain the better transdermal transport of both hydrophilic and hydrophobic permeants
from o/w MEs [77].
Short-chain alcohols such as ethanol, isopropyl alcohol (IPA), and polyethylene glycol (PEG) have
a pronounced permeation enhancement ability compared to medium chain alcohols, such as n-propanol,
n-butanol, and n-pentanol, which are more suitable as co-surfactants than PEs [78]. Investigators gave
ethanol great attention in transdermal delivery regarding its mechanism of permeation enhancement.
Researchers compared the effectiveness of ethanol as a PE with lauric acid and sodium tauroglycocholate
to help facilitate the passage of aminophylline through skin. Lauric acid showed the best enhancing
ability in the first hour, while after four hours ethanol leaped ahead, resulting in a 60% permeation
enhancement, which is explained by ethanol’s ability to temporarily modify the SC to increase
permeation [45].
The enhancement ability of ethanol tends to have an optimum range, where in high concentrations
drug penetration is reduced [79]. At low levels (<25%), ethanol has little or no effect on the pore
pathway. It has been proposed that the ethanol enhancement effect at low levels may be interpreted in
terms of increasing fluidity in the transport rate-limiting lipid domains [80]. In other words, low ethanol
content (<50%) may be effective in fluidizing the SC lipid bilayer at or near the polar head plane,
but not in the bilayer hydrocarbon interiors. The polar/ionic permeants were transported via the pore
pathway at all ethanol concentrations. In contrast, at high ethanol levels (>50%), there was a significant
increase in new pore formation in the SC, while at very high ethanol levels (≥75%), the pore pathway
appeared to dominate the transport of all permeants, including the lipophilic permeants, irrespective
of polarity [81].

5. Biocidal Property of NE
Surfactant NEs have been proven to have a broad-spectrum biocidal activity against a variety
of microorganisms, including gram-positive and gram-negative bacteria, spores, and enveloped
viruses [82]. Soybean oil nanodroplets, when stabilized by a detergent and solvent, selectively fuse
Sci. Pharm. 2019, 87, 17 7 of 34

with the bacterial membrane or viral envelope, destabilizing lipids and initiating the disruption of the
pathogen. Myc et al. demonstrated an effective anti-fungal activity through a novel NE consisting of
oil, three non-ionic detergents, solvents, and water [82].
Previously, such a combination of properties against microbes could only be achieved with
antibiotics and disinfectants [83]. This capability shows great selectivity as a standalone mode
of action and as an added feature to formulations, without the need of preservatives to maintain
stability. Moreover, this particular property implies the direct application of the NE on the skin surface,
even without the need of disinfecting the area of application. Despite this antimicrobial activity, NEs
do not exert any toxicity to skin layers or to normal cells.
As reported, these antimicrobial NEs were composed of detergents, oils, and 20% water, forming
a w/o system. Eucalyptus oil, which has a broad-spectrum antibacterial activity, was used as an oil
phase in a NE applied to a Listeria monocytogenes strain. Here, the NE showed a sporicidal activity,
mediated by both Triton X-100 and tri-n-butyl phosphate components. This unique sporicidal action
of the emulsion is interesting because Bacillus spores are generally resistant to most disinfectants,
including many commonly used detergents [84,85]. NE also possesses a broad-spectrum activity
against a variety of bacterial strains, such as Salmonella, E. coli, S. aureus [76,86]. It has been reported
that a novel NE composed of tributyl phosphate, soybean oil, and Triton X-100, has the potential to
be used as a topical antimicrobial agent against several pathogens [87]. Although all the previous
studies done on antimicrobial NEs were intended for topical use, this property could be exploited for
transdermal applications.

6. Methods of NE Preparation
Depending on the desired formulation, the appropriate preparation method should be carefully
selected to optimize the droplet size distribution, since it strongly affects the stability behavior of the
NE [33]. Below, methods of NE preparation are divided into high and low energy methods.

6.1. High Energy Methods

High energy NE formation involves the use of devices that reduce the size of the inner phase
droplet into a unified and reproducible range. This method is sophisticated, consumes large amount of
energy, and is not suitable for thermolabile components such as proteins, enzymes, nucleic acid, and
retinoids [88].

6.1.1. High-Pressure Homogenization

The high-pressure homogenization method uses many techniques to achieve the required particle
size (PS). Among these techniques are hydraulic shear, cavitation, and intense turbulence [89], usually
under a pressure of 500 to 5000 psi [90]. The surfactant and co-surfactant are passed through a small
opening (orifices) where dilution is performed later.
This method can be done on multiple cycles, passing the liquids through the homogenizer multiple
times to unify and reduce the droplet size down to 1–10 nm. Optimally, a NE with an oil content of
20% is used for this method, as a large amount of oil in the formulation reduces the productivity of this
method significantly. This is one of the most frequently used methods for the preparation of NEs using
a high energy technique [88].

6.1.2. Microfluidization
Microfluidization is a patented mixing technology that involves a high pressure positive
displacement pump with a pressure of 500–20,000 psi [89]. This pressure forces the phases to
pass through micro-channels. The two phases are mixed and proceed as a coarse emulsion, made to
pass through micro-channels, which leads to a massive amount of shear. The liquids are then passed
through an interaction chamber microfluidizer unit until the desired droplet size is achieved [88].
Sci. Pharm. 2019, 87, 17 8 of 34

This method is expensive and is not suitable to produce NE in large quantities, so up-scaling using this
method is difficult and not viable [91].

6.1.3. Ultra-Sonication
A sonicator probe introduces a vibration of a certain wavelength which produces cavitation,
producing a small droplet size, hence preventing the NE from coalescing [92]. This method can only
be used in a laboratory setting, as large volumes of production are not feasible [90].

6.1.4. Jet Disperser

Similar to microfluidization but with no moving parts, this method is able to handle extreme
pressures of up to 400 MPa [88]. Two jets facing each other pour two different liquids through the
nozzles, causing them to collide with each other. With the help of laminar elongation flow, the formed
NE is collected through an orifice plate that coordinates the energy dispersion. This method is not as
efficient as microfluidization, but it is less expensive and more energy efficient [92].

6.2. Low Energy Methods

6.2.1. Phase Inversion Temperature

Also known as the condensation method, the phase inversion temperature method was first
carried out in 1968 [93]. They concluded that the increase in temperature results in the chemical
changes of polyoxyethylene surfactants by degradation of the polymer chain with temperature [89,94].
This method involves the transitioning of the liquid phases by the spontaneous curvature of the
surfactant [93]. This could be achieved by keeping the composition of the emulsion fixed while changing
the temperature of the setting (phase inversion temperature, PIT) [95]. It can also be performed by
keeping the temperature constant and changing the composition of the system (emulsion inversion
point, EIP) [96].

6.2.2. Spontaneous Emulsification

This method is the simplest technique to produce NEs, it does not require any special equipment
or devices. The oil phase, surfactant, co-surfactant, and the aqueous phase are added together in
a stepwise manner, where gentle stirring is sufficient to produce the NE. The limitation of this method
is the rigorous process of selection of the correct ingredients to produce a stable NE with a small droplet
size [91,97,98].

6.2.3. Solvent Displacement Method

This method involves the dissolving of the oil phase in a solvent such as ethanol or acetone,
then adding them both to the aqueous phase containing the surfactant and co-surfactant. This method
does not require energy as it happens spontaneously at room temperature [93]. The viscosity of the
emulsion, surfactant type, and concertation are the defined parameters that must be obtained to acquire
the required NE [99,100].

7. Characterization of Transdermal NE
The characterization of a NE includes an array of testing steps that guarantee the successful
formation of the NE. The droplet size confirmation, stability, and compatibility of all the formulation
components, skin irritation testing, and successful transdermal delivery of the drug play an important
role in the biological activity of the NE [101,102]. The most common characterization tests are collected
and summarized in Table 1.
Sci. Pharm. 2019, 87, 17
Tests
Visual inspection
Viscosity
Transmission electron microscopy (TEM)
Scanning electron microscope (SEM)
Atomic force microscopy (AFM)
Morphology
Ultrasonic resonator technology
Particle size
Photon correlation spectroscopy (PCS)
Polydispersity index (PDI)
Dynamic light scattering (DLS)
Zeta potential (ZP)
Electro-conductivity
Refractive index
In vitro skin permeation
In vivo
dermato-pharmacokinetics
and pharmacodynamics
Skin irritation
Table 1. Characterization tests performed for transdermal nanoemulsion (NE).
Equipment
Naked eye
Rotational viscometer
Neutron scattering
Cryo-electron microscope
A conductivity meter
A refractometer
Franz diffusion cell apparatus
Intact live animals
HPLC
Live animals (rats or rabbits)
Significance
To determine the successful formation of the NE.
Low viscosity NE has faster release and rapid
skin penetration than high viscosity NE, o/w NE
are usually low in viscosity compared to the w/o
NE.
To verify the droplets fabricated with enough
consistency in their shape and size being in
the nano-range.
To quantify the homogeneity and dispersion of
the NE as well as to estimate the broadness and
range of the droplet size. The lower the PDI
value (<0.2) and the higher the ZP, the better NE
stability is against onward ripping and other
destabilizing forces.
An indication of preliminary change in NE
droplet size change although the relationship
between electrical conductivity and NE
instability is not linear.
A strong indication of uniformity and the
formation of an isotropic NE.
To assess transcutaneous penetration or
membrane retention.
To establish a plasma drug concentration-time
profile or to assess a pharmacological drug effect.
To determine whether NE produced irritation
or not.
9 of 34
Description Ref
Visual observation of sudden turbidity followed by the production of clear and
[103]
transparent NE.
Measurement of the amount of torque needed to rotate the paddle in the NE. [104–106]
The NE sample is negatively stained with a 1% solution of phosphotungstic acid
and then applied to the copper or carbon coated grid, depending on the model of
the TEM or SEM used. The accelerating voltage used is usually 20 kV, and by [107–109]
using the appropriate software and magnification, a quantitative measurement
can be achieved, along with the quality and consistency of the NE drops.
Size and size distribution are measured from the collected measurements of the
scattering of dynamic light of the NE droplets.
[110–112]
ZP is measured as the potential charge difference between the particles and the
continuous phase.
The meter measures the amount of electrical current or conductance in the NE
sample. The meter, equipped with a probe, is placed into the sample to be
measured. The meter applies a voltage between two electrodes inside the probe. [113,114]
Electrical resistance from the total dispersed particles in sample causes a drop in
voltage, which is read by the meter.
Comparing the refractive index (RI) of the NE with water (RI = 1.333), where the
closer the NE value to the water value, the more uniform and transparent the NE [115,116]
is.
A sample of NE is placed into the donor compartment after placing a variety of
membranes, either synthetic or excised skin from an animal model. The receiver
compartment of the Franz diffusion cell is filled with a phosphate buffer saline
with a pH of 7.4, which simulates the blood stream. It is then stirred at 100 rpm at
a temperature of 37 ◦ C. A sample of 1 mL is taken either manually or [19,117]
automatically, filtered, then analyzed using UV spectroscopy or HPLC. Once the
value of the released drug has been determined for every hour, the steady state
flux (Jss) is calculated with the formula Jss = P.CD , where P is the permeability
coefficient and CD is the donor chamber concentration.
Administering NE to a shaved animal skin. Blood samples are withdrawn at
different intervals, centrifuged, and then the plasma is analyzed using HPLC to
determine the amount of drug that reached circulation. Furthermore, the [36,84,118–120]
pharmacodynamics properties of the NE are assessed depending on the
pharmacological effect of the drug.
The healthy experimental animals were divided into groups and the formulation
was applied on the hair-free skin of animals by uniform spreading within a
specific area. The experiment was usually carried out for 7 days and the
[121–124]
application sites were graded according to a visual scoring scale. The sites tested
were put under observation for 48 h to detect if any erythema or edema was
formed after application. Skin irritation was scored following the Draize method.
Sci. Pharm. 2019, 87, 17 10 of 34

8. NEs for Transdermal Delivery of Hydrophilic Drugs

8.1. Transdermal w/o NEs Containing Hydrophilic Drugs

From the literature, several experiments have explored the efficacy of w/o NEs on the transdermal
permeation of hydrophilic drugs through both animal and human skin (Table 2). However, there is
still a surprising lack of data on NEs optimized for the delivery of hydrophilic drugs [125]. When the
water-soluble metoprolol was formulated in o/w NE, it exhibited higher in vitro release compared to
w/o NE. However, there was no significant difference between the 2 types of NE regarding rat skin
permeation and no mechanism was suggested [126].
The loading of ropinirole hydrochloride (RHCL), a selective non-ergoline dopamine D2 receptor
agonist used to treat Parkinson’s disease in transdermal NEs, was investigated. RHCL has poor
bioavailability (50%) due to the extensive first pass effect. Transdermal w/o NE was compared to the
aqueous solution. The results were very promising, where the w/o NE did indeed penetrate rat skin at
a rate of 63.23 mg/cm2 /h, while the aqueous solution failed to facilitate the transport of RHCL through
the skin. These results showed a very promising prospect for w/o NE to be used for the delivery of
hydrophilic drugs that show poor bioavailability through conventional routes of administration [127].
Previous studies reported similar results explaining that ME and NE could modify the surface electrical
charge of an ionic drug and then enhance the permeability of a hydrophilic drug [128,129].
The potency of w/o NE, to facilitate inulin permeation, a diagnostic aid for renal function that
is considered a large and water-soluble molecule, was tested. The ability to permeate hairless and
hairy mice skin and also hairy rat skin was investigated. The drug flux was dependent on the HLB
of surfactant mixture, rather than any other factors, including the drug molecular weight (Mw) and
animal skin characteristics, such as the SC thickness and follicle-type. Using in vitro testing and
comparing the penetration results of the NE with a micellar solution and aqueous control, the results
revealed that the transdermal permeation of the micellar system was lower than the NE, due to the high
viscosity and homogeneity with the follicular sebum [130]. This hypothesis was further supported by
the low penetration result of “low-oil” NE that failed to enter the follicular pathway to successfully
facilitate the release of inulin from the inner water phase. The combined results suggested that w/o NE,
which is compatible with the lipophilic sebum of hair follicle, enhanced the transport of hydrophilic
solutes, implying that such transport is transfollicular [130].
The presence of sebum in the hair follicle greatly hinders the delivery of hydrophilic
compounds [131–133]. Caffeine was detected in blood only 5 min after application on the skin
of healthy volunteers for open follicles and 20 min for closed hair follicles, emphasizing the importance
of the pore pathway in hydrophilic drug delivery. Experimentally, an in vivo application showed
better penetration through the hair follicle than the in vitro experiment regarding the transfollicular
route [134,135].
The NE with soybean oil, span 80 as a surfactant, and isopropyl alcohol as a co-surfactant,
at concentrations of 46.5, 32.4, and 10%, respectively, was developed for incorporating glycyrrhizin,
which is used for a variety of health complications, such as allergies, ulcers, chronic hepatitis,
and arthritis. The developed w/o NE has enhanced penetration properties on human cadaver skin,
where the optimum formula showed the most linear and sustained release profile due to a low
droplet size and reduced viscosity [136]. The topical application of a single dose of w/o NE that
was composed of olive oil, span 80 as non-ionic surfactant, and containing plasmid DNA, mediated
transgene expression. The deposition of plasmid DNA was primarily in the hair follicles. In contrast,
none of cationic liposomes mediated transgenic protein expression. Thus, w/o NE was suggested to be
a suitable candidate in gene therapy to facilitate the transfection of follicular keratinocytes [137].
As reported, thiocolchicoside (TCC), with a very poor oral bioavailability due to an extensive
first pass metabolism, was incorporated into a w/o NE consisting of linseed and Sefsol as the oil
phase, span 80 as the surfactant, and Transcutol P as a co-surfactant, having an average PS of 117 nm.
The results showed a 5-fold improvement in transdermal delivery compared to the aqueous solution
Sci. Pharm. 2019, 87, 17 11 of 34

in vitro using porcine skin. These promising data serves as a good indicator to carry out future in vivo
studies for TCC NE to further prove TCC efficiency [138].
Caffeine NE was developed using OA and eucalyptol, which both serve a dual function of being
an oil phase and a PE, showing a 51% and 54% passage of caffeine, compared to 27% penetration
of a topical caffeine solution when the follicles were opened. On the other hand, the percentage of
penetrated drug was only 18.9% through blocked follicles [139]. Follicular delivery is tricky in vitro,
since human skin follicles tend to collapse, becoming blocked with dry sputum, and secretion pathways
become inactive. In vivo results always show better results in live skin than dead skin [140].
Sci. Pharm. 2019, 87, 17 12 of 34

Table 2. Nanoemulsions (w/o) for the transdermal delivery of hydrophilic drugs.

Nanoemulsion Components
Permeant Underlying Mechanism of Penetration Ref.
Oil Phase Surfactant Co-Surfactant Aqueous Phase
Enhanced transdermal delivery is mainly attributed to the
thermodynamic activity of the drug along with the low viscosity,
Ropinirole Isopropyl alcohol
Brij 30 Brij 35 + Brij 30 Water resulting in a shortened release lag time from 12 to 2.7 h. [127]
hydrochloride (IPA)
Increasing the ethanol content from 20 to 30% showed a slight
increase in flux from 20.25 to 25.94 µg/cm2 .
Using a low hydrophile-lipophile balance (HLB) surfactant
mixture showed better penetration compared to aqueous and
Inulin Olive oil Tween 80 No co-surfactant used Water other micellar formulations. This enhancement in follicular [130]
penetration is attributed to the solubilization of sebum by
NE components.
The selected optimum formula showed enhanced and sustained
Glycyrrhizin Soybean oil Span 80 Brij 35 + IPA Water release profile on human skin. Results are attributed to low [136]
droplet size and viscosity.
When DNA plasmid was loaded into w/o NE, a more condensed
DNA plasmid Olive oil Tween 80 No co-surfactant used Water state of DNA was formed, resulting in higher gene expression [137]
levels due to its deposition in hair follicles.
A 5-fold increase in the penetration of thiocolchicoside (TCC)
was reported. The small droplet size as well as the NE
Thiocolchicoside Linseed oil Span 80 Transcutol P Water [138]
components acting as penetration enhancers are the main driving
factors for transdermal enhancement.
Caffeine loaded NE formulations showed the transport of 51%
Caffeine OA/EU Volpo-N10 Ethanol Water and 54% of the drug, compared to 27% in case of a topical caffeine [139]
solution. This is attributed to the transfollicular route by NE.
The o/w 5-aminolevulinic acid (ALA) showed a high flux rate,
but not the w/o NE, which is mostly attributed to NE
5-Aminolevulinic components. The addition of α-terpineol, which is a penetration
Soybean oil Span 80 α-terpineol Water [141]
acid enhancer, did not yield any further improvement in penetration.
The thermodynamic activity of the drug is also one of the
contributing factors.
Sci. Pharm. 2019, 87, 17 13 of 34

8.2. NE Mechanisms for Enhanced Transdermal Delivery of Hydrophilic Drugs

The mechanism by which a hydrophilic drug is transported through skin from a w/o NE, is indeed
different from a hydrophobic drug. As the passage through skin requires an increased lipophilicity [12],
it is then important to study the transdermal transport of a hydrophilic compound delivered from
NE systems. It was hypothesized that a hydrophilic drug would not be available for percutaneous
transport from NEs, unless water from the NE freely permeates transdermally. Therefore, the sufficient
mobility of water within NE vehicle and the sufficient percutaneous transport of water across the skin
barrier are both required [142]. Moreover, the active compound’s PS and the droplet size play a more
important role here than in o/w [143]. Different reported mechanistic theories are discussed as follows
and illustrated in Figure 2.

Figure 2. Transdermal enhancement of hydrophilic drugs from NE: (A) Increasing drug thermodynamic
activity. (B) Modification of surface electrical charge of ionic drugs. (C) Solubilizing of sebum by NE
Figure 2. Transdermal enhancement of hydrophilic drugs from NE: (A) Increasing drug
components to facilitate follicular delivery. (D) Pore pathway of large water-soluble molecules loaded
thermodynamic activity. (B) Modification of surface electrical charge of ionic drugs. (C) Solubilizing
in w/o NE. (E) Carrying of small water-soluble molecules into o/w NE for follicular delivery.
of sebum by NE components to facilitate follicular delivery. (D) Pore pathway of large water-soluble
molecules loaded
8.2.1. Increasing Drug in w/o NE. (E) Carrying
Thermodynamic of small water-soluble molecules into o/w NE for follicular
Activity
delivery.
Drug thermodynamic activity is a major factor in skin permeation, where the drug moves from the
9. NEsphase
internal for thetoTransdermal
the external Delivery
phase, thenof Hydrophobic
depositing intoDrugs
the skin layers. This provides a continuous
process which has a major role on the extent to which the drug can diffuse transdermally, especially
for9.1. Transdermal
hydrophilic O/W NEs
drugs. ThisContaining
phenomenon Hydrophobic Drugs with glycyrrhizin, which has a large Mw of
was observed
822.94 g/mol, where having
The incorporation of alipophilic
low concentration of theNEs
drugs in o/w surfactant/co-surfactant
is reported to be a tool mixture (Smix) in
to enhance thethe
formula showed a better release through human skin than with a large concentration,
transdermal permeation of such drugs, despite the variable explained mechanisms (Table 3). hypothesizing
that theAceclofenac
low viscositywas dueformulated
to the low Smix
in o/w content contributed
NE and compared to to
give the drug molecules
conventional gel. Themore freedom
optimized
to formula, consisting
move through phasesof 10%
and Labrafil,
eventually 35.33% Tween
penetrate 80,SC.
the 17.6% Transcutol P, activity
Thermodynamic 5% Triacetin,
is alsoand 32%
enhanced
water, showed improved permeation enhancement and acceptable irritation results.
with the reversal of altering the skin’s natural barrier resistance and modifying the drug SC partition The in vivo
efficacy showed
coefficient 40% more anti-inflammatory
by NE components as surfactants andresults
PEsin[136].
comparison to thebeen
It has also aceclofenac
suggestedgel [151].
that ME, by
The spontaneous
continuously fluctuatingemulsification
interfaces, may technique
increasewas
the used
drug to preparehence
mobility, the NE formulation
aiding containing
its penetration [144].
clozapine, a drug that exhibits only 27% oral bioavailability due to the extensive hepatic
In addition, the high solubilization capacity of ME increases the thermodynamic activity of permeants, metabolism.
OA was used as the oil phase, Tween 20 as the surfactant, and Transcutol P as the co-surfactant (Smix
3:1), with 1% clozapine. The NE was incorporated into a gel to extend the contact time with the skin.
Overall, 70% of the drug penetrated the skin over a period of 10 h, exhibiting both a rapid release
profile over the first 2 h and an extended release profile over the 8 h of skin contact [10].
A tamoxifen o/w NE was investigated for cancer suppression performance in tumor-induced
mice. The relative tumor volume after the 23 days testing period showed promising results and
Sci. Pharm. 2019, 87, 17 14 of 34

which is one of the driving forces of drug transport [144]. The drug in this energy-rich system can
diffuse across the flexible interfacial surfactant film between the phases. This is a thermodynamic
process that increases partitioning and diffusion into the SC [145].

8.2.2. Modification of Surface Electrical Charge of Ionic Drugs

The hydrophilic drug RHCL, with low Mw, when topically applied as an aqueous solution,
showed no penetration properties, despite the drug’s hydrophilicity and small molecular size. Based
on these findings, NE formulation was developed with isopropyl myristate (IPM) as the oil phase (5%),
Brij® 35 + Brij® 30 (20–30%) as the surfactant, IPA as the co-surfactant (20–30%), and double-distilled
water (34.5–50%).The developed NE exhibited a significant increase in drug permeation rate [105].
It was explained that NEs could modify the surface electrical charge of an ionic drug and then enhance
the permeability of a hydrophilic drug, as reported in literature [146,147].

8.2.3. Solubilization of Sebum by NE Components

SC layers tend to be thinner closer to follicular orifices, which are rich in terms of their capillary
network. Areas of the human body with higher follicular density like the forehead show good follicular
drug delivery compared to other areas like the forearm [136]. However, follicular delivery depends on
presence of sebum and the blockage of pores. Otberg et al. [136] reported that the artificial blocking
of hair follicles by a varnish wax mixture significantly reduced skin penetration of the hydrophilic
permeant caffeine.
It was found that a w/o NE with a low HLB surfactant mixture showed better penetration
compared to aqueous and other micellar formulations, where compatibility with the lipophilic nature
of the sebum plays a role in solubilizing it by the surfactant mixture, thus clearing the way for the drug
to penetrate more easily [130]. There seems to be an ongoing limitation concerning in vitro release
studies of the follicular pathway due to follicular shaft collapse and the cessation of perifollicular
circulation. Moreover, the animal skin model used to evaluate this route is not always consistent with
the release profile when applied to human skin [140]. However, certain in vivo human studies with
the hydrophilic molecule caffeine NE showed that the transfollicular route contributed to over a third
of the total penetration [139,148,149].

8.2.4. Pore Pathway for the Transport of Large Water-Soluble Molecules Loaded into W/O NEs
Large water-soluble drugs are unable to cross the lipid corneocyte domain. The pore pathway or
hair follicle route, a primary path which w/o NEs containing large molecules use to reach the main
circulation, is an important transdermal mode of delivering water-soluble permeants. In the gene
delivery approach, hair follicles might be a portal toward the entrance of topically applied negatively
charged DNA [150].
Recently, scientists researched the need to deliver and transfect plasmid DNA to follicular
keratinocytes. They found that transdermal w/o NE allowed DNA expression into therapeutic
transgenic proteins [137]. The prepared plain NE had a mean PS of 42.3 nm, while after the addition of
a DNA plasmid, a smaller NE with a mean PS of 32 nm was produced. The levels of gene expression
after the topical application of the DNA NE were significantly higher than with the plasmid DNA
aqueous solution. The authors hypothesized that forming a stable w/o NE may drive the conformation
of DNA into a more condensed state. They supported their hypothesis by highlighting the absence of
aggregates and large particles during the PS analysis [137].

8.2.5. Carrying of Small Water-Soluble Molecules into O/W NE for Follicular Delivery
The skin permeation of the hydrophilic small molecule 5-aminolevulinic acid (ALA) in both w/o
and o/w NE was studied. Low in vitro drug release of w/o NEs was detected and attributed to their
high viscosity. Surprisingly, o/w NEs showed the highest permeation flux through nude mice skin,
meanwhile, w/o NEs did not show any improvement over the aqueous solution. This result was
Sci. Pharm. 2019, 87, 17 15 of 34

explained by the required process of drug partitioning from the inner phase to the external phase in w/o
NE, while the release from o/w requires diffusion through the external phase [141]. The in vivo high
permeation from the o/w NE suggested that other mechanisms than the lipid pathway predominate
the delivery. Among these suggested mechanisms are the follicular/sebaceous route, the enhancer
effect, and an increase of thermodynamic activity, as suggested by Zhang el al [151].
The importance of hair follicles for o/w NE penetration was verified by in vivo confocal laser
scanning microscopy in mice models. Representative fluorescence staining indicated enhanced ALA
penetration into deeper skin layers after the application of the o/w nanocarrier systems compared to the
control. However, the effect of permeation enhancement for ALA was not detected in the in vitro skin
permeation study. The given explanation for this was that, after skin excision, the follicular volume
was reduced by the contraction of the elastic fibers, such that the follicles were less receptive to the
applied substances [141]. The extended release properties obtained after 48 h address the following
question: Is retaining hydrophilic small molecules within skin layers for longer periods a possible
mechanism of o/w NEs? From our point of view, w/o NEs, for small hydrophilic molecules, may
facilitate their transepidermal permeation. In contrast, the trans-appendageal route is the accepted
route of permeation in the case of incorporation in w/o NEs.

9. NEs for the Transdermal Delivery of Hydrophobic Drugs

9.1. Transdermal O/W NEs Containing Hydrophobic Drugs

The incorporation of lipophilic drugs in o/w NEs is reported to be a tool to enhance the transdermal
permeation of such drugs, despite the variable explained mechanisms (Table 3).
Aceclofenac was formulated in o/w NE and compared to conventional gel. The optimized formula,
consisting of 10% Labrafil, 35.33% Tween 80, 17.6% Transcutol P, 5% Triacetin, and 32% water, showed
improved permeation enhancement and acceptable irritation results. The in vivo efficacy showed 40%
more anti-inflammatory results in comparison to the aceclofenac gel [151].
The spontaneous emulsification technique was used to prepare the NE formulation containing
clozapine, a drug that exhibits only 27% oral bioavailability due to the extensive hepatic metabolism.
OA was used as the oil phase, Tween 20 as the surfactant, and Transcutol P as the co-surfactant (Smix
3:1), with 1% clozapine. The NE was incorporated into a gel to extend the contact time with the skin.
Overall, 70% of the drug penetrated the skin over a period of 10 h, exhibiting both a rapid release
profile over the first 2 h and an extended release profile over the 8 h of skin contact [10].
A tamoxifen o/w NE was investigated for cancer suppression performance in tumor-induced
mice. The relative tumor volume after the 23 days testing period showed promising results and
increased bioavailability in comparison to the orally administered drug, due to the avoidance of the
first pass metabolism related to the transdermal route and consequently a better penetration to the
target site [152].
Cumin was loaded in an o/w NE, exploring its capacity to reduce the oxidative damage related to
a variety of diseases. The NE formulations showed good permeation results and high zeta potential,
which is an indication of good stability. The highest permeation was observed after 6 h. The formulation
was able to restore hepatic enzyme plasma levels after the extensive hepatotoxic effect induced by
paracetamol after 7 days. Furthermore, a prolonged effect was witnessed, lasting up to 7 days after the
single topical application of the NE. This indicated that a weekly application for patients suffering
from chronic disease is convenient, with long term health benefits [5].
The secondary effect of tricycle antidepressants (imipramine and doxepin) which possess
an analgesic effect on the peripheral nerve endings was investigated in o/w NE formulations. The NE
formulation application increased the paw withdrawal time by 13% and 28% for the imipramine NE
and the doxepin NE, respectively [153].
A hyaluronic acid-based NE was used as a carrier for vitamin E [154]. The effect on the stability of
the NE was investigated by altering the zeta potential, degree of substitution, pH, and crosslinking
Sci. Pharm. 2019, 87, 17 16 of 34

agents. The encapsulation efficiency results, small size, and improved stability showed that the
investigated NE was a potential transdermal carrier for lipophilic drugs [154].
An antifungal agent (amphotericin B) was incorporated into an o/w NE to enhance the transdermal
delivery and effectiveness against Candida albicans and Aspergillus niger. The study revealed that
the NE’s synergistic ability allows a reduction of the antifungal dose, with increasing therapeutic
efficiency [155]. The prepared NE (Capmul PG8 as the oil phase, with Labrasol and PEG-400 as the
surfactant and co-surfactant, respectively, and DMSO as the transdermal enhancer) was compared to
the commercially available Fungizome® gel and drug solution as well. NE amphotericin B exhibited
enhanced transdermal penetration and exhibited a stronger antifungal ability, coupled with longer
a shelf life when compared to the conventional amphotericin B topical dose [155].
Singh et al. developed a NE for Carvedilol, a drug that has a short plasma half-life and extensive
first pass metabolism. Upon conducting the in vitro and in vivo experimentation, good results were
obtained, indicating a great potential for Carvedilol NE to be used in the future [156]. In another
study, a significant increase in the Carvedilol permeability coefficient in the NE vehicle was obtained,
compared to control film [157].
Finally, human skin permeation studies showed that MEs even enable iodide ions to diffuse
through the skin [158]. A high drug amount was released from the NE in comparison to the nano-lipid
carrier, which promoted the penetration into the skin, as previously reported [159].
Sci. Pharm. 2019, 87, 17 17 of 34

Table 3. Nanoemulsion (o/w) for the transdermal delivery of hydrophobic drugs.

NE Components
Permeant Underlying Mechanism of Penetration Ref.
Oil Phase Surfactant Co-Surfactant PE Aq. Phase
Compared to aceclofenac gel, the developed NE formulation showed an
Aceclofenac Labrafil Tween 80 Transcutol P —- Water increased anti-inflammatory effect and better penetration. The suggested [151]
mechanism is the low droplet size and viscosity.
The optimized formulation showed the highest permeation value, with an
inhibition of 70.8% of inflammation area. The permeation efficiency of
Sefsol-218 and Cremophor-EL/Tween
Celecoxib Transcutol P —- Water celecoxib is attributed to the small droplet size, low viscosity, and [160]
Triacetin 80
permeation enhancement factor of both the hydrophilic and hydrophobic
domains in the NE phases.
The NE formulation exhibited a prolonged Tmax with a large AUC value
giving the olmesartan NE great bioavailability compared to its oral
Olmesartan Clove oil Tween 20 PEG — Water [161]
counterpart. The small droplet size and NE components enhancing skin
penetration were the underlying mechanisms here.
The NE formulation showed better permeation results in comparison with
the plain drug gel, drug solution, and the marketed formulation,
Transcutol P. The permeation enhancing ability played a major role.
Ketoprofen OA Tween 80 Transcutol P — Water [162]
Incorporating the NE into a gel showed no improvement in flux,
indicating the NE components, along with the small droplet size, is what
led to such a profound skin permeation ability.
The bioavailability of GLBD was enhanced by 3.92 times compared to the
Labrafac and Diethylene glycol
Glibenclamide Tween 80 — Water oral formulation. The NE small droplet size and penetration enhancement [163]
tricetin monoethyl ether
ability of NE components are the underlying mechanisms.
The combination of OA along with Tween 20 and Transcutol P showed
great penetration enhancing ability, allowing a 3-fold improvement in
Clozapine OA Tween 20 Transcutol P — Water transdermal release compared to the traditional emulsion. Here, the [10]
enhancing ability of the NE components is the mechanism that
successfully aided the delivery of the drug transdermally.
Good penetration results were reported with the selected formulation
Dill essential
Tamoxifen OA Chromophore RH40 Ethanol Water which had the smallest droplet size, and the addition of 5% Dill essential [152]
oil
oil showed a profound effect on tamoxifen flux.
Using human skin in this study, it was demonstrated that both caffeine
and naproxen showed enhanced human epidermal permeation, mostly
Caffeine and
OA and EU Volpo N10 Ethanol —- Water through the solubilizing of the active elements in the NE components, [164]
Naproxen
followed by NE components ability to disrupt the lipid bilayer of the
SC cells.
The plasma total antioxidant capacity reached maximum efficiency after 7
days of transdermal application of cumin NE. The enhancement ability of
Cumin OA Tween 20 Ethanol —- Water [5]
these three components in the NE played a big role in raising the
antioxidant activity systemically using cumin.
Sci. Pharm. 2019, 87, 17 18 of 34

Table 3. Cont.

NE Components
Permeant Underlying Mechanism of Penetration Ref.
Oil Phase Surfactant Co-Surfactant PE Aq. Phase
For both drugs, the resulted NE droplet size was below 28 nm. The
selection of OA as an oil phase was due to its ability to fluidize the lipid
Imipramine and bilayer of the SC, along with 5% terpene (limonene). This gave promising
OA Labrasol Plural oleique Limonene Water [153]
doxepin results, showing both the local and systemic analgesic activity of the drugs.
Low droplet size and disruption of SC lipid bilayer was the underlying
mechanism of penetration.
Droplet size reduction and positive charge NE binding to the skin are
Methylene
Vitamin E Tween 80 —- Water important influencers in the successful transdermal delivery of vitamin E [154]
chloride
from hyaluronic acid-based NE.
Using a CLSM to determine the depth of penetration for a capsaicin
loaded NE, fluorescence intensity was detected through all skin layers,
Tween 80 and Span
Capsaicin Oleoresin — — Water indicating the successful penetration of the NE formulation. Droplet sizes [165]
80
of 20 to 62 nm and the optimization of the HLB value of Smix were the
main contributors to such successful results.
Antifungal NE was developed and compared to formulation in the market
Amphotericin B NE showed greater penetration results than aqueous dye
Amphotericin B CPG8 LAB PEG 400 — Water [155]
and commercial product using CLSM. Results are attributed to NE
components ability to enhance transdermal delivery through SC.
An improvement of 1.72-fold in the AUC was reported with the carvedilol
Carvedilol OA and IPM Tween20 Carbitol — Water NE over the oral formulation. Such results were mainly attributed to the [156,157]
optimum droplet size and low viscosity.
The synergetic properties of the NE components contributed towards the
Meloxicam Caprylic acid Tween 80 PEG 400 — Water enhanced penetration of MLX into the skin, altering the tightly packed [111]
nature of the SC.
Ethanol,
3,5-Dihydroxy- n-butanol, The usage of four types of co-surfactants and oil droplet nano-sizing were
IPM Cremophor EL 40 — Water [166]
4-isopropylstilbene n-propanol, what led to good in vitro drug release results.
1,2propanediol
A piroxicam NE was developed and incorporated into an emulgel. Even
though incorporating the NE into a gel form eased applicability to the skin,
Piroxicam OA Tween 80 Ethanol — Water [167]
flux was noticeably reduced compared to normal the NE. By using 35%
ethanol, low viscosity of the formulation was the leading mechanism here.
AUC: Area under the curve, CLSM: Confocal laser scanning microscope, CXB: Celecoxib, CPG8: Capmul PG8, EU: Eucalyptol, GLBD: Glibenclamide, HLB: Hydrophilic-lipophilic balance,
IPM: Isopropyl myristate, LAB: Labrasol, MLX: Meloxicam, NE: Nanoemulsion, OA: Oleic acid, PE: Permeation enhancer, PEG: Polyethylene glycol, SC: Stratum corneum, Tmax: The time
required to reach maximum plasma concentration.
 Despite follicular delivery not being the usual pathway of hydrophobic molecules, such drugs
when formulated in o/w NEs, accumulate near hair follicles. Fluorescent dye (Nile red, 0.1% w/w)
was incorporated in a capsaicin o/w NE to study skin penetration. Capsaicin is an interesting lipid
soluble active ingredient with multiple therapeutic uses, such as in cancer treatment, inflammation,
Sci. Pharm. 2019, 87, 17 19 of 34
and treating cardiovascular system (CVS) diseases. Being able to deliver capsaicin while bypassing
its first pass effect would definitely be of outmost importance. The optimized NE has shown deep
permeation
9.2. through
NE Mechanisms forpig
theskin layers.Transdermal
Enhanced Strong fluorescence
Delivery ofintensity wasDrugs
Hydrophobic witnessed across all of the skin
layers, where the tested NE showed red fluorescence deep to the dermis, reaching up to 700 μm. The
In many studies, NEs have proven to be a superior novel drug delivery system, not only due to the
strongest fluorescent spot was observed near the hair follicles. Capsaicin o/w NEs can penetrate and
advantages mentioned above (ease of preparation, cost effectiveness, and stability) but most importantly
accumulate in the skin well through the hair follicle [165]. Similar results were obtained from ME and
due to how efficient and effective NEs are in delivering hydrophobic drugs transdermally to the
demonstrated that o/w and not w/o ME is a promising vehicle for the transfollicular delivery of the
systemic circulation [168]. Several mechanisms have been proposed by various studies, as mentioned
lipophilic adapalene [200].
below and presented in Figure 3.
.

Figure 3. Mechanisms of transdermal enhancement of hydrophobic drugs from NE: (A) Disruption of
lipid bilayer of the SC. (B) Enhancement of transdermal permeation through oil droplet nano-sizing.
(C) Binding of positively charged NE to negatively charged skin. (D) Changing drug partition into
skin layers. (E) Hydrating skin and the dilation of the SC intercellular channels. (F) Changing the
permeation pathway of lipophilic permeants to follicular delivery with an o/w NE.

9.2.1. Disruption of the SC Lipid Bilayers

The hydrophobicity of the SC bilayer allowed the entry of PEs and surfactants containing fatty
acids. They disrupt the bilayer by creating separate domains, inducing highly permeable pathways
in the SC [37]. The enhanced skin permeation and penetration of meloxicam (MLX) from the NE gel
was reported, which can be attributed to its compositions in which the ingredients such as caprylic
acid (short-chain fatty acid), Tween 80, and PG act as permeation enhancers, potentially significantly
reducing the barrier properties of the SC through the extraction of SC lipids [111]. In contrast, the very
low permeation rate of MLX from the Tween 80 and propylene glycol solution across hairless mouse
skin over 36 h was reported [169]. The difference in the results here may be attributed to the fact that
the NE components act synergistically and significantly disrupt the barrier properties of the SC [170].
MLX transport might be attributed to the formation of the pores in the lipid bilayer, therefore creating
an increased effective volume inside the SC lipid domain available for drug diffusion [171].
Sci. Pharm. 2019, 87, 17 20 of 34

From the improved penetration results of the caffeine and naproxen NEs, it was hypothesized
that OA and eucalyptol act as enhancers that disrupt the SC of the skin through the dissolution of
lipids layers, allowing the active compound to pass through [164].
The fennel essential oil NE showed high potential for reducing the plasma glucose levels of rats
when delivered via the transdermal route, due to the presence of OA and PEG, as both of them act
as PEs for dermal delivery, since they increase the fluidity of the liquid portion of the SC. Moreover,
Tween 20 (a surfactant) enhanced the flux of the materials permeating through biological membranes,
resulting in the better penetration of oil and hence improving its therapeutic activity [172].
An enhanced permeation profile of the lipophilic celecoxib (CXB) in an o/w formulation was
reported [20]. This result was explained by the disruption and induction of a temporary denaturation
state of the keratin filaments in the SC in its various layers, resulting in a transport pathway in the lipid
bilayers. An optimized NE was prepared by dissolving 2% w/w of CXB in a 10% w/w combination
of Sefsol 218 and triacetin (1:1). Then, a 50% w/w mixture of Tween 80 and Transcutol P was slowly
added in the oil phase.

9.2.2. Enhancement of Transdermal Permeation through the Nano-Sizing of Oil Droplets

In order to investigate the effect of the oil droplet size on transdermal permeation, 1% curcumin
was added to an o/w NE. Two homogenization techniques were applied, using either a high-speed
homogenizer adjusted at 24,000 rpm or a high-pressure homogenizer accustomed to 1500 bar.
The high-speed method resulted in a PS around 600 nm, while the high-pressure technique resulted
in a rather smaller size (less than 100 nm). Studies have revealed that anti-inflammatory activity is
superior with a smaller NE droplet size [173]. In addition, small droplets have a better chance to
adhere to membranes, along with covering a large surface area, transporting bioactive NE compounds
in a more controlled way [37]. A larger droplet size with rigid architecture may not readily penetrate.
The small droplet size of the NE also provides a very large surface area for drug transfer into the
skin [111].
An anti-inflammatory compound, 3,5-dihydroxy-4-isopropylstilbene (DHPS), was integrated into
an o/w NE, dramatically improving the transdermal effect of the DHPS. The obtained enhancement was
explained by the NE providing better dispersion and a larger contact surface area for the drug molecules,
compared to drug suspension, leading to elevated solubility and permeability [166]. Reducing the
droplet size from large (5 microns) to sub-micron emulsion (100 nm) increased diazepam transdermal
penetration. This submicron size resulted in satisfactory drug delivery for up to 6 h, with only one
single topical application of diazepam cream onto mice skin [174]. A possible explanation of the
enhanced transdermal delivery of lipophilic progesterone and hydrophilic adenosine compared to
the drug solutions was the large surface area of the NE, which is associated with the low interfacial
tension and the small droplet size [40].
Glibenclamide was incorporated into a transdermal NE gel, due to its low oral bioavailability
and its severe and fatal hypoglycemic potential. A NE gel was formulated successfully and tested for
compatibility and release. The results of the study indicated that NE gel formulations enhanced the
drug permeation, and this was attributed to nano-sizing [163]. The large surface area was one possible
explanation for this, however, the authors agreed that the role of droplet size was less important than
previously assumed by other authors [175,176]. In another study, data revealed that a negatively
charged NE with a smaller PS (136 nm) had a higher in vitro release and increased penetration when
compared to the positively charged formulation (157 nm). The results were explained by the fact that
when the particles are smaller, the surface and the solubility pressure of the particles is bigger [177,178].
Su et al. experimented with the effective parameters for the PS of an octyldodecanol NE.
They suggested that the water percentage played the biggest part on NE droplet size, and that the
more water content, the bigger the droplet size. In their opinion, the effective parameters for droplet
size are the mixing rate (higher rotation speeds resulted in a smaller oil droplet size), temperature,
and NE surfactants used in the NE [179].
Sci. Pharm. 2019, 87, 17 21 of 34

9.2.3. Binding of the Positively Charged NE to Negatively Charged Skin

Aiming to evaluate the effect of charge on skin penetration, fludrocortisone acetate and
flumethasone pivalate NEs were experimented with, with a pre-impregnated aqueous phase of
either a negative or positive charge. Observing the charge effect through release studies revealed that
the positively charged NE was more efficient in transporting the active molecules through skin than
the negatively charged one. The explanation for this enhanced ability is the binding of the positive NE
onto the negatively charged skin, leading to an increase in contact time which eventually allows the
NE oily core to pass steadily successfully pass through the various skin layers [180]. The inclusion of
a positive charged carrier in the formulation and the subsequent adherence to the negative charged
skin was aimed to increase the NE retention time and consequently the drug bioavailability [163].
The skin accumulation of miconazole (MCZ) from positively charged droplets is higher than that from
negatively charged ones. This is attributed to the binding affinity of droplets to negatively charged
skin. Researchers proposed that positively charged vehicles appear to be promising for topical drug
delivery [181]. NEs containing chitosan were demonstrated to be more adequate as a vehicle for
substances intended to act on the skin, such as sunscreens, as photoprotective products are topically
administrated and they should not permeate through the skin. Since chitosan is positively charged,
with a degree of deacetylation of 79%, it could explain the attractive interaction of the polymer with
the skin which is negatively charged, thus preventing skin permeation [182].
Regarding stability, the positive surface charge of nanodroplets induced by phytosphingosine and
the pH provided good stability for the NE, as the repulsive forces prevented the collision of NE droplets
by a steric shield [183,184]. Regarding skin permeation, it has been shown that a positively charged
NE significantly promoted the diffusion rate of both econazole and MCZ nitrate through rat skin when
compared to the negatively charged preparations. This was done through increasing the amount
of cumulated drug and consequently the transdermal flux [143]. On the other hand, the increased
penetration of the positively charged NE is explained by the increased interaction and adsorption of
particles with the negatively charged keratinized corneocytes of the SC, the main barrier of the skin.
Here, the results showed that there is a clear relation between the NE charge and skin penetration,
however, the authors revealed that there was no detectable skin permeation due to drug accumulation
in the skin layers [177,178]. Finally, we can conclude from the literature that NE charge influence is
more relevant than droplet size regarding skin retention, while the droplet size effect dominates in the
context of skin permeation.

9.2.4. Enhancement Transdermal Permeation by Reducing the NE Viscosity

The successful passage of the drug through the SC was related to NE’s low viscosity, along with
the presence of an aqueous portion in the formulation. Researchers observed the impact of viscosity
when they compared their NE with a gel formulation, hypothesizing that low viscosity, working in
conjunction with other factors, such as droplet size, increases the contact area with skin, resulting in
easier penetration [163].
The permeation enhancement of celecoxib (CBX) was attributed to the NE’s low viscosity and
formulation stability, where both the oil phase and the water phase work in conjunction, leading to
more efficient drug permeation compared to conventional gel [160]. A piroxicam NE exhibited better
skin flux compared to the NE gel preparation. The obtained results were attributed to the low viscosity
of the NE, which facilitates the passage of NE components through the SC [167]. The small globule size
of the terbinafine NE and its low viscosity were the main factors for enhanced drug permeation [110].
As the proportion of the Carbopol 934 matrix increased in the terbinafine NE gel, the drug permeation
rate declined, along with increased drug deposition in the epidermis and dermis. Here, the authors
concluded that increasing the NE viscosity tends to change the formulation from transdermal delivery
to topical delivery [185]. Similar results were obtained for valsartan transdermal permeation in
Carbopol gel, where the gelling agent reduced drug penetration due to the transformation of the NE to
a highly ordered nanostructure [186].
Sci. Pharm. 2019, 87, 17 22 of 34

A lower inulin skin permeation rate was detected from micellar formulations, regardless of the
surfactant composition. The given explanation for this was that the high viscosity of all micellar
systems possibly played a role in lowering the permeation rates of inulin across hairy mouse skin into
the Franz cell receiver compartment [130].

9.2.5. Changing the Drug Partition into Skin Layers

Scientists compared the difference in penetration profile between the copaiba oil NE with Tween
20 as the surfactant and the oil alone by measuring the amount of the lipophilic drug β-caryophyllene.
Upon conducting permeation and retention studies, it was found that when copaiba oil was used
alone, the lipophilic drug was only found in the SC layer, failing to penetrate into the epidermis and
dermis, while the copaiba oil NE was able to be detected successfully in the SC, epidermis, and dermis,
demonstrating NE’s inherent ability to provide a more efficient penetration and retention in skin layers
than the drug alone [187]. A study researched the importance of ME structure on drug uptake into the
skin. In particular, cyclosporine skin retention data suggested that the surfactant type, co-surfactant
type, water content, and the presence of a rheological modifier could enhance drug mobility and
modify its tendency to partition into the SC [188]. Most likely, both o/w and w/o MEs are valuable
systems that preferentially retain the drug into skin, but the mechanism still requires more studies and
explanations [189].

9.2.6. Hydrating the Skin and Dilating the SC Intercellular Channels

Hydration disrupts the SC structure. Overall, 15% to 20% of the SC weight is composed of
water [190]. Skin hydration leads to SC swelling and the opening of its tight brick structures,
consequently increasing permeability [38]. It has been reported that extended skin hydration makes the
intercellular spaces distend, dilating the lacunar networks that link the connections of the water pool
system in SC interstices, which are disconnected under normal conditions [191]. Actually, the existing
water in the intercellular lipid region act as water pools and are not continuous toward the deeper layers
of the SC. However, small channels are formed when the skin is hydrated, reducing the diffusional
resistance through the rough SC structures. Both lipophilic and hydrophilic compounds benefit from
this feature. Small hydrophilic molecule penetration enhancers such as azone seem to benefit the most
from hydration, regarding absorption in corneocytes at least [192]. Even though such a mechanism
seems to increase both the penetration of both hydrophilic and lipophilic compounds, researchers
argue that hydrophilic compounds may not benefit greatly from skin hydration, since the formation of
such water pool channels in the SC is short lived and can only aid the lipophilic compounds in o/w
NEs. They explained that such NEs containing water as an external phase saturate SC layers faster
and hence lead to an enhanced permeability into the dermis [65,193]. In addition, the aqueous part of
NE enters through the appendageal pathway and enlarges the interlamellar volume of the SC lipid
bilayer, allowing the lipophilic drug to pass more easily through the various sheets of the SC [173].
An additional mechanism was suggested, where hydration and increasing water content accelerates
permeant thermodynamic activity and consequently penetration through the skin [194].
These observations suggested that the transport properties can be regulated by the water content
in the SC [195]. The NE components may change the water gradient in upper skin layers by
avoiding evaporation, exhibiting an occlusive effect, and consequently affecting skin permeation [196].
Fourier transform infrared spectroscopy detected that by increasing water concentration in MEs,
the ratio of the amide I/II band gradually increases, suggesting an increase in the hydration of the
SC [197,198]. In general, increased tissue hydration appears to increase the transdermal delivery of
both hydrophilic and lipophilic permeants [199].

9.2.7. Changing the Permeation Pathway of Lipophilic Permeants to Follicular Delivery

Despite follicular delivery not being the usual pathway of hydrophobic molecules, such drugs
when formulated in o/w NEs, accumulate near hair follicles. Fluorescent dye (Nile red, 0.1% w/w)
Sci. Pharm. 2019, 87, 17 23 of 34

was incorporated in a capsaicin o/w NE to study skin penetration. Capsaicin is an interesting lipid
soluble active ingredient with multiple therapeutic uses, such as in cancer treatment, inflammation,
and treating cardiovascular system (CVS) diseases. Being able to deliver capsaicin while bypassing
its first pass effect would definitely be of outmost importance. The optimized NE has shown deep
permeation through pig skin layers. Strong fluorescence intensity was witnessed across all of the
skin layers, where the tested NE showed red fluorescence deep to the dermis, reaching up to 700 µm.
The strongest fluorescent spot was observed near the hair follicles. Capsaicin o/w NEs can penetrate
and accumulate in the skin well through the hair follicle [165]. Similar results were obtained from ME
and demonstrated that o/w and not w/o ME is a promising vehicle for the transfollicular delivery of
the lipophilic adapalene [200].

10. Conclusions
Many hydrophilic and hydrophobic drugs were successfully loaded in w/o or o/w NEs, respectively,
demonstrating their ability to pass into general circulation. Accordingly, the NE components have
to be optimally selected to suit the drug psychochemical properties and simultaneously predict NE
penetration behavior through the skin.
For the first time, different penetration mechanisms of each NE type have been explored and
studied. For hydrophilic drugs, different mechanisms have explained their mode of penetration
for w/o NEs. The facilitated transport of these permeants was attributed to the effect of the NE
in increasing the drug thermodynamic activity or modifying the surface electrical charge of ionic
drugs. The same enhancement was sometimes explained by the solubilization of sebum by the NE
components, facilitating the follicular delivery of hydrophilic drugs. Most of these mechanisms focus
more on the nature of the NE than the physicochemical properties of the drug. However, other
transdermal strategies focus on permeant properties, namely either large or small Mw water-soluble
molecules loaded into w/o NEs, triggering follicular delivery, the normal dominant pathway of
hydrophilic permeants.
On the other hand, the penetration of hydrophobic drugs focuses more on the effect of the
NE components on the skin barrier. The disruption of the lipid bilayer of the SC was still the
most popular mechanism of transdermal enhancement, despite the reported enhancement through
oil droplet nano-sizing and reducing viscosity during the preparation of the NE. The binding of
a positively charged NE to negatively charged skin was reported as a controversial mechanism, as‘some
researchers ascribed the acceleration in penetration to this mechanism while others revealed that
it causes an increase in drug accumulation and retention in skin layers, and hence a reduction in
permeation. Hydrating the skin and dilating the SC intercellular aqueous and lipid channels were
reported to enhance the permeation of lipophilic permeants. Moreover, changing the permeation
pathway of lipophilic permeants to follicular delivery using o/w NEs was reported, despite the
well-known facts that oily drugs pass through the lipid pathway and that the area of the pore pathway
is neglected. The question is: Do we need to incorporate lipophilic drugs in o/w NEs? Does changing
the pathway to hair follicles reduce the transdermal passage of lipophilic permeants? We suggest that
o/w NEs have low potential in enhancing the permeation of hydrophobic molecules, therefore, further
studies are needed.
Most likely, the overall combination of such different mechanisms is the real cause of the skin
penetration enhancement effect for both types of permeants. It seems that no one specific mechanism
could provide a satisfying explanation for the superiority of NEs compared to other formulations.
Finally, the mechanisms reported in the literature are highly diverse, to the extent that a definite
mechanism is not very conclusive, highlighting the need for further investigation.
The compatibility with a wide range of hydrophobic and hydrophilic drugs, depending on its
type (o/w and w/o), makes NE a very encouraging transdermal dosage form. The diverse preparation
techniques, high drug encapsulation capacity (either in oil or water droplets), stability, safety, and
cost effectiveness motivate researchers to carry out further studies. This review suggests future
Sci. Pharm. 2019, 87, 17 24 of 34

research should be focused on more understanding of the penetration mechanisms, the development of
a ready-to-use NE, and targeting the CNS via a transdermal NE. From the literature, NEs are considered
a useful tool for by passing the P-glycoprotein (P-gp) pump through the general ‘masking’ concept
of the active agents, where the agents prevent the droplets from being discovered by the transporter
while they enter the blood brain barrier. Moreover, surfactants such as polysorbate 80 (P80) in NE
droplets are well-known P-gp inhibitors.
Despite all the trials made in understanding the mechanisms of how NEs are able to successfully
transport hydrophilic and hydrophobic compounds through skin layers into the systemic circulation and
the various proposed mechanisms hypothesized and summarized in this review article, more focused
research is needed to fully study, confirm, and understand the exact interaction that happens between
the skin layers and NE components through physical observation and real time monitoring of the NE
penetration of the skin. In other words, the drug molecule pathway through the SC lipoid or pore
domain needs to be fully understood.
Certain NE components seem to show a wide compatibility range with many drugs, such as OA,
labrasol, ethanol, and Tween, as seen in the publications shared in this review. This could be developed
into a concept of formulating a general-purpose NE carrier that is pre-formulated and manufactured as
a ready-to-use NE, allowing formulators to only add the active constituent. The industrial development
and production of NEs could be also very promising, being a time-saving and cost-effective process
based on the simplicity in NE preparation as well as the cheapness of their constituents. Within a
specific HLB range, pharmacists would be able to prepare transdermal NEs for patients who show
low compliance or inability to tolerate the side effects associated with taking the drug orally. The low
toxicity of many of NE components makes this a very promising concept for future work in transdermal
delivery systems.
A lot of research is still required for the development of novel NE formulations, especially in
drugs that target the central nervous system. Such drugs are most often very lipophilic in nature,
passing through the blood brain barrier to exert their action and usually requiring patients to take
them chronically for months or years. This drug group is also extensively metabolized by the liver.
Delivering these drugs transdermally, through bypassing the hepatic metabolism, sustains a steady
plasma concentration and potentially provides a more localized transdermal application for a fast
onset of action.

Funding: This research received no external funding.

Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations
ALA 5-aminolevulinic acid
CVS cardiovascular system
DHPS 3,5-Dihydroxy-4-isopropylstilbene
DMSO Dimethyl sulfoxide
EIP emulsion inversion point
GIT gastrointestinal tract
HLB hydrophile-lipophile balance
IPA Isopropyl alcohol
IPM Isopropyl myristate
MCZ Miconazole
ME Microemulsion
MLX Meloxicam
MPa Mega Pascal
Mw molecular weight
NE Nanoemulsion
NMP N-methyl pyrrolidone
Sci. Pharm. 2019, 87, 17 25 of 34

NSAIDs Non-steroidal anti-inflammatory drugs

o/w oil-in-water
OA Oleic acid
PE Permeation enhancer
PEG Polyethylene glycol
PIT Phase inversion temperature
PS Particle size
psi Pounds square inch
RHCL Ropinirole hydrochloride
SC Stratum corneum
SLS sodium lauryl sulfate
Smix surfactant/co-surfactant mixture
TCC Thiocolchicoside
CBX Celecoxib
w/o water-in-oil

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