CHAPTER 1

INTRODUCTION
 CHAPTER 1

1 Introduction
1.1 Colloidal drug delivery systems 1
1.2 Microemulsions
1.2.1 Introduction 3
1.2.2 Difference from emulsion, 5
1.2.3. Advantages of Microemulsion based systems 5
1.2.4 Disadvantages of Microemulsion Based Systems 7
1.2.5 Microemulsion formation and phase behaviour 7
1.2.6 Formulation Consideration 14
1.2.7 Method of preparation 16
l

1.2.8 Microemulsion characterization 17

L2.9 Applications of Microemulsion 18
1.3 Acne
1.3.1 Introduction 19
1.3.2 Pathophysiology of Acne 20
1.3.2 Treatment 26
1.4 References 31
1. INTRODUCTION
1.1 COLLOIDAL DRUG DELIVERY SYSTEMS
With high development costs of new therapeutic molecules, there is an
increasing trend for newer drug delivery formulations to facilitate the
controlled release of existing therapeutic molecules. Among many drug
delivery systems (DDS), the colloidal drug delivery systems are
becoming more interesting since they enable the release of drugs to be
controlled and/or bioavailability improved. When developing these
formulations, the goal is to obtain systems with optimized drug loading
and release properties, long shelf life and low toxicity (1, 2). Colloidal
science is often defined as the science of materials with size less than
microns. In terms of drug delivery, the flexibility in the internal structure
and surface has led to the adaptation of the colloidal drug carriers as the
platform for wide range of existing products as a means to achieve new
delivery modes for existing drugs and improve their therapeutic profiles.
The benefits of using colloidal drug delivery systems (CDDS) lies in the
fact that the internal structure of the particle is important, as the carrier is
often used to solubilize the lipophilic drugs that have limited water
i

solubility and are otherwise difficult to formulate. From

biopharmaceutical perspective, they offer the opportunity to design
features into the surface of the carrier to enable it to transfer through
biological barriers. The benefits associated with the CDDS includes
solubilization, increased dissolution rates, protection of labile agents,
controlled and sustained release,; high surface area, targeting,
extravascular and transdermal transport etc. CDDS are increasingly
providing the pharmaceutical industry with solutions to formulation and
clinical problems, & opportunities for line extensions for existing drugs

i
through improved therapeutic outcomes or alternative routes of
administration (3).
The fundamentals of a successful pharmaceutical formulation are to
enable delivery of the active substance to the target organ at
therapeutically relevant levels, with negligible discomfort and side effects
to the patient. In this respect, the route of administration is of major
influence. Topical administration offers several attractions compared to
the traditional routes. However, the main indicator for topical
administration is when the skin itself is the target organ. Despite the
substantial potential of transdermal and dermal drug delivery, only
relatively few drugs are yet commercially available as topical
formulations. The main limitation lies in the barrier function of the skin,
which is considered one of the most impermeable epithelia of the human
body to exogenous substances. Therefore, the major challenge for topical
formulations today is to provide a sufficient increase in drug penetration
into the skin, without inducing significant irreversible alterations to the
skin barrier function. Penetration enhancement with special formulation
approaches is mainly based on the usage of colloidal carriers. Colloidal
carriers have attracted the main interest because they are promising
systems having localized effect. The carriers accumulate in stratum
comeum or other upper skin layers are not expected to penetrate into
viable skin. The common characteristic of all colloidal carriers is the
submicron sized particles which are intended to transport entrapped
active molecules into the skin (4).
A few of the most widely examined ODDS are micelles, emulsions (often
termed as macroemulsions), microemulsions, liposomes, niosomes and
nanoparticles. Recently, much attention has been given to the use of
microemulsions for topical and transdermal delivery as they show
excellent biocompatibility and capability of incorporating drugs of

2
varying solubility. Microemulsions represent pharmaceutically versatile
formulations for drug delivery to and through the skin. This novel vehicle
I
has a potential of increasing percutaneous delivery of both hydrophilic
I
and lipophilic drugs compared to conventional formulations (5,6,7, 8).
I
A typical literature search using the keyword “microemulsion” would
return more than 1000 references perjyear in the past decades. Numerous
books and reviews appeared concerning the structure of microemulsions,
their phase behavior and applications. Hence, it is obvious that everything
known about microemulsions cannot ibe said within a reasonable number
of pages. In the following a selection has been made describing only the
most important facts and concepts having an intersection with the topic of
the thesis.
1.2. MICROEMULSIONS
1.2.1 Introduction
The concept of microemulsion was introduced, as early as 1940, by Hoar
I

and Schulman who generated a clear, single-phase solution by titrating a

milky emulsion with hexanol (9). Schulman and coworkers (1959) coined
the term “microemulsion” (10) and it has since been defined and indeed
redefined on many occasions. Different workers have defined the term
differently. Danielsson and Lindman, in 1981, described microemulsion
as “a system of water, oil and amphiphile which is a single optically
isotropic and thermodynamically I stable liquid solution” (11).
Microemulsion, as defined by Attwood, is a system of water, oil,
surfactant and co-surfactant, which | is a transparent, single optically
isotropic, and thermodynamically stable liquid (12).
In other words “Microemulsions are I liquid dispersions of water and oil
that are made homogenous, transparent (or translucent) and
thermodynamically stable by the addition of relatively large amounts of a

3
 surfactant and a cosurfactant and having diameter of the droplets in the
range of 100- 1000 A0 (1 - 100 nm).”(Fig 1 and 2)

MICROEMULSION
Hydrophilic
Hydrophobic Phase
Phase

Cosurfactant
Surfactant
Surfactant: Forms the interfacial film
Cosui factnat: Ensures flexibility of interfacial layer
Reduces the interfacial tension

Figure 1. Microemulsion

Bicontinuous microemulsion

Figure 2. Microemulsion structures.

They appear to represent a state intermediate between thermodynamically

stable solubilized solutions i.e. micelles containing solubilized oils and
ordinary' emulsions which are relatively unstable (13).
Three types (Fig 3)
1 O/W Microemulsion
2 W/O Microemulsion
3 Bicontinuous Microemulsion

4
 Figure 3. OAV, W/O and Bicontinuous Microemulsions.
1.2.2. Difference from emulsion
The term microemulsion implies a close relationship to ordinary emulsion
(macro emulsion). Microemulsion state embraces a number of different
microstructures, most of which have little in common with classical two-
phase emulsions. Microemulsions are readily distinguished from normal
emulsion by their particle size, transparency, low viscosity and more
fundamentally, their thermodynamic stability. Although systems
containing low concentration of dispersed phase (less than 25%) may,
like emulsions, be composed of droplets of either oil dispersed in water
(o/w) or water dispersed in oil (w/o), they are essentially stable, single
phase swollen micellar solution rather than unstable two phase
dispersions (14).
1.2.3. Advantages of Microemulsion based systems (14)
(a) Improvement in oral bioavailability: Dissolution rate dependence
absorption is a major factor that limits the bioavailability of poor water-
soluble drugs. Microemulsion is a novel approach to improve the water

5
solubility and ultimately, bioavailability of lipophilic drugs.
Microemulsion presents the drug to gastrointestinal tract in solubilised
and mieroemulsified form; subsequent increase in specific surface area
enables more efficient drug transport through intestinal aqueous boundary
layer and through the absorptive brush border membrane leading to
improved bioavailability.
(b) Reduction in Inter-Subject and Intra-Subject Variability: There are
several drugs which show large inter-subject and intra-subject variation in
absorption, leading to decrease performance of drug and patient non-
compliance. Microemulsion drug delivery system is a proven approach to
overcome inter and intra subject variation.
(c) Reduction of Food Effects: Food is a major factor affecting the
therapeutic performances of the drug in the body. There are several
research papers specifying that performance of microemulsion is
independent of food and that microemulsion offer reproducibility of
plasma profile.
(d) Ease of Manufacturing and Scale Up: This is one of the most
important advantages that makes microemulsion a unique delivery system
when compared to others like solid dispersions, liposome, nanoparticles
etc. dealing with improvement of bioavailability. Microemulsion requires
very simple and economical manufacturing facilities like simple mixer
with agitator and volumetric liquid filling equipment for large-scale
manufacturing. This explains the interest of industry for microemulsion
based drug delivery systems.
(e) Ability to Deliver Peptides that are Prone to Enzymatic Hydrolysis in
GIT: One unique property of microemulsion is the ability to deliver
macromolecules like peptides, hormones, enzyme substrate and inhibitors
and the ability to offer protection from enzymatic hydrolysis.

6
(f) No Influence of Lipid Digestion Process: Unlike other lipid-based
drug delivery systems, the performance of microemulsion is not
influenced by the lipolysis, emulsification by the bile salts, action of
pancreatic lipases and mixed micellar formation. Microemulsions are not
necessarily digested before the drug is absorbed, as they present the drug
in micrpemulsified form, which can easily penetrate the mucin, and water
unstirred layer.
(g) Improvement of bioavailability of antifungal and anti-inflammatory
drug by topical microemulsion.
(h) As Solid Dosage Form for Oral Administration: Microemulsion can
be converted into the various solid dosage forms by adsorbing onto the
solid surface.
1.2.4. Disadvantages ofMicroemulsion Based Systems (13)
1 Use of a large concentration of surfactant and cosurfactant
necessary for stabilizing the nanodroplets.
2 ’ Limited solubilizing capacity for high-melting substances
3 The surfactant must be nontoxic for using pharmaceutical
applications
4 Microemulsion stability is influenced by environmental
parameters such as temperature and pH. These parameters
change upon microemulsion delivery to patients.
1.2.5 Microemulsion formation and phase behaviour
Theories ofmicroemulsion formation (15, 16)
Different theories have been put forward to explain the formation and
stability of microemulsion. The three widely accepted theories include
mixed-film theories, solubilization theories, and thermodynamic
treatments.
Mixed-Film Theories

7
These theories emphasized the formation of interfacial film and ultralow
interfacial tension. The spontaneous formation of microemulsion droplets
was considered to be due to the formation of a complex film at the oil-
water interface by the surfactant and cosurfactant resulting in the
reduction of oil-water interfacial tension to very low values
(approximating zero to negative). The mixed film as per the theory was
assumed to be liquid and duplex in nature (having different characteristics
at the oil and water faces) with a two-dimensional spreading pressure m
which in turn influenced the interfacial tension Ai as per the following
equation.
Ai = Xo / w- ni
where Ao / w denotes oil/water interfacial tension in the absence of the
film. Due to absorption of large amounts of surfactant and co- surfactant,
the spreading pressure may become larger than Ao / w resulting in a
negative1 interfacial tension and thereby providing the energy to increase
the interfacial area. This increase in interfacial area causes reduction in
droplet sizes. The production of negative interfacial tension is a transient
phenomenon.
Solubilization Theories
These theories considered microemulsion to be thermodynamically stable
monophasic solutions of water-swollen (w/o) or oil-swollen (o/w)
spherical micelles. For example, while explaining the relationship
between;oil-in-water microemulsion and the isotropic aqueous micellar
region, the solubilization of oil in normal micelles was found to be small
and concentration of all the component were found to be critical for an
aqueous imicelle to solubilise large amounts of hydrocarbon and swell
directly into oil droplet without forming a large number of intermediate
structures of low curvature.
Thermodynamic Treatments

8
According to the theory, the process of formation of oil droplets form a
bulk oil .phase is accompanied by an increase in the interfacial area AA,
and hence an interfacial energy yAA. The entropy of dispersion of the
droplets is equal to TAS and hence the free energy of the formation of the
system is given by the expression:
AGf = yAA - TAS
Where AGf is the free energy of formation, y is the surface tension of the
oil-water interface and T is the temperature. Formation of microemulsion
is accompanied by a very large change in AA due to the formation of
large number of very small droplets. During the formation of a
microemulsion, the value of y remains positive all the times as against
believed in the earlier theories, but it is very small and hence can be
easily offset by the entropic component. The entropic component
becomes dominant due to the large dispersion entropy arising from the
mixing of the one phase in the other in the form of large number of small
droplets.
Thus, a negative free energy of formation is obtained and hence in such
cases the process of microemulsification is spontaneous and the resulting
system is thermodynamically stable.
Phase Behaviour (13)
The phase behaviour of simple microemulsion systems composing oil,
water and surfactant can be studied with the aid of ternary phase diagram
(at fixed pressure and temperature) in which each comer of the diagram
represents 100% concentration of the particular component.
Generally, pharmaceutical microemulsions contain additional
components such as a cosurfactant and/or dmg. The co-surfactant is also
amphiphilic with an affinity for both the oil and aqueous phases and
partitions to an appreciable extent into the surfactant interfacial
monolayer present at the oil-water interface.

9
A large number of drug molecules are by themselves surface active and
they are expected to influence phase behaviour. For four or more
components, pseudo ternary phase diagrams are used to study the phase
behaviour. In this diagram a comer will typically represent a binary
mixture of two components such as surfactant/co-surfactant, water/drug
or oil/drug. The number of different phases present for a particular
mixture can be visually assessed.
Hypothetical Phase Regions of Microemulsion Systems

S
(Surfactant + cosurfactant)

Water Oil

Figure 4. Hypothetical Phase Diagram.

From above figure 4, we can see that,
> With oil concentration surfactant forms reverse micelles capable of
solubilizing water molecules in their hydrophilic interior.
> Continued addition of water in this system may result in the
formation of W/O microemulsion in which water exists as droplets

10
 surrounded and stabilized by interfacial layer of the surfactant / co­
surfactant mixture.
> At a limiting water content, the isotropic clear region changes to a
turbid, bireffingent one.
> Upon further dilution with water, a liquid crystalline region may be
formed in which the water is sandwiched between surfactant
double layers.
> Finally, as amount of water increases, this lamellar structure will
break down and water will form a continuous phase containing
droplets of oil stabilized by a surfactant / co-surfactant (O/W
microemulsions)
Microemulsion structure
Microemulsions are very interesting systems, because the oil + surfactant
+ water interface forms a wide variety of structures to avoid the direct oil
/ water contact (17).
Micelles are the simplest structures; spherical or cylindrical objects
formed by surfactant molecules, separating oil and water. Micelles (Fig
5A) are like drops of oil in water and reverse micelles (Fig 5B) are like
drops of water in oil.

REVESE
MICELLES MICELLES
11
 Figure 5. Micelle (A) and reverse micelle (B)
Another microemulsion structure is the Lamellae Water and oil
consecutive layers separated by surfactant layers conveniently oriented.
The lamellae structure (Fig 6) is similar to the smectic thermotropic
phase, it presents birefringence and maintains the order even at diluted
concentrations.

Figure 6. Lamellae Structure

The lamellae structure is related with Spherulite structure (onion
structure; Fig 7), its thermodynamic stability is still under study.

Figure 7. Spherulite Structure

12
The Bicontinuous Structure or sponge phase is a quite intricate structure.
Thinking that the sponge surface is the surfactant, a bicontinuous
structure (Fig 6) is shown below:

Figure 8. Bicontinuous Structure

Other microemulsion structures possible are:

Figure 9. Interconnected Rod-iike Micelles

Figure 10. Vesicles

13
The principal factors for explaining microemulsion structure changes are
1. Surfactant Shape
2. Entropie Energy Terms
3. Solvent Properties as Ionic Force and pH

1.2.6. Formulation Consideration:

Surfactant
The surfactant chosen must be able to lower the interfacial tension to a
very small value which facilitates dispersion process during the
preparation of the microemulsion and provide a flexible film that can
readily deform around the droplets and be of the appropriate lipophilic
character to provide the correct curvature at the interfacial region. It is
generally accepted that low HLB ! surfactants are favoured for the
formulation of w/o microemulsion, whereas surfactants with high HLB
(>12) are preferred for the formation j of o/w microemulsion. Surfactants
having HLB greater than 20 often require the presence of cosurfactants to
reduce their effective HLB to a value within the range required for
microemulsion formation (14).
Co-surfactant
In most cases, single-chain surfactants^ alone are unable to reduce the o/w
interfacial tension sufficiently to enable a microemulsion to form. The
presence of cosurfactants allows the interfacial film sufficient flexibility
to take up different curvatures required to form microemulsion over a
i

wide range of composition. If a single surfactant film is desired, the

lipophilic chains of the surfactant should be sufficiently short, or contain
fluidizing groups (e.g. unsaturated bonds). Short to medium chain length
alcohols (C3-C8) are commonly added as cosurfactants which further
reduce the interfacial tension and increase the fluidity of the interface
(14).

14
Oils
The oil component influences curvature by its ability to penetrate and
hence swell the tail group region of tlje surfactant monolayer. Short chain
oils penetrate the tail group region to a greater extent than long chain
alkanes, and hence swell this region]to a greater extent, resulting in an
increased negative curvature (and reduced effective HLB). Various long
and medium chain triglycerides like labrafac, lauroglycol, labrafil M
1944CS and olive oil, have also been reported (14).
Temperature
Temperature plays an important role jin the formation of microemulsion
when a non-ionic surfactant is used inimicroemulsion formulation. At low
temperature, non-ionic surfactants are hydrophilic and form o/w
microemulsion, but at higher temperature, they are lipophilic and form
w/o microemulsion. At an intermediate temperature, called the HLB
temperature, hydrophilic-lipophilic j interactions just balance and
microemulsions may have bicontinuous structure. The effect of
temperature in microemulsion containing non-ionic surfactants such as
polyoxyethylene alkyl ethers as the polyoxyethylene group is dehydrated
with increasing temperature. This has the effect of substantially altering
the packing ratio and in extremity, is manifested by phase separation or
phase inversion. Ionic surfactants are not strongly influenced by
temperature; a temperature rise doesl not cause an increase in positive
curvature due to counter ion dissolution (14).
Surfactant- Co-surfactant ratio
The surfactant and co-surfactant ratio is a key factor influencing the
phase properties. Attwood et al showed how size and location of
microemulsion is changed on changing the mass ratio of polysorbate 40 /
sorbitol from 1: 1 to 1: 3.5. Similar studies using polysorbate 80 and
Polysorbate 60 have shown a change in the optimum polysorbate /

15
sorbitol mass ratio (i.e., that producing the largest microemulsion region)
from 1: 2.5 for polysorbate 80 to 1: 2 for polysorbate 60 to 1: 1.5 for
polysorbate 40. Such effects were attributed to differences in the packing
of surfactant and co-surfactant at the oil/water interface. Gasco et al have
i
used microemulsions of similar composition prepared using polysorbate
60 but with phosphate buffer rather tlian water, to study the in vitro drug
release of propranolol (14).

1.2.7 Method ofpreparation

Titration Method
Appropriate quantities of surfactant, j co-surfactant and oil are weighed
and then mixed to form a monophasic; system (if required a little heat can
also be applied for blending oil and emulsifier). The mixture is then
titrated with double distilled water at constant temperature with mild
agitation until a clear, isotropic and tljermodynamically stable dispersion

with low viscosity is obtained. Volume of double distilled water required

to produce W/O microemulsions must be noted. The order of mixing for
O/W microemulsions is more flexible! One of the simplest methods is to
blend oil, emulsifier and co-surfactant and then pour this into water with
a little stirring (13).
Alternative to Titration Method
Appropriate quantities of surfactant, oil and water are taken and mixed
together to form milky emulsion. The mixture is then titrated with a
fourth component, the co-surfactant until the mixture becomes clear. If
more oil is added to a w/o system, the system becomes cloudy, but the
addition of more co-surfactant again gives a clean transparent
microemulsion.
Both O/W and W/O types are possible and may be converted one to the
other, by adding more of the internal phase or by altering the type of

16
emulsifier. As the internal phase is added, the emulsion will pass through
visco-elastic gel stage; with further addition, an emulsion of the opposite
type will occur (13).
Cook et ah reported a method of forming microemulsion which involved
forming a mixture of liquids so as to obtain a microemulsion-forming
liquid system and then dividing the mixture into two streams. Each of the
streams was then pressurized to a pressure of at least 4000 psi and
ejected.
1.2.8. Microemulsion characterization
The characterization of these systems is highly challenging due to their
small droplet size with fluctuating boundaries and complex structure.
Basic components in a physicochemical characterization of
microemulsion systems are:
1 Phase stability and phase behavior,
2 Microstructure, dimension (size and size distribution), shape and
surface\ features (specific area, charge and distribution),
3 Local molecular arrangements, interactions and dynamics.
Among these properties, particle size; interactions, and dynamics are of
fundamental importance since they control many of the general properties
of microemulsions. In particular, the size distributions of microemulsions
give essential information for a reasonable understanding of the
mechanism governing both the stability and penetration into the
membrane (18, 19). Many technologies like dynamic light scattering
(DLS) (20, 21), small angle neutron scattering (SANS) and small angle
X-ray scattering (SAXS) (24, 25, 26) as well as cryo transmission
electron microscopy (27,28, 29) and pulsed field gradient spin echo (self­
diffusion) NMR (30, 31, 32) has been in growing use in particle size
characterization. Other methods, e.g. electrokinetic chromatography,
conductance, viscosity, electrical birefringence, infrared spectroscopy and

17
calorimetry are also employed for investigating the internal
physicochemical states of microemulsions. Viscosity measurement can
indicate the presence of rodlike or worm-like reverse micelles while
conductivity measurement provides a means of determining whether a
microemulsion is oil-continuous or water-continuous as well provide a
means of monitoring phase inversion phenomena. Dielectric
measurements are a powerful means of probing both the structural and
dynamic features of microemulsion systems. Pulsed field gradient NMR
is also used extensively to measure self-diffusion coefficients of the
various components and yields information on the mobility and
microenvironment (33,34,35).
1.2.9. Applications ofMicroemulsion
Pharmaceutical Applications (14)
■ Parenteral delivery
* Oral drug delivery
■ Topical drug delivery
■ Ocular and pulmonary delivery
■ Perflouro microemulsions
■ Microemulsions in biotechnology
■ Solubilization of drugs in microemulsions
Other Applications (14)
■ Microemulsion in enhanced oil recovery
* Microemulsions as fuels
■ Microemulsions as lubricants, cutting oils and corrosion inhibitors
" Microemulsions as coatings and textile finishing
■ Microemulsions in detergency
■ Microemulsions in cosmetics
■ Microemulsions in agrochemicals
■ Microemulsions in food

18
 * Microemulsions in environmental remediation and detoxification
I
* Microporous media synthesis (inicroemulsion gel technique)
* Microemulsions in analytical applications
■ Microemulsions as liquid membranes
■ Novel crystalline colloidal arrays as chemical sensor materials

1.3. ACNE
1.3.1. Introduction
Acne is a chronic inflammatory dermatosis of the pilosebaceous unit
(PSU) and is characterized by several;abnormalities in sebum production,
follicular epithelial desquamation, bacterial proliferation, inflammation
and immunologic host reaction. Recent findings have shown the influence
of genetic factors in the follicular environment that optimize
Propionibacterium acne proliferation. Acne is the most common family of
skin disorders and in cases of extreme disfiguration can cause a great deal
of embarrasement and anxiety in affected patients. Acne accounts for a
tremendous number of office visits to both family physicians anf
dermatologists. According to the statistics, nearly 85% of people aged 12-
25 years old, approximately 8% of adults ages 25-34 years old and 3% of
adults aged 35-44 years old experience some degree of acne (36).
The earlier the symptoms start the more severe is the course of disease.
The prevalence of acne in adolscents can reach upto 80% and it produces
clinical conditions that can carry over into adulthood creating a negative
impact on quality of life. Acne vulgaris affects virtually all youngsters
beginning at the age of about 13 years and continuing through the age of
23 years. In females the onset may occur at an age as early as 10 or later
on in 20’s and 30’s and it has been speculated that hormonal influence,
stress and increased cosmetic consciousness all play a role. Although

19
acne is not a life-threatening disease, it has significant physical and
psychological ramifications such as permanent scarring, poor self-image,
social inhibition, depression, anxiety and suicidal tendency. Therefore,
acne should be regarded as a serious medical disorder (37).
1.3.1. Pathophysiology of Acne
Understanding the pathophysiology of acne can help the physician tailor
therapy to the individual patient. Acne lesions arise from pilosebaceous
units, which consist of sebaceous glands and small hair follicles. These
units are found everywhere on the body except the palms and soles.
Pilosebaceous density is greatest on the face, upper neck and chest, at
roughly nine times the concentration found elsewhere on the body.
Pilosebaceous units are present and active at birth as a reaction to
maternal hormones. Obstruction of the pilosebaceous canal is the primary
cause of acne and occurs because of a variety of factors (38).
Factors promoting the development of acne are as follow (fig-11).

20
 INCREASED SEBUM ♦I PROINFLA MALA.T ORY INFLAMMATION
HORMONES
1) Adrenal/Ovarian LIPIDS t IL-lcif, TN Fat, HLA-DRf,
Androgen Production T
Earl)' lymphocytic infiltration
led LINO LEI C CD4+> CDs*
Androgen
Testosterone < DHT ACID Langerhans cells HLA-DR+
Basal keratinocytes HLA-DR+
2) Sebocyte hormone
rx Comedones caontain IL-laT
Production Cell wallsubstances
DUCTAL

i l
0
n
%

o
1
u
IMPAIRED peptidogfycan polysaccharide
Steroil-5a-reductase T HYPER- fragments
EPIDERMAL CORNIFI CATION

\
Androgen receptor f BARRIER Macrophage activation: IL-St,
TNFat, ICAM -lt.
Cortkotrophin
releasing hormone Recruitment of
WnmELDvaMdoud neutrophils/leu cos into
Cortkotrophin INFLUX OF MDV epithelial wall of sebaceous
releasing receptors !, 2 WATER follicle
Complement activation
Neutrophilic hydrolases:
disruption of sebaceous follicale
Hypothesis KERATINOCUTES 1 Cytokine release Immune granuloma type
DAMAGE I reaction
Fact Humoral imm line responseT
Heat shock protein
Superan tigen s/Mitogen

Figure 11. Factors promoting the development of acne, (modified from ref-39)

21
Increased sebum production
Increased sebum excretion is a major factor involved in the
pathophysiology of acne. Sebum is a mixture of relatively nonpolar
lipids, most of which are synthesized, de novo by the sebaceous gland to
i

coat the fur as a hydrophobic protection against overwetting and for heat
insulation in mammals.The functions of the sebaceous gland, which are
possibly associated with the development of acne are (40):
• Production of sebum
• Regulation of cutaneous steroidogenesis
• Regulation of local androgen synthesis
• Interaction with neuropeptides
• Synthesis of specific lipids with antimicrobial activity
• Exhibition of pro- and anti-inflammatory properties
Ductal cornification
It is hypothesized that a local follicular deficiency of linoleic acid, effects
of IL-la and androgens as potential factors involved in follicular
hyperkeratinization causing an apparent early cornification of the
keratinocytes and scaling. Earliest changes in the hair follicle occur when
the follicular canal becomes blocked with abnormally keratinized
desquamating cells. This plug starts above the opening of the sebaceous
gland into the follicular canal and causes gradual expansion of cells and
i
sebum within the canal. The plug becomes visible at the skin surface as a
white papule ("whitehead," or closed comedo). If the opening of the
follicular canal dilates, this plug protrudes from the canal and turns a dark
color ("blackhead," or open comedo) (39).
Bacterial colonization of the pilosebaceous ducts: Acne is not an
infectious disease and, therefore, not contagious. Among the bacteria
species that colonize normal skin as resident flora, only those able to

22
colonize the follicular duct and multiply there can be pathogenic for acne.
Only three species of microorganism^ could therefore be associated with
the development of acne lesions: propionibacteria, coagulase-negative
staphylococci, and yeasts of the species Malassezia. However, acne did
not improve after antifungal treatment, so yeasts could not be associated
with the pathogenesis of acne. Staphylococci could also be excluded,
because these develop antibiotic resistance during the first weeks of
treatment in most patients and the numbers quickly rise. Scientific
interest has therefore been focused on propionibacteria. Propionibacteria
are Gram-positive, non-motile, pleomorphic rod-shaped cells that ferment
sugars to yield propionic acid as one of the end products in this metabolic
process. P. acnes are the predominant resident microorganism on
sebaceous gland-rich areas of skin in adults. On human skin,
propionibacteria can be found from; birth until death. Bacteriological
analysis and sebum production investigated in multiple body areas
demonstrated a high association between P. acnes levels and sebum
production. The pathogenicity of propionibacteria is thought to be due to,
first, the production of exocellular enzymes and other bioactive
exocellular products, which could act as virulence determinants, and,
second, on the microorganism's interaction with the immune system.
Propionibacteria resist phagocytosis and can persist intracellularly within
macrophages for prolonged periods (39).
Inflammation
Bacteria, most importantly P. acnes, are present in increased numbers in
persons who have acne. Much of the inflammation that eventually occurs
arises from the action of enzymes produced by the bacteria. These
enzymes hydrolyze sebum into free) fatty acids, which stimulate the
inflammatory process. Chemotactic factors are released by this reaction,
attracting neutrophils. As the follicular wall becomes inflamed, an

23
erythematous papule appears at the skin surface. With increased sebum
production, obstruction and bacterial colonization, the follicular unit
ruptures, spilling its.contents into the dermis. The inflow of neutrophils
causes the formation of pustules. Continuation of severe inflammation
leads to formation of nodules and subsequent cysts (41).
The different stages of acne are illustrated below (figure 12) :

Figure 12: Stages of acne. (1) Normal follicle; (2) open comedo
(blackhead); (3) closed comedo (whitehead); (6) papule; (5) pustule,
(adopted from ref- 41)
Acne may be triggered or worsened by external factors such as
mechanical obstruction (i.e., helmets, shirt collars), occupational
exposures, or medications. Common medications that may cause or affect
acne are listed in Table -1 (42).

24
Table 1. Medication that trigger or exacerbate acne.

More commonly Less commonly

Anabolic steroids (eg. Danzol Azathioprine

(Danocrine), testosterone)

Bromides Cyclosporine

Corticosteroids (eg. Prednisone Disulfiram

(Deltasone))

Corticotrophin (H. P. Acthar) Phenobarbital

Isoniazid (Nydrazid) Quinidine

Lithium Tetracycline

Phenytion (Dilantin) Vitamins Bi, Be, Bn, and D2

Visible signs of acne (41)

Comedones
An early acne blemish is caused by blocked pores, and at first the skin
does not look red and inflamed. These early blemishes are called open
comedones (blackspecks/blackheads) and closed comedones (whiteheads)
(Figure 13).

Comedones Papules & Pustules Modules (cysts or bolls)

(pimples, zits)
Black Head White Head Papule Pustule Nodule Cyst
Co*p*do (nflanaj Modus —-V
Fol'do ■» - me*m«d
’ 1 *•,■*—“ WfM
.. wCum , r- , 1 .Tnwpe*
MBum l ^. • nuu« iKtJ-,
-Duua
-yr-Kma* j liasue
-RiCUrt
-r-
oJMcT&L*^
...
V
. r, •—-Ke«*in r -— Karan KstaLn Kamn - IBM I • I 1<

7^ ■ L
PreWendon
afusacaoi*
l
ft
■raj pus

8i*nd

A B
Figure 13: Visible signs of acne, Comedomes (A), Papules & pustules
(B) and nodules (C).

25
Papules and pustules
As these early spots get larger and inflamed, they become papules and
pustules (pimples or zits).
Nodules
Very large and deep lumps can also develop in some people, these are
called nodules and cysts (like boils), and can be painful.
Oily skin
The sebum production increases so that your skin looks and feels oily.
Hyperpigmentation
After the inflammation subsides; the skin can be discolored by brown
acne stains (called hyperpigmentation) and damaged by scars. Acne scars
are common and may occur even in mild acne.
In 1990, the American Academy of Dermatology developed a
classification scheme for primary acne vulgaris. This grading scale
delineates three levels of acne: mild, moderate, and severe. Mild acne is
characterized by the presence of few to several papules and pustules, but
no nodules. Patients with moderate acne have several to many papules
and pustules, along with a few to several nodules. With severe acne,
patients have numerous or extensive papules and pustules, as well as
many nodules. Acne also is classified by type of lesion— comedonal,
papulopustular, and nodulocystic. Pustules and cysts are considered
inflammatory acne.
1.3.2. Treatment
Acne is a treatable disease, but treatment usually lasts for years.
Compliance is very important in the treatment and non-compliance is the
main reason for treatment failure. The aims of treatment of acne are to
improve the disfiguring the psychological distress and to prevent scarring.
The pathopysiological goal of acne treatment includes normalization of
keratiniation, the reduction of interfollicular P.acnes, the reduction of

26
 i

inflammation and the reduction of sebaceous gland activity. Our

therapeutic menu includes a variety of drugs, both topical and systemic,
that can affect areas of pathophysiology (42).
The main part of acne treatment uses the topical route. More than 50% of
acne patients belong to the group presenting with acne comedonica and
papulopustulosa. Topical treatment affects at least three of the four main
pathogenetic factors responsible for the development of acne, i.e.
hyperseborrhea, hyperkeratosis, microbial colonization and inflammation.
Topical therapy is inevitable in acne treatment and is mainly indicated in
the mild to moderate acne. In more severe forms, a combined topical and
systemic therapy is recommended. Table 1 takes account of various
topical agents that are available for acne treatment. The available topical
agents have a direct or indirect infuence on the pathogenetic factors and
are selected according to the predominant type of acne lesions (43).

27
Table: 2. Overview of topical antia cne agents.
CATEGORY AGENTS
Retinoids Tretinoin, isotretinoin, adapalene,
tazarotene, moltretinide, al l-trans-
i
retinoyl- p - glucuronide
a - Hydroxy and p - hydroxy acids Glycolic acid, mandelic acid, lactic
acid,
Salicylic acid, lipohydroxy acid
Antibacterial agents Erythromycin, azithromycin,
clindamycin,
nadifl oxacin, azelaic acid,
chloroxylenol,
sodium sulfacetamide, benzoyl
peroxide,
T-3912, tea tree oil, 5-amino
levulinic acid
Antiandrogens Cyproterone acetate
Miscellaneous Nicotinamide,
j
sodium ascorbyl
phosphate,
pyruvic acid, sulfur

Topical retinoids, as monotherapy, are mainly used in patients with non­

inflammatory comedones, in combination with other topical and systemic
drugs in mild, moderate and several inflammatory acne. The major
disadvantage of topical therapy for the treatment of acne is that many of
the agents used cause tolerability; problems. Significant erythema,
dryness, peeling, scaling, and irritation may result in discontinuation of
treatment, and in patients who continue treatment, irritation may lead to
compliance problems. It is increasingly been recognized by the
28
pharmaceutical industry that for these molecules drug delivery systems
(vehicle) play an important role (44).
With this in mind, pharmaceutical research programs have focused on
methods of minimizing the irritation caused by topical retinoids while
preserving or improving their therapeutic benefits. The results of these
programs have yielded several new developments. Novel drug delivery
strategies can play a pivotal role in1 improving the topical delivery of
antiache agents by enhancing their dermal localization with a
concomitant reduction in their side effects (43).
Novel Colloidal drug delivery systems for topical treatment
While conventional formulations containing lower concentrations of
antiacne agents can lead to less significant results, the novel delivery
systems present the potential to reduce the side effects without reducing
the efficacy. Therefore, with novel delivery systems outstanding
advantages can be noticed in comparison to conventional formulations.
The application of these systems to the skin distributes the topical agents
gradually and, in some cases, has demonstrated the ability to reduce the
irritancy of some antiacne drugs, yet it maintains a similar efficacy when
compared with conventional formulations. In addition to the controlled
release of the drug into the epidermis, these systems can also promote
follicular targeting, creating high local concentrations of the active
compounds in the PSU. The colloidal drug delivery systems have the
advantage of penetrating more efficiently into the hair follicles than do
non-particulate systems, such as conventional formulations, so long as the
size is selected in an appropriate manner. This provides a high local
concentration over a prolonged period of time (45).

29
An algorithm for the suggested management of the acne is presented in the following figure.
Lesion typefs)
J t
Comedonal lesions ------ r
Mixed comedenal lesions T
Papules and pustules

b
— Cystic lesions

cc

C
a
9

T5
%
J5
tt

ft.
£%*
ft.
o—

—►
Prescribe one of the Prescribe one of the following; Prescribe Benzoyl peroxide.
t Prescribe course of oral
following; Retinoid+topical antibiotic antibiotic + mixed
Topical retinoid Retincrd+benzoyl peroxide ccmedonal-papulopustular
Azeiaic acid Retinofd+benzoyl If resultsare topical therapy.
t
Salicylic acid peroxide+topical antibiotic unsatisfactory, switch to or
Azelaic acid+benzoyl add t opical antibiotic.
peroxide If resultsare
Azelaic acid-rtopfeat antibiotic unsatisfactory, consider
whether patient is a
candidate for oral
isotretinoin (Acutane)
therapy.
t
If results are If results are If results are If results are
unsatisfactory, unsatisfactory, prescribe unsatisfactory, add course unsatisfactory, consider
increase strength or retinoid + course of oral of oral antibiotic. possibility of
change medication. antibiotic. endocrinopathy.
1
If endocrinopathy If no sign of
is identified, treat. endocrinopathy is
found, consider repeat
course of oral
isotretinoin.

Figure 14. Algorithm for the management of acne

o
m
1.4. REFERENCES
1. Kaparissides C, Alexandridou S, Kotti K, Chaitidou S. Recent
advances in novel drug delivery systems. Azojono J
Nanotechnol. 2006; 2:1-11.
2. Bagwe RP, Kanicky JR, Palla BJ, Patanjali PK, Shah DO.
Improved drug delivery using microemulsions: Rationale,
recent progress and new horizons. Crit Rev Ther Drug Carrier
Syst. 2001; 18: 77-140.
3. Westesen K, Bunjes S, Hammer G, Siekmann B. Novel
colloidal drug delivery systems. PDA J Pharm Sci Tech. 2001;
55:240-247.
4. Kreilgaard M. Influence of microemulsions on cutaneous drug
delivery. Adv Drug Del Rev. 2002; 54: S77-S98.
5. Boltri L, Morel S, Trotta M, Gasco, MR. In vitro transdermal
permeation of nifedipine from thickened microemulsions. J
Pharm Belg. 1994; 49: 3151-320.

6. Bonina FP, Carelli V, Di Colo G, Montenegro L, Nannipieri

E. Vehicle effects on in vitro skin permeation of and stratum
I

comeum affinity for model drugs caffeine and testosterone. Int

J Pharm. 1993\ 100:41-48.
7. Delgado-Charro MB, Iglesias-Vilas G, Blanco-Mendez J,
Lopez-Quintela MA, Guy RH. Delivery of a hydrophilic
solute through the skin from novel microemulsion systems.
Eur J Pharm Biopharm. 1997; 43: 37-42.
8. Kreilgaard M, Pedersen EJ, Jaroszewski JW. NMR
characterization and transdermal drug delivery potential
microemulsion systems. J Control Release. 2000; 69: 421-
433.

31
9. Hoar TP, Schulman JH. Transparent water-in-oil disper sions:
the oleopathic hydro-micelle. Nature. 1943; 152: 102-103.
10. Schulman JH, Stoeckenius W, Prince LM. Mechanism of
formation and structure of micro emulsions by electron
microscopy. J Phys Chem. 1959; 63: 1677-1680.
11. Danielsson I, Lindman B. The definition of a microemulsion.
Colloid Surf. 1981; 3: 391-392.
12. Kreuter J. Microemulsions; In: colloidal drug delivery system
New York, Marcel Dekker. 1994; 31-71.
13. Patel MR, Patel RB, Parikh JR, Bhatt KK, Kundawala AJ.
Microemulsions: as novel drug delivery vehicle. Pharma Rev.
2007; 5: www.pharmainfo.net/reviews/microemulsions-novel-
drug-delivery-vehicle. Accessed on 30 Nov 2009.
14. Ghosh PK, Murthy RSR. Microemulsions: a potential drug
delivery system. Current Drug Del. 2006; 3: 167-180.
i

15. Swarbrick J, Boylan JC. Microemulsions; In: Encyclopedia of

Pharmaceutical Technology. New York, Marcel Dekker 1994;
375- 421.
16. Lawrence MJ, Rees GD. Microemulsion-based media as novel
drug delivery systems. Adv Drug Deliv Rev. 2000; 45: 89-
121.
17. Hellweg T. Phase structure of microemulsions. Curr Opin
Colloid Interface Sci. 2002; 7: 50-56.
18. Constantinides PP, Yiv SH. Particle size determination of
phaseinverted water-in-oil microemulsions under different
dilution and storage conditions. Int J Pharm. 1995; 115: 225-
234.

32
19. Mueller BW, Mueller RH. Particle size distributions and
particle size alterations in microemulsions. J Pharm Sci. 1984;
73: 919-922.
20. Kang BK, Chon SK, Kim SH, et al. Controlled release of
paclitaxel from microeimulsion containing PLGA and
evaluation of antitumour activity in vitro and in vivo. Int J
Pharm. 2004; 286: 147- 156.
21. Porras M, Solans C, Gonzalez, C, Martinez A, Guinart A,
Gutierrez, JM. Studies of formation of w/o nanoemulsions.
Colloid Surf A: Physicochem Eng Asp. 2004; 249: 115-118.
22. Silas JA, Kaler EW. Effect of multiple scattering on SANS
spectra from bicontinuous microemulsions. J Colloid Interface
Sci. 2003;257:291-298.
23. Pedersen JS. Analysis of small-angle scattering data from
micelles and microemulsions: Free-form approaches and
model fitting. Curr Opin Colloid Interface Sci. 1999; 4: 190-
196.
24. Podlogar F, Gasperlin M, Tomsic M, Jamnik A, Bester Rogac
M. Structural characterisation of water-Tween 40®/Inwitor
308®- isopropyl myristate microemulsions using different
experimental methods. Int J Pharm. 2004; 276: 115-128.
25. Kawakami K, Yoshikawa T, Moroto Y. Microemulsion
i

formulation for enhanced absorption of poorly soluble drugs:

I. Prescription design. J Control Release. 2002; 81: 65-74.
i

26. Glatter O, Orthaber D, Stradner A. Sugar-ester nonionic

microemulsion: Structural characterization. J Colloid Interface
Sci. 2001;241:215-225.
27. Danino D, Talmon Y, Zana R. Cryo-TEM of thread like
micelles: on the grid microstructural transformations induced

33
 during specimen preparation. Colloid Surf A: Physicochem
Eng Asp. 2002; 169: 67-73.
28. Jian X, Ganzuo L, Zhiqiang Z, Guowei Z, Kejian J. A study of
the microstructure of CTAB/l-butanol/oetane/water system by
PGSENMR,conductivity and cryo-TEM. J Colloid Surf A:
Physicochem Eng Asp. 2001; 191:269-278.
29. Magdassi S, Ben Moshe M, Talmon Y, Danino D.
Microemulsion based on anionic gemini surfactant. Colloid
Surf A: Physicochem Eng Asp. 2003; 212: 1-7.
30. Andersson M, Lofroth J-E. Small particles of a
heparin/chitosan complex prepared from a pharmaceutically
acceptable microemulsion. Int J Pharm. 2003; 257: 305-309.
31. Kreilgaard M, Pedersen EJ? Jaroszewski JW. NMR
characterization and transdermal drug delivery potential
microemulsion systems. J Control Release. 2000; 69:421-433.
32. Lopez F, Cinelli G, Ambrosone L, Colafemmina G, Ceglie A,
Palazzo G. Role of the cosurfactant in water-in-oil
microemulsion: interfacial properties tune the enzymatic
activity of lipase. Colloid Surf A: Physicochem Eng Asp.
2004; 237: 49-59.
33. Djordjevic L, Primorac M, Stupar M, Krajisnik D.
Characterization of caprylocaproyl macrogolglycerides based
microemulsion drug delivery vehicles for an amphiphilic drug.
Int J Pharm. 2004; 271: 1149.
34. Mrestani Y, El-Mokdad N, Ruttinger HH, Neubert RHH.
Characterization of partitioning behaviour of cephalosporins
using microemulsion and micellar electrokinetie
chromatography. Electrophoresis. 1998; 19: 2895-2899.

34
35. Mrestani Y, Neubert RHH, Krause A. Partition behaviour of
drugs in microemulsions measured by electrokinetic
chromatography. Pharm Res. 1998; 15: 799-801.
36. Feldman S, Carecci R, Barham K, Hancox J. Diagnosis and
i

treatment of acne. American Fam Phy. 2004; 69: 2123-2130.

37. Gollnick BP, Zouboulis j CC, Akamatsu H, Kurokawa I,
Schulte A. Pathogenesis and pathogenesis related treatment of
acne. J Dermatol. 1991; 18:489-99.
38. Holmes RL, Williams M,: Cunliffe WJ. Pilo-sebaceous duct
obstruction and acne. Br J Dermatol. 1972; 87: 327-32.
39. Jappe U. Pathological mechanisms of acne with special
emphasis on propionibacterium acnes and related therapy.
Acta Derm Venereol. 2003;; 83: 241-248.
40. Zouboulis CC. Acne and sebaceous gland function. Clin
Dermatol. 2004; 22: 360-366.
41. Russell JJ. Topical therapy for acne. Am Fam Phys. 2000; 61:
1-13. i

42. Feldman S, Careccia RE, Barham KL, Hancox J. Diagnosis

and treatment of acne. Am Fam Phys. 2004; 69:2123-2130.
43. Leyden JJ. Emerging topical retinoid therapies. J Am Acad
Dermatol. 1998; 38: S1-S4.
44. Zaenglein AL. Topical retinoids in the treatment of acne
vulgaris. Semin Cutan Med Surg. 2008; 27: 177-182.
45. Date AA, Naik B, Nagarsenker MS. Novel drug delivery
systems: Potential in improving topical delivery of Antiacne
agents. Skin Pharmacol Physiol. 2006; 19: 2-16.

35
TV.5J5