European Journal of Pharmaceutical Sciences 167 (2021) 105989

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European Journal of Pharmaceutical Sciences

journal homepage: www.elsevier.com/locate/ejps

Evaluation of cellulose based films comprising tea tree oil against

dermatophytes and yeasts
Flavia Laffleur a, *, Martin Ataii a, Magdalena Nagler b
a
Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, Innsbruck 6020, Austria
b
Institute of Microbiology, University of Innsbruck, Technikerstraße 25 d, Innsbruck 6020, Austria

A R T I C L E I N F O A B S T R A C T

Keywords: Background: Onychomycosis is defined as infection caused by nondermatophytic molds and yeasts: tinea
Adhesiveness unguium is caused by dermatophytes. Purpose: Within this study, hydroxyethyl cellulose (HEC) as an important
Antifungal non-ionic, water-soluble cellulose derivative was chosen to develop formulations containing tea tree oil as active
Inhibition zone assay
antifungal agent were developed and evaluated for their potential in the treatment of onychomycosis.
Nail
Tea tree oil
Methods: Two polymeric films based on HEC (HEC-B-04 and HEC-E-10) were obtained by solvent evaporation
method and characterized in terms of appearance, disintegration, stickiness, elongation, rheological behavior
and adhesiveness. Moreover, different strains of dermatophytes such as Trichophyton rubrum and yeasts as
Candida albicans were treated with polymeric films containing tea tree oil (0.5 – 2 % v/v) in order to determine
their antifungal potential by the inhibition zone assay.
Results: HEC-B-04 and HEC-E-10 were investigated by SEM measurements resulting in confluent surface
morphology. HEC-B-04 and HEC-E-10 showed disintegration after 32.7 min and 34.0 min, respectively.
Furthermore, HEC-E-10 revealed a moisture index of 1.74 and underpinned adhesive properties in terms of
required detachment force with 4.86 N. HEC-E-10 pointed to the most antifungal one among the others against
Trichophyton rubrum and Candida albicans.
Conclusion: Taking these findings in consideration, promising adhesive onychial formulations were developed as
forthcoming approach in treatment of nail infections.

1. Introduction
Onychomycosis is infection caused by nondermatophytic molds and
yeasts: tinea unguium is caused by dermatophytes, which affects 20 % of
the population across the world and constituting 50 % of all the nail
diseases (Lipner and Scher, 2019). Onychomycosis affects patients with
human immunodeficiency virus and further strikes one third of patients
with diabetes increasing the severity of foot disorders (Piraccini et al.,
2010).Success in treating onychomycosis is overshadowed by several
limitations. Limitations of onychomycosis cure is represented by slow
growth of nails (I), the resistance against antifungals (II) and inappro­
priate time periods of treatment (III) (Elewski, 1998). Available anti­
fungals on the market are listed in form of creams, ointments, lacquers,
solutions, lotions and powders. So far, commercially used formulations
are not specifically suitable for the nail because of their undesired
removal due to rubbing and washing (Akhtar et al., 2016). Thus, there is
an unmet need for adhesive and long lasting formulations to overcome
these drawbacks.
Formulations such as films have gained importance in the pharma­
ceutical field as novel, patient friendly, convenient products convincing
with their small size and thickness (Peh and Wong, 1999). Within this
study, conventional polymeric excipients such as cellulose derivates
were chosen in order to prepare novel dosage forms following the sol­
vent evaporation method. Hydroxyethylcellulose, as one of the signifi­
cant cellulosic ether derivatives, is approved by the U.S. Food and Drug
Administration (FDA). Due to its biocompatibility and low toxicity, HEC
is used as thickener, binder or as a protective suspension and colloid
stabilizer in many applications such as coating and biomedical appli­
cations (El Fawal et al., 2020). Tea tree oil known for its antibacterial,
antifungal and anti-inflammatory properties is an essential oil being
obtained from the leaves of Melaleuca alternifolia. Its composition is
based on a complex mixture of compounds, mostly sesquiterpene and

* Corresponding author at: Department of Pharmaceutical Technology, Institute of Pharmacy, Center for Molecular Biosciences Innsbruck, University of Innsbruck,
Innrain 80/82, 6020 Innsbruck, Austria.
E-mail address: Flavia.Laffleur@uibk.ac.at (F. Laffleur).

https://doi.org/10.1016/j.ejps.2021.105989
Received 14 July 2021; Received in revised form 27 August 2021; Accepted 31 August 2021
Available online 11 September 2021
0928-0987/© 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
F. Laffleur et al.
Table 1
Composition of formulations containing polymers.
Ingredients Ethanol [mL] HEC [g] HPMC [g] PPG [g] PVP [g]
HEC-B-04 20 0.4 - 0.23 0.4
HEC-E-10 20 1.0 0.2 0.23 0.4
monoterpene hydrocarbons a well as their associated alcohols (Flores
et al., 2013). Tea tree oil was embedded in the polymeric films in several
concentrations (0.5-2% v/v) and tested on Trichophyton rubrum and
Candida albicans by agar disk diffusion assay.
These solid formulations were evaluated with respect to physical
appearance, disintegration, water uptake, rheological behavior, surface
pH and the adhesiveness on the nail plate. By this study, adhesive films
might be the next step in the direction of a prolonged residence time on
the site of action contributing less frequency of application (I), higher
patient’s compliance (II) and the faster cure of onychomycosis due to an
intimate contact of the dosage form (III).
2. Materials and methods
2.1. Materials
2-Hydroxyethyl-cellulose (HEC) (average Mw 90.000 Da),
(Hydroxypropyl)methyl cellulose (HPMC) were all received from Sigma
Aldrich (Steinheim, Germany). Polyvinylpyrrolidone (PVP) was
received from BASF (Ludwigshafen, Germany), Propylene glycol (PPG)
and tea tree oil were obtained from Gatt-Koller (Absam, Austria).
Candida albicans ATCC® 36232 and Trichophyton rubrum ATCC® 28188
were purchased at Huecker GmbH, Germany. All other solvents and
reagents used were of analytical grade and received from commercial
sources.
2.2. Preparation of films
HEC-B-04 and HEC-E-10 were prepared by solvent evaporation
method using film forming polymers. For the preparation of formulation
HEC-B-04, firstly, 0.4 g HEC was completely dissolved in 20 mL of
ethanol filled in a beaker (Duran, Wertheim, Germany). Secondly, 0.4 g
PVP was added slowly to the mixture and finally 0.23 g PPG as plasti­
cizer was subjoined. The mixture was stirred for 15 min. Afterwards, the
polymeric solution was poured in a 100 × 21 mm petri dish (Fisher
scientific, Vienna, Austria). Furthermore, an inverted funnel was placed
over the petri dish to avoid sudden evaporation. The solvent was
allowed to evaporate overnight. HEC-E-10 was prepared following the
same procedure as described above with addition of 1.0 g HEC instead of
0.4 g as well as admixing 0.2 g of HPMC as shown in Table 1.
2.3. Characterization of films
Physical appearance was determined in regards of texture especially
smoothness, dryness, stickiness, wrinkles and transparency. HEC-B-04
and HEC-E-10 (diameter of 14 mm) were weighed, respectively, for
weight uniformity (n=3). Thickness of each formulation was ascertained
with a digital caliper (MarCal 16EWRI, Mahr GmbH, Göttignen, Ger­
many) (Semalty et al., 2008). Folding endurance was evaluated by
repeated folding of the films at the same place till breaking or folding
over 300 times without breaking was observed (Obaidat et al., 2011).
The surface pH was determined by adding a drop of water to the for­
mulations and measuring the pH with a pH micro electrode (VWR,
Austria).
2.4. Scanning electron microscopy measurements
Surface morphology of HEC-B-04 and HEC-E-10 was recorded by
scanning electron microscopy (SEM) using a ZEISS EVO LS 10 connected
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European Journal of Pharmaceutical Sciences 167 (2021) 105989
with SmartSEM program (Oberkochen, Germany) instrument. HEC-B-04
and HEC-E-10 were cut into 1 cm2 pieces and layered on the SEM stub.
The films were then coated with gold metal using a sputter coater (SC
7640; Polaron Quorum Technologies, LOT-QuantumDesign GmbH,
Darmstadt, Germany) (Khan et al., 2015).
2.5. Rheological measurements
The rheological properties of HEC-B-04 and HEC-E-10 were analyzed
with a thermostatically controlled plate–plate rheometer (Thermo
Haake Mars, Haake GmbH, Karlsruhe, Germany). 1 % (w/v) of HEC-B-
04 and HEC-E-10 was prepared and 0.5 mL samples were transferred
to the rheometer. For all samples the apparent viscosity (η) was
measured immediately after equilibration. The shear stress was set at a
range of 0.5–500 Pa and the gap between two plates was set 0.5 mm
(Laffleur and Messirek, 2016).
2.6. Surface tension
The measurement of the surface tension is based on interaction be­
tween formulations and interfaces. Herein, DuNouy ring was utilized
while the interaction of a platinum ring with the surface was tested with
a tensiometer (KSV, KSV Instruments LTD, Helsinki, Finland). DuNouy
ring was immersed in 10 mL of HEC-B-04 and HEC-E-10, respectively.
The upward movement of the ring achieves a meniscus of the liquid. The
measurement of the tension determines the maximum force until the
lamella rupture (Zelkó et al., 2002).
2.7. Moisture uptake
Percentage of moisture content was ascertained by weighing the 14
mm of diameter HEC-B-04 and HEC-E-10 individually and keeping those
in desiccators containing CaCl2 at room temperature for 24 h. After­
wards, films were weighed until constant weight. The percentage
moisture content was determined as the difference between initial and
final weight with regard to initial weight (de Kumar et al., 2011).
Furthermore, moisture absorption capacity of the films was determined
while weighing accurately films and placing in desiccator containing
saturated solution of KCl keeping the humidity inside the desiccator at
84 %. After 24 h, films were taken out, reweighed and percentage
moisture was calculated according following equation (El-Houssieny
et al., 2016):
Moisture absorption [%] = [(final weight − initial weight)/initial weight]
× 100
2.8. Stability and disintegration study
In order to determine stability and disintegration behavior two as­
sessments were performed. Firstly disintegration time according to the
United States Pharmacopeia 30, volume 1, and year 2007 was recorded.
For this purpose, films with 14 mm of diameter were placed into the
disintegration tester (ERWEKA ZT 223, Heusenstamm, Germany).
Briefly, HEC-B-04 and HEC-E-10 was placed into the six tubes filled with
artificial sweat fluid (composed of 1 L H2O, 5.0 g NaCl, 1.0 g urea and
1.0 g lactic acid), the pH was adjusted to 4.5-5.5 with NH4OH (Marques
et al., 2011). The experimental set up was at an oscillating frequency of
0.5 s− 1 as well as a temperature of 37 ± 0.5◦ C. The disintegration time of
films was recorded (Laffleur and Messirek, 2016). In the second test,
HEC-B-04 and HEC-E-10 of 14 mm in diameter were mounted in be­
tween to clamps. Both clamps were pulled apart by adding weights.
Elongation was determined by disruption point of the films, respec­
tively. Measurements were performed in triplicate for each formulation
(Shah et al., 2011). Percentage elongation was calculated according to
following equation:
F. Laffleur et al.
Table 2
Parameter and physico-chemical characteristics of HEC based films.
Formulations Weight [g] Folding endurance [times]
HEC-B-04 0.0366 ± 0.0015 >300
HEC-E-10 0.0487 ± 0.0008 154 ± 5
Elongation at break [%] = Increase in length [mm]/initial length [mm] × 100
2.9. Adhesive investigations
2.9.1. Preparation of nail samples
Horn substance was collected from bovine hoof provided by a local
slaughterhouse. After cleaning the hoof, connective tissue was removed
from the horn material. The nails were cut into small pieces of 2 × 2 cm.
Nail samples were kept in the freezer at – 20◦ C until further use.
2.9.2. Ex vivo adhesive assay
Adhesiveness of HEC-B-04 and HEC-E-10 was determined by
detachment force. Films were cut into 14 mm in diameter samples,
respectively. Nail samples were incubated with HEC-B-04 and HEC-E-10
for 15 min, respectively. Films were glued on their upper part to a
hanging device from a laboratory stand connected to weights. The
addition of weights was conducted until the required force to detach the
formulation from the nail was achieved (Govindasamy et al., 2013) k.
The adhesive force was calculated according following equation:
/ hesive properties. Furthermore HEC-E-10 was designed with higher
Adhesive force[kg ∗ m / s] = adhesive strength [kg] × 9.8 m s− 1
2.9.3. Agar based susceptibility testing method: disk diffusion assay
In order to test the susceptibility of important dermatophytes like
Trichophyton rubrum and yeast as Candida albicans to various concen­
trations of the active component, a disk diffusion approach was con­
ducted. First, the maximum applicable tea tree oil concentration not
interfering with the formulation properties was tested visually by
exclusion of precipitation and found to be 2 %. Then, formulations HEC-
B-04 and HEC-E-10 were prepared with 0, 0.5, 1 and 2 % of tea tree oil
and discs of 6 mm diameter were blanked. Pure cultures from Tricho­
phyton rubrum and Candida albicans were cultivated on malt extract agar
(Carl Roth, Karlsruhe, Germany) until conidia formation for up to 7 days
at 37◦ C. Subsequently, spores were yielded with sterile ringer solution,
spore concentration was measured with a cell counter (Countess,
Thermo Fisher Scientific, Massachusetts, USA) and adjusted to 106
conidia/mL. In triplicates for each formulation and each tested species, a
total of 105 conidia/ml were plated on sabouraud dextrose agar plates
(Becton Dickson, Heidelberg, Germany), the formulation patches of
various concentrations were placed on the plates and incubated at 37◦ C.
Fig. 1. (a) and (b) Scanning Electronic microscope (SEM) micrograph of testing polymeric films HEC-B-04 (Fig. 1a) and HEC-E-10 (Fig. 1b), respectively.
European Journal of Pharmaceutical Sciences 167 (2021) 105989
Flatness [mm] Dry Flexible Smooth Wrinkled pH
0.1833 ± 0.0153 Yes Yes No Yes 5.7
0.2767 ± 0.0231 Yes Yes No Yes 5.7
As a positive control, sterile filter paper was dipped in tea tree oil (100
%) and placed onto separate plates. Finally, diameters of the inhibition
zones around the patches were measured after 24 h (C. albicans), and 72
h (T. rubrum) of incubation.
2.9.4. Statistical analysis
The statistical data analysis was performed with GraphPad Prism
5.01. The analysis of variance (ANOVA) was used to compare the means.
A probability of less than 0.05 (P<0.05) was considered statistically
significant. All results are presented as mean ± SD.
3. Results and discussion
3.1. Preparation and characterization of films
In the present study, HEC-B-04 and HEC-E-10 were prepared ac­
cording to the composition as shown in Table 1. Polymeric excipients
were chosen due to their solubility properties and suitability for film
building features. HEC was chosen as the basis due to its reported ad­
amount of HEC and additional HPMC as synergistic effect for enhancing
adhesive features.
HEC-B-04 and HEC-E-10 were characterized in regards of their
physico-chemical properties and other parameters such as transparency,
dryness, smoothness, flatness, stickiness and flexibility as shown in
Table 2.
The range of the flatness varied from 0.1833 ± 0.0153 mm (HEC-B-
04) to 0.2767 ± 0.0231 mm (HEC-E-10) and weight between 0.0366 ±
0.0015 g (HEC-B-04) and 0.0487 ± 0.0008 g (HEC-E-10). The surface pH
of HEC-B-04 and HEC-E-10 was 5.7, respectively. HEC-B-04 and HEC-E-
10 showed folding endurance over 150 times. All prepared films were
dry and flexible.
In respect to the manufacture process, solvent evaporation method
exhibits benefits such as ease of the process. The process consists of four
main procedures, namely, preparation of the solution, transferring to a
mold, drying the solvent, and cutting the final form. The reproducibility
was given due to the ease of processing. Furthermore, SEM measure­
ments were conducted in order morphologically characterize HEC-B-04
and HEC-E-10 according to the surface and texture (Khan et al., 2015).
Results were shown in Fig. 1. All formulations exhibited suitable texture
3
F. Laffleur et al.
Fig. 2. Surface tension measured by KSV-Instrument with a DuNouy Ring.
Samples were measured in triplicate (n=3 ± SD).
for nail application exhibiting smoothness and confluency.
3.2. Rheological investigations
Rheology was evaluated as a determinant of the drying rates and
uniformity in terms of the physical appearance of the films (Morales and
McConville, 2011). The rheological investigations provide disclosures
about stress sweep and adhesive connection with regard of increased
viscosity (η). η was given at a constant angular frequency of 6.283 rad/s
(=1Hz) and was plotted as a function of time. The enhancement ratios in
Fig. 3. Moisture uptake study of tested formulations, indicated values are means of at least 3 experiments (± SD).
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European Journal of Pharmaceutical Sciences 167 (2021) 105989
viscosity at the beginning of the study and after 120 min showed 1.4-fold
and 2.7-fold improvement in terms of formulations HEC-B-04 and
HEC-E-10, respectively. Consequently, the added amount of HEC
showed an impact in terms of viscosity behavior of HEC-B-04 and
HEC-E-10.
3.3. Surface tension and moisture uptake
Surface tension measures the cohesive energy presented at an
interface. Within this study, the two phases were determined to be air as
light phase and aqueous phase as heavy phase. Moreover, it was re­
ported that a high surface energy correlates with a lower contact angle
bearing an impact in adhesion properties. The herein found results,
shown in Fig. 2, exhibiting the surface tension values for HEC-B-04 and
HEC-E-10 with 67.63 ± 3.57 (mN/m) and 64.85 ± 2.10 (mN/m),
respectively.
Taking these findings in consideration, the forces in this study
depend on factors such as contact angle of liquid interaction and probe
properties.
Moisture content and moisture uptake studies were accomplished
showing the findings in Fig. 3. The moisture index is given 1.09 (HEC-B-
04) and 1.74 (HEC-E-10), respectively. The moisture content of the
designed formulations was menial resulting in stability and reduce of
brittleness during long-term storage. The moisture uptake of the for­
mulations was low leading in reduced bulkiness (Ubaidulla et al., 2007).
3.4. Stability evaluation
In polymeric systems, the surficial part of the carrier initially hy­
drates while dissolving in order to generate an outer viscous layer.
Within this process a matrix bulk hydration, swelling and erosion phase
are connected. Due to this connection, dissolution rate and availability
of embedded drug is depending on the processes as reported by Roy et al.
(Sinha Roy and Rohera, 2002). The intermolecular and intramolecular
forces, the water uptake and the cohesiveness of a polymer exhibit a
great impact on its adhesion.
As a consequence, the disintegration profile of tested formulations is
shown as a suitable indicator for their cohesive properties (Shahnaz
et al., 2010). Disintegration profiles were shown in Fig. 4 to elucidate
the cohesiveness of the formulations HEC-B-04 (32.67 min) and
HEC-E-10 (34.00 min), respectively.
These findings are correlating with the original properties of the used
polymers regarding swelling behavior, namely, tested polymers
F. Laffleur et al.
Fig. 4. Overview of disintegration behavior of testing formulations. The indicated disintegration time represents an average of at least 3 experiments ± SD.
Fig. 5. Evaluation of adhesive strength represents the force required to detach
the formulations. Indicated values are the means of at least 3 experiments
(± SD).
exhibited high swelling properties leading to pronounced intermolec­
ular action and therefore strong cohesiveness of the films.
3.5. Adhesive study and elongation evaluation
The suitability for nail application was seen in appropriate nail
adhesion. In order to determine the adhesive force formulations HEC-E-
10 and HEC-B-04 with 4.86 and 0.8 N, respectively, were evaluated as
shown in Fig. 5. The combination of HPMC and higher amount of HEC to
improved adhesiveness.
Besides the crucial parameter of adhesive force, the mechanical
property plays an important key factor on the physical integrity of the
dosage form. Therefore, percentage of elongation was determined as
displayed in Fig. 6. Formulation HEC-E-10 was the most promising
representative with 7.08 % followed by HEC-B-04 with 3.70 %. Gener­
ally, elongation will increase with amount of plasticizer. This correlates
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European Journal of Pharmaceutical Sciences 167 (2021) 105989
Fig. 6. Elongation study of tested HEC-B-04 and HEC-E-10. Samples were
measured in triplicate (n=3 ± SD).
with the literature given that plasticizers affects the increase in adhesion
(Perumal et al., 2008).
3.6. Susceptibility testing
Two fungal strains (T.rubrum and C.albicans), which are very often
blamed for nail mycoses, were used in this test series. Both strains were
treated with films comprising tea tree oil in different concentrations. Tea
tree oil served as ingredient with antifungal effect. The effect of HEC-B-
04 and HEC-E-10 containing different concentrations of tea tree oil was
tested on different fungus strains. Both films without tea tree oil served
as controls.
It could be shown that HEC-E-10 without tea tree oil also had a
F. Laffleur et al.
Fig. 7. Antifungal assay with T.rubrum and C.albicans. HEC-B-04 and HEC-E-10 containing tea tree oil in different concentrations. Indicated values are the means of
at least 3 experiments (± SD).
certain inhibitory effect on the strains. It was shown, however, that this
was dependent on the strain tested. Thus HEC-E-10 inhibited without
ingredient T.rubrum and C.albicans. HEC-E-10 without tea tree oil
showed an inhibitory zone diameter (IZD) of 0.23 cm on T.rubrum and
0.2 cm on C. albicans compared to HEC-B-04 (0 cm IZD for both strains).
This could be due to the polymer components per se, a possible residual
content of ethanol or a handling the disk to the agar plate.
On the basis of the films with active ingredient, however, it could
also be shown that the different fungal strains were inhibited differently
by the formulations. Clearly, both HEC-B-04 and HEC-E-10 were ex­
pected to have greater IZD at both fungal strains at higher concentra­
tions of tea tree oil. It should be pointed out that T.rubrum of HEC-E-10
was inhibited more strongly in all three concentrations than C.albicans,
and an IZD of about 0.7 cm had already been reached from a concen­
tration of 0.5% v/v.
HEC-B-04 showed appropriate potent inhibition in T.rubrum (0.93
cm IZD with 2 % tea tree oil, 0.8 cm with 1% v/v and 0.23 cm with 0.5%
v/v, respectively) followed by C.albicans (0.93 cm IZD with 2% v/v tea
tree oil, 0.77 cm with 1% v/v and 0.17% v/v with 0.5%, respectively).
HEC-E-10 achieved more promising inhibitory effect on C.albicans (0.97
cm IZD with 2% v/v tea tree oil, 0.8 cm with 1% v/v and 0.5 cm with
0.5% v/v, respectively) at all concentrations followed by T.rubrum (1.13
cm IZD with 2% v/v tea tree oil, 0.9 cm with 1% v/v and 0.7 cm with
0.5% v/v, respectively) compared to HEC-B-04. As expected, concen­
trations of 2% v/v tea tree oil of HEC-E-10 are the strongest inhibition
with 0.97 cm IZD of C.albicans and 1.13 cm IZD in T.rubrum. The data are
shown graphically in Fig. 7.
4. Conclusion
This study reports on adhesive polymeric nail delivery systems. HEC-
B-04 and HEC-E-10 were formulated based on cellulose derivatives like
hydroxyethyl cellulose and hydroxypropylmethyl cellulose according to
the solvent evaporation method. The stability, elongation and in vitro
disintegration studies revealed that the amount of HEC as well as the
combination with PVP influences the release profiles of the drug which
is in accordance with the formulation’s antifungal activity. In this
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European Journal of Pharmaceutical Sciences 167 (2021) 105989
regards, use of hydroxyethyl cellulose in combination with PVP in nail
formulation convinced in their suitability for nail application. Thus, it
can be concluded that HEC/PVP-based nail films are promising candi­
dates for nail application.
CRediT authorship contribution statement
Flavia Laffleur: Conceptualization, Methodology, Investigation,
Writing – original draft, Writing – review & editing. Martin Ataii:
Investigation, Visualization. Magdalena Nagler: Investigation,
Visualization.
Declaration of Competing Interest
The authors have no relevant affiliations or financial involvement
with any organization or entity with a financial interest in or financial
conflict with the subject matter or materials discussed in the manuscript.
This includes employment, consultancies, honoraria, stock ownership or
options, expert testimony, grants or patents received or pending, or
royalties. No writing assistance was utilized in the production of this
manuscript.
Acknowledgement
This research received funding under the grant AP 740040 by Tiroler
Wissenschaftsförderung, Austria.
References
Akhtar, N., Sharma, H., Pathak, K., 2016. Onychomycosis: potential of nail lacquers in
transungual delivery of antifungals. Scientifica. https://doi.org/10.1155/2016/
1387936.
de Kumar, P., Mallick, S., Mukherjee, B., Sengupta, S., Pattnaik, S., Chakraborty, S.,
2011. Optimization of in-vitro permeation pattern of ketorolac tromethamine
transdermal patches. Iran. J. Pharm. Res. 10, 193–201. https://doi.org/10.22037/
ijpr.2011.927.
El-Houssieny, B., El-Dein, E., El-Messiry, H., 2016. Formulation and evaluation of
dexibuprofen transdermal films in rabbits. Br. J. Pharm. Res. 9, 1–13. https://doi.
org/10.9734/BJPR/2016/18988.
F. Laffleur et al.
El Fawal, G., Hong, H., Song, X., Wu, J., Sun, M., He, C., Mo, X., Jiang, Y., Wang, H.,
2020. Fabrication of antimicrobial films based on hydroxyethylcellulose and ZnO for
food packaging application. Food Packag. Shelf Life 23. https://doi.org/10.1016/j.
fpsl.2020.100462.
Elewski, B.E., 1998. Onychomycosis: pathogenesis, diagnosis, and management. Clin.
Microbiol. Rev. https://doi.org/10.1128/cmr.11.3.415.
Flores, F.C., de Lima, J.A., Ribeiro, R.F., Alves, S.H., Rolim, C.M.B., Beck, R.C.R., da
Silva, C.B., 2013. Antifungal activity of nanocapsule suspensions containing tea tree
oil on the growth of Trichophyton rubrum. Mycopathologia 175, 281–286. https://
doi.org/10.1007/s11046-013-9622-7.
Govindasamy, P., Kesavan, B.R., Narasimha, J.K., 2013. Formulation of unidirectional
release buccal patches of carbamazepine and study of permeation through porcine
buccal mucosa. Asian Pac. J. Trop. Biomed. 3, 995–1002. https://doi.org/10.1016/
S2221-1691(13)60192-6.
Khan, S., Boateng, J.S., Mitchell, J., Trivedi, V., 2015. Formulation, characterisation and
stabilisation of buccal films for paediatric drug delivery of omeprazole. AAPS
PharmSciTech 16, 800–810. https://doi.org/10.1208/s12249-014-0268-7.
Laffleur, F., Messirek, A., 2016. Development of mucoadhesive thio-carboxymethyl
cellulose for application in buccal delivery of drugs. Ther. Deliv. 7 https://doi.org/
10.4155/tde.15.91.
Lipner, S.R., Scher, R.K., 2019. Onychomycosis: treatment and prevention of recurrence.
J. Am. Acad. Dermatol. https://doi.org/10.1016/j.jaad.2018.05.1260.
Marques, M.R.C., Loebenberg, R., Almukainzi, M., 2011. Simulated biological fluids with
possible application in dissolution testing. Dissolution Technol. https://doi.org/
10.14227/DT180311P15.
Morales, J.O., McConville, J.T., 2011. Manufacture and characterization of
mucoadhesive buccal films. Eur. J. Pharm. Biopharm. 77, 187–199. https://doi.org/
10.1016/j.ejpb.2010.11.023.
Obaidat, R.M., Bader, A., Al-Rajab, W., Sheikha, G.A., Obaidat, A.A., 2011. Preparation
of mucoadhesive oral patches containing tetracycline hydrochloride and carvacrol
European Journal of Pharmaceutical Sciences 167 (2021) 105989
for treatment of local mouth bacterial infections and candidiasis. Sci. Pharm. 79,
197–212. https://doi.org/10.3797/scipharm.1004-18.
Peh, K.K., Wong, C.F., 1999. Polymeric films as vehicle for buccal delivery: swelling,
mechanical, and bioadhesive properties. J. Pharm. Pharm. Sci. Publ. Can. Soc.
Pharm. Sci. Soc. 2, 53–61 doi:10952770.
Perumal, V.A., Lutchman, D., Mackraj, I., Govender, T., 2008. Formulation of
monolayered films with drug and polymers of opposing solubilities. Int. J. Pharm.
358, 184–191. https://doi.org/10.1016/j.ijpharm.2008.03.005.
Piraccini, B.M., Sisti, A., Tosti, A., 2010. Long-term follow-up of toenail onychomycosis
caused by dermatophytes after successful treatment with systemic antifungal agents.
J. Am. Acad. Dermatol. 62, 411–414. https://doi.org/10.1016/j.jaad.2009.04.062.
Semalty, M., Semalty, A., Kumar, G., 2008. Formulation and characterization of
mucoadhesive buccal films of glipizide. Indian J. Pharm. Sci. 70, 43–48. https://doi.
org/10.4103/0250-474X.40330.
Shah, S., Prabhu, P., Gundad, S., 2011. Formulation development and investigation of
domperidone transdermal patches. Int. J. Pharm. Investig. 1, 240. https://doi.org/
10.4103/2230-973x.93008.
Shahnaz, G., Perera, G., Sakloetsakun, D., Rahmat, D., Bernkop-Schnürch, A., 2010.
Synthesis, characterization, mucoadhesion and biocompatibility of thiolated
carboxymethyl dextran-cysteine conjugate. J. Control. Release 144, 32–38. https://
doi.org/10.1016/j.jconrel.2010.01.033.
Sinha Roy, D., Rohera, B.D., 2002. Comparative evaluation of rate of hydration and
matrix erosion of HEC and HPC and study of drug release from their matrices. Eur. J.
Pharm. Sci. 16, 193–199. https://doi.org/10.1016/S0928-0987(02)00103-3.
Ubaidulla, U., Reddy, M.V.S., Ruckmani, K., Ahmad, F.J., Khar, R.K., 2007. Transdermal
therapeutic system of carvedilol: effect of hydrophilic and hydrophobic matrix on in
vitro and in vivo characteristics. AAPS PharmSciTech 8. https://doi.org/10.1208/
pt0801002.
Zelkó, R., Orbán, Á., Süvegh, K., Riedl, Z., Rácz, I., 2002. Effect of plasticizer on the
dynamic surface tension and the free volume of Eudragit systems. Int. J. Pharm. 244,
81–86. https://doi.org/10.1016/S0378-5173(02)00317-4.