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Dipartimento di Scienze Farmaceutiche, Universit`a degli Studi di Perugia, Perugia 06123, Italy
Dipartimento di Chimica, Biologia e Biotecnologie, Universit`a degli Studi di Perugia, Perugia 06123, Italy
INTRODUCTION
Acne vulgaris, acne infantum, acne fulminans, and induced
acne, deriving from exposition to cosmetics, pharmaceutical
treatment with steroids or face autoinduced trauma, are the
main cutaneous chronic inflammations affecting the sebaceous
follicles.1 Acne vulgaris, the most frequent form of these diseases, is a multifactorial chronic dermatosis that appears generally during puberty as well as in adult age.
Acne has four main pathogenetic mechanisms: comedogenesis (sebaceous follicle obstruction), increased sebum production, Propionibacterium acne colonization, and inflammation.2
It is characterized by various clinical signs as seborrhoea,
comedones, erythematous papules and pustules, less frequently
nodules, deep pustules, or pseudocysts.3,4 Comedones, deriving
from an excessive keratinization in the sebaceous follicle, represent the starting point for the inflammation process beginning.
The hyperproliferation of keratinocytes is stimulated by the
androgenic hormone testosterone, able to bind specific membrane receptors of this kind of cells, and the conversion of such
hormone in dihydrotestosterone is particularly enhanced (from
two to 20 times more) in the acneic skin.5,6
The androgenic hormones provoke sebaceous gland hypertrophy responsible for the increased sebum production. A microbial flora, a stagnant cap of lipids and desquamed cells
are present in the follicular canal. These components represent
an optimal microenvironment for anaerobic bacteria growth,
Correspondence to: Luana Perioli (Telephone: +39-075-5855133; Fax: +39075-5855163; E-mail: luana.perioli@unipg.it)
This article contains supplementary material available from the authors upon
request or via the Internet at http://wileylibrary.com.
Journal of Pharmaceutical Sciences, Vol. 104, 39043912 (2015)
C 2015 Wiley Periodicals, Inc. and the American Pharmacists Association
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3905
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MgAl-RET Gel
ZnAl-RET Gel
RET
0.05 g
Ethanol 96%
Propylene glycol
HEC
Deionized water
20 g
25 g
5g
49.95 g
0.084 g
Corresponding
to 0.05 g of RET
(loading 58.9%)
20 g
25 g
5g
49.91 g
0.110 g
Corresponding
to 0.05 g of RET
(loading 45.5%)
20 g
25 g
5g
49.89 g
between the excitation and the emission light. The spectra were
corrected for the response of instrument components at each
wavelength. The traces of fluorescence intensity as function of
irradiation time, which gave information on the photochemical
stability of the samples, were recorded fixing the excitation and
emission wavelengths and monitoring the emission intensity
during the irradiation. Steady-state irradiation experiments
were performed using a xenon lamp as light source, selecting
the excitation wavelength (366 nm) by a monochromator and
setting a band-pass slit of 35 nm.
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Figure 1. Scanning electron micrographs of MgAl-HTlc nitrate (a, b) and MgAl-HTlc-RET (c, d).
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Figure 2. Scanning electron micrographs of ZnAl-HTlc nitrate (a, b) and ZnAl-HTlc-RET (c, d).
DOI 10.1002/jps.24612
Stability Studies
Retinoic acids, such as tretinoin and isotretinoin, are photolabile compounds, whose photostability strongly depends on
the environment.28 This instability occurs even in commercially available product. Absorption spectra of an isotretinoin
formulation before and after steady-state irradiation were in
fact carried out (figure can be found in Supporting Information, SI 6). Absorption intensity decrease (almost 70%) was
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To evaluate the effects of the inorganic matrices on the photochemical stability of RET and its gel formulations, fluorescence methods were used. Generally, fluorescence is a very sensitive and selective parameter, whose intensity is proportional
to the concentration of the emitting species, thus quite useful to compare the relative photostability of samples measured
in the same experimental conditions. In Figure 7, the fluorescence spectra of RET, MgAl-RET, and ZnAl-RET are reported;
all the samples presented a broad spectrum with maximum
around 550 nm. The effect of steady-state irradiation on the
concentration of isotretinoin samples was quantitatively monitored acquiring the fluorescence intensity at the maximum
upon irradiation and 366 nm. The traces, reported in Figure 7,
indicate that intercalation compounds are more stable than the
DOI 10.1002/jps.24612
Figure 6. Absorption spectra of crystalline RET (squares), MgAlHTlc-RET (circles) and ZnAl-HTlc-RET (triangles).
free RET sample. These data clearly reveal that upon intercalation, both in MgAl-HTlc and ZnAl-HTlc, the drug under investigation improves its photostability, likely because the anchoring
to the matrices reduces the geometrical rearrangements, which
are responsible for the degradation of RETs.
The gel formulations, obtained by homogeneous dispersion
of RET, MgAl-HTlc-RET, and ZnAl- HTlc-RET in gel, have
been tested with the same methods and conditions to evaluate whether their dispersion can modify the sample solidity.
RET and MgAl-RET gels maintain the reached photostability,
as shown by the fluorescence traces reported in Figure 8. On
the contrary, ZnAl-RET gel presents a lower solidity to light
exposure, which needs to be further investigated.
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CONCLUSIONS
Retinoic acid has been intercalated into HTlc lamellar structure with the aim to increase its photostability. It showed
high affinity for both MgAl-HTlc and ZnAl-HTlc and its intercalation has successfully occurred giving rise to hybrids,
MgAl-HTlc-RET and ZnAl-HTlc-RET, having high drug loading (58.9% and 45.5%, respectively). In this supramolecular arrangement, the active ingredient loses its crystalline structure
and its molecules are homogeneously dispersed and immobilized into the HTlc nanogalleries.
Intercalated compound crystals are flat and have micrometric size; they can be directly used for the preparation of a
formulation, safe and no-abrasive, suitable to carry the drug
on acneic skin.
Photochemical studies highlighted interesting aspects and
the most important information was obtained from fluorescence
measurements. The fluorescence traces were recorded to have
information about the effect of intercalation into HTlc on RET
photostability. The results clearly reveal that upon intercalation in MgAl-HTlc or ZnAl-HTlc, the active ingredient shows
improved photostability. The formulation obtained by dispersing MgAl-HTlc-RET in gel preserved the obtained degree of
photostability.
Rheology studies showed that the formulations containing
the hybrids are characterized by suitable viscosity and extrudibility, better than commercial formulation and better for MgAlRET. HTlc presence in the formulation furnishes a positive contribution because it confers pseudo-plastic-tixotropic behavior
to the gel, more evident in the case of MgAl-RET. This phenomenon is undoubtedly important for the success and acceptability of final formulation because the increased viscosity at
25C assures product homogeneity with good stability during
the shelf-life (e.g., reduction of sedimentation rate or no sedimentation), whereas the increase of fluidity at 32C can certainly guarantee a good spreadability on the acneic skin injured
and often inflamed.
Perioli et al., JOURNAL OF PHARMACEUTICAL SCIENCES 104:39043912, 2015
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ACKNOWLEDGMENTS
The Authors wish to thank Dr. Michele Sisani for collaboration
in the measurements of XRPD and Mr. Marco Marani for the
precious collaboration and technical assistance.
A part of this work was presented at XLIX Symposium AFI,
Rimini (Italy).29
The authors declare that they have no personal financial or
nonfinancial conflicts of interest in this research.
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