ORIGINAL RESEARCH ARTICLE
`
`Am J Clin Dermatol 2011; 12 (2): 133-141
`1175-0561/11/0002-0133/$49.95/0
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Resveratrol-Containing Gel for the
`Treatment of Acne Vulgaris
`A Single-Blind, Vehicle-Controlled, Pilot Study
`
`Gabriella Fabbrocini, Stefania Staibano, Giuseppe De Rosa, Valeria Battimiello, Nunzio Fardella, Gennaro Ilardi,
`Maria Immacolata La Rotonda, Amelia Longobardi, Marialuisa Mazzella, Maria Siano, Francesco Pastore,
`Valerio De Vita, Maria Luisa Vecchione and Fabio Ayala
`
`Department of Systematic Pathology, Division of Clinical Dermatology, University of Naples Federico II, Naples, Italy
`
`Abstract
`
`Background: Acne vulgaris is a complex, chronic, and common skin disorder of pilosebaceous units. The
`major pathogenic factors involved are ductal hyperkeratinization, obstruction of sebaceous follicles re-
`sulting from abnormal keratinization of the infundibular epithelium, stimulation of sebaceous gland se-
`cretion by androgens, and microbial colonization of pilosebaceous units by Propionibacterium acnes, which
`promotes perifollicular inflammation.
`Aim: The aim of the study was to investigate the therapeutic effects of resveratrol, a natural phytoalexin
`produced by some spermatophytes, such as grapes and other plants, on acneic skin.
`Methods: Resveratrol was incorporated in a carboxymethylcellulose-based gel. The chemical stability of
`resveratrol after storage at 4°C for 30 days was investigated by high-performance liquid chromatography
`(HPLC). The resveratrol-containing hydrogel was administered to 20 patients affected by acne vulgaris
`enrolled in this single-blind study. The resveratrol-containing formulation was applied daily as a solo
`treatment on the right side of the face for 60 days, while the hydrogel vehicle was applied to the left side of the
`face as a control. To objectively evaluate the results, a digital photographic database was used to collect
`images. The number and type of lesions were recorded for each patient, to compare the Global Acne Grading
`System (GAGS) score before treatment with that obtained at the end of the study. Moreover, with the
`innovative technique of follicular biopsy, areas of acneic skin were prepared for histopathology. The average
`area occupied by microcomedones at baseline was compared with that at the end of treatment.
`Results: HPLC analysis demonstrated that resveratrol, upon incorporation into the gel, did not convert to its
`cis-isomer when stored at 4°C for 30 days. All patients were satisfied with the active treatment and none
`experienced adverse effects. Clinical evaluation showed a 53.75% mean reduction in the GAGS score on the
`resveratrol-treated sides of the face compared with 6.10% on the vehicle-treated sides of the face. These data
`were supported by histologic analysis, which showed a 66.7% mean reduction in the average area of micro-
`comedones on the resveratrol-treated sides of the face. The comparison with the vehicle-treated side of
`the face (9.7% reduction) showed a clinically relevant and statistically significant decrease of lesions in areas
`treated with resveratrol-containing hydrogel.
`Conclusion: This pilot study showed positive results for resveratrol gel in acne, and should be considered a
`valid starting point for further testing of the effectiveness of this molecule in different concentrations and
`formulations and in a larger group of patients.
`
`Background
`
`Acne vulgaris is a complex, chronic, and common skin dis-
`order of pilosebaceous units that occurs predominantly on the
`skin of the face, the upper back, and the upper chest. This
`
`disease usually begins at the time of the sharp increase in an-
`drogen production that occurs during adolescence, but may
`affect individuals of any age.[1] In recent years, the multi-
`factorial nature of acne has been elucidated but much remains
`to be learned. The major pathogenic factors involved are ductal
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`134
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`Fabbrocini et al.
`
`hyperkeratinization, obstruction of sebaceous follicles resulting
`from abnormal keratinization of the infundibular epithelium,
`stimulation of sebaceous gland secretion by androgens, and
`microbial colonization of pilosebaceous units by Propioni-
`bacterium acnes, which promotes perifollicular inflammation.
`Acne begins when the pilosebaceous ducts become plugged
`with keratinocytes to form comedones, sebum builds up and
`distends the follicles, and the anerobe P. acnes proliferates
`in the sebum. If the comedo ruptures into the dermis, in-
`flammation results and a pustule or papule forms.[2]
`Currently, inflammatory acne is treated with antibacterials
`(both oral and topical) or oral isotretinoin. Although widely
`used and often effective, antibacterials have a significant limi-
`tation, which is the development of resistant strains of P. acnes.
`Oral isotretinoin is by far the most effective treatment for severe
`inflammatory acne, routinely delivering dramatic improve-
`ments.[3-6] However, because of its established teratogenicity,
`isotretinoin use has been restricted. Furthermore, the recently
`publicized, controversial risk of depression/suicidal ideation
`has made isotretinoin use even more problematic.[7,8] Current
`topical treatment options for inflammatory acne are sub-
`optimal. Rational development of new treatments for acne is
`based on a molecular description of the mechanisms involved
`in acne.
`in-
`Overstimulation of the initiation of the preclinical
`flammatory process or defective negative feedback regulation
`may be major reasons for the interruption of the normal cycling
`of the sebaceous follicle and be responsible for the initiation of
`the clinical inflammatory process in acne. Hereditary factors
`and excess androgen activity, for example, in puberty, may
`cause overstimulation, thus triggering sterile inflammatory
`phenomena. Neuroendocrinologic regulation and environ-
`mental factors, such as dietary lipids and smoking, have also
`been suggested to represent trigger mechanisms.[9] Hyper-
`proliferation of the follicular epithelium leads to formation of
`the first acne lesions that can be found in normal-looking skin,
`the microcomedones (comedones with an area between 0.016
`and 0.042 mm2), characterized by the accumulation of abnor-
`mally desquamated corneocytes and excess sebum[10] and which
`are believed to be the precursor lesions of acne. The micro-
`comedones may evolve into clinically visible comedones
`(blackheads and whiteheads) or inflammatory lesions (papules,
`pustules, or cysts). The very early stage of acne lesion devel-
`opment is associated with vascular endothelial cell activation
`and involvement of inflammatory events,[11] which corrobo-
`rates the suggestion that acne may represent a genuine in-
`flammatory disorder without involvement of bacteria in its
`initiation.
`
`Recently, it has been demonstrated that, in inflammatory
`acne lesions, there is activation of nuclear factor-kB signaling
`(a transcription factor critical for upregulation of many proin-
`flammatory cytokine genes). As a result, inflammatory cyto-
`kine genes (e.g. tumor necrosis factor [TNF]-a and interleukin
`[IL]-1X) are activated. These primary cytokines cause in-
`flammation by acting on endothelial cells to prepare adhesion
`molecules (e.g. intercellular adhesion molecule-1) to facilitate
`recruitment of all inflammatory cells into the skin. TNFa and
`IL-1X also help to stimulate the proliferation of secondary
`cytokines, such as IL-8, which can cooperate in the movement
`of the inflammatory cells towards the increased concentration
`of these particular chemicals.[12]
`P. acnes seems to be involved in both the initiation of acne
`lesions as a precursor of inflammation and in later events.
`P. acnes is a Gram-positive, non-motile rod, non-spore-
`forming, anerobic bacillus. As it is a member of the normal
`flora and is a harmless commensal, it is largely incapable of
`tissue invasion or serious infection. The organism metabolizes
`sebaceous triglycerides, consuming the glycerol fraction and
`discarding free fatty acids. As a consequence of P. acnes proli-
`feration and metabolism in the obstructed follicles, P. acnes pro-
`duces neutrophil chemoattractants. P. acnes also activates the
`complement system and is generally inflammatory when brought
`into contact with the immune system.[13] It has been shown that
`P. acnes, through activation of toll-like receptor 2 expressed in
`human monocytes, induces chemokine/cytokine synthesis in
`these cells.[14] These findings in combination with the expression
`of active toll-like receptors 2 and 4 and of CD14 in human ker-
`atinocytes have implicated P. acnes and toll-like receptors in acne
`inflammation. Recently, it has been shown that keratinocytes
`secrete selective human b-defensin-2 (hBD2) and IL-8 in res-
`ponse to P. acnes and that these molecules are potent chemo-
`tactic factors for leukocyte and neutrophil infiltration as well as
`for keratinocyte growth potential and differentiation at the site
`of P. acnes infections. This evidence shows that P. acnes is im-
`portant for maintaining the inflammation process in acne.[15]
`Because of the pathophysiology of the disease, the ideal
`treatment would be a single drug capable of suppressing the
`inflammatory response and also inhibiting keratinocytes and
`P. acnes proliferation. A potential candidate to fulfill these
`criteria is the phytoalexin resveratrol (3,4,5-trihydroxy-trans-
`stilbene). Resveratrol is a natural compound that exists in
`nature as cis- and trans-isomers. Most of the recorded health
`benefits are attributed to the trans-isomer.[16] The aim of this
`pilot cosmetic study was to evaluate the effects of a new topical
`formulation of trans-resveratrol in patients affected by in-
`flammatory acne vulgaris.
`
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`
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`Resveratrol-Containing Gel for Acne Vulgaris
`
`135
`
`Materials and Methods
`
`Patients
`
`The study group consisted of 20 patients (12 men and
`8 women) with facial acne vulgaris involving both sides of their
`faces with nearly symmetric appearance, as determined by
`clinical criteria. Their age at entry to the trial ranged from 18 to
`23 years.
`At the time the study started, patients had been in ther-
`apeutic wash-out for at least 4 weeks for both topical and sys-
`temic acne therapies. The exclusion criteria were a history of
`allergy, diabetes mellitus, and severe systemic disease.
`
`Materials
`
`Trans-resveratrol,
`sodium carboxymethylcellulose, and
`polysorbate 60 were purchased from Sigma-Aldrich Corp.
`(St Louis, MO, USA). Analytic grade 96% ethanol, and high-
`performance liquid chromatography (HPLC) grade acetoni-
`trile, methanol, and glacial acetic acid were obtained from
`Carlo Erba Reagenti (Milan, Italy). Glycerol (glycerin) was
`purchased from Polichimica (Bologna, Italy).
`
`Methods
`
`Resveratrol Analysis
`Trans-resveratrol analysis and detection of cis-resveratrol
`were carried out using an HPLC system by Shimadzu (Kyoto,
`Japan), consisting of a LC-10AD pump equipped with a Luna
`C18 (250 · 4.6 mm, 5 mm) column (Phenomenex, Torrance,
`CA, USA), a Rheodyne 7725i injection valve, and a SPV-10A
`UV-vis detector set at the wavelength of 303 nm. The system
`was controlled by a SCL-10A VP System Controller (Shi-
`madzu) connected with a computer. Chromatograms were ac-
`quired and analyzed by a Class VP Client/Server 7.2.1 program
`(Shimadzu). The analysis was performed with a mobile phase of
`methanol/water (50 : 50) with 0.5% volume in volume acetic
`acid in isocratic conditions at a flow rate of 1 mL/min. Cis-
`resveratrol was prepared from the trans-isomer by sunlight
`irradiation for 3 days, as reported elsewhere.[16]
`
`Preparation of the Resveratrol-Containing Gel
`The gel was prepared by dissolving 5 g of sodium carboxy-
`methylcellulose and 10 g of 85% glycerol in 85 g of deionized
`water under stirring. Separately, in the absence of light, res-
`veratrol was dissolved in deionized water at a final concen-
`tration of 0.01% weight in volume. Then, 1 g of this aqueous
`solution was incorporated into 10 g of the vehicle gel under
`
`stirring in the absence of light. The vehicle gel and resveratrol-
`containing gel were stored at 4°C.
`The stability of trans-resveratrol in the gel was evaluated
`soon after preparation and after different time frames of stor-
`age at 4°C. For each analysis, 0.5 g of the preparation was taken
`from the sample and resveratrol was extracted with 5 mL of
`methanol; then, the suspension was centrifuged at 4°C for
`30 minutes at 60 · g and the supernatant was analyzed by HPLC.
`
`Cyanoacrylate Follicular Biopsy
`Cyanoacrylate follicular biopsy, also known as skin surface
`biopsy, is a simple noninvasive method that removes only dead
`tissue and is used to study the stratum corneum, the many types
`of pathology within this compartment, and a vast array of
`microorganisms that may colonize or invade the layer.[17]
`It is the most reliable tool to sample the follicular contents of
`facial skin[18] such as hair and keratin. In the acneic or nearly
`acneic skin, keratin material may be particularly abundant
`(microcomedones) prior to the appearance of clinically evident
`comedones (macrocomedones). Cyanoacrylate follicular bi-
`opsy was performed by distributing three drops of cyanoacry-
`late evenly on a slide holder, which was then applied with a
`slight pressure to the chosen area of the face. After about
`3 minutes, the slide was removed, stripping an area of skin
`surface with a slow and undulating movement and being careful
`not to damage the stratum corneum sticker with its comedones.
`Ematossilin-eosin was used to stain each sample and the
`micrometric scale of the Zeiss optical microscope was used to
`determine the density of microcomedones, i.e. the number
`present per mm2, and the area occupied by each micro-
`comedone. From these calculations, we then determined the
`average area of microcomedones. The parameter used to test
`the treatment was the average area occupied by micro-
`comedones before and after treatment.
`
`Resveratrol Protocol and Patient Monitoring
`At baseline (T0), patients underwent a careful dermatologic
`examination and received accurate information about the stu-
`dy. All patients signed the informed consent, which contained
`the study’s description, aims, methods, and possible adverse
`effects. The study was submitted to a cosmeceutic committee,
`according to Italian cosmeceutic rules.
`In order to carry out a comparative analysis, three digital
`photographs (front view, right side of the face, left side of the
`face) were taken and gathered in a database. The severity of the
`lesions was evaluated using the Global Acne Grading System
`(GAGS)[19] that considers six sites (five facial, one truncal) by
`giving each a specific numerical factor based on the surface area
`
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Am J Clin Dermatol 2011; 12 (2)
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`136
`
`Fabbrocini et al.
`
`Table I. Global Acne Grading System (GAGS)
`
`Statistical Analysis
`
`Localization
`I = forehead
`II = right cheek
`III = left cheek
`IV = nose
`V = chin
`VI = chest and back
`
`Factor per grade (0–4)
`2 · grade
`2 · grade
`2 · grade
`1 · grade
`1 · grade
`3 · grade
`
`Grade
`0 = no lesions
`1 = ‡1 comedo
`2 = ‡1 papule
`3 = ‡1 pustule
`4 = ‡1 nodule
`
`The digital photographic data were analyzed using a test for
`nonparametric data (Wilcoxon signed-rank test for paired data).
`The null hypothesis (H0) was that the median of the differences is
`zero (P+ =P -) and the alternative hypothesis (HA) was that the
`median of the differences is negative (P+ < P-), a = 0.001. The
`result is given by computing the binomial probability.
`
`Results
`
`The first step of the study was the development of an HPLC
`method to allow the detection of both trans- and cis-isomers of
`resveratrol. The chromatographic analysis of the aqueous sol-
`ution containing trans-resveratrol gave a well defined peak with
`the retention time of 6.4 minutes. The sunlight irradiation of the
`trans-resveratrol solution for 3 days resulted in two distinct
`chromatographic peaks that were attributed, based on a pre-
`vious report,[20] to trans- and cis-resveratrol, respectively (data
`not shown).
`The second step of the study was the preparation of a der-
`matologic formulation containing resveratrol in which trans-
`resveratrol was incorporated at the final concentration of 1 mg/g
`of preparation. The stability of resveratrol in the preparation
`was evaluated soon after preparation and after 1, 10, and
`30 days of storage at 4°C by HPLC analysis. Figure 1 shows the
`chromatogram obtained from the analysis of the resveratrol-
`containing gel after 30 days of storage. In this figure, only the
`chromatographic peak attributed to trans-resveratrol was ob-
`served, while there were no peaks at a higher retention time, in-
`dicating the absence of cis-resveratrol. The chromatographic peaks
`with a retention time between 2 and 4 minutes (figure 1) were
`not attributed to resveratrol or to chemical compounds derived
`from it, because they were also found upon analysis of the vehi-
`cle gel.
`It is noteworthy that, in this study, in order to avoid inter-
`ference with resveratrol activity on the skin, preservatives were
`
`1500
`
`1000
`
`500
`
`0
`
`m Volts
`
`0
`
`2
`
`6
`4
`Time (min)
`
`8
`
`10
`
`Fig. 1. Chromatogram from the analysis of the resveratrol-containing gel
`after storage at 4°C for 30 days.
`
`of each and the distribution and density of pilosebaceous units.
`This factor is multiplied by the known grade to give the value of
`the local score. The sum of the local scores gives the global score
`(table I).
`At the end of the study, patients were asked to rate the clinical
`results on a 10-point satisfaction scale (1 = lowest to 10 = highest)
`including compliance, smell, texture, results, and quality of life.
`Before the treatment was started, the GAGS score was cal-
`culated and all patients with a score between 8 and 24 were
`included in the study. In order to obtain an objective evaluation
`of the efficacy of the treatment, the number of acne lesions
`before beginning treatment and at the end of the study was
`recorded for each patient.
`Moreover, to better define clinical results and to correctly
`evaluate the efficacy of the therapy, for each patient, two cyano-
`acrylate follicular biopsies (on the right and left sides of the face)
`were executed. Furthermore, digital photographs were captured
`while performing cyanoacrylate follicular biopsies: this guar-
`anteed the reproducibility of the technique and better diagnostic
`accuracy.
`The study was single blinded; each patient received two
`identical containers, marked by a letter of reference: ‘C’ for the
`formulation containing the vehicle gel and ‘R’ for the medi-
`cated formulation. Each patient was instructed to apply the gel
`in container R to the right side of the face, and the gel in con-
`tainer C to the left side once a day, preferably in the evening.
`Patients were told to store the gels in the refrigerator at a stable
`temperature.
`All patients attended the first follow-up visit 1 month later
`(T1) and each had three digital photographs taken. By analyz-
`ing digital photographs collected at T0, two follicular biopsies
`were obtained from the same sites of each patient’s face. Each
`patient returned the questionnaire that was given out during the
`first visit and since no patient had adverse effects, treatment was
`continued and the patients were asked to return for the last
`control check after a further 30 days.
`Therefore, the second and final follow-up occurred 60 days
`(T2) after the baseline visit, with the same evaluations repeated.
`
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Am J Clin Dermatol 2011; 12 (2)
`
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`Resveratrol-Containing Gel for Acne Vulgaris
`
`137
`
`Table II. Global Acne Grading System (GAGS) score before and after treatment with resveratrol and vehicle gels
`
`Patient no.
`
`Resveratrol-treated side
`
`Vehicle-treated side
`
`GAGS score before
`treatment
`
`GAGS score after
`treatment
`
`reduction
`(%)
`
`GAGS score before
`treatment
`
`GAGS score after
`treatment
`
`reduction
`(%)
`
`1
`
`2
`
`3
`
`4
`
`5
`
`6
`
`7
`
`8
`
`9
`
`10
`
`14
`
`12
`
`33
`
`14
`
`16
`
`9
`
`22
`
`14
`
`15
`
`18
`
`7
`
`2
`
`8
`
`10
`
`12
`
`6
`
`12
`
`7
`
`3
`
`5
`
`50
`
`83
`
`76
`
`29
`
`25
`
`33
`
`45
`
`50
`
`80
`
`72
`
`14
`
`12
`
`33
`
`14
`
`16
`
`9
`
`22
`
`14
`
`15
`
`18
`
`12
`
`12
`
`28
`
`18
`
`14
`
`15
`
`16
`
`12
`
`11
`
`16
`
`15
`
`14
`
`0
`
`15
`-28
`
`12
`-67
`
`27
`
`14
`
`27
`
`11
`
`6
`
`11
`
`12
`
`13
`
`14
`
`15
`
`16
`
`17
`
`18
`
`19
`
`20
`
`16
`
`26
`
`15
`
`19
`
`29
`
`23
`
`17
`
`18
`
`16
`
`22
`
`4
`
`7
`
`6
`
`8
`
`15
`
`16
`
`10
`
`11
`
`9
`
`8
`
`75
`
`73
`
`60
`
`58
`
`48
`
`30
`
`41
`
`39
`
`44
`
`64
`
`16
`
`26
`
`15
`
`19
`
`29
`
`23
`
`17
`
`18
`
`16
`
`22
`
`24
`
`14
`
`18
`
`28
`
`22
`
`17
`
`17
`
`15
`
`21
`
`8
`
`7
`
`5
`
`3
`
`4
`
`0
`
`5
`
`6
`
`5
`
`Mean
`
`18.40
`
`8.30
`
`53.75
`
`18.40
`
`17.25
`
`6.10
`
`not added. Indeed, for time frames longer than 1 month, the
`presence of microbiologic contamination was sometimes ob-
`served. For this reason, although resveratrol transformation
`was not observed at the timepoints considered, the resveratrol-
`containing formulations were used for no more than 30 days,
`after which time new preparations were provided to the patients.
`Data obtained using the GAGS at T0, T1, and T2 showed an
`average 53.75% reduction in clinical lesions on the resveratrol-
`treated sides of the face compared with 6.10% on the vehicle-
`treated sides of the face (table II).
`Data collected from digital photographs at times T0, T1, and
`T2 and analyzed with the sign test for paired data (p < 0.001)
`showed that for the lesions on the resveratrol-treated side of the
`face the median of the differences in GAGS scores was negative,
`in contrast to what was observed on the vehicle-treated side of
`the face. Based on this result, it could be argued that the re-
`duction in the degree of acne severity on the treated side of the
`face was statistically significant (figures 2 and 3).
`The analysis of cyanoacrylate follicular biopsies showed a
`66.7% mean decrease from baseline in the total area and a de-
`
`crease in the density of microcomedones (area between 0.016
`and 0.042 mm2) on the resveratrol-treated side of the face,
`versus a 9.7% decrease on the vehicle-treated side of the face
`(table III). Furthermore, there was a total disappearance of
`macrocomedones in some patients (figures 4 and 5).
`According to the patients’ subjective self-evaluation, using
`the 10-point scale, the mean satisfaction rate was 8 for com-
`pliance, 9 for smell, 8 for texture, 8 for results, and 9 for quality of
`life.
`
`Discussion
`
`The etiology of acne is not yet fully clarified but it is widely
`accepted that its pathogenesis is multifactorial, with abnormal
`follicular differentiation and increased cornification, enhanced
`sebaceous gland activity and hyperseborrhea, and bacterial
`hypercolonization, as well as inflammation and immunologic
`host reactions being the major contributors.
`New research into acne pathogenesis has identified several
`novel pharmacologic targets to interrupt the inflammatory
`
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Am J Clin Dermatol 2011; 12 (2)
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`

`138
`
`Fabbrocini et al.
`
`a
`
`b
`
`is a natural compound produced by some
`Resveratrol
`spermatophytes, such as grapes and other plants, and is present
`at concentrations of up to 100 mmol/L in red wines and at much
`lower concentrations in white wines. It is reported to have
`cardiovascular-protective, cancer-chemopreventive, antioxidant,
`and anti-inflammatory properties.[23,24]
`The keratinocytic hyperproliferation process, which is the
`basis of the follicular obstruction in acne, appears to be inhibited
`by resveratrol. In fact, even at submicromolar concentrations,
`resveratrol inhibits the proliferation of normal human keratino-
`cytes in vitro and at higher concentrations (40–100 mmol/L)
`is cytotoxic to these cells.[25] Our previous studies of HaCaT cells
`(a model for skin keratinocytes) indicated that resveratrol
`
`a
`
`b
`
`Fig. 2. A patient (no. 1) before (a) and after (b) treatment with resveratrol-
`containing gel.
`
`acne process. Indeed, some anti-inflammatory activity ob-
`served with the use of oral antibacterials may relate to their
`action in the inflammatory cascade.[21] Based on the patho-
`physiology of the disease, researchers have targeted therapy to
`try to suppress hyperseborrhea and the inflammatory response,
`and also inhibit keratinocytes and P. acnes proliferation. From
`this point of view, resveratrol seems to be an interesting mole-
`cule because it is able to interfere with the hyperproliferation
`of keratinocytes, the inflammatory response, and P. acnes
`growth.[22]
`
`Fig. 3. A patient (no. 3) before (a) and after (b) treatment with resveratrol-
`containing gel.
`
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Am J Clin Dermatol 2011; 12 (2)
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`139
`
`Table III. The average area of microcomedones before and after treatment with resveratrol and vehicle gels
`
`Patient no.
`
`Resveratrol-treated side
`
`Vehicle-treated side
`
`before treatment
`
`after treatment
`
`before treatment
`
`after treatment
`
`1
`
`2
`
`3
`
`4
`
`5
`
`6
`
`7
`
`8
`
`9
`
`10
`
`0.022
`
`0.016
`
`0.016
`
`0.024
`
`0.017
`
`0.022
`
`0.024
`
`0.025
`
`0.020
`
`0.024
`
`0.015
`
`0
`
`0.012
`
`0
`
`0.013
`
`0
`
`0.01
`
`0.01
`
`0
`
`0
`
`0.025
`
`0.010
`
`0.019
`
`0.021
`
`0.021
`
`0.014
`
`0.023
`
`0.021
`
`0.022
`
`0.025
`
`0.020
`
`0.010
`
`0.016
`
`0.025
`
`0.018
`
`0.011
`
`0.018
`
`0.022
`
`0.019
`
`0.022
`
`0.023
`
`change (%)
`-20
`
`change (%)
`-32
`-100
`-25
`-100
`-24
`-100
`-96
`-60
`-100
`-100
`-76
`-59
`-64
`-72
`-88
`-45
`-56
`-50
`-59
`-41
`–66.7
`
`0.025
`
`0.024
`
`0.021
`
`0.020
`
`0.024
`
`0.025
`
`0.017
`
`0.018
`
`0.021
`
`0.016
`
`0.021
`
`0.023
`
`0.020
`
`0.018
`
`0.022
`
`0.023
`
`0.015
`
`0.015
`
`0.018
`
`0.014
`
`0.019
`
`0
`-16
`+19
`-14
`-21
`-22
`+5
`-14
`-12
`-8
`-4
`-5
`-10
`-8
`-8
`-12
`-17
`-14
`-13
`–9.7
`
`11
`
`12
`
`13
`
`14
`
`15
`
`16
`
`17
`
`18
`
`19
`
`20
`
`Mean
`
`0.021
`
`0.022
`
`0.018
`
`0.018
`
`0.024
`
`0.022
`
`0.018
`
`0.020
`
`0.022
`
`0.017
`
`0.021
`
`0.005
`
`0.009
`
`0.006
`
`0.005
`
`0.003
`
`0.012
`
`0.008
`
`0.010
`
`0.009
`
`0.010
`
`0.007
`
`acts on an adaptive protein, p66, which at elevated doses
`leads to apoptosis, while in physiologic conditions promotes
`senescence.[26]
`Furthermore, resveratrol interferes with inflammation be-
`cause it regulates numerous intracellular signaling cascades
`converging with the activation of nuclear factor-kB and acti-
`vator protein-1, which act independently or in combination to
`control the expression of target genes, such as TNFa, IL-1b,
`and metalloproteinases.[27] In vivo experiments in mice showed
`a reduction in TNFa in splenic cells after treatment with re-
`sveratrol from 2 to 4 weeks, blocking the early stage of the
`inflammatory process.[28] It has been shown that resveratrol is
`active against several bacteria, including Neisseria gonorrhoeae,
`Neisseria meningitidis,[29] Helicobacter pylori,[30] Staphylococcus
`aureus, Enterococcus faecalis, Pseudomonas aeruginosa,[31] and
`Proteus mirabilis.[32] Recently, it has been reported that resver-
`atrol inhibits P. acnes replication in vitro at the lowest con-
`centration tested (50 mg/L) and is bactericidal at the highest
`concentration tested (200 mg/L).[33]
`Because of its anti-inflammatory properties, its antiproliferative
`effects, and its ability to inhibit P. acnes, we believed that re-
`
`sveratrol could be considered a potentially interesting molecule
`for acne treatment.
`In this study, we applied resveratrol in a carboxymethyl-
`cellulose-based gel. We demonstrated that resveratrol was stable
`in its trans-form upon storage of the gel at 4°C for the time neces-
`sary for the study. No preservative was added to the formulation
`in order to avoid interference with the activity of resveratrol.
`Future studies will be focused on the development of a resveratrol-
`containing gel suitable for long-term storage. Moreover, the use of
`nanotechnologies to improve the resveratrol penetration into the
`skin will be investigated. Therefore, further studies are planned to
`evaluate new formulations, with the aim of developing a com-
`mercial preparation.
`
`Conclusion
`
`At the end of the study, all patients had a visible clinical
`improvement on the resveratrol-treated side of the face, in-
`cluding a remarkable decrease in inflammation and pustular
`lesions. This clinical improvement was well documented by
`digital photographs. Our results indicate a clinically relevant
`
`ª 2011 Adis Data Information BV. All rights reserved.
`
`Am J Clin Dermatol 2011; 12 (2)
`
`7 of 9
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`

`

`140
`
`Fabbrocini et al.
`
`a
`
`b
`
`and statistically significant decrease of microcomedones and
`macrocomedones in facial areas treated with the gel for-
`mulation containing resveratrol. The treatment was well tol-
`erated, with no adverse effects reported on the resveratrol- or
`vehicle-treated sides of the face. All patients were satisfied with
`the treatment in terms of compliance, smell, texture, results,
`and quality of life. Our study provides only preliminary evi-
`dence for an alternative treatment for patients affected by acne
`vulgaris but it should be considered a valid starting point for
`further research into the effectiveness of resveratrol at different
`concentrations, in innovative formulations, in larger groups of
`patients, and for more widespread use on the body.
`
`a
`
`b
`
`Fig. 5. A follicular biopsy before (a) and after (b) treatment with resveratrol-
`containing gel (patient no. 3).
`
`Acknowledgments
`
`This study was achieved thanks to the contribution of the ‘‘Assessorato
`all’Agricoltura e alle Attivita` Produttive della Regione Campania.’’ The
`authors have no conflicts of interest that are directly relevant to the content
`of this study.
`
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`ª 2011 Adis Data Information BV. All rights reserved.
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`Am J Clin Dermatol 2011; 12 (2)
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