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High-frequency devices effect in vitro: promissing approach in the treatment of acne vulgaris?
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"tienePdf" => "pt" "tieneTextoCompleto" => "pt" "tieneResumen" => "pt" "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "729" "paginaFinal" => "734" ] ] "contieneResumen" => array:1 [ "pt" => true ] "contieneTextoCompleto" => array:1 [ "pt" => true ] "contienePdf" => array:1 [ "pt" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figura 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 2280 "Ancho" => 3341 "Tamanyo" => 374676 ] ] "descripcion" => array:1 [ "pt" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Efeitos antimicrobianos da terapia de AF. (A) Tratamento com AF mostra diminuição significativa da contagem de bactérias e (B) fungos <span class="elsevierStyleItalic">in vitro</span>. (C) UFC de <span class="elsevierStyleItalic">P. acnes</span> são significativamente reduzidas após 1 minuto, e a continuação do tratamento leva a redução adicional.</p>" ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Leonie Frommherz, Markus Reinholz, Anne Gürtler, Pia‐Charlotte Stadler, Till Kaemmerer, Lars French, Benjamin M. Clanner‐Engelshofen" "autores" => array:7 [ 0 => array:2 [ "nombre" => "Leonie" "apellidos" => "Frommherz" ] 1 => array:2 [ "nombre" => "Markus" "apellidos" => "Reinholz" ] 2 => array:2 [ "nombre" => "Anne" "apellidos" => "Gürtler" ] 3 => array:2 [ "nombre" => "Pia‐Charlotte" "apellidos" => "Stadler" ] 4 => array:2 [ "nombre" => "Till" "apellidos" => "Kaemmerer" ] 5 => array:2 [ "nombre" => "Lars" "apellidos" => "French" ] 6 => array:2 [ "nombre" => "Benjamin M." "apellidos" => "Clanner‐Engelshofen" ] ] ] ] ] "idiomaDefecto" => "pt" "Traduccion" => array:1 [ "en" => array:9 [ "pii" => "S0365059622001945" "doi" => "10.1016/j.abd.2021.09.015" "estado" => "S300" "subdocumento" => "" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:1 [ "total" => 0 ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0365059622001945?idApp=UINPBA00008Z" ] ] "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2666275222002053?idApp=UINPBA00008Z" "url" => "/26662752/0000009700000006/v4_202410140645/S2666275222002053/v4_202410140645/pt/main.assets" ] ] "itemSiguiente" => array:17 [ "pii" => "S0365059622002008" "issn" => "03650596" "doi" => "10.1016/j.abd.2021.09.016" "estado" => "S300" "fechaPublicacion" => "2022-11-01" "aid" => "628" "documento" => "article" "crossmark" => 1 "subdocumento" => "fla" "abierto" => array:3 [ "ES" => 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"at0045" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Agarose gel electrophoresis images for IL-17A genotypes, GG genotype 3, 4, 5, 6, 7 and 9 lanes, AG genotype 2, 10 and 11 lanes, AA genotype 1 and 8 lanes.</p>" ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Azza Gaber Antar Farag, Azza Zagloul Labeeb, Amany Nagy Abdalla Gerges, Mustafa Elsayed Elshaib" "autores" => array:4 [ 0 => array:2 [ "nombre" => "Azza Gaber Antar" "apellidos" => "Farag" ] 1 => array:2 [ "nombre" => "Azza Zagloul" "apellidos" => "Labeeb" ] 2 => array:2 [ "nombre" => "Amany Nagy Abdalla" "apellidos" => "Gerges" ] 3 => array:2 [ "nombre" => "Mustafa Elsayed" "apellidos" => "Elshaib" ] ] ] ] ] "idiomaDefecto" => "en" "Traduccion" => array:1 [ "pt" => array:9 [ "pii" => "S2666275222002120" "doi" => "10.1016/j.abdp.2022.09.024" "estado" => "S300" "subdocumento" => "" "abierto" => array:3 [ "ES" => 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Article</span>" "titulo" => "Effect of crosstalk between Th17 and Th9 cells on the activation of dermal vascular smooth muscle cells in systemic scleroderma and regulation of tanshinone IIA" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => "en" "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "716" "paginaFinal" => "728" ] ] "contieneResumen" => array:1 [ "en" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:8 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 3020 "Ancho" => 2508 "Tamanyo" => 449969 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at0025" "detalle" => "Figure " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "<p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">IL-17 neutralizing antibody and Tanshinone IIA inhibit the differentiation of immature T-lymphocytes into Th9. (A and B) The proportion of immature T-lymphocytes differentiated into Th9 in each group was detected using flow cytometry. (C). The content of IL-9 secreted by immature T-lymphocytes in each group was detected by ELISA. IL-17 concentration 100 ng/mL, anti-IL-17 concentration 8 μg/mL, Tanshinone IIA concentration 50 μM. * vs. control group, p < 0.05, *** vs. control group, p < 0.001, <span class="elsevierStyleSup">###</span> vs. IL-9 group, p < 0.001.</p>" ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Mengguo Liu" "autores" => array:1 [ 0 => array:2 [ "nombre" => "Mengguo" "apellidos" => "Liu" ] ] ] ] ] "idiomaDefecto" => "en" "Traduccion" => array:1 [ "pt" => array:9 [ "pii" => "S2666275222002132" "doi" => "10.1016/j.abdp.2022.09.025" "estado" => "S300" "subdocumento" => "" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:1 [ "total" => 0 ] "idiomaDefecto" => "pt" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S2666275222002132?idApp=UINPBA00008Z" ] ] "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S036505962200201X?idApp=UINPBA00008Z" "url" => "/03650596/0000009700000006/v2_202306261757/S036505962200201X/v2_202306261757/en/main.assets" ] "en" => array:19 [ "idiomaDefecto" => true "cabecera" => "<span class="elsevierStyleTextfn">Original Article</span>" "titulo" => "High-frequency devices effect in vitro: promissing approach in the treatment of acne vulgaris?" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "729" "paginaFinal" => "734" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Leonie Frommherz, Markus Reinholz, Anne Gürtler, Pia-Charlotte Stadler, Till Kaemmerer, Lars French, Benjamin M. Clanner-Engelshofen" "autores" => array:7 [ 0 => array:3 [ "nombre" => "Leonie" "apellidos" => "Frommherz" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 1 => array:4 [ "nombre" => "Markus" "apellidos" => "Reinholz" "email" => array:1 [ 0 => "Markus.Reinholz@med.uni-muenchen.de" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">*</span>" "identificador" => "cor0005" ] ] ] 2 => array:3 [ "nombre" => "Anne" "apellidos" => "Gürtler" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 3 => array:3 [ "nombre" => "Pia-Charlotte" "apellidos" => "Stadler" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 4 => array:3 [ "nombre" => "Till" "apellidos" => "Kaemmerer" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 5 => array:3 [ "nombre" => "Lars" "apellidos" => "French" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 6 => array:3 [ "nombre" => "Benjamin M." "apellidos" => "Clanner-Engelshofen" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Department of Dermatology and Allergy, Ludwig Maximilian University, Munich, Germany" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Dr Phillip Frost Department of Dermatology & Cutaneous Surgery, University of Miami, Miller School of Medicine, Miami, USA" "etiqueta" => "b" "identificador" => "aff0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:8 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2690 "Ancho" => 3341 "Tamanyo" => 729556 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at0005" "detalle" => "Fig. " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Plasma source and mechanism effects. (a) Plasma device consisting of a handheld with intensity adjust and different glass tubes to plug in. Formation of orange colored plasma when turned on. (b) Pictures taken with a thermal imaging camera after 0, 1, 2 and 4 minutes without any sign of heat development. (c) Ozone formation during time measured by an O<span class="elsevierStyleInf">3</span> detector demonstrating an increase of O<span class="elsevierStyleInf">3</span> concentration after 10 minutes already. The dashed line indicates the odor threshold concentration for ozone (20‒40 µg/m<span class="elsevierStyleSup">3</span>; 0.01‒0.02 ppm).</p>" ] ] ] "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Acne vulgaris is frequently affecting approximately 80% of adolescents and is, therefore, one of the most common reasons for consultation with a dermatologist.<a class="elsevierStyleCrossRef" href="#bib0005"><span class="elsevierStyleSup">1</span></a> The pathogenesis of this inflammatory skin disorder is multifactorial including increased sebum production, altered follicular keratinization, bacterial colonization with <span class="elsevierStyleItalic">Propionibacterium acnes,</span> and inflammation.<a class="elsevierStyleCrossRefs" href="#bib0010"><span class="elsevierStyleSup">2–4</span></a> It is divided into four grades from mild (including comedones and a few inflammatory papules) to severe (along with inflammatory nodules and scarring).<a class="elsevierStyleCrossRef" href="#bib0025"><span class="elsevierStyleSup">5</span></a> Furthermore, acne is considered a stigmatizing disease leading to low self-esteem and social withdrawal, also reflected by a high Dermatology Life Quality Index (DLQI) resembling those of psoriasis vulgaris patients.<a class="elsevierStyleCrossRef" href="#bib0030"><span class="elsevierStyleSup">6</span></a> Thus, the demand for effective treatments is particularly high. A multitude of treatment options exists ranging from over-the-counter medication to oral therapies including retinoids and antibiotics. Recently, high-frequency devices have gained increasing attention claiming to support lymph drainage, prevent both hair loss and the development of wrinkles, and ultimately, to improve acne lesions. Those high-frequency devices colloquially termed Violet Wand (VW), deliver Cold Atmospheric Pressure Plasma (CAPP). CAPP has multiple bioactive properties e.g., by the release of charged particles and reactive species (O<span class="elsevierStyleInf">3</span>, NO, NO<span class="elsevierStyleInf">2</span>).<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> In the early last century, High Frequency (HF) therapy was already known as a versatile treatment option in patients with infectious diseases of the skin, eczema, and wounds. Additionally, even migraines, neuralgia, tuberculosis, and many more diseases were treated by this method.<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a> Probably due to the development of antibiotics and lacking data on efficacy, it became less important in the middle of the last century. Not only because of its antimicrobial effects as a “physical antiseptic” but also because of the global increase of multi-resistant bacterial strains, plasma medicine is currently an emerging field and ongoing studies are ubiquitous. Recently, it was shown that HF devices are more effective than common antiseptics in targeting wound pathogens <span class="elsevierStyleItalic">in vitro</span>,<a class="elsevierStyleCrossRefs" href="#bib0035"><span class="elsevierStyleSup">7,8</span></a> arising the question of whether HF therapy may be also beneficial in acne-prone skin and how it affects common skin flora.</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Plasma source</span><p id="par0010" class="elsevierStylePara elsevierViewall">In this study, <span class="elsevierStyleItalic">Signstek Portable High Frequency Machine</span> (Signstek, Full Rise LLC, Wilmington, DE/USA) carrying CE labeling (approved for use in humans in Europe) was used as a technical device consisting of a handheld with an intensity regulator on the bottom and an output for plugging in different glass tubes filled with helium below atmospheric pressure. Four different glass tubes are supplied: a comb tube for treating the scalp, a tongue tube for sensitive areas (such as dark circles beneath the eyes, nose, or lips), a mushroom tube for large areas (forehead, back), and a bent tube with a spherical tip for regular areas like cheeks (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>a).</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0015" class="elsevierStylePara elsevierViewall">In all experiments here, the bent tube was used at a working frequency of 50 Hz. The intensity was regulated at 50%.</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Technical measurements</span><p id="par0020" class="elsevierStylePara elsevierViewall">Current strength was measured for three conditions (low, medium, and full power) using a standard ammeter (Multimeter ABB Metrawatt 2012). Applied voltage corresponding to the electric outlet (in Germany: 230 V).</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Evaluation of heat development</span><p id="par0025" class="elsevierStylePara elsevierViewall">For further investigations on the precise mechanism of action of our plasma source, a thermal imaging camera (FLIR MS Technology C3, FLIR® Systems, Inc., Wilsonville, Oregon, USA) was utilized to detect potential heat development. Pictures were taken after 1, 2, and 4 minutes of HF treatment of an agar plate to resemble an experiment on bacteria.</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Ozone formation</span><p id="par0030" class="elsevierStylePara elsevierViewall">Ozone (O<span class="elsevierStyleInf">3</span>) concentration was measured using an O<span class="elsevierStyleInf">3</span> detector (Leory PM 2.5 O<span class="elsevierStyleInf">3</span> Ozondetektor TVOC, Thingnovation, Shenzhen, China). Our plasma source was switched on to 100% in a room with locked windows and doors of approximately 10.5 cubic meters. Measurement time points were 10, 20, 30, 40 and 50 minutes.</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Microbiology</span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Bacteriology</span><p id="par0035" class="elsevierStylePara elsevierViewall">To study the effects of HF therapy on bacteria, swabs were taken from healthy skin in three different body areas (forehead, forearm, and genital area) from a healthy individual. In a second run, <span class="elsevierStyleItalic">Propionibacterium acnes</span> (ATCC® 6919™) was tested separately.</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Mycology</span><p id="par0040" class="elsevierStylePara elsevierViewall">To further evaluate the effects on fungi, clinical isolates of <span class="elsevierStyleItalic">Trichophyton</span> (T.) <span class="elsevierStyleItalic">mentagrophytes</span>, <span class="elsevierStyleItalic">T. violaceum</span>, <span class="elsevierStyleItalic">T. mentagrophytes</span>, <span class="elsevierStyleItalic">T. benhamiae</span>, <span class="elsevierStyleItalic">T. rubrum</span> and <span class="elsevierStyleItalic">Microsporum</span> (M.) <span class="elsevierStyleItalic">canis</span> determined by medical-laboratory assistants were treated as mentioned below.</p></span></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Cultivation of bacteria/fungi</span><p id="par0045" class="elsevierStylePara elsevierViewall">Skin flora from three different body areas was isolated on blood agar plates and incubated for 24 h at 37 °C before HF treatment.</p><p id="par0050" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">P. acnes</span>, as an anaerobic bacterium, was cultivated in an anaerobic vial (BD BACTEC™ Lytic/10 Anaerobic/F Culture Vial) until an approximate cell density of 3.0 × 10<span class="elsevierStyleSup">8</span> CFU/mL referring to McFarland Standard nº 1 was reached, then following HF treatment on a blood agar plate. After treatment, plates were incubated for 7d at 37 °C in anaerobic conditions using BD GasPak™. Fungal species were cultivated on BD Dermatophyte Agar for 7d at 37 °C before treatment.</p></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">In vitro model for high frequency testing in bacteria/fungi</span><p id="par0055" class="elsevierStylePara elsevierViewall">After incubation, each investigated colony was resuspended in approximately 0.5 mL of sterile 0.9% saline solution. For skin flora, the authors chose five morphologically different appearing colonies for further evaluation. Thereafter, 1 µL of the microbial solution was applied centrally to the appropriate agar plates in pairs. Next, the HF device with an attached, wipe-disinfected bent tube was fixed on a stand at a distance of 5 mm above the plate. Of each pair, one plate was treated for 1 minute, and the other one served as a control to determine the relative difference. In the case of <span class="elsevierStyleItalic">P. acnes</span>, treatment periods varied between 1 and 8 minutes (1, 2, 4 and 8 minutes of HF treatment). Subsequently, the microbial solution of treated and untreated plates was spread using sterilized glass beads followed by an incubation period of 1d for skin flora and 7d for <span class="elsevierStyleItalic">P. acnes</span> and fungi at 37 °C. After incubation, on plates with more than approximately 1000 colonies, three 1 cm² areas on each agar plate were counted, and mean values were multiplied by the size of the agar plate (57 cm²) to obtain the absolute number of colonies per plate. Plates with 100 to 1000 colonies were divided into quarters and one counted quarter was multiplied by four, plates with less than 100 colonies were counted completely.</p></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">16S sequencing</span><p id="par0060" class="elsevierStylePara elsevierViewall">Bacterial colonies of each plate pair were further characterized by performing a PCR (HotStarTaq Master Mix Kit, Qiagen) as described previously.<a class="elsevierStyleCrossRef" href="#bib0045"><span class="elsevierStyleSup">9</span></a></p><p id="par0065" class="elsevierStylePara elsevierViewall">Primers 27f (5’-AGA GTT TGA TCA TGG CTC A-3’) and 1492 r (5’-TAC GGT TAC CTT GTT ACG ACT T-3’) partially amplified the 16S rRNA gene using a thermocycler (Biometra Professional Basic, Analytik Jena GmbH, Jena, Germany).</p><p id="par0070" class="elsevierStylePara elsevierViewall">The DNA products were visualized by 1.0% agarose gel electrophoresis to confirm a successful amplification. Sanger sequencing was performed by Eurofins Genomics (Ebersberg, Germany) using the 27f primer. Obtained sequences were aligned using the online tool BLASTn to identify the species (<a href="https://blast.ncbi.nlm.nih.gov/Blast.cgi">https://blast.ncbi.nlm.nih.gov/Blast.cgi</a>; NCBI).<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a></p></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Statistical analysis</span><p id="par0075" class="elsevierStylePara elsevierViewall">For visualization and statistical analyses (unpaired Student’s <span class="elsevierStyleItalic">t</span>-test and row test), GraphPad Prism (version 9.0.0, GraphPad Software Inc, San Diego, CA/USA) was used and data was visualized in logarithmically scaled graphs. All experiments were performed in triplicates. The following values are indicated by mean ± standard deviation. The significance level was set at p < 0.05 (*), p < 0.01 (**), and p < 0.001 were considered highly significant (***).</p></span></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Results</span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Assessment of technical data and mechanism of action</span><p id="par0080" class="elsevierStylePara elsevierViewall">To evaluate electrical current strength during treatment, a multimeter was used showing a current ranging from 0.28 (full power) to 0.32 ampere (low power) at a voltage of 230 V.</p><p id="par0085" class="elsevierStylePara elsevierViewall">Heat development after 1, 2, and 4 minutes of HF treatment were not observed. Pictures taken with a thermal imaging camera did not show changes in temperature during the investigated period (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>b).</p><p id="par0090" class="elsevierStylePara elsevierViewall">Ozone formation with a concentration of 0.015 ppm was observed after 10 minutes of a fully switched-on device. After 20 minutes, O<span class="elsevierStyleInf">3</span> concentrations almost doubled (c = 0.027 ppm). In the further course, concentration levels increased (c = 0.032 after 30 minutes, c = 0.040 after 40 minutes, c = 0.090 after 50 minutes) (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>c).</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Effects of high frequency therapy against skin flora</span><p id="par0095" class="elsevierStylePara elsevierViewall">To assess the efficacy of HF therapy against common skin flora, blood agar plates with and without HF treatment were counted after the incubation period followed by a PCR to specify bacterial species. In broad terms, for almost every identified species a significant reduction in colony count was observed. More precisely, on the forehead, the authors detected <span class="elsevierStyleItalic">Micrococcus yunnanensis</span> twice, <span class="elsevierStyleItalic">Aerococcus viridans</span>, <span class="elsevierStyleItalic">Staphylococcus epidermidis</span> and <span class="elsevierStyleItalic">Staphylococcus capitis</span>. <span class="elsevierStyleItalic">M. yunnanensis</span> was reduced by 67.5% (31267.0 ± 81.7 vs. 1016.3 ± 57.1; p < 0.001) and on the second plate by 12.5% (6405.0 ± 18.5 vs. 5605.3 ± 19.4; p < 0.001), <span class="elsevierStyleItalic">A. viridans</span> by 26.1% (9339.0 ± 72.1 vs. 6906.3 ± 58.8; p < 0.001), <span class="elsevierStyleItalic">S. epidermidis</span> by 83.43% (3785.7 ± 68.7 vs. 627.3 ± 18.2; p < 0.001) and <span class="elsevierStyleItalic">S. capitis</span> by 41.9% (1488.0 ± 70.4 vs. 864.3 ± 30.3; p < 0.001).</p><p id="par0100" class="elsevierStylePara elsevierViewall">Likewise for bacteria of the forearm: <span class="elsevierStyleItalic">B. cereus</span> was reduced by 75.9% (11396.3 ± 757.7 vs. 2750.7 ± 127.9; p < 0.001), <span class="elsevierStyleItalic">S. capitis</span> by 63.9% (2700.3 ± 104.0 vs. 973.7 ± 25.1; p < 0.001), <span class="elsevierStyleItalic">A. urinaeequi</span> by 95.1% (16201.0 ± 757.9 vs. 800.0 ± 35.0; p < 0.001), <span class="elsevierStyleItalic">M. yunnanensis</span> by 99.0% (7209.3 ± 344.8 vs. 75.0 ± 12.8; p < 0.001) and <span class="elsevierStyleItalic">M. luteus</span> by 3.1% (6280.0 ± 443.3 vs. 6087.7 ± 119.3; p = 0.5).</p><p id="par0105" class="elsevierStylePara elsevierViewall">Regarding bacterial species of the genital region, <span class="elsevierStyleItalic">S. capitis</span> was reduced by 81.6% (31699.3 ± 763.7 vs. 5839.7 ± 111.3; p < 0.001), <span class="elsevierStyleItalic">S. lugdunensis</span> by 97.6% (15214.3 ± 405.1 vs. 363.3 ± 16.2; p < 0.001), <span class="elsevierStyleItalic">M. luteus</span> 17.9% (957.0 ± 37.6 vs. 785.3 ± 22.3; p = 0.002), <span class="elsevierStyleItalic">S. haemolyticus</span> 99.0% (6684.0 ± 273.0 vs. 70.0 ± 7.2; p < 0.001), <span class="elsevierStyleItalic">M. yunnanensis</span> 70.3% (4322.0 ± 252.3 vs. 1284.3 ± 86.4; p < 0.001) (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>a).</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Effects of high frequency therapy against dermatophytes</span><p id="par0110" class="elsevierStylePara elsevierViewall">The next step of this study was to further evaluate the effects of HF treatment against a random selection of dermatophytes. The total number of all dermatophytes’ colonies was significantly reduced by 58.0% after HF treatment (55.8 ± 33.6 vs. 18.2 ± 6.5; p = 0.040). More precisely, in <span class="elsevierStyleItalic">T. mentagrophytes</span> colonies were reduced by 61.5% (69.3 ± 4.0 vs. 26.7 ± 3.1; p < 0.001), in <span class="elsevierStyleItalic">T. violaceum</span> by 78.6% (103.0 ± 6.6 vs. 22.0 ± 3.0; p < 0.001), in <span class="elsevierStyleItalic">T. benhamiae</span> by 45.2% (17.7 ± 1.5 vs. 9.7 ± 1.5; p = 0.003), in <span class="elsevierStyleItalic">M. canis</span> by 69.1% (65.7 ± 2.1 vs. 20.3 ± 1.5; p < 0.001) and in <span class="elsevierStyleItalic">T. rubrum</span> by 61.6% (31.3 ± 1.5 vs. 12.0 ± 2.0; p < 0.001) (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>b).</p></span><span id="sec0090" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0130">Effects of high frequency therapy against <span class="elsevierStyleItalic">P. acnes</span></span><p id="par0115" class="elsevierStylePara elsevierViewall">To further address the question of whether HF treatment is beneficial against <span class="elsevierStyleItalic">P. acnes</span>, <span class="elsevierStyleItalic">P. acnes</span> was treated by HF over time (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>c). CFU on different blood agar plates was counted before and after 1, 2, 4, and 8 minutes of HF treatment. Significant results were reached after 1 minute of treatment (7288.0 ± 145.9 vs. 30.0 ± 3.6; p < 0.001). Continuing the treatment for 2 minutes, 4- and 8-minutes lead to a further decrease in the amount of CFU. When comparing the values after 2 and 4 minutes of treatment a further significant reduction of <span class="elsevierStyleItalic">P. acnes</span> CFU was observed (4.7 ± 0.6 vs. 1.3 ± 1.5; p = 0.020). Prolonged treatment over 8 minutes seemed not to have further benefits (1.3 ± 1.5 vs. 1 ± 0; p = 0.700) (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>c).</p></span></span><span id="sec0095" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0135">Discussion</span><p id="par0120" class="elsevierStylePara elsevierViewall">In this study, HF treatment demonstrated a microbicidal effect on skin flora and pathogens in vitro with a significant decrease in bacterial and fungal counts after a short treatment period.</p><p id="par0125" class="elsevierStylePara elsevierViewall">While the authors show that HF devices do not work through heat development, we observed increasing O<span class="elsevierStyleInf">3</span> formation during the time of application suggesting its main antimicrobial effects are likely mediated by the development of ozone gas and therefore oxidative stress for the microbes. O<span class="elsevierStyleInf">3</span> is known to have an anti-inflammatory effect with antimicrobial properties.<a class="elsevierStyleCrossRefs" href="#bib0055"><span class="elsevierStyleSup">11,12</span></a> In high concentrations (usually >200 µg/m<span class="elsevierStyleSup">3</span>, >0.1 ppm) over prolonged periods of time, O<span class="elsevierStyleInf">3</span> might contribute to the development of lung diseases, such as COPD or emphysema,<a class="elsevierStyleCrossRefs" href="#bib0065"><span class="elsevierStyleSup">13,14</span></a> but the amount of O<span class="elsevierStyleInf">3</span> in HF treatment is too small and time of application too short to cause serious health issues. The authors were able to show that skin flora from three different body areas including different aerobic bacteria was significantly reduced after 1 minute of HF treatment. In accordance with that, a previous study demonstrated a complete eradication of bacterial wound isolates in vitro.<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a> Furthermore, the present results are concordant with the findings of a study from 2015 showing that CAPP and VW can reduce a variety of bacteria, including Multi-Resistant Strains like oxacillin-resistant <span class="elsevierStyleItalic">Staphylococcus Aureus</span> (MRSA) and vancomycin-resistant <span class="elsevierStyleItalic">Enterococci</span> (ESBL) as well as fungi, like the yeast <span class="elsevierStyleItalic">Candida albicans</span>. The antimicrobial effects of VW did not relevantly differ from modern HF devices.<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a> The present study and previous studies clearly highlighted the antiseptic properties of HF therapies, thus suggesting that HF instead of antibiotics might be an alternative antimicrobial treatment option for easily accessible superficial infections, especially since multi-drug resistant bacterial strains are increasing. Moreover, the antimicrobial effect is also important for the reduction of <span class="elsevierStyleItalic">P. acnes</span> colonization as the authors were able to show a significant reduction after 1 minute of treatment with the HF device. Since <span class="elsevierStyleItalic">P. acnes</span> is a key player in acne vulgaris pathogenesis its reduction might display an important complemental treatment option.</p></span><span id="sec0100" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0140">Conclusion</span><p id="par0130" class="elsevierStylePara elsevierViewall">The results collected in this study, although in vitro, provide a mechanistic basis for HF as a complementary treatment option for patients with acne. It might also have a beneficial effect on patients with superficial infectious skin disorders. As the in vitro results are promising, further in vivo studies are needed to prove efficacy and tolerability in a real-life setting with long-term follow-ups to monitor possible side effects.</p></span><span id="sec0105" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0145">Financial support</span><p id="par0135" class="elsevierStylePara elsevierViewall">None.</p></span><span id="sec0110" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0150">Authors’ contributions</span><p id="par0140" class="elsevierStylePara elsevierViewall">Leonie Frommherz: Collected and analysed the data and drafted the manuscript.</p><p id="par0145" class="elsevierStylePara elsevierViewall">Markus Reinholz: Collected and analysed the data and drafted the manuscript.</p><p id="par0150" class="elsevierStylePara elsevierViewall">Anne Gürtler: Collected data, reviewed them and the manuscript.</p><p id="par0155" class="elsevierStylePara elsevierViewall">Pia-Charlotte Stadler: Collected data, reviewed them and the manuscript.</p><p id="par0160" class="elsevierStylePara elsevierViewall">Till Kaemmerer: Collected data, reviewed them and the manuscript.</p><p id="par0165" class="elsevierStylePara elsevierViewall">Lars French: Collected data, reviewed them and the manuscript.</p><p id="par0170" class="elsevierStylePara elsevierViewall">Benjamin M. Clanner-Engelshofen: Collected and analyzed the data and drafted the manuscript.</p></span><span id="sec0115" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0155">Conflict of interest</span><p id="par0175" class="elsevierStylePara elsevierViewall">None declared.</p></span></span>" "textoCompletoSecciones" => array:1 [ "secciones" => array:11 [ 0 => array:3 [ "identificador" => "xres1921646" "titulo" => "Abstract" "secciones" => array:6 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Objectives" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Study limitations" ] 5 => array:2 [ "identificador" => "abst0030" "titulo" => "Conclusions" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec1657554" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:9 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Plasma source" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Technical measurements" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Evaluation of heat development" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Ozone formation" ] 4 => array:3 [ "identificador" => "sec0035" "titulo" => "Microbiology" "secciones" => array:2 [ 0 => array:2 [ "identificador" => "sec0040" "titulo" => "Bacteriology" ] 1 => array:2 [ "identificador" => "sec0045" "titulo" => "Mycology" ] ] ] 5 => array:2 [ "identificador" => "sec0050" "titulo" => "Cultivation of bacteria/fungi" ] 6 => array:2 [ "identificador" => "sec0055" "titulo" => "In vitro model for high frequency testing in bacteria/fungi" ] 7 => array:2 [ "identificador" => "sec0060" "titulo" => "16S sequencing" ] 8 => array:2 [ "identificador" => "sec0065" "titulo" => "Statistical analysis" ] ] ] 4 => array:3 [ "identificador" => "sec0070" "titulo" => "Results" "secciones" => array:4 [ 0 => array:2 [ "identificador" => "sec0075" "titulo" => "Assessment of technical data and mechanism of action" ] 1 => array:2 [ "identificador" => "sec0080" "titulo" => "Effects of high frequency therapy against skin flora" ] 2 => array:2 [ "identificador" => "sec0085" "titulo" => "Effects of high frequency therapy against dermatophytes" ] 3 => array:2 [ "identificador" => "sec0090" "titulo" => "Effects of high frequency therapy against P. acnes" ] ] ] 5 => array:2 [ "identificador" => "sec0095" "titulo" => "Discussion" ] 6 => array:2 [ "identificador" => "sec0100" "titulo" => "Conclusion" ] 7 => array:2 [ "identificador" => "sec0105" "titulo" => "Financial support" ] 8 => array:2 [ "identificador" => "sec0110" "titulo" => "Authors’ contributions" ] 9 => array:2 [ "identificador" => "sec0115" "titulo" => "Conflict of interest" ] 10 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2021-08-01" "fechaAceptado" => "2021-09-21" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec1657554" "palabras" => array:4 [ 0 => "Acne vulgaris" 1 => "Bacterial infection" 2 => "High-frequency" 3 => "Microbiome" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:3 [ "titulo" => "Abstract" "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Acne vulgaris is an inflammatory skin disorder leading to an impairment of quality of life and is therefore not only a cosmetic issue. Its pathogenesis is multifactorial – of particular importance is the colonization with the bacterium <span class="elsevierStyleItalic">Propionibacterium acnes</span>. A wide range of different treatment options exists including topical and systemic treatments depending on severity. High Frequency (HF) therapy, historically developed in the 19<span class="elsevierStyleSup">th</span> century, claims antimicrobial effects on acne skin, but solid data on its efficacy and mechanism of action is lacking.</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Objectives</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">The main objective of this study was to determine the efficacy of HF therapy on skin flora and <span class="elsevierStyleItalic">P. acnes</span> in vitro using a commercial device as well as to review studies on the mechanism of action.</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">The plasma source was investigated regarding electrical settings, heat, and ozone development. Bacterial skin flora, fungal isolates, and <span class="elsevierStyleItalic">P. acnes</span> were exposed to HF in vitro and compared to unexposed controls by evaluating the number of colonies on agar plates. To further analyze bacterial species from normal skin flora, 16S-sequencing was performed. Statistical analyses were carried out using row analysis and unpaired <span class="elsevierStyleItalic">t</span>-test.</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">HF treatment led to a significant reduction of almost every bacterial and fungal species investigated in this study. Moreover, the number of colonies forming units was significantly decreased in <span class="elsevierStyleItalic">P. acnes</span> after HF treatment compared to controls in vitro.</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Study limitations</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">The experiments were performed in vitro only. To assess clinical effects further in vivo experiments are necessary.</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Conclusions</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">The results collected in this study, although in vitro, provide a mechanistic basis for HF as a complementary treatment option for patients with acne. It might also have a beneficial effect on patients with superficial infectious skin of the skin.</p></span>" "secciones" => array:6 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Objectives" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Study limitations" ] 5 => array:2 [ "identificador" => "abst0030" "titulo" => "Conclusions" ] ] ] ] "NotaPie" => array:1 [ 0 => array:2 [ "etiqueta" => "☆" "nota" => "<p class="elsevierStyleNotepara" id="npar0005">Study conducted at the Department of Dermatology and Allergy, University Hospital, LMU Munich, Munich, Germany.</p>" ] ] "multimedia" => array:2 [ 0 => array:8 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2690 "Ancho" => 3341 "Tamanyo" => 729556 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at0005" "detalle" => "Fig. " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Plasma source and mechanism effects. (a) Plasma device consisting of a handheld with intensity adjust and different glass tubes to plug in. Formation of orange colored plasma when turned on. (b) Pictures taken with a thermal imaging camera after 0, 1, 2 and 4 minutes without any sign of heat development. (c) Ozone formation during time measured by an O<span class="elsevierStyleInf">3</span> detector demonstrating an increase of O<span class="elsevierStyleInf">3</span> concentration after 10 minutes already. The dashed line indicates the odor threshold concentration for ozone (20‒40 µg/m<span class="elsevierStyleSup">3</span>; 0.01‒0.02 ppm).</p>" ] ] 1 => array:8 [ "identificador" => "fig0010" "etiqueta" => "Fig. 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 2409 "Ancho" => 3341 "Tamanyo" => 369520 ] ] "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at0010" "detalle" => "Fig. " "rol" => "short" ] ] "descripcion" => array:1 [ "en" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Antimicrobial effects of HF therapy. (a) HF treatment shows a significant decrease of bacterium and (b) fungus count in vitro. (c) CFU of <span class="elsevierStyleItalic">P. acnes</span> are significantly reduced after 1 minute, continuing the treatment leads to further reduction.</p>" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:14 [ 0 => array:3 [ "identificador" => "bib0005" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Epidemiology of acne vulgaris" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "K. Bhate" 1 => "H.C. 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