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commonly symmetrical&#44; hyperchromic&#44; well-defined&#44; and located in anatomical sites exposed to the sun&#44; especially the face&#46;<a class="elsevierStyleCrossRef" href="#bib0015"><span class="elsevierStyleSup">3</span></a> Its pathophysiology is still not completely understood&#44; but there is an interaction between genetics &#40;autosomal dominant inheritance&#41;&#44; hormonal factors&#44; and exposure to solar radiation&#44; leading to greater focal melanogenesis&#46;<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#8211;7</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Phenotypic changes in the skin with melasma result from both morphological &#40;enlarged melanocytes and prominent dendrites&#41; and dysfunctional epidermal alterations&#44; resulting from the complex interaction between keratinocytes&#44; melanocytes&#44; and components of the upper dermis&#46;<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">There is a clear relationship between sun exposure and melasma worsening&#44; 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UVB&#44; or VL in an <span class="elsevierStyleItalic">ex vivo</span> model&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Material and methods</span><p id="par0030" class="elsevierStylePara elsevierViewall">This was a <span class="elsevierStyleItalic">quasi</span>-experimental <span class="elsevierStyleItalic">ex vivo</span> study of skin fragments of 22 patients with facial melasma&#44; diagnosed by an experienced dermatologist&#46; Participants were selected among patients attended at the dermatology service of Hospital das Cl&#237;nicas&#44; Faculty of Medicine de Botucatu&#44; SP&#44; Brazil&#44; between September&#47;2018 and January&#47;2019&#46; The project was approved by the research ethics committee &#40;No&#46; 2&#44;700&#44;889&#41; and all participants signed an informed consent form before inclusion&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">Adult patients &#40;&#62;18 years old&#41;&#44; of both sexes&#44; with phototypes III&#8211;V&#44; with facial melasma and without treatment&#44; except for sunscreen for at least 30 days&#44; were eligible for the study&#46; The phototype restriction was based on the fact that phototypes I and II represent less than 10&#37; of cases of melasma and do not show satisfactory melanogenesis after experimental irradiation&#46;<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#44;11</span></a> There is no description of melasma in phototype VI&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">Patients with other facial dermatoses&#44; photosensitive dermatoses&#44; collagenoses&#44; blood dyscrasias&#44; users of anticoagulant medication&#44; those who were immunosuppressed&#44; and pregnant or lactating women were not included in the study&#46; Sampling was carried out by convenience&#44; recruiting consecutive consenting patients during medical consultations at the institution&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Sample material collection occurred in a standardized way and under a sterile procedure&#46; Two samples from each participant were collected&#58; skin with melasma and normal adjacent skin&#44; up to 2&#8239;cm apart&#44; under local infiltrative anesthesia with 2&#37; lidocaine with vasoconstrictor&#46; The removal included the reticular dermis&#44; with a 3&#8239;mm punch and 6&#8211;0 nylon monofilament suture&#46; Peripheral areas of the face were preferred&#44; in order to minimize unsightly scars&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Samples were sectioned longitudinally in two parts and stored in 10&#8239;mL of DMEM &#40;Dulbecco&#8217;s Modified Eagle&#8217;s Medium&#44; high glucose &#8211; D5796&#44; Sigma Inc&#46;&#44; United Kingdom&#41; in a sterile transparent plastic bottle and kept at room temperature&#44; according to the protocol set forth by Olivatti&#46;<a class="elsevierStyleCrossRef" href="#bib0060"><span class="elsevierStyleSup">12</span></a> A part of the fragment was placed immediately in the dark&#44; while the other was exposed to radiation&#44; for a total UVB&#44; UVA&#44; and VL dose &#40;effective in the tissue&#41; of 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span>&#44; 1&#46;524&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; and 40&#8239;J&#47;cm<span class="elsevierStyleSup">2</span> &#40;blue-violet light component&#41;&#44; respectively&#46; The fragments were irradiated with artificial sources of UVB &#40;230&#8239;&#181;W&#47;cm<span class="elsevierStyleSup">2</span>&#59; source&#58; FS72T12&#47;UVB&#47;HO&#41; for 12&#8239;min&#44; UVA &#40;1270&#8239;&#181;W&#47;cm<span class="elsevierStyleSup">2</span>&#59; source&#58; Phillips TL 100W&#47;10R&#41; for 6&#8239;min&#44; and LED light &#40;110&#8239;mW&#47;cm<span class="elsevierStyleSup">2</span> in the blue-violet band&#59; source&#58; GBRLUX 200W&#41; for 20&#8239;min&#44; at a standardized distance of 10&#8239;cm&#46; The absorbance of the plastic vials was around 50&#37; for all radiations&#46; The allocation of participants in each type of irradiation was consecutive &#40;not randomized&#41;&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">After irradiation&#44; both fragments &#40;irradiated and non-irradiated&#41; were cultured in a dark setting&#46; After 72&#8239;hours&#44; the samples were removed from the culture medium and preserved in 10&#37; buffered formalin for 36&#8239;hours&#46; Subsequently&#44; they were dehydrated in 70&#37; alcohol for at least 12&#8239;hours before embedding in paraffin and Fontana-Masson &#40;FM&#41; staining&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall">To estimate the melanization of the fragments&#44; four photographs of central and interfolicular areas of the samples stained by FM were captured&#44; favoring hotspot areas&#59; the images were saved in&#46; JPG format &#40;1264&#8239;&#215;&#8239;681 pixels&#41;&#46; During the entire capture and analysis process&#44; the investigators were blinded to the anatomical sites or type and irradiation regime&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">The selected images were analyzed using ImageJ 1&#46;43&#44; from the manual selection of the basal layer&#44; followed by thresholding using the &#8220;red&#8221; channel&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a></p><p id="par0070" class="elsevierStylePara elsevierViewall">The main outcome of the study was the percentage of melanization of the basal layer&#44; compared between groups &#40;melasma <span class="elsevierStyleItalic">vs&#46;</span> adjacent normal skin&#41; for each radiation &#40;UVB&#44; UVA&#44; or VL&#41;&#46; The percentage of melanin in the superficial dermis&#44; dendrite count&#44; and melanosome granulation were also evaluated &#40;score&#58; 1&#43;&#44; fine granulation&#59; 2&#43;&#44; moderate granulation&#59; 3&#43;&#44; coarse granulation&#41; in the sampled areas&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">The mean value of the percentage of melanin in the basal layer and in the upper dermis&#44; dendrite count&#44; and granulation scores were tabulated for each specimen&#46; The comparison groups were assessed by a generalized linear mixed effects model&#44; adjusted for phototype &#40;robust analysis&#41; with an unstructured covariance matrix and &#352;id&#225;k&#8217;s <span class="elsevierStyleItalic">post-hoc</span> correction&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> The probability distributions varied for the type of variable analyzed &#40;ordinal and gamma&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">The sample was sized after a pretest with radiation dosimetry that led to the expectation of a 10&#37; increase in the percentage of melanization in the basal layer evaluated by digital image analysis of the slides stained by FM&#44; considering alpha of 5&#37; and power of 20&#37;&#46;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">15</span></a></p><p id="par0085" class="elsevierStylePara elsevierViewall">The data were tabulated in MSExcel 2013 and analyzed in IBM SPSS v&#46; 25&#46; Two-tailed <span class="elsevierStyleItalic">p</span>-values &#8804;0&#46;05 were considered significant&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Results</span><p id="par0090" class="elsevierStylePara elsevierViewall">A total of 89 skin fragments from 22 patients with facial melasma were evaluated &#40;one participant volunteered four samples in order to test all irradiations&#41;&#46; The mean age &#40;standard deviation&#41; of the participants was 42&#46;1 &#40;10&#46;4&#41; years&#44; 21 of whom were female&#44; with the following phototypes&#58; III &#40;46&#37;&#41;&#44; IV &#40;27&#37;&#41;&#44; and V &#40;27&#37;&#41;&#46; Three culture fragments were contaminated &#40;UVA&#44; normal adjacent skin&#44; pre-irradiation&#59; UVB&#44; melasmic skin&#44; post-irradiation&#59; and UVB&#44; normal adjacent skin&#44; post-irradiation&#41; and were not included in the analysis of the results&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">Patients whose samples were submitted to UVB&#44; UVA&#44; and VL irradiation had the following phototypes&#44; respectively&#58; III &#40;33&#37; <span class="elsevierStyleItalic">vs</span>&#46; 37&#37; vs&#46; 0&#37;&#41;&#44; IV &#40;44&#37; <span class="elsevierStyleItalic">vs</span>&#46; 37&#37; <span class="elsevierStyleItalic">vs</span>&#46; 71&#37;&#41;&#44; and VL &#40;22&#37; <span class="elsevierStyleItalic">vs</span>&#46; 25&#37; <span class="elsevierStyleItalic">vs</span>&#46; 28&#37;&#41; &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;35&#41;&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">Melanogenesis was observed in melasma and in normal adjacent skin &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#60;&#8239;0&#46;01&#41; after all irradiations &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1&#8211;3</a>&#44; <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#58; median UVB increment &#40;4&#46;7&#37; <span class="elsevierStyleItalic">vs</span>&#46; 8&#46;5&#37;&#41;&#44; UVA &#40;9&#46;5&#37; <span class="elsevierStyleItalic">vs</span>&#46; 9&#46;9&#37;&#41;&#44; and VL &#40;6&#46;8&#37; <span class="elsevierStyleItalic">vs</span>&#46; 11&#46;7&#37;&#41;&#44; with no significant difference in the comparison between anatomical sites&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0105" class="elsevierStylePara elsevierViewall">When adjusted for phototype&#44; the dendrite count increased &#40;68&#46;8&#37;&#41; only after irradiation with UVB in both anatomical sites &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; An increase in melanin granulation &#40;coarser melanosomes&#41; was observed only after irradiation with UVA and only in the skin with melasma &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;05&#41;&#46; An increase was observed in the percentage of melanin in the upper dermis in the adjacent skin after irradiation with UVB&#44; and in melasmic skin after irradiation with UVA &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#8804;&#8239;0&#46;05&#41;&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Discussion</span><p id="par0110" class="elsevierStylePara elsevierViewall">Effective epidermal melanogenesis was observed in skin models &#40;<span class="elsevierStyleItalic">ex vivo</span>&#41; submitted to UVB&#44; UVA&#44; and VL&#44; with no quantitative difference between the anatomical sites &#40;facial melasma and normal adjacent skin&#41;&#44; supporting the importance of broad spectrum photoprotection in the treatment of pigmentary dermatoses&#44; such as melasma&#46;<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">16</span></a></p><p id="par0115" class="elsevierStylePara elsevierViewall">The radiation doses tested&#44; although independent&#44; were equivalent to approximately 6&#8211;12&#8239;min of sun exposure at midday&#44; in the summer&#44; in countryside Brazil &#40;latitude&#58; 22&#186;53&#8242;09&#8243;S&#59; longitude&#58; 48&#186;26&#8242;42&#8243;W&#59; altitude&#58; 804&#8239;m&#41;&#44; on a cloudless day&#44; as measured on December 9&#44; 2018&#44; by the researchers&#46; However&#44; the levels of solar UVA and VL reach an energy plateau from 9 am to 6&#8239;pm&#44; while the UVB peaks between 11 am and 3&#8239;pm&#44; both in summer and in winter&#46; This may justify the report of melasma worsening after brief unprotected exposures outside peak UVB hours&#44; or even after long exposures with a photoprotector &#40;<span class="elsevierStyleItalic">e&#46;g&#46;</span>&#44; during a traffic jam&#41;&#44; since sunscreens do not completely block solar radiation&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> UVB&#44; UVA&#44; and VL radiation measured in closed environments &#40;<span class="elsevierStyleItalic">e&#46;g</span>&#46;&#44; from cell phone and computer screens&#44; or fluorescent lamps&#41; are negligible for the purposes of melanogenesis &#40;data not shown&#41;&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">VL has a wide spectrum of wavelengths &#40;400&#8211;700&#8239;nm&#41; that do not act homogeneously on pigmentation&#46; A study carried out in France tested the spectrum of blue-violet light &#40;400&#8211;500&#8239;nm&#41; in comparison with the spectrum of red light &#40;620&#8211;750&#8239;nm&#41;&#59; those authors observed no pigmentation induced by the latter&#44; justifying the choice of the blue-violet band for the present study&#46;<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">18</span></a></p><p id="par0125" class="elsevierStylePara elsevierViewall">Only opaque sunscreens have effective protection against VL&#59; the majority of them also perform better in UVA protection&#46; In a Brazilian survey with 41 opaque sunscreens&#44; 63&#37; blocked &#62;99&#46;9&#37; of UVA&#44; and 63&#37; blocked &#62;99&#46;9&#37; of blue-violet light transmittance&#46; While the percentage was the same&#44; it was not overlapped&#58; 31&#37; of opaque filters that blocked &#62;99&#46;9&#37; of the VL did not present the same performance for UVA&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> The present study is the first to highlight the role of VL in the melanogenesis of patients with facial melasma&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">No differential melanogenesis was observed between melasma and adjacent normal skin at the doses tested&#44; which does not exclude the possibility that melasmic skin is sensitive to lower doses of irradiation or responds differently to repeated exposures&#46; The greater basal pigmentation of melasma itself can also present some resistance to melanogenesis induced by chronic exposure to UVR and VL&#46; Furthermore&#44; there is greater epidermal retention of melanin in melasma&#44; and its melanogenesis depends on several other dermal changes&#44; which favor the maintenance of pigmentation to the detriment of melanogenic induction by UVR and VL&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a></p><p id="par0135" class="elsevierStylePara elsevierViewall">Melanogenesis occurs unevenly&#44; depending on the type and dose of radiation&#46; UVB causes erythema that persists for days to weeks &#40;depending on the phototype&#41; and pigmentation that evolves to tanning&#46; UVA causes erythema in four hours and immediate pigmentation that evolves with lasting tanning&#46; In VL&#44; erythema is less prominent and may occur early &#40;within two hours&#41;&#46; Pigmentation is sustained with progression to tanning&#44; but only in higher phototypes&#46;<a class="elsevierStyleCrossRefs" href="#bib0095"><span class="elsevierStyleSup">19&#8211;21</span></a></p><p id="par0140" class="elsevierStylePara elsevierViewall">Every melanogenic response occurs in a dose-dependent manner&#46; However&#44; it is essential to consider the phototypes when analyzing the performance of different radiations&#44; since melanogenesis is different in the higher phototypes &#40;III&#8211;V&#41;&#44; causing greater pigmentation with evolution to tanning in a dose-dependent manner&#46; In turn&#44; in lower phototypes &#40;I&#8211;I&#41; irradiation promotes less stimulation&#46; In an experimental study&#44; UVA and VL did not induce melanogenesis in patients with phototype II&#46;<a class="elsevierStyleCrossRefs" href="#bib0055"><span class="elsevierStyleSup">11&#44;22</span></a></p><p id="par0145" class="elsevierStylePara elsevierViewall">Human melanogenesis is a complex process in which different interactions occur in an organized manner to protect skin structures and to regulate cycles that are essential for survival&#44; such as the circadian cycle&#46; Melanocytes are a network with their own sense of ambient light&#44; responding according to the aforementioned objectives&#46;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">23</span></a></p><p id="par0150" class="elsevierStylePara elsevierViewall">Different characteristics were observed in the melanogenic response when comparing anatomical sites and radiations&#46; The formation of dendrites occurs in the G2 phase of melanocyte growth and is an important sign of melanogenic response&#59; in the present study&#44; it was mainly induced by the irradiation with the greatest mutagenic potential &#40;UVB&#41;&#46; Although melanogenesis is perceived in a similar way in both anatomical sites&#44; the formation of dendrites can indicate a greater persistence of the phenomenon and greater efficiency in the transfer of melanosomes in the epidermis&#46;<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a></p><p id="par0155" class="elsevierStylePara elsevierViewall">Melasmic skin is also characterized by the presence of more mature melanosomes&#44; equivalent to what occurs in more pigmented populations&#46; In the present experiment&#44; UVA irradiation promoted more evident granulation than the other tested radiations exclusively on skin with melasma&#44; showing that&#44; in melasma&#44; melanogenesis is more intense and resembles the most pigmented phenotypes&#46; In fact&#44; African ancestry was associated with an increased risk of melasma in the Brazilian population&#44; and ancestral genes related to melanogenesis may justify the differentiated pigment response between anatomical sites&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a></p><p id="par0160" class="elsevierStylePara elsevierViewall">The role of dermal melanin remains unclear in melasma&#59; one of the hypotheses is that the architectural dysfunction of the basement membrane and the autophagy of the melanocytes of the dermis may cause the presence of dermal melanophages&#44; similar to what occurs in photoaging&#46;<a class="elsevierStyleCrossRefs" href="#bib0120"><span class="elsevierStyleSup">24&#44;25</span></a> In the present study&#44; UVA and UVB promoted a higher rate of melanin in the upper dermis&#59; however&#44; this density was approximately 100 times smaller than that of the epidermis in both anatomical sites&#44; which likely little contributes to the clinical phenotype&#44; but may signal underlying alterations in the upper dermis&#44; basement membrane&#44; and basal layer&#46;</p><p id="par0165" class="elsevierStylePara elsevierViewall">Finally&#44; despite the fundamental role of solar radiation in worsening melasma&#44; no differential expressions of p53 between the melasma and the normal adjacent skin were observed&#44; suggesting that the maintenance of melanogenesis may be due to underlying changes in the upper dermis and epidermis&#44; which induce melanocytic hyperfunction&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a> Furthermore&#44; the exclusive use of sunscreen&#44; despite its preventive effect&#44; is not sufficient for the complete remission of the condition&#44; which indicates the need for a continuous pathophysiological study in search of treatments that lead to the reversal of the entire process&#46;<a class="elsevierStyleCrossRefs" href="#bib0045"><span class="elsevierStyleSup">9&#44;26</span></a></p><p id="par0170" class="elsevierStylePara elsevierViewall">The present study promoted a preliminary analysis of the independent effects of UVB&#44; UVA&#44; and VL radiation&#46; Further investigations should be conducted in order to elucidate the effect of combinations and dosages of irradiations that mirror environmental conditions&#46; The interaction between UVA and VL is still unknown&#59; they are believed to interact in the same melanin precursor during melanogenesis&#46; This association may result in a more evident behavior of melanogenesis in natural situations&#44; where VL and UVA are expressed together&#46;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">20</span></a></p><p id="par0175" class="elsevierStylePara elsevierViewall">This study has limitations inherent to the technique in an <span class="elsevierStyleItalic">ex vivo</span> model&#44; which may differ depending on collection and storage&#46; However&#44; this limitation did not prevent the identification of a melanogenic response&#46; Another limitation can be pointed out by the fact that irradiations in <span class="elsevierStyleItalic">ex vivo</span> skin models occur with independent wavelengths &#40;UVB&#44; UVA and VL&#41;&#44; which is not the case in environmental exposure&#46; In addition to the factors presented above&#44; <span class="elsevierStyleItalic">ex vivo</span> models are dissociated from the organism&#44; limiting the interaction between the hormonal&#44; neural&#44; and vasomotor systems&#46;</p><p id="par0180" class="elsevierStylePara elsevierViewall">New experiments should consider low-intensity repeated irradiation models&#44; as well as associated irradiation&#44; to better reproduce the daily exposure of patients&#46; In addition&#44; the inclusion of substances such as tranexamic acid and antioxidants in the prevention of melanogenesis in melasma can be tested&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Conclusions</span><p id="par0185" class="elsevierStylePara elsevierViewall">An epidermal melanogenic response induced by UVB&#44; UVA&#44; or VL was observed both in skin with melasma and in normal adjacent skin&#46; The morphological patterns related to the photobiology of melanogenesis suggest that different radiations promote individualized responses in the skin with melasma&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Financial support</span><p id="par0190" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleGrantSponsor" id="gs0005">CNPq</span> &#40;PIBIC No&#46; <span class="elsevierStyleGrantNumber" refid="gs0005">148501&#47;2018-4</span>&#41;&#59; <span class="elsevierStyleGrantSponsor" id="gs0010">FUNADERM</span> &#40;<span class="elsevierStyleGrantNumber" refid="gs0010">001&#47;2019</span>&#41;&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Authors&#8217; contributions</span><p id="par0195" class="elsevierStylePara elsevierViewall">Giovana Piteri Alcantara&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0200" class="elsevierStylePara elsevierViewall">Ana Cl&#225;udia Cavalcante Esposito&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#44; elaboration and writing of the manuscript&#59; effective participation in research orientation&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0205" class="elsevierStylePara elsevierViewall">Thain&#225; Oliveira Felicio Olivatti&#58; Approval of the final version of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the manuscript&#46;</p><p id="par0210" class="elsevierStylePara elsevierViewall">Melissa Mari Yoshida&#58; Elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the manuscript&#46;</p><p id="par0215" class="elsevierStylePara elsevierViewall">H&#233;lio Amante Miot&#58; Statistical analysis&#59; approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; effective participation in research orientation&#59; intellectual participation in propaedeutic and&#47;or therapeutic conduct of studied cases&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Conflicts of interest</span><p id="par0220" class="elsevierStylePara elsevierViewall">None declared&#46;</p></span></span>"
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      "en" => array:3 [
        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">The independent role of solar radiation in the differential melanogenesis between melasma and adjacent skin is unknown&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Objectives</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">To assess the melanogenic responses of skin with facial melasma and of the adjacent skin to UVB&#44; UVA&#44; and visible light&#44; in an <span class="elsevierStyleItalic">ex vivo</span> model&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">This was a quasi-experimental study involving 22 patients with melasma&#46; Facial melasma and adjacent skin samples were collected and stored in DMEM medium&#44; at room temperature&#46; One fragment was placed under the protection from light&#44; while another was exposed to UVB&#44; UVA&#44; and visible light &#40;blue-violet component&#41;&#58; 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span>&#44; 1&#46;524&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; and 40&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; respectively&#46; Subsequently&#44; all samples were kept for 72&#8239;hours in a dark environment and stained by Fontana-Masson to assess basal layer pigmentation&#44; dendrites&#44; and melanin granulation&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Effective melanogenesis was observed in the basal layer in melasma and in the normal adjacent skin after all irradiations &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#60;&#8239;0&#46;01&#41;&#44; with the following median increment&#58; UVB &#40;4&#46;7&#37; <span class="elsevierStyleItalic">vs</span>&#46; 8&#46;5&#37;&#41;&#44; UVA &#40;9&#46;5&#37; <span class="elsevierStyleItalic">vs</span>&#46; 9&#46;9&#37;&#41;&#44; and visible light &#40;6&#46;8&#37; <span class="elsevierStyleItalic">vs</span>&#46; 11&#46;7&#37;&#41;&#44; with no significant difference between anatomical sites&#46; An increase in melanin granulation &#40;coarser melanosomes&#41; was observed only after irradiation with UVA and only in the skin with melasma &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;05&#41;&#46; An increase in the melanocyte dendrite count induced by UVB radiation was observed in both anatomical sites &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#8804;&#8239;0&#46;05&#41;&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Study limitations</span><p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Use of an <span class="elsevierStyleItalic">ex vivo</span> model&#44; with independent irradiation regimes for UVB&#44; UVA&#44; and visible light&#46;</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Conclusions</span><p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">Melanogenesis induced by UVB&#44; UVA&#44; and visible light was observed both in melasma and in the adjacent skin&#46; The morphological patterns suggest that different irradiations promote individualized responses on the skin with melasma&#46;</p></span>"
        "secciones" => array:6 [
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            "identificador" => "abst0005"
            "titulo" => "Background"
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          1 => array:2 [
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            "titulo" => "Objectives"
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            "identificador" => "abst0015"
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            "titulo" => "Study limitations"
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            "identificador" => "abst0030"
            "titulo" => "Conclusions"
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    "NotaPie" => array:2 [
      0 => array:2 [
        "etiqueta" => "&#8902;"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0010">How to cite this article&#58; Alcantara GP&#44; Esposito ACC&#44; Olivatti TOF&#44; Yoshida MM&#44; Miot HA&#46; Evaluation of <span class="elsevierStyleItalic">ex vivo</span> melanogenic response to UVB&#44; UVA&#44; and visible light in facial melasma and unaffected adjacent skin&#46; An Bras Dermatol&#46; 2020&#59;95&#46; <span class="elsevierStyleInterRef" id="intr0005" href="https://doi.org/10.1016/j.abd.2020.02.015">https&#58;&#47;&#47;doi&#46;org&#47;10&#46;1016&#47;j&#46;abd&#46;2020&#46;02&#46;015</span></p>"
      ]
      1 => array:2 [
        "etiqueta" => "&#8902;&#8902;"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0015">Study conducted at the Department of Dermatology and Radiotherapy&#44; Faculty of Medicine&#44; Universidade Estadual Paulista&#44; Botucatu&#44; SP&#44; Brazil&#46;</p>"
      ]
    ]
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          "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Histological sections of skin with melasma and normal adjacent skin samples stained with Fontana-Masson before and after irradiation with 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span> UVB&#44; revealing an increase in the basal melanin density in the different samples&#46; &#40;A&#41; Normal adjacent skin before UVB irradiation&#59; &#40;B&#41; normal adjacent skin after UVB irradiation&#59; &#40;C&#41; skin with melasma before UVB irradiation&#59; &#40;D&#41; skin with melasma after UVB irradiation&#46;</p>"
        ]
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      1 => array:8 [
        "identificador" => "fig0010"
        "etiqueta" => "Figure 2"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr2.jpeg"
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        "detalles" => array:1 [
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        "descripcion" => array:1 [
          "en" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Histological sections of skin with melasma and normal adjacent skin samples stained with Fontana-Masson before and after irradiation with 1&#46;524&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span> UVB&#44; revealing an increase in the basal melanin density in the different samples&#46; &#40;A&#41;&#44; normal adjacent skin before UVB irradiation&#59; &#40;B&#41;&#44; normal adjacent skin after UVB irradiation&#59; &#40;C&#41;&#44; skin with melasma before UVB irradiation&#59; &#40;D&#41;&#44; skin with melasma after UVB irradiation&#46;</p>"
        ]
      ]
      2 => array:8 [
        "identificador" => "fig0015"
        "etiqueta" => "Figure 3"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
            "imagen" => "gr3.jpeg"
            "Alto" => 653
            "Ancho" => 1255
            "Tamanyo" => 279996
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        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "at0015"
            "detalle" => "Figure "
            "rol" => "short"
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Histological sections of skin with melasma and normal adjacent skin samples stained with Fontana-Masson before and after irradiation with 40&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span> visible light &#40;VL&#41;&#44; revealing an increase in the basal melanin density between the samples&#46; &#40;A&#41;&#44; normal adjacent skin before VL irradiation&#59; &#40;B&#41;&#44; normal adjacent skin after VL irradiation&#59; &#40;C&#41;&#44; skin with melasma before VL irradiation&#59; &#40;D&#41;&#44; skin with melasma after VL irradiation&#46;</p>"
        ]
      ]
      3 => array:8 [
        "identificador" => "tbl0005"
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        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
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        "detalles" => array:1 [
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        "tabla" => array:3 [
          "leyenda" => "<p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">MelBasal&#44; percentage of melanin in the basal layer of the epidermis&#59; MelUD&#44; percentage of melanin in the upper dermis &#40;&#215;100&#41;&#59; GranMelanos&#58; granulation score of melanosomes of the epidermis &#40;1&#43;&#44; fine granulation&#59; 2&#43;&#44; moderate granulation&#59; 3&#43;&#44; coarse granulation&#41;&#44; mean of four fields&#46;</p>"
          "tablatextoimagen" => array:1 [
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col">Irradiation&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col">Variable&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
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                  \t\t\t\t  " colspan="3" align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Facial melasma</th><th class="td" title="\n
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                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Normal adjacent skin</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col"><span class="elsevierStyleItalic">p</span> &#40;time&#8239;&#215;&#8239;group&#41;<a class="elsevierStyleCrossRef" href="#tblfn0010">&#42;&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr><tr title="table-row"><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Pre&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Post&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">p</span><a class="elsevierStyleCrossRef" href="#tblfn0005">&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Pre&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Post&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">p</span><a class="elsevierStyleCrossRef" href="#tblfn0005">&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " rowspan="4" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">UVB</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelBasal&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">64 &#40;60&#8211;65&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">65 &#40;65&#8211;68&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">54 &#40;53&#8211;56&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">56 &#40;55&#8211;62&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">0&#46;09&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelUD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">44 &#40;30&#8211;77&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">45 &#40;38&#8211;97&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">44 &#40;32&#8211;71&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">67 &#40;41&#8211;87&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;04&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;84&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Dendrites&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6 &#40;2&#8211;8&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">10 &#40;8&#8211;18&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6 &#40;3&#8211;14&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">11 &#40;7&#8211;16&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;59&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">GranMelanos&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
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                  \t\t\t\t ; entry_with_role_rowhead " rowspan="4" align="left" valign="\n
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                  \t\t\t\t">UVA</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelBasal&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">62 &#40;54&#8211;66&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">70 &#40;57&#8211;72&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">56 &#40;49&#8211;61&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">57 &#40;50&#8211;63&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelUD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">41 &#40;18&#8211;88&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">86 &#40;36&#8211;144&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">45 &#40;27&#8211;90&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">74 &#40;34&#8211;204&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Dendrites&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">8 &#40;4&#8211;11&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">8 &#40;2&#8211;22&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">3 &#40;2&#8211;5&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">5 &#40;3&#8211;9&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">GranMelanos&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">1&#46;0 &#40;1&#46;0&#8211;1&#46;8&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;03&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " rowspan="4" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Visible light</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelBasal&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">73 &#40;59&#8211;74&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">75 &#40;69&#8211;80&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">64 &#40;56&#8211;66&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">70 &#40;61&#8211;75&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelUD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">62 &#40;54&#8211;72&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">52 &#40;45&#8211;83&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">66 &#40;59&#8211;83&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">75 &#40;53&#8211;127&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Dendrites&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">2 &#40;1&#8211;4&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">4 &#40;2&#8211;14&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1 &#40;0&#8211;3&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">4 &#40;1&#8211;5&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;06&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">GranMelanos&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">1&#46;4 &#40;1&#46;0&#8211;2&#46;0&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1&#46;5 &#40;1&#46;0&#8211;2&#46;3&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0&#46;11&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1&#46;5 &#40;1&#46;0&#8211;2&#46;0&#41;&nbsp;\t\t\t\t\t\t\n
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Investigation
Evaluation of ex vivo melanogenic response to UVB, UVA, and visible light in facial melasma and unaffected adjacent skin
Giovana Piteri Alcantaraa, Ana Cláudia Cavalcante Espositob, Thainá Oliveira Felicio Olivattic, Melissa Mari Yoshidac, Hélio Amante Miota,
Autor para correspondência
heliomiot@gmail.com

Corresponding author.
a Department of Dermatology and Radiotherapy, Faculty of Medicine, Universidade Estadual Paulista, Botucatu, SP, Brazil
b Graduate Program in Pathology, Faculty of Medicine, Universidade Estadual Paulista, Botucatu, SP, Brazil
c Faculty of Medicine, Universidade Estadual Paulista, Botucatu, SP, Brazil
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          "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Histological sections of skin with melasma and normal adjacent skin samples stained with Fontana-Masson before and after irradiation with 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span> UVB&#44; revealing an increase in the basal melanin density in the different samples&#46; &#40;A&#41; Normal adjacent skin before UVB irradiation&#59; &#40;B&#41; normal adjacent skin after UVB irradiation&#59; &#40;C&#41; skin with melasma before UVB irradiation&#59; &#40;D&#41; skin with melasma after UVB irradiation&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall">Melasma is a chronic and acquired hypermelanosis resulting from a dysfunction of melanogenesis&#58; increased production&#44; transfer&#44; and greater maturity of melanosomes&#46;<a class="elsevierStyleCrossRef" href="#bib0005"><span class="elsevierStyleSup">1</span></a> It is a frequent condition in the dermatological routine&#59; according to a study by the Brazilian Society of Dermatology published in 2018&#44; it accounts for 3&#46;6&#37; of dermatological consultations&#46;<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> It affects mainly women of childbearing age &#40;25&#8211;50 years&#41; and intermediate phototypes &#40;III&#8211;V&#41;&#46; The clinical lesions of melasma are macules&#44; commonly symmetrical&#44; hyperchromic&#44; well-defined&#44; and located in anatomical sites exposed to the sun&#44; especially the face&#46;<a class="elsevierStyleCrossRef" href="#bib0015"><span class="elsevierStyleSup">3</span></a> Its pathophysiology is still not completely understood&#44; but there is an interaction between genetics &#40;autosomal dominant inheritance&#41;&#44; hormonal factors&#44; and exposure to solar radiation&#44; leading to greater focal melanogenesis&#46;<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#8211;7</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Phenotypic changes in the skin with melasma result from both morphological &#40;enlarged melanocytes and prominent dendrites&#41; and dysfunctional epidermal alterations&#44; resulting from the complex interaction between keratinocytes&#44; melanocytes&#44; and components of the upper dermis&#46;<a class="elsevierStyleCrossRef" href="#bib0040"><span class="elsevierStyleSup">8</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">There is a clear relationship between sun exposure and melasma worsening&#44; due to its exclusive occurrence in photo exposed areas&#44; prevalence in professionals who are exposed to the sun&#44; and better therapeutic response associated with photoprotection&#46;<a class="elsevierStyleCrossRefs" href="#bib0045"><span class="elsevierStyleSup">9&#44;10</span></a> Even with evidence of the role of solar radiation in the clinical history of the disease&#44; the individualized response pattern to the different bands of solar radiation &#40;UVB&#44; UVA&#44; and visible light &#91;VL&#93;&#41; of melasma in comparison with normal adjacent skin is not known&#46;</p><p id="par0020" class="elsevierStylePara elsevierViewall">Knowledge of the different influences of solar radiation in the melanogenesis of melasma can lead to a better understanding of the pathophysiology of the disease&#44; as well as to the development of new treatment options and optimized photo protection strategies&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">This study aimed to determine the melanogenic response of facial melasma in comparison with the normal adjacent skin after irradiation with UVA&#44; UVB&#44; or VL in an <span class="elsevierStyleItalic">ex vivo</span> model&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Material and methods</span><p id="par0030" class="elsevierStylePara elsevierViewall">This was a <span class="elsevierStyleItalic">quasi</span>-experimental <span class="elsevierStyleItalic">ex vivo</span> study of skin fragments of 22 patients with facial melasma&#44; diagnosed by an experienced dermatologist&#46; Participants were selected among patients attended at the dermatology service of Hospital das Cl&#237;nicas&#44; Faculty of Medicine de Botucatu&#44; SP&#44; Brazil&#44; between September&#47;2018 and January&#47;2019&#46; The project was approved by the research ethics committee &#40;No&#46; 2&#44;700&#44;889&#41; and all participants signed an informed consent form before inclusion&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">Adult patients &#40;&#62;18 years old&#41;&#44; of both sexes&#44; with phototypes III&#8211;V&#44; with facial melasma and without treatment&#44; except for sunscreen for at least 30 days&#44; were eligible for the study&#46; The phototype restriction was based on the fact that phototypes I and II represent less than 10&#37; of cases of melasma and do not show satisfactory melanogenesis after experimental irradiation&#46;<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3&#44;11</span></a> There is no description of melasma in phototype VI&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">Patients with other facial dermatoses&#44; photosensitive dermatoses&#44; collagenoses&#44; blood dyscrasias&#44; users of anticoagulant medication&#44; those who were immunosuppressed&#44; and pregnant or lactating women were not included in the study&#46; Sampling was carried out by convenience&#44; recruiting consecutive consenting patients during medical consultations at the institution&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Sample material collection occurred in a standardized way and under a sterile procedure&#46; Two samples from each participant were collected&#58; skin with melasma and normal adjacent skin&#44; up to 2&#8239;cm apart&#44; under local infiltrative anesthesia with 2&#37; lidocaine with vasoconstrictor&#46; The removal included the reticular dermis&#44; with a 3&#8239;mm punch and 6&#8211;0 nylon monofilament suture&#46; Peripheral areas of the face were preferred&#44; in order to minimize unsightly scars&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Samples were sectioned longitudinally in two parts and stored in 10&#8239;mL of DMEM &#40;Dulbecco&#8217;s Modified Eagle&#8217;s Medium&#44; high glucose &#8211; D5796&#44; Sigma Inc&#46;&#44; United Kingdom&#41; in a sterile transparent plastic bottle and kept at room temperature&#44; according to the protocol set forth by Olivatti&#46;<a class="elsevierStyleCrossRef" href="#bib0060"><span class="elsevierStyleSup">12</span></a> A part of the fragment was placed immediately in the dark&#44; while the other was exposed to radiation&#44; for a total UVB&#44; UVA&#44; and VL dose &#40;effective in the tissue&#41; of 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span>&#44; 1&#46;524&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; and 40&#8239;J&#47;cm<span class="elsevierStyleSup">2</span> &#40;blue-violet light component&#41;&#44; respectively&#46; The fragments were irradiated with artificial sources of UVB &#40;230&#8239;&#181;W&#47;cm<span class="elsevierStyleSup">2</span>&#59; source&#58; FS72T12&#47;UVB&#47;HO&#41; for 12&#8239;min&#44; UVA &#40;1270&#8239;&#181;W&#47;cm<span class="elsevierStyleSup">2</span>&#59; source&#58; Phillips TL 100W&#47;10R&#41; for 6&#8239;min&#44; and LED light &#40;110&#8239;mW&#47;cm<span class="elsevierStyleSup">2</span> in the blue-violet band&#59; source&#58; GBRLUX 200W&#41; for 20&#8239;min&#44; at a standardized distance of 10&#8239;cm&#46; The absorbance of the plastic vials was around 50&#37; for all radiations&#46; The allocation of participants in each type of irradiation was consecutive &#40;not randomized&#41;&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">After irradiation&#44; both fragments &#40;irradiated and non-irradiated&#41; were cultured in a dark setting&#46; After 72&#8239;hours&#44; the samples were removed from the culture medium and preserved in 10&#37; buffered formalin for 36&#8239;hours&#46; Subsequently&#44; they were dehydrated in 70&#37; alcohol for at least 12&#8239;hours before embedding in paraffin and Fontana-Masson &#40;FM&#41; staining&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall">To estimate the melanization of the fragments&#44; four photographs of central and interfolicular areas of the samples stained by FM were captured&#44; favoring hotspot areas&#59; the images were saved in&#46; JPG format &#40;1264&#8239;&#215;&#8239;681 pixels&#41;&#46; During the entire capture and analysis process&#44; the investigators were blinded to the anatomical sites or type and irradiation regime&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">The selected images were analyzed using ImageJ 1&#46;43&#44; from the manual selection of the basal layer&#44; followed by thresholding using the &#8220;red&#8221; channel&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">13</span></a></p><p id="par0070" class="elsevierStylePara elsevierViewall">The main outcome of the study was the percentage of melanization of the basal layer&#44; compared between groups &#40;melasma <span class="elsevierStyleItalic">vs&#46;</span> adjacent normal skin&#41; for each radiation &#40;UVB&#44; UVA&#44; or VL&#41;&#46; The percentage of melanin in the superficial dermis&#44; dendrite count&#44; and melanosome granulation were also evaluated &#40;score&#58; 1&#43;&#44; fine granulation&#59; 2&#43;&#44; moderate granulation&#59; 3&#43;&#44; coarse granulation&#41; in the sampled areas&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">The mean value of the percentage of melanin in the basal layer and in the upper dermis&#44; dendrite count&#44; and granulation scores were tabulated for each specimen&#46; The comparison groups were assessed by a generalized linear mixed effects model&#44; adjusted for phototype &#40;robust analysis&#41; with an unstructured covariance matrix and &#352;id&#225;k&#8217;s <span class="elsevierStyleItalic">post-hoc</span> correction&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">14</span></a> The probability distributions varied for the type of variable analyzed &#40;ordinal and gamma&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">The sample was sized after a pretest with radiation dosimetry that led to the expectation of a 10&#37; increase in the percentage of melanization in the basal layer evaluated by digital image analysis of the slides stained by FM&#44; considering alpha of 5&#37; and power of 20&#37;&#46;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">15</span></a></p><p id="par0085" class="elsevierStylePara elsevierViewall">The data were tabulated in MSExcel 2013 and analyzed in IBM SPSS v&#46; 25&#46; Two-tailed <span class="elsevierStyleItalic">p</span>-values &#8804;0&#46;05 were considered significant&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Results</span><p id="par0090" class="elsevierStylePara elsevierViewall">A total of 89 skin fragments from 22 patients with facial melasma were evaluated &#40;one participant volunteered four samples in order to test all irradiations&#41;&#46; The mean age &#40;standard deviation&#41; of the participants was 42&#46;1 &#40;10&#46;4&#41; years&#44; 21 of whom were female&#44; with the following phototypes&#58; III &#40;46&#37;&#41;&#44; IV &#40;27&#37;&#41;&#44; and V &#40;27&#37;&#41;&#46; Three culture fragments were contaminated &#40;UVA&#44; normal adjacent skin&#44; pre-irradiation&#59; UVB&#44; melasmic skin&#44; post-irradiation&#59; and UVB&#44; normal adjacent skin&#44; post-irradiation&#41; and were not included in the analysis of the results&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">Patients whose samples were submitted to UVB&#44; UVA&#44; and VL irradiation had the following phototypes&#44; respectively&#58; III &#40;33&#37; <span class="elsevierStyleItalic">vs</span>&#46; 37&#37; vs&#46; 0&#37;&#41;&#44; IV &#40;44&#37; <span class="elsevierStyleItalic">vs</span>&#46; 37&#37; <span class="elsevierStyleItalic">vs</span>&#46; 71&#37;&#41;&#44; and VL &#40;22&#37; <span class="elsevierStyleItalic">vs</span>&#46; 25&#37; <span class="elsevierStyleItalic">vs</span>&#46; 28&#37;&#41; &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;35&#41;&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">Melanogenesis was observed in melasma and in normal adjacent skin &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#60;&#8239;0&#46;01&#41; after all irradiations &#40;<a class="elsevierStyleCrossRefs" href="#fig0005">Figs&#46; 1&#8211;3</a>&#44; <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#58; median UVB increment &#40;4&#46;7&#37; <span class="elsevierStyleItalic">vs</span>&#46; 8&#46;5&#37;&#41;&#44; UVA &#40;9&#46;5&#37; <span class="elsevierStyleItalic">vs</span>&#46; 9&#46;9&#37;&#41;&#44; and VL &#40;6&#46;8&#37; <span class="elsevierStyleItalic">vs</span>&#46; 11&#46;7&#37;&#41;&#44; with no significant difference in the comparison between anatomical sites&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0105" class="elsevierStylePara elsevierViewall">When adjusted for phototype&#44; the dendrite count increased &#40;68&#46;8&#37;&#41; only after irradiation with UVB in both anatomical sites &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; An increase in melanin granulation &#40;coarser melanosomes&#41; was observed only after irradiation with UVA and only in the skin with melasma &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;05&#41;&#46; An increase was observed in the percentage of melanin in the upper dermis in the adjacent skin after irradiation with UVB&#44; and in melasmic skin after irradiation with UVA &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#8804;&#8239;0&#46;05&#41;&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Discussion</span><p id="par0110" class="elsevierStylePara elsevierViewall">Effective epidermal melanogenesis was observed in skin models &#40;<span class="elsevierStyleItalic">ex vivo</span>&#41; submitted to UVB&#44; UVA&#44; and VL&#44; with no quantitative difference between the anatomical sites &#40;facial melasma and normal adjacent skin&#41;&#44; supporting the importance of broad spectrum photoprotection in the treatment of pigmentary dermatoses&#44; such as melasma&#46;<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">16</span></a></p><p id="par0115" class="elsevierStylePara elsevierViewall">The radiation doses tested&#44; although independent&#44; were equivalent to approximately 6&#8211;12&#8239;min of sun exposure at midday&#44; in the summer&#44; in countryside Brazil &#40;latitude&#58; 22&#186;53&#8242;09&#8243;S&#59; longitude&#58; 48&#186;26&#8242;42&#8243;W&#59; altitude&#58; 804&#8239;m&#41;&#44; on a cloudless day&#44; as measured on December 9&#44; 2018&#44; by the researchers&#46; However&#44; the levels of solar UVA and VL reach an energy plateau from 9 am to 6&#8239;pm&#44; while the UVB peaks between 11 am and 3&#8239;pm&#44; both in summer and in winter&#46; This may justify the report of melasma worsening after brief unprotected exposures outside peak UVB hours&#44; or even after long exposures with a photoprotector &#40;<span class="elsevierStyleItalic">e&#46;g&#46;</span>&#44; during a traffic jam&#41;&#44; since sunscreens do not completely block solar radiation&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> UVB&#44; UVA&#44; and VL radiation measured in closed environments &#40;<span class="elsevierStyleItalic">e&#46;g</span>&#46;&#44; from cell phone and computer screens&#44; or fluorescent lamps&#41; are negligible for the purposes of melanogenesis &#40;data not shown&#41;&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">VL has a wide spectrum of wavelengths &#40;400&#8211;700&#8239;nm&#41; that do not act homogeneously on pigmentation&#46; A study carried out in France tested the spectrum of blue-violet light &#40;400&#8211;500&#8239;nm&#41; in comparison with the spectrum of red light &#40;620&#8211;750&#8239;nm&#41;&#59; those authors observed no pigmentation induced by the latter&#44; justifying the choice of the blue-violet band for the present study&#46;<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">18</span></a></p><p id="par0125" class="elsevierStylePara elsevierViewall">Only opaque sunscreens have effective protection against VL&#59; the majority of them also perform better in UVA protection&#46; In a Brazilian survey with 41 opaque sunscreens&#44; 63&#37; blocked &#62;99&#46;9&#37; of UVA&#44; and 63&#37; blocked &#62;99&#46;9&#37; of blue-violet light transmittance&#46; While the percentage was the same&#44; it was not overlapped&#58; 31&#37; of opaque filters that blocked &#62;99&#46;9&#37; of the VL did not present the same performance for UVA&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">17</span></a> The present study is the first to highlight the role of VL in the melanogenesis of patients with facial melasma&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">No differential melanogenesis was observed between melasma and adjacent normal skin at the doses tested&#44; which does not exclude the possibility that melasmic skin is sensitive to lower doses of irradiation or responds differently to repeated exposures&#46; The greater basal pigmentation of melasma itself can also present some resistance to melanogenesis induced by chronic exposure to UVR and VL&#46; Furthermore&#44; there is greater epidermal retention of melanin in melasma&#44; and its melanogenesis depends on several other dermal changes&#44; which favor the maintenance of pigmentation to the detriment of melanogenic induction by UVR and VL&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a></p><p id="par0135" class="elsevierStylePara elsevierViewall">Melanogenesis occurs unevenly&#44; depending on the type and dose of radiation&#46; UVB causes erythema that persists for days to weeks &#40;depending on the phototype&#41; and pigmentation that evolves to tanning&#46; UVA causes erythema in four hours and immediate pigmentation that evolves with lasting tanning&#46; In VL&#44; erythema is less prominent and may occur early &#40;within two hours&#41;&#46; Pigmentation is sustained with progression to tanning&#44; but only in higher phototypes&#46;<a class="elsevierStyleCrossRefs" href="#bib0095"><span class="elsevierStyleSup">19&#8211;21</span></a></p><p id="par0140" class="elsevierStylePara elsevierViewall">Every melanogenic response occurs in a dose-dependent manner&#46; However&#44; it is essential to consider the phototypes when analyzing the performance of different radiations&#44; since melanogenesis is different in the higher phototypes &#40;III&#8211;V&#41;&#44; causing greater pigmentation with evolution to tanning in a dose-dependent manner&#46; In turn&#44; in lower phototypes &#40;I&#8211;I&#41; irradiation promotes less stimulation&#46; In an experimental study&#44; UVA and VL did not induce melanogenesis in patients with phototype II&#46;<a class="elsevierStyleCrossRefs" href="#bib0055"><span class="elsevierStyleSup">11&#44;22</span></a></p><p id="par0145" class="elsevierStylePara elsevierViewall">Human melanogenesis is a complex process in which different interactions occur in an organized manner to protect skin structures and to regulate cycles that are essential for survival&#44; such as the circadian cycle&#46; Melanocytes are a network with their own sense of ambient light&#44; responding according to the aforementioned objectives&#46;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">23</span></a></p><p id="par0150" class="elsevierStylePara elsevierViewall">Different characteristics were observed in the melanogenic response when comparing anatomical sites and radiations&#46; The formation of dendrites occurs in the G2 phase of melanocyte growth and is an important sign of melanogenic response&#59; in the present study&#44; it was mainly induced by the irradiation with the greatest mutagenic potential &#40;UVB&#41;&#46; Although melanogenesis is perceived in a similar way in both anatomical sites&#44; the formation of dendrites can indicate a greater persistence of the phenomenon and greater efficiency in the transfer of melanosomes in the epidermis&#46;<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a></p><p id="par0155" class="elsevierStylePara elsevierViewall">Melasmic skin is also characterized by the presence of more mature melanosomes&#44; equivalent to what occurs in more pigmented populations&#46; In the present experiment&#44; UVA irradiation promoted more evident granulation than the other tested radiations exclusively on skin with melasma&#44; showing that&#44; in melasma&#44; melanogenesis is more intense and resembles the most pigmented phenotypes&#46; In fact&#44; African ancestry was associated with an increased risk of melasma in the Brazilian population&#44; and ancestral genes related to melanogenesis may justify the differentiated pigment response between anatomical sites&#46;<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a></p><p id="par0160" class="elsevierStylePara elsevierViewall">The role of dermal melanin remains unclear in melasma&#59; one of the hypotheses is that the architectural dysfunction of the basement membrane and the autophagy of the melanocytes of the dermis may cause the presence of dermal melanophages&#44; similar to what occurs in photoaging&#46;<a class="elsevierStyleCrossRefs" href="#bib0120"><span class="elsevierStyleSup">24&#44;25</span></a> In the present study&#44; UVA and UVB promoted a higher rate of melanin in the upper dermis&#59; however&#44; this density was approximately 100 times smaller than that of the epidermis in both anatomical sites&#44; which likely little contributes to the clinical phenotype&#44; but may signal underlying alterations in the upper dermis&#44; basement membrane&#44; and basal layer&#46;</p><p id="par0165" class="elsevierStylePara elsevierViewall">Finally&#44; despite the fundamental role of solar radiation in worsening melasma&#44; no differential expressions of p53 between the melasma and the normal adjacent skin were observed&#44; suggesting that the maintenance of melanogenesis may be due to underlying changes in the upper dermis and epidermis&#44; which induce melanocytic hyperfunction&#46;<a class="elsevierStyleCrossRef" href="#bib0020"><span class="elsevierStyleSup">4</span></a> Furthermore&#44; the exclusive use of sunscreen&#44; despite its preventive effect&#44; is not sufficient for the complete remission of the condition&#44; which indicates the need for a continuous pathophysiological study in search of treatments that lead to the reversal of the entire process&#46;<a class="elsevierStyleCrossRefs" href="#bib0045"><span class="elsevierStyleSup">9&#44;26</span></a></p><p id="par0170" class="elsevierStylePara elsevierViewall">The present study promoted a preliminary analysis of the independent effects of UVB&#44; UVA&#44; and VL radiation&#46; Further investigations should be conducted in order to elucidate the effect of combinations and dosages of irradiations that mirror environmental conditions&#46; The interaction between UVA and VL is still unknown&#59; they are believed to interact in the same melanin precursor during melanogenesis&#46; This association may result in a more evident behavior of melanogenesis in natural situations&#44; where VL and UVA are expressed together&#46;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">20</span></a></p><p id="par0175" class="elsevierStylePara elsevierViewall">This study has limitations inherent to the technique in an <span class="elsevierStyleItalic">ex vivo</span> model&#44; which may differ depending on collection and storage&#46; However&#44; this limitation did not prevent the identification of a melanogenic response&#46; Another limitation can be pointed out by the fact that irradiations in <span class="elsevierStyleItalic">ex vivo</span> skin models occur with independent wavelengths &#40;UVB&#44; UVA and VL&#41;&#44; which is not the case in environmental exposure&#46; In addition to the factors presented above&#44; <span class="elsevierStyleItalic">ex vivo</span> models are dissociated from the organism&#44; limiting the interaction between the hormonal&#44; neural&#44; and vasomotor systems&#46;</p><p id="par0180" class="elsevierStylePara elsevierViewall">New experiments should consider low-intensity repeated irradiation models&#44; as well as associated irradiation&#44; to better reproduce the daily exposure of patients&#46; In addition&#44; the inclusion of substances such as tranexamic acid and antioxidants in the prevention of melanogenesis in melasma can be tested&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Conclusions</span><p id="par0185" class="elsevierStylePara elsevierViewall">An epidermal melanogenic response induced by UVB&#44; UVA&#44; or VL was observed both in skin with melasma and in normal adjacent skin&#46; The morphological patterns related to the photobiology of melanogenesis suggest that different radiations promote individualized responses in the skin with melasma&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Financial support</span><p id="par0190" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleGrantSponsor" id="gs0005">CNPq</span> &#40;PIBIC No&#46; <span class="elsevierStyleGrantNumber" refid="gs0005">148501&#47;2018-4</span>&#41;&#59; <span class="elsevierStyleGrantSponsor" id="gs0010">FUNADERM</span> &#40;<span class="elsevierStyleGrantNumber" refid="gs0010">001&#47;2019</span>&#41;&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Authors&#8217; contributions</span><p id="par0195" class="elsevierStylePara elsevierViewall">Giovana Piteri Alcantara&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0200" class="elsevierStylePara elsevierViewall">Ana Cl&#225;udia Cavalcante Esposito&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#44; elaboration and writing of the manuscript&#59; effective participation in research orientation&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0205" class="elsevierStylePara elsevierViewall">Thain&#225; Oliveira Felicio Olivatti&#58; Approval of the final version of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the manuscript&#46;</p><p id="par0210" class="elsevierStylePara elsevierViewall">Melissa Mari Yoshida&#58; Elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; critical review of the manuscript&#46;</p><p id="par0215" class="elsevierStylePara elsevierViewall">H&#233;lio Amante Miot&#58; Statistical analysis&#59; approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analyzing&#44; and interpreting the data&#59; effective participation in research orientation&#59; intellectual participation in propaedeutic and&#47;or therapeutic conduct of studied cases&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Conflicts of interest</span><p id="par0220" class="elsevierStylePara elsevierViewall">None declared&#46;</p></span></span>"
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    "fechaRecibido" => "2019-10-27"
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            2 => "Ultraviolet rays"
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        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">The independent role of solar radiation in the differential melanogenesis between melasma and adjacent skin is unknown&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Objectives</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">To assess the melanogenic responses of skin with facial melasma and of the adjacent skin to UVB&#44; UVA&#44; and visible light&#44; in an <span class="elsevierStyleItalic">ex vivo</span> model&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">This was a quasi-experimental study involving 22 patients with melasma&#46; Facial melasma and adjacent skin samples were collected and stored in DMEM medium&#44; at room temperature&#46; One fragment was placed under the protection from light&#44; while another was exposed to UVB&#44; UVA&#44; and visible light &#40;blue-violet component&#41;&#58; 166&#8239;mJ&#47;cm<span class="elsevierStyleSup">2</span>&#44; 1&#46;524&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; and 40&#8239;J&#47;cm<span class="elsevierStyleSup">2</span>&#44; respectively&#46; Subsequently&#44; all samples were kept for 72&#8239;hours in a dark environment and stained by Fontana-Masson to assess basal layer pigmentation&#44; dendrites&#44; and melanin granulation&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Effective melanogenesis was observed in the basal layer in melasma and in the normal adjacent skin after all irradiations &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#60;&#8239;0&#46;01&#41;&#44; with the following median increment&#58; UVB &#40;4&#46;7&#37; <span class="elsevierStyleItalic">vs</span>&#46; 8&#46;5&#37;&#41;&#44; UVA &#40;9&#46;5&#37; <span class="elsevierStyleItalic">vs</span>&#46; 9&#46;9&#37;&#41;&#44; and visible light &#40;6&#46;8&#37; <span class="elsevierStyleItalic">vs</span>&#46; 11&#46;7&#37;&#41;&#44; with no significant difference between anatomical sites&#46; An increase in melanin granulation &#40;coarser melanosomes&#41; was observed only after irradiation with UVA and only in the skin with melasma &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#61;&#8239;0&#46;05&#41;&#46; An increase in the melanocyte dendrite count induced by UVB radiation was observed in both anatomical sites &#40;<span class="elsevierStyleItalic">p</span>&#8239;&#8804;&#8239;0&#46;05&#41;&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Study limitations</span><p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Use of an <span class="elsevierStyleItalic">ex vivo</span> model&#44; with independent irradiation regimes for UVB&#44; UVA&#44; and visible light&#46;</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Conclusions</span><p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">Melanogenesis induced by UVB&#44; UVA&#44; and visible light was observed both in melasma and in the adjacent skin&#46; The morphological patterns suggest that different irradiations promote individualized responses on the skin with melasma&#46;</p></span>"
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        "nota" => "<p class="elsevierStyleNotepara" id="npar0010">How to cite this article&#58; Alcantara GP&#44; Esposito ACC&#44; Olivatti TOF&#44; Yoshida MM&#44; Miot HA&#46; Evaluation of <span class="elsevierStyleItalic">ex vivo</span> melanogenic response to UVB&#44; UVA&#44; and visible light in facial melasma and unaffected adjacent skin&#46; An Bras Dermatol&#46; 2020&#59;95&#46; <span class="elsevierStyleInterRef" id="intr0005" href="https://doi.org/10.1016/j.abd.2020.02.015">https&#58;&#47;&#47;doi&#46;org&#47;10&#46;1016&#47;j&#46;abd&#46;2020&#46;02&#46;015</span></p>"
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                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Facial melasma</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Normal adjacent skin</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col"><span class="elsevierStyleItalic">p</span> &#40;time&#8239;&#215;&#8239;group&#41;<a class="elsevierStyleCrossRef" href="#tblfn0010">&#42;&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr><tr title="table-row"><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Pre&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Post&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">p</span><a class="elsevierStyleCrossRef" href="#tblfn0005">&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Pre&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Post&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">p</span><a class="elsevierStyleCrossRef" href="#tblfn0005">&#42;</a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " rowspan="4" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">UVB</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelBasal&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">64 &#40;60&#8211;65&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">65 &#40;65&#8211;68&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">54 &#40;53&#8211;56&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">56 &#40;55&#8211;62&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;09&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelUD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">44 &#40;30&#8211;77&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">45 &#40;38&#8211;97&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;24&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">44 &#40;32&#8211;71&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">67 &#40;41&#8211;87&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;04&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;84&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Dendrites&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6 &#40;2&#8211;8&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">10 &#40;8&#8211;18&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">6 &#40;3&#8211;14&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">11 &#40;7&#8211;16&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;59&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">GranMelanos&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">2&#46;0 &#40;1&#46;5&#8211;2&#46;1&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;6 &#40;1&#46;3&#8211;2&#46;0&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;11&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;9 &#40;1&#46;4&#8211;2&#46;0&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&#46;9 &#40;1&#46;6&#8211;2&#46;0&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;80&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;98&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " rowspan="4" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">UVA</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelBasal&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">62 &#40;54&#8211;66&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">70 &#40;57&#8211;72&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">56 &#40;49&#8211;61&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">57 &#40;50&#8211;63&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&#60;0&#46;01&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;06&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">MelUD&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">41 &#40;18&#8211;88&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">86 &#40;36&#8211;144&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;05&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">45 &#40;27&#8211;90&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">74 &#40;34&#8211;204&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;50&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;88&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Dendrites&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">8 &#40;4&#8211;11&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">8 &#40;2&#8211;22&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;92&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">3 &#40;2&#8211;5&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">5 &#40;3&#8211;9&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;53&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
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ISSN: 03650596
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