The Impact of the Circadian Clock on Skin Physiology and Cancer Development.

Skin cancers are growing in incidence worldwide and are primarily caused by exposures to ultraviolet (UV) wavelengths of sunlight. UV radiation induces the formation of photoproducts and other lesions in DNA that if not removed by DNA repair may lead to mutagenesis and carcinogenesis. Though the factors that cause skin carcinogenesis are reasonably well understood, studies over the past 10-15 years have linked the timing of UV exposure to DNA repair and skin carcinogenesis and implicate a role for the body's circadian clock in UV response and disease risk. Here we review what is known about the skin circadian clock, how it affects various aspects of skin physiology, and the factors that affect circadian rhythms in the skin. Furthermore, the molecular understanding of the circadian clock has led to the development of small molecules that target clock proteins; thus, we discuss the potential use of such compounds for manipulating circadian clock-controlled processes in the skin to modulate responses to UV radiation and mitigate cancer risk.

Efficient Picosecond Laser for Tattoo Removal in Rat Models.

BACKGROUND Tattoos are popular in modern times. Due to the occurance of adverse effects such as poor aesthetic value, scar hyperplasia, and abnormal pigments, there is a high demand for uniform operation standards as well as standards for tattoo technologies. In the present study we used Sprague-Dawley rats to assess the tattoo removal efficacy of use of a picosecond laser at various energy values. MATERIAL AND METHODS Tattoos were made on the backs of rats, then we used a picosecond laser set at various energy parameters to remove the tattoos. After performing the removal procedure in multiple groups, we selected the most suitable energy levels with corresponding parameters for the tattoo removal. We recruited human volunteers who wanted their tattoos removed and used the energy level found to perform best during tattoo removal experiments. The tattoo removal effects were evaluated and verified. Four tattoo volunteers were treated by using the optimal energy parameters for picosecond laser technology. RESULTS Through characterization observation and pathological staining results, it was demonstrated that the 1.9 mJ/µbeam energy laser had the best hollowing effect and the most complete pigment particle crushing effect in the rat skin, and had the best tattoo removal effect. CONCLUSIONS We leveraged the evaluation standard to choose the most suitable energy value of the picosecond laser, which had a good tattoo removal effect and could be employed as a reference for clinical removal of tattoos. This process provides criteria for tattoo removal evaluations as well as alternatives for tattoo removal in clinical practice.

Effects of 308 nm excimer light treatment on the skin microbiome of atopic dermatitis patients.

BACKGROUND: The skin microbiome has been implicated in the pathophysiology of atopic dermatitis (AD). Although 308 nm excimer light treatment is an effective phototherapy for AD, its effects on the skin microbiome currently remain unclear. Therefore, we investigated the effects of the excimer light treatment on the skin bacterial and fungal microbiome of lesional skin of AD. METHODS: Swab samples were collected from 11 healthy controls, non-lesional and lesional skin of 11 AD patients. The excimer light treatment was administered to the lesional skin. The composition of the skin microbiome, the clinical score and skin barrier function of the lesional skin were examined before and after the treatment. The composition of the skin microbiome was determined by sequencing bacterial 16S and fungal internal transcribed spacer regions. RESULTS: The excimer light treatment significantly changed the composition of the bacterial microbiome in the lesional skin of AD, as well as improved the clinical score and skin barrier function. The treatment increased the relative abundance of the phylum Cyanobacteria and decreased that of the phylum Bacteroidetes in lesional skin. At the species level, the treatment significantly decreased the relative abundance of Staphylococcus aureus (S aureus) in lesional skin. There was also a significant correlation between the reduction of S aureus and improvement of the clinical outcomes. CONCLUSION: Our findings suggest that alterations of the skin microbiome with excimer light treatment, specifically the decrease in the abundance of S aureus, are partly involved in the improvement of AD lesions.

Nonsurgical Treatment of Postpartum Lower Abdominal Skin and Soft-Tissue Laxity Using Microfocused Ultrasound With Visualization.

BACKGROUND: Microfocused ultrasound with visualization (MFU-V) is a well-established treatment modality for skin tightening. There is a paucity of evidence for its use in body treatments, such as the lower abdomen. OBJECTIVE: To investigate the effectiveness and safety of MFU-V in treating lower abdominal skin and soft-tissue laxity in postpartum women. METHODS: The lower abdomen of 20 female patients between 6 and 24 months postpartum are treated with MFU-V using 1.5-, 3.0-, and 4.5-mm transducers. Data are prospectively collected and analyzed at 3 and 6 months using subject-reported and investigator-reported outcome measures. One additional patient underwent planned abdominoplasty 6 weeks after MFU-V treatment with tissue assessed intraoperatively and histologically. RESULTS: There was a mean improvement of 1.0 and 1.3 grades at 6 months using the investigator-reported and patient-reported skin laxity scale, respectively (p < .001). Patient-reported outcomes and satisfaction survey showed consistent improvement at 6 months. Histological examination of pretreated tissue showed increased total collagen, increased number and thickness of fibrous septae, and no change in fat cells within pretreated tissue compared with the control. No significant adverse events were recorded. CONCLUSION: MFU-V is an effective and safe treatment modality for lower abdominal skin laxity in postpartum patients.

Histology of ex vivo skin after treatment with fractionated picosecond Nd:YAG laser in high and low-energy settings.

Background: The fractionated picosecond laser produces microscopic lesions in the epidermis and dermis, which are known as laser-induced optical breakdown (LIOB) and intra-dermal laser-induced cavitation (LIC). There have been multiple histological reports on these phenomena, although some have been challenged on the grounds of similarity to artifacts. Asian skins, with a higher melanin content, may react differently to this treatment, and present literature is also lacking in this area.Purpose: To observe and report the histological effect of different energy levels and parameters of the fractional 532 nm/1064 nm picosecond laser on Asian skin ex vivo.Methods: Six skin samples were taken from clinically normal-looking perilesional areas and treated with different energy levels and parameters of the fractional 532 nm/1064 nm picosecond laser. The specimens were then sent to the lab for H&E staining, and the slides were reviewed by a dermatopathologist.Results: Superficial, intra-epidermal LIOBs were seen in skin treating at higher laser energies; deep, intra-dermal LICs were seen in skin treated at lower energies. Lesion sizes and depths were consistent with previously reported values on Caucasian skins, and lesions were spaced in 600-µm intervals or its multiple.Conclusions: The histological findings are consistent with results from other ethnicities, and the spacing of lesions is a strong indication of their validity as LIOBs or LICs.

Sunscreen or simulated sweat minimizes the impact of acute ultraviolet radiation on cutaneous microvascular function in healthy humans.

NEW FINDINGS: What is the central question of this study? Are ultraviolet radiation (UVR)-induced increases in skin blood flow independent of skin erythema? Does broad-spectrum UVR exposure attenuate NO-mediated cutaneous vasodilatation, and does sunscreen or sweat modulate this response? What are the main findings and their importance? Erythema and vascular responses to UVR are temporally distinct, and sunscreen prevents both responses. Exposure to UVR attenuates NO-mediated vasodilatation in the cutaneous microvasculature; sunscreen or simulated sweat on the skin attenuates this response. Sun over-exposure may elicit deleterious effects on human skin that are separate from sunburn, and sunscreen or sweat on the skin may provide protection. ABSTRACT: Exposure to ultraviolet radiation (UVR) may result in cutaneous vascular dysfunction independent of erythema (skin reddening). Two studies were designed to differentiate changes in erythema from skin vasodilatation throughout the 8 h after acute broad-spectrum UVR exposure with (+SS) or without SPF-50 sunscreen (study 1) and to examine NO-mediated cutaneous vasodilatation after acute broad-spectrum UVR exposure with or without +SS or simulated sweat (+SW) on the skin (study 2). In both studies, laser-Doppler flowmetry was used to measure red cell flux, and cutaneous vascular conductance (CVC) was calculated (CVC = flux/mean arterial pressure). In study 1, in 14 healthy adults (24 ± 4 years old; seven men and seven women), the skin erythema index and CVC were measured over two forearm sites (UVR only and UVR+SS) before, immediately after and every 2 h for 8 h post-exposure (750 mJ cm-2 ). The erythema index began to increase immediately post-UVR (P < 0.05 at 4, 6 and 8 h), but CVC did not increase above baseline for the first 4-6 h (P ≤ 0.01 at 6 and 8 h); +SS prevented both responses. In study 2, in 13 healthy adults (24 ± 4 years old; six men and seven women), three intradermal microdialysis fibres were placed in the ventral skin of the forearm [randomly assigned to UVR (450 mJ cm-2 ), UVR+SS or UVR+SW], and one fibre (non-exposed control; CON) was placed in the contralateral forearm. After UVR, a standardized local heating (42°C) protocol quantified the percentage of NO-mediated vasodilatation (%NO). The UVR attenuated %NO compared with CON (P = 0.01). The diminished %NO was prevented by +SS (P < 0.01) and +SW (P < 0.01). Acute broad-spectrum UVR attenuates NO-dependent dilatation in the cutaneous microvasculature, independent of erythema. Sunscreen protects against both inflammatory and heating-induced endothelial dysfunction, and sweat might prevent UVR-induced reductions in NO-dependent dilatation.

UVB- and NGF-induced cutaneous sensitization in humans selectively augments cowhage- and histamine-induced pain and evokes mechanical hyperknesis.

Exaggerated itch responses to pruritic chemical provocations and mechanical stimuli are evident in patients with chronic itch, for example, in atopic dermatitis. Currently used human models of itch do not account for such itch sensitization features, and the mechanisms underlying clinical itch sensitization are unknown. This study utilized two established human models of cutaneous nociceptive sensitization to explore how pre-established inflammatory hyperalgesia (ultraviolet-B-irradiation; "UVB") and non-inflammatory neurotrophic pain sensitization (nerve growth factor; "NGF") alter sensitivity to chemical and mechanically evoked itch. Twenty healthy volunteers participated in the UVB experiment. Six volar forearm areas (2 cm diameter) were UVB irradiated with ≤2 × minimal erythemal dose, and two non-irradiated areas were used as controls. Sixteen healthy volunteers participated in the NGF experiment and had 2 µg intradermally injected (4 × 50 µL in 2 cm diameter areas) into both volar forearms. Isotonic saline was applied as control. Pain sensitivity measurements (mechanical and heat pain thresholds) were conducted to validate the models. Subsequently, itch was evoked using histamine and cowhage spicules in the sensitized skin areas, and itch/pain was rated using visual analogue scales. Mechanical hyperknesis (increased itch to punctuate stimuli) was probed with von Frey filaments before/after each itch provocation. Both UVB- and NGF models induced robust primary mechanical hyperalgesia (P < .01) and hyperknesis (P < .05). Neither of the models augmented itch in response to chemical itch provocations but significant increases specifically for pain ratings were observed for both histamine and cowhage (P < .05). This suggests that these models are of limited value as proxies for itch sensitization to pruritogens observed, e.g., in inflammatory dermatoses.

Temperature elevation in the human brain and skin with thermoregulation during exposure to RF energy.

BACKGROUND: Two international guidelines/standards for human protection from electromagnetic fields define the specific absorption rate (SAR) averaged over 10 g of tissue as a metric for protection against localized radio frequency field exposure due to portable devices operating below 3-10 GHz. Temperature elevation is suggested to be a dominant effect for exposure at frequencies higher than 100 kHz. No previous studies have evaluated temperature elevation in the human head for local exposure considering thermoregulation. This study aims to discuss the temperature elevation in a human head model considering vasodilation, to discuss the conservativeness of the current limit. METHODS: This study computes the temperature elevations in an anatomical human head model exposed to radiation from a dipole antenna and truncated plane waves at 300 MHz-10GHz. The SARs in the human model are first computed using a finite-difference time-domain method. The temperature elevation is calculated by solving the bioheat transfer equation by considering the thermoregulation that simulates the vasodilation. RESULTS: The maximum temperature elevation in the brain appeared around its periphery. At exposures with higher intensity, the temperature elevation became larger and reached around 40 °C at the peak SAR of 100 W/kg, and became lower at higher frequencies. The temperature elevation in the brain at the current limit of 10 W/kg is at most 0.93 °C. The effect of vasodilation became notable for tissue temperature elevations higher than 1-2 °C and for an SAR of 10 W/kg. The temperature at the periphery was below the basal brain temperature (37 °C). CONCLUSIONS: The temperature elevation under the current guideline for occupational exposure is within the ranges of brain temperature variability for environmental changes in daily life. The effect of vasodilation is significant, especially at higher frequencies where skin temperature elevation is dominant.

Skin thermophysiological effects of 448 kHz capacitive resistive monopolar radiofrequency in healthy adults: A randomised crossover study and comparison with pulsed shortwave therapy.

Radiofrequency-based electrophysical agents (EPA) have been used in therapy practice over several decades (e.g., shortwave therapies). Currently, there is insufficient evidence supporting such devices operating below shortwave frequencies. This laboratory-based study investigated the skin physiological effects of 448 kHz capacitive resistive monopolar radiofrequency (CRMRF) and compared them to pulsed shortwave therapy (PSWT). In a randomised crossover study, seventeen healthy volunteers received four treatment conditions - High, Low and Placebo dose conditions receiving 15-min CRMRF treatment and a Control condition receiving no intervention. Fifteen participants also received high dose PSWT for comparison. Treatment was applied to the right lower medial thigh. Pre, post and 20-min follow-up measurements of skin temperature (SKT), skin blood flow (SBF) and nerve conduction velocity (NCV) were obtained using Biopac MP150 system. Group data were compared using the ANOVA model. Statistical significance was set at p ≤ 0.05 (0.8P, 95%CI). Significant increase and sustenance of SKT with both high and low dose CRMRF was demonstrated over the other groups (p < 0.001). PSWT increased SKT significantly (p < 0.001) but failed to sustain it over the follow-up. However, among the five conditions, only high dose CRMRF significantly increased and sustained SBF (p < 0.001). Overall, the CRMRF physiological responses were significantly more pronounced than that of PSWT. No significant changes in NCV were noted for any condition. Physiological changes associated with CRMRF were more pronounced when compared to PSWT, placebo or control. Any potential stronger therapeutic benefits of CRMRF need to be confirmed by comparative clinical studies.

Noninvasive red and near-infrared wavelength-induced photobiomodulation: promoting impaired cutaneous wound healing.

The innumerable intricacies associated with chronic wounds have made the development of new painless, noninvasive, biophysical therapeutic interventions as the focus of current biomedical research. Red and near-infrared light-induced photobiomodulation therapy appears to emerge as a promising drug-free approach for promoting wound healing, reduction in inflammation, pain and restoration of function owing to penetration power in conjunction with their ability to positively modulate the biochemical and molecular responses. This review will describe the physical properties of red and near-infrared light and their interaction with skin and highlight their efficacy of wound repair and regeneration. Near-infrared (800-830 nm) was found to be the most effective and widely studied wavelength range followed by red (630-680 nm) and 904 nm superpulsed light exhibiting beneficial photobiomodulatory effects on impaired dermal wound healing.

Rutin increases critical wavelength of systems containing a single UV filter and with good skin compatibility.

BACKGROUND/PURPOSE: Ultraviolet (UV) radiation is responsible for sunburns, skin cancer, photoaging, and the production of reactive oxygen species (ROS). The awareness on preventing these deleterious effects made the use of anti-UVB formulations an important part of population habits; however, despite the availability of several antioxidants capable of ROS scavenging, the pharmaceutical market lacks products associating UV filters with natural compounds of proven efficacy. Here, we investigated the effect of rutin, a flavonoid with antioxidant activity, associated with UVB filters in dermocosmetic preparations. METHODS: Formulations were assessed through its antioxidant activity, in vitro photoprotective effectiveness, photostability, and in vivo skin tolerance (hydration, transepidermal water loss, and erythema). RESULTS: Samples containing rutin were compatible with the human skin and presented a pronounced antioxidant potential, with scavenging activity values 75% higher than the ones containing only UVB filters. Although rutin could not prevent the sunscreens photodegradation post-irradiation, the bioactive compound significantly increased the formulations critical wavelengths, showing a photoprotective gain, especially in the UVA range. CONCLUSION: In conclusion, the absorption in the UVA range, coupled with ROS scavenging potential, proved the positive effect of rutin applied to anti-UVB formulations, making this bioactive compound a promising candidate for photoprotection improvement.

Design and Fabrication of Double-Focused Ultrasound Transducers to Achieve Tight Focusing.

Beauty treatment for skin requires a high-intensity focused ultrasound (HIFU) transducer to generate coagulative necrosis in a small focal volume (e.g., 1 mm³) placed at a shallow depth (3-4.5 mm from the skin surface). For this, it is desirable to make the F-number as small as possible under the largest possible aperture in order to generate ultrasound energy high enough to induce tissue coagulation in such a small focal volume. However, satisfying both conditions at the same time is demanding. To meet the requirements, this paper, therefore, proposes a double-focusing technique, in which the aperture of an ultrasound transducer is spherically shaped for initial focusing and an acoustic lens is used to finally focus ultrasound on a target depth of treatment; it is possible to achieve the F-number of unity or less while keeping the aperture of a transducer as large as possible. In accordance with the proposed method, we designed and fabricated a 7-MHz double-focused ultrasound transducer. The experimental results demonstrated that the fabricated double-focused transducer had a focal length of 10.2 mm reduced from an initial focal length of 15.2 mm and, thus, the F-number changed from 1.52 to 1.02. Based on the results, we concluded that the proposed double-focusing method is suitable to decrease F-number while maintaining a large aperture size.

Epidermis-dermis junction as a novel location for bone marrow-derived cells to reside in response to ionizing radiation.

Bone marrow-derived cells (BMDCs) can migrate into the various organs in the mice irradiated by ionizing radiation (IR). However, it may not be the case in the skin. While IR is used for bone marrow (BM) transplantation, studying with the epidermal sheets demonstrated that the BMDC recruitment is extraordinarily rare in epidermis in the mouse. Herein, using the chimera mice with BM from green fluorescent protein (GFP) transgenic mice, we simply examined if BMDCs migrate into any layers in the total skin, as opposed to the epidermal sheets, in response to IR. Interestingly, we identified the presence of GFP-positive (GFP(+)) cells in the epidermis-dermis junction in the total skin sections although the epidermal cell sheets failed to have any GFP cells. To examine a possibility that the cells in the junction could be mechanically dissociated during separating epidermal sheets, we then salvaged such dissociated cells and examined its characteristics. Surprisingly, some GFP(+) cells were found in the salvaged cells, indicating that these cells could be derived from BM. In addition, such BMDCs were also associated with inflammation in the junction. In conclusion, BMDCs can migrate to and reside in the epidermis-dermis junction after IR.

Evaluation of light-emitting diodes (LED) effect on skin biology (in vitro study).

BACKGROUND: Interest in anti-aging approaches has grown significantly in recent years. The most popular are the non invasive methods to decrease the signs of aging. One such method is LED-based therapy. METHODS: This study investigated the potential of two different wavelengths, 590 nm and 630 nm, combined or not, in the photobiomodulation of proteins involved in the slowdown of the skin aging. RESULTS: These in vitro results on cell viability, cell shape, and mitochondrial function support and build on previous studies suggested that LED treatment is safe. Regarding its biological functions, our data indicated that the combination of two different wavelengths acted in synergy to enhance the impact of each irradiation alone. Combined, the LED wavelengths could improve in vitro the cell shape, the cell proliferation, and the level of major proteins involved in the healing process. CONCLUSION: These benefits may lead to reinforcement of the skin organization and structure. This hypothesis will be checked in future clinical studies.

Surface dose characterisation of the Varian Ir-192 HDR conical surface applicator set with a vertically orientated source.

BACKGROUND: Conical surface applicators with an Ir-192 high-dose-rate brachytherapy source are a common modality for the treatment of non-melanomatous skin cancer with high tumour control rates. Surface dose characterisation of the Varian Varisource GammaMed+ IX afterloader vertical type surface applicators is performed two dimensionally using high-resolution film dosimetry. AIM: The focus of this study was to determine if Varian surface applicators with a vertical source suffer from the dose distribution irregularities reported for comparable applicators. Our goal was to evaluate if the irregularities found affected treatment and dose output verification procedures. METHODS: Ionisation chamber-based verification of applicator output was established according to guidelines provided by the manufacturer. For additional measurement of surface dose Gafchromic EBT3 film dosimetry was used. The term "therapeutic dose" was defined as 85% of the prescribed dose level. RESULTS: For the 10 different applicator inserts evaluated, cold spots were observed. Mean cold spot size was 2.0 mm × 3.6 mm (± 0.6 mm). The cold spots were dosimetrically well below 85% of the prescribed dose. The cold spot was situated 2.2 mm (1.4-2.7 mm) unilaterally from the central axis and caused general asymmetry in the dose profiles intersecting the cold spot area. A source tilt of approximately 8° (± 1°) was determined for the source used for irradiation. CONCLUSIONS: A central underdosed area exceeding 15 % of the prescribed dose has not been previously reported. Source tilt was observed and found to affect clinical use and possibly treatment outcome in applicators using a vertically arranged source. Surface applicators with a vertically orientated source were subject to dose irregularities that could impact on chamber-based applicator output verification procedures. We recommend film dosimetry-backed applicator commissioning to avoid systematic errors.

Grp1-associated scaffold protein regulates skin homeostasis after ultraviolet irradiation.

Grp1-associated scaffold protein (Grasp), the product of a retinoic acid-induced gene in P19 embryonal carcinoma cells, is expressed primarily in brain, heart, and lung of the mouse. We report herein that Grasp transcripts are also found in mouse skin in which the Grasp gene is robustly induced following acute ultraviolet-B (UVB) exposure. Grasp(-/-) mice were found to exhibit delayed epidermal proliferation and a blunted apoptotic response after acute UVB exposure. Immunohistochemical analyses revealed that the nuclear residence time of the tumor suppressor protein p53 was reduced in Grasp(-/-) mice after UVB exposure. Taken together, our results suggest that a physiological role of Grasp may be to regulate skin homeostasis after UVB exposure, potentially by influencing p53-mediated apoptotic responses in skin.

Entrance and propagation pattern of high-frequency electrical currents in biological tissues as applied to fractional skin rejuvenation using penetrating electrodes.

BACKGROUND: Fractional resurfacing of the skin using radiofrequency devices has been used for collagen remodeling and rejuvenation. OBJECTIVES: To determine how radiofrequency current enters and propagates through tissue, and the pattern of the resulting effect. MATERIALS AND METHODS: An electrosurgical device with a 0.4 MHz frequency output was used as the source of radiofrequency current. Current was applied via a metallic needle introduced into a large piece of cow liver, with different amounts of energy delivered at multiple points. Cross-sections of the liver were then studied for tissue effect. RESULTS: Thermal coagulation of tissue started from the tip of the electrode. With higher energy, a rim of coagulated tissue formed around the entire length of the needle. This rim of coagulated tissue was thicker around the tip of the electrode. CONCLUSION: Radiofrequency currents have a tendency to move toward the center of the bulk of tissue. When an electrode of a fractional radiofrequency device enters the skin, maximum heating effect will be around the tip of the electrode in the dermis. This phenomenon can preserve epidermis from injury during dermal heating, reducing post-procedural skin surface side effects seen with many skin rejuvenation procedures.

Impact of 60-GHz millimeter waves and corresponding heat effect on endoplasmic reticulum stress sensor gene expression.

Emerging high data rate wireless communication systems, currently under development, will operate at millimeter waves (MMW) and specifically in the 60 GHz band for broadband short-range communications. The aim of this study was to investigate potential effects of MMW radiation on the cellular endoplasmic reticulum (ER) stress. Human skin cell lines were exposed at 60.4 GHz, with incident power densities (IPD) ranging between 1 and 20 mW/cm(2) . The upper IPD limits correspond to the ICNIRP local exposure limit for the general public. The expression of ER-stress sensors, namely BIP and ORP150, was then examined by real-time RT-PCR. Our experimental data demonstrated that MMW radiations do not change BIP or ORP150 mRNA basal levels, whatever the cell line, the exposure duration or the IPD level. Co-exposure to the well-known ER-stress inducer thapsigargin (TG) and MMW were then assessed. Our results show that MMW exposure at 20 mW/cm(2) inhibits TG-induced BIP and ORP150 over expression. Experimental controls showed that this inhibition is linked to the thermal effect resulting from the MMW exposure.

Effects of polarization and apodization on laser induced optical breakdown threshold.

We investigated the influence of polarization and apodization on laser induced optical breakdown threshold in transparent and diffuse media using linearly and radially polarized light. We demonstrate a lower irradiance threshold for optical breakdown using radially polarized light. The dominance of radial polarization in higher-order multiphoton ionization has important medical applications where a lower irradiance threshold may allow reaching deeper layers inside the skin with less risk of collateral damage and thereby improving safety and efficacy of treatment.