Illuminating microflora: shedding light on the potential of blue light to modulate the cutaneous microbiome.

Cutaneous diseases (such as atopic dermatitis, acne, psoriasis, alopecia and chronic wounds) rank as the fourth most prevalent human disease, affecting nearly one-third of the world's population. Skin diseases contribute to significant non-fatal disability globally, impacting individuals, partners, and society at large. Recent evidence suggests that specific microbes colonising our skin and its appendages are often overrepresented in disease. Therefore, manipulating interactions of the microbiome in a non-invasive and safe way presents an attractive approach for management of skin and hair follicle conditions. Due to its proven anti-microbial and anti-inflammatory effects, blue light (380 - 495nm) has received considerable attention as a possible 'magic bullet' for management of skin dysbiosis. As humans, we have evolved under the influence of sun exposure, which comprise a significant portion of blue light. A growing body of evidence indicates that our resident skin microbiome possesses the ability to detect and respond to blue light through expression of chromophores. This can modulate physiological responses, ranging from cytotoxicity to proliferation. In this review we first present evidence of the diverse blue light-sensitive chromophores expressed by members of the skin microbiome. Subsequently, we discuss how blue light may impact the dialog between the host and its skin microbiome in prevalent skin and hair follicle conditions. Finally, we examine the constraints of this non-invasive treatment strategy and outline prospective avenues for further research. Collectively, these findings present a comprehensive body of evidence regarding the potential utility of blue light as a restorative tool for managing prevalent skin conditions. Furthermore, they underscore the critical unmet need for a whole systems approach to comprehend the ramifications of blue light on both host and microbial behaviour.

Highlighting nuances of blue light phototherapy: Mechanisms and safety considerations.

The efficacy of blue light therapy in dermatology relies on numerous clinical studies. The safety remains a topic of controversy, where potentially deleterious effects were derived from in vitro rather than in vivo experiments. The objectives of this work were (1) to highlight the nuances behind "colors" of blue light, light propagation in tissue and the plurality of modes of action; and (2) to rigorously analyze studies on humans reporting both clinical and histological data from skin biopsies with focus on DNA damage, proliferation, apoptosis, oxidative stress, impact on collagen, elastin, immune cells, and pigmentation. We conclude that blue light therapy is safe for human skin. It induces intriguing skin pigmentation, in part mediated by photoreceptor Opsin-3, which might have a photoprotective effect against ultraviolet irradiation. Future research needs to unravel photochemical reactions and the most effective and safe parameters of blue light in dermatology.

Cryptochrome 1 is modulated by blue light in human keratinocytes and exerts positive impact on human hair growth.

Photoactivation of cryptochrome-family proteins by blue light is a well-established reaction regulating physiology of plants, fungi, bacteria, insects and birds, while impact of blue light on cryptochrome synthesis and/or activity in human non-visual cells remains unknown. Here, we show that 453 nm blue light induces cryptochrome 1 (CRY1) accumulation in human keratinocytes and the hair follicle. CRY1 is prominently expressed in the human anagen hair follicle, including epithelial stem cells. Specific silencing of CRY1 promotes catagen, while stimulation of CRY1 by KL001 prolongs anagen ex vivo by altering the expression of genes involved in apoptosis and proliferation. Together, our study identifies a role for CRY1 in sustaining human hair growth. Previously, we demonstrated positive effects of 453 nm blue light on hair growth ex vivo. Taken all together, our study suggests that CRY1 might mediate blue light-dependent positive effects on hair growth.

Method for Investigation of Photobiological Effects of Light on Human Skin Cells Mediated by Low Doses of Light.

Driven by evolution, human skin cells have developed an extraordinary ability both to sense and to respond to the photons of sunlight through a plethora of photobiological interactions, activating intracellular signalling cascades and regulating skin cells homeostasis. It has recently been reported that some of these photobiological responses triggered by low levels of light (or the so-called photobiomodulation) could initiate beneficial therapeutic effects. Identification of these effective light-based therapeutic solutions requires in-depth understanding of the parameter space. The physical, biological, and chemical conditions that need to be fulfilled to facilitate such positive photobiological effects are to be carefully deciphered. Here, we provide the protocols that were specifically developed to investigate multidimensional parameter space driving photobiological interactions triggered by light (photobiomodulation) in the skin cells. The approach is based on the so-called design of experiment (DoE), a statistical method, which allows for the investigation of multidimensional parameters landscapes. This goes hand in hand with sharing practical tips for the design of light-based devices inducing these effects. To exemplify practical applications of the developed methods and light-based devices, we disclose experimental data sets and emphasize robustness and reproducibility of the results.

Dose-response of human follicles during laser-based hair removal: Ex vivo photoepilation model with classification system embracing morphological and histological features.

OBJECTIVES: Photoepilation is a commonly used technology in home-use devices (HUDs) and in professional systems to remove unwanted body hair using pulses of laser or intense pulsed light (IPL). Albeit HUDs and professional systems operate at different fluences and treatment regimes, both demonstrate high hair reduction. The underlying mechanisms, however, remain unknown partly due to high divergence of the existing literature data. The objective of this study was to develop an ex vivo photoepilation model with a set of criteria evaluating response to light pulses; and to investigate dose-response behavior of hair follicles (HFs) subjected to a range of fluences. METHODS: After ex vivo treatment (single pulse, 810 nm, 1.7-26.4 J/cm2 , 4-64 ms pulse) human anagen HFs were isolated and maintained in culture for 7-10 days. Response to light was evaluated based on gross-morphology and histological examination (H&E and TUNEL stainings). RESULTS: HFs treated ex vivo demonstrated a dose-dependent response to light with five distinct classes defined by macroscopic and microscopic criteria. Fluences below 13.2 J/cm2 provoked catagen-like transition, higher fluences resulted in coagulation in HF compartments. CONCLUSION: Observed changes in the HF organ culture model were reflected by clinical efficacy. The developed photoepilation model provides an easy and fast method to predict clinical efficacy and permanency of light-based hair removal devices. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Light-based home-use devices for hair removal: Why do they work and how effective they are?

OBJECTIVES: This review has the following objectives: Firstly, it provides an explanation of the evolution of laser/intense pulsed light (IPL) hair reduction modalities from high fluence professional devices to low fluence home-use appliances. Secondly, it summarises published literature reviews on home-use devices (HUDs) as evidence of their growing credibility. Thirdly, it proposes mechanistic differences in light delivery regimes and the resulting divergences in mode of action. MATERIALS AND METHODS: An extensive literature search was performed to review the progress of laser/IPL-induced hair reduction and determine what evidence is available to explain the mode of action of professional and HUDs for hair removal. Establishing the likely biological mode of action of professional high-fluence systems versus home-use low-fluence appliances was performed by combining data obtained using ex vivo hair follicle (HF) organ culture and the clinical results involving human participants. RESULTS: Significant basic science and clinical evidence has been published to confirm the clinical efficacy and technical safety of many laser and IPL home-use devices for hair removal. Clearly, HUDs are different compared to professional systems both in terms of fluence per pulse and in terms of biological mechanisms underlying hair removal. Here we presented data showing that a single low fluence pulse of both 810 nm laser (6.6 J/cm2 , 16 ms) and IPL (9 J/cm2 , 15 ms and 6.8 J/cm2 , 1.9 ms) leads to induction of catagen transition. Catagen transition was characterized by morphological changes similar to what occurs in vivo with occasional detection of apoptosis in the dermal papilla and outer root sheath cells. This suggests that high hair reduction can be expected in vivo and longer-term treatment might result in HF miniaturization due to a cumulative effect on the dermal papilla and outer root sheath cells. In line with this hypothesis, in this review we demonstrate that long-term application of a commercially-available home-use IPL appliance resulted in persistent hair reduction (80%) one year after last treatment. These data are in line with what was previously reported in the literature, where clinical studies with home-use IPL appliances demonstrated high efficacy of hair reduction on female legs, armpits and bikini zones, with full hair regrowth after four treatments following cessation of IPL administration. Limitations of HUDs include lack of hair clearance for very dark skin types and low speed of treatment compared with professional devices. Numerous uncontrolled and controlled clinical efficacy studies and technical safety investigations on consumer-use appliances support many of the leading manufacturers' claims. ANALYSIS & CONCLUSIONS: Manufacturers make consumer appliances safe and easy to use by considering "human factors," needs and capabilities of a variety of users. Safety is of primary concern to manufacturers, regulators and standards bodies as these appliances may be accessible to children or their use attempted on unsuitable skin types without full awareness of potential side effects. Consumer cosmetic appliances are provided with warnings and obvious safety notices describing the nature of any ocular or dermal hazard and precautions for reducing risk of accidental injury, infection, etc. HUDs employing optical energy are provided with design and engineering controls such as safety switches, alarms and sensors to prevent their incorrect operation or eye exposure. In-vivo studies demonstrated that low fluence home-use hair removal devices can result in high hair reduction efficacy after a short treatment regime, while prolonged and less frequent (once in six weeks) maintenance treatment over a year can lead to high and sustained hair reduction even one year after cessation of treatment. Home-use hair removal devices can be a useful adjunct to professional in-office treatments with high professional awareness. There are sufficient positive arguments for practitioners to make the case to patients for HUDs as "companion" products to professional treatments. In addition, devices for hair removal can be used effectively as stand-alone products by the consumer if they are willing to adopt a regime of regular or frequent use. Further clinical studies involving dynamic observation of HF cycle stage and type (terminal vs. vellus) over the total duration of treatment, for example, using biopsies or non-invasive imaging are necessary to confirm the proposed mode of action of low fluence pulses in a combination with treatment and maintenance regimes. Lasers Surg. Med. 51:481-490, 2019. © 2019 Wiley Periodicals, Inc.

Development of a Novel Approach to Studying Corneodesmosomes and Stratum Corneum Adhesion: Extending Knowledge on the Pathophysiology of Sensitive Skin.

BACKGROUND/AIMS: Aberrant skin barrier and intercorneocyte adhesion are potential contributors to the pathomechanism of sensitive skin (SS). Here we aimed to develop a novel and easy-to-apply method to analyze corneodesmosomes and to interrogate potential differences between corneocytes of subjects with SS and non-SS (NSS). METHODS: Corneocytes of the volar forearm and upper outer quadrant of the left buttock of SS (n = 10) and NSS (n = 8) subjects were extracted as a function of depth using adhesive tape and stained with anti-desmoglein 1 (DSG1) antibody. The total area of corneocytes and the number and average size of cells per tape was estimated using image processing. RESULTS: The total area of extracted corneocytes and the quantity of DSG1 decreased with depth. The level of decrease, total area of corneocytes, and average area of individual cells differed between anatomical locations. In SS, a larger total area of extracted corneocytes and a larger average cell size per tape was found at all inspected depths. CONCLUSION: The developed novel and easy-to-apply approach allows investigation of corneodesmosome components. We confirm a role of altered corneocytes in the pathomechanism of SS. The disclosed protocol can further be optimized in studies of skin conditions with strongly affected corneodesmosomes.

Does blue light restore human epidermal barrier function via activation of Opsin during cutaneous wound healing?

BACKGROUND AND OBJECTIVE: Visible light has beneficial effects on cutaneous wound healing, but the role of potential photoreceptors in human skin is unknown. In addition, inconsistency in the parameters of blue and red light-based therapies for skin conditions makes interpretation difficult. Red light can activate cytochrome c oxidase and has been proposed as a wound healing therapy. UV-blue light can activate Opsin 1-SW, Opsin 2, Opsin 3, Opsin 4, and Opsin 5 receptors, triggering biological responses, but their role in human skin physiology is unclear. MATERIALS AND METHODS: Localization of Opsins was analyzed in situ in human skin derived from face and abdomen by immunohistochemistry. An ex vivo human skin wound healing model was established and expression of Opsins confirmed by immunohistochemistry. The rate of wound closure was quantitated after irradiation with blue and red light and mRNA was extracted from the regenerating epithelial tongue by laser micro-dissection to detect changes in Opsin 3 (OPN3) expression. Retention of the expression of Opsins in primary cultures of human epidermal keratinocytes and dermal fibroblasts was confirmed by qRT-PCR and immunocytochemistry. Modulation of metabolic activity by visible light was studied. Furthermore, migration in a scratch-wound assay, DNA synthesis and differentiation of epidermal keratinocytes was established following irradiation with blue light. A role for OPN3 in keratinocytes was investigated by gene silencing. RESULTS: Opsin receptors (OPN1-SW, 3 and 5) were similarly localized in the epidermis of human facial and abdominal skin in situ. Corresponding expression was confirmed in the regenerating epithelial tongue of ex vivo wounds after 2 days in culture, and irradiation with blue light stimulated wound closure, with a corresponding increase in OPN3 expression. Expression of Opsins was retained in primary cultures of epidermal keratinocytes and dermal fibroblasts. Both blue and red light stimulated the metabolic activity of cultured keratinocytes. Low levels of blue light reduced DNA synthesis and stimulated differentiation of keratinocytes. While low levels of blue light did not alter keratinocyte migration in a scratch wound assay, higher levels inhibited migration. Gene silencing of OPN3 in keratinocytes was effective (87% reduction). The rate of DNA synthesis in OPN3 knockdown keratinocytes did not change following irradiation with blue light, however, the level of differentiation was decreased. CONCLUSIONS: Opsins are expressed in the epidermis and dermis of human skin and in the newly regenerating epidermis following wounding. An increase in OPN3 expression in the epithelial tongue may be a potential mechanism for the stimulation of wound closure by blue light. Since keratinocytes and fibroblasts retain their expression of Opsins in culture, they provide a good model to investigate the mechanism of blue light in wound healing responses. Knockdown of OPN3 led to a reduction in early differentiation of keratinocytes following irradiation with blue light, suggesting OPN3 is required for restoration of the barrier function. Understanding the function and relationship of different photoreceptors and their response to specific light parameters will lead to the development of reliable light-based therapies for cutaneous wound healing. Lasers Surg. Med. © 2018 Wiley Periodicals, Inc.

Differential response of human dermal fibroblast subpopulations to visible and near-infrared light: Potential of photobiomodulation for addressing cutaneous conditions.

BACKGROUND OBJECTIVES: The past decade has witnessed a rapid expansion of photobiomodulation (PBM), demonstrating encouraging results for the treatment of cutaneous disorders. Confidence in this approach, however, is impaired not only by a lack of understanding of the light-triggered molecular cascades but also by the significant inconsistency in published experimental outcomes, design of the studies and applied optical parameters. This study aimed at characterizing the response of human dermal fibroblast subpopulations to visible and near-infrared (NIR) light in an attempt to identify the optical treatment parameters with high potential to address deficits in aging skin and non-healing chronic wounds. MATERIALS AND METHODS: Primary human reticular and papillary dermal fibroblasts (DF) were isolated from the surplus of post-surgery human facial skin. An in-house developed LED-based device was used to irradiate cell cultures using six discrete wavelengths (450, 490, 550, 590, 650, and 850 nm). Light dose-response at a standard oxygen concentration (20%) at all six wavelengths was evaluated in terms of cell metabolic activity. This was followed by an analysis of the transcriptome and procollagen I production at a protein level, where cells were cultured in conditions closer to in vivo at 2% environmental oxygen and 2% serum. Furthermore, the production of reactive oxygen species (ROS) was accessed using real-time fluorescence confocal microscopy imaging. Here, production of ROS in the presence or absence of antioxidants, as well as the cellular localization of ROS, was evaluated. RESULTS: In terms of metabolic activity, consecutive irradiation with short-wavelength light (⇐530 nm) exerted an inhibitory effect on DF, while longer wavelengths (>=590 nm) had essentially a neutral effect. Cell behavior following treatment with 450 nm was biphasic with two distinct states: inhibitory at low- to mid- dose levels (<=30 J/cm2 ), and cytotoxic at higher dose levels (>30 J/cm2 ). Cell response to blue light was accompanied by a dose-dependent release of ROS that was localized in the perinuclear area close to mitochondria, which was attenuated by an antioxidant. Overall, reticular DFs exhibited a greater sensitivity to light treatment at the level of gene expression than did papillary DFs, with more genes significantly up- or down- regulated. At the intra-cellular signaling pathway level, the up- or down- regulation of vital pathways was observed only for reticular DF, after treatment with 30 J/cm2 of blue light. At the cellular level, short visible wavelengths exerted a greater inhibitory effect on reticular DF. Several genes involved in the TGF-ß signaling pathway were also affected. In addition, procollagen I production was inhibited. By contrast, 850 nm near-infrared (NIR) light (20 J/cm2 ) exerted a stimulatory metabolic effect in these cells, with no detectable intracellular ROS formation. Here too, reticular DF were more responsive than papillary DF. This stimulatory effect was only observed under in vivo-like low oxygen conditions, corresponding to normal dermal tissue oxygen levels (approximately 2%). CONCLUSION: This study highlights a differential impact of light on human skin cells with upregulation of metabolic activity with NIR light, and inhibition of pro-collagen production and proliferation in response to blue light. These findings open-up new avenues for developing therapies for different cutaneous conditions (e.g., treatment of keloids and fibrosis) or differential therapy at distinct stages of wound healing. Lasers Surg. Med. 50:859-882, 2018. © 2018 Wiley Periodicals, Inc.

Effects of blue light on inflammation and skin barrier recovery following acute perturbation. Pilot study results in healthy human subjects.

BACKGROUND/PURPOSE: While growing evidence supports the therapeutic effect of 453 nm blue light in chronic inflammatory skin diseases, data on its effects on acutely perturbed human skin are scarce. In this study, we investigated the impact of 453 nm narrow-band LED light on healthy skin following acute perturbation. METHODS: Tape stripping and histamine iontophoresis were performed on the forearm of 22 healthy volunteers on 2 consecutive weeks. In 1 week, challenges were followed by irradiation for 30 minutes. In the other week (control), no light was administered. Reactions were evaluated up to 72 hours thereafter by transepidermal water loss (TEWL), diffuse reflectance spectroscopy, and skin surface biomarkers. RESULTS: Skin barrier disruption resulted in upregulation of IL-1α at 24 hours after tape stripping (P = .029). In contrast, irradiation abrogated this effect (P > .05). Irradiation also resulted in higher TEWL at 24 hours and in higher b* value at 72 hours after tape stripping compared to the control (P = .034 and P = .018, respectively). At 30 minutes following histamine iontophoresis and irradiation, a trend toward a higher a* value compared to the control was observed (P = .051). CONCLUSION: We provide the first in vivo evidence that blue light at 453 nm exerts biological effects on acutely perturbed healthy human skin.

A new path in defining light parameters for hair growth: Discovery and modulation of photoreceptors in human hair follicle.

BACKGROUND AND OBJECTIVE: Though devices for hair growth based on low levels of light have shown encouraging results, further improvements of their efficacy is impeded by a lack of knowledge on the exact molecular targets that mediate physiological response in skin and hair follicle. The aim of this study was to investigate the expression of selected light-sensitive receptors in the human hair follicle and to study the impact of UV-free blue light on hair growth ex vivo. MATERIAL AND METHODS: The expression of Opsin receptors in human skin and hair follicles has been characterized using RT-qPCR and immunofluorescence approaches. The functional significance of Opsin 3 was assessed by silencing its expression in the hair follicle cells followed by a transcriptomic profiling. Proprietary LED-based devices emitting two discrete visible wavelengths were used to access the effects of selected optical parameters on hair growth ex vivo and outer root sheath cells in vitro. RESULTS: The expression of OPN2 (Rhodopsin) and OPN3 (Panopsin, Encephalopsin) was detected in the distinct compartments of skin and anagen hair follicle. Treatment with 3.2 J/cm2 of blue light with 453 nm central wavelength significantly prolonged anagen phase in hair follicles ex vivo that was correlated with sustained proliferation in the light-treated samples. In contrast, hair follicle treatment with 3.2 J/cm2 of 689 nm light (red light) did not significantly affect hair growth ex vivo. Silencing of OPN3 in the hair follicle outer root sheath cells resulted in the altered expression of genes involved in the control of proliferation and apoptosis, and abrogated stimulatory effects of blue light (3.2 J/cm2 ; 453 nm) on proliferation in the outer root sheath cells. CONCLUSIONS: We provide the first evidence that (i) OPN2 and OPN3 are expressed in human hair follicle, and (ii) A 453 nm blue light at low radiant exposure exerts a positive effect on hair growth ex vivo, potentially via interaction with OPN3. Lasers Surg. Med. 49:705-718, 2017. © 2017 Wiley Periodicals, Inc.

Sensitive Skin: Assessment of the Skin Barrier Using Confocal Raman Microspectroscopy.

BACKGROUND/AIMS: Sensitive skin (SS), a frequently reported condition in the Western world, has been suggested to be underlined by an impaired skin barrier. The aim of this study was to investigate the skin barrier molecular composition in SS subjects using confocal Raman microspectroscopy (CRS), and to compare it with that of non-SS (NSS) individuals as well as atopic dermatitis (AD) and allergic rhinoconjunctivitis (AR) subjects, who frequently report SS. METHODS: Subjects with SS (n = 29), NSS (n = 30), AD (n = 11), and AR (n = 27) were included. Stratum corneum (SC) thickness, water, ceramides/fatty acids, and natural moisturizing factor (NMF) were measured by CRS along with transepidermal water loss and capacitance on the ventral forearm, thenar, and cheek. Sebum levels were additionally measured on the forearm and cheek. RESULTS: No differences between SS and NSS subjects were found regarding SC thickness, water, and NMF content, yet a trend towards lower ceramides/fatty acids was observed in the cheek. Compared to AD subjects, the SS group showed higher ceramides/fatty acid content in the forearm, whereas no differences emerged with AR. The correlation of macroscopic biophysical techniques and CRS was weak, yet CRS confirmed the well-known lower content of NMF and water, and thinner SC in subjects with filaggrin mutations. CONCLUSION: The skin barrier in SS is not impaired in terms of SC thickness, water, NMF, and ceramides/fatty acid content. The failure of biophysical techniques to follow alterations in the molecular composition of the skin barrier revealed by CRS emphasizes a strong need in sensitive and specific tools for in vivo skin barrier analysis.

Sensitive skin and the influence of female hormone fluctuations: results from a cross-sectional digital survey in the Dutch population.

Sensitive skin is a widespread condition, which is most frequently reported by women. Changing hormone levels during the menstrual cycle and menopause have been suggested among the stimuli triggering sensitive skin. To investigate the perceived influence of fluctuating hormone levels on self-assessed sensitive skin, including symptoms and stimuli linked to skin sensitivity, as well as potential changes in facial and body skin and sensitive body parts, depending on hormonal status. A digital questionnaire was distributed to a population of women aged 20-65 years old. A total of 278 women were included in the analysis. About 42% premenopausal women declared a perception of (increased) skin sensitivity just before and during the menstrual cycle, while this was reported by almost 32% of peri- and postmenopausal women following the menopause. The majority of reported symptoms included the presence of bumps/pimples, dryness, itching, and redness, and the majority of reported stimuli were shaving, weather, toiletries, and emotions. No differences emerged regarding characteristics of facial and body skin across different hormonal status. Significant differences in sensitivity of body parts emerged for the face and feet, reported by a larger percentage of premenopausal women and peri- and postmenopausal women, respectively. The prevalence of the perceived effects of fluctuating hormone levels on self-assessed sensitive skin in women is high. These effects should be taken into consideration in skin testing and dermatological practice, and support the need for selecting personal care routine or treatment during the menstrual cycle and menopause.

Photobiomodulation devices for hair regrowth and wound healing: a therapy full of promise but a literature full of confusion.

Photobiomodulation is reported to positively influence hair regrowth, wound healing, skin rejuvenation and psoriasis. Despite rapid translation of this science to commercial therapeutic solutions, significant gaps in our understanding of the underlying processes remain. The aim of this review was to seek greater clarity and rationality specifically for the selection of optical parameters for studies on hair regrowth and wound healing. Our investigation of 90 reports published between 1985 and 2015 revealed major inconsistencies in optical parameters selected for clinical applications. Moreover, poorly understood photoreceptors expressed in skin such as cytochrome c oxidase, cryptochromes, opsins etc. may trigger different molecular mechanisms. All this could explain the plethora of reported physiological effects of light. To derive parameters for optimal clinical efficacy of photobiomodulation, we recommend a more rational approach to underpin clinical studies, with research on molecular targets and pathways using well-defined biological model systems to enable translation of optical parameters from in vitro to in vivo. Furthermore, special attention needs to be paid when conducting studies for hair regrowth, aiming for double-blind, placebo-controlled randomized clinical trials as the gold standard for quantifying hair growth.

Clinical, biophysical, immunohistochemical, and in vivo reflectance confocal microscopy evaluation of the response of subjects with sensitive skin to home-use fractional non-ablative photothermolysis device.

BACKGROUND: Fractional photothermolysis using professional devices is a well-accepted and a widely used technique for skin rejuvenation. Recently, the technology has also been implemented in devices for home-use. Yet, a subpopulation of consumers exists that reacts excessively to this stimulation and reports "sensitive skin" (SS). OBJECTIVE: The goal of this study was to evaluate the response of subjects with SS and NSS to fractional non-ablative photothermolysis to provide additional insights in the pathophysiology of SS. METHODS: Subjects with SS and non-sensitive skin (NSS), selected using a proprietary questionnaire were stimulated by applying a home-use fractional non-ablative photothermolysis device. Self-reported perceptions and objective effects were evaluated after 0.5, 8, 24, and 72 hours by clinical, biophysical and immunohistochemical assessment, and in vivo reflectance confocal microscopy (RCM). RESULTS: Significantly fewer mast cells were found in SS compared to NSS subjects, 0.5 and 72 hours after stimulus based on tryptase staining, and SS subjects report discomfort more frequently. Immunohistochemical biomarkers revealed new insights in the effects of fractional non-ablative photothermolysis, which were supported by RCM: peri- and interlesional epidermal proliferation, and changes in keratinocyte differentiation. CONCLUSION: Previously, we have already reported that SS could be elicited by mechanical and chemical stimuli. Thus, mild yet excessive self-reported perceptions described here supports the hypothesis about existence of generalized skin sensitivity. Furthermore, it supports a view point suggesting involvement of TRPV1 receptors in this phenomenon. While histological evaluation, in line with our previous results points to the role of mast cells in SS, overall, however, fractional non-ablative photothermolysis causes only mild damage, nearly equal in SS and NSS and could be used as an in vivo model for skin regeneration without manipulating the skin barrier. Lasers Surg. Med. 48:474-482, 2016. © 2016 Wiley Periodicals, Inc.

Microspectroscopic Confocal Raman and Macroscopic Biophysical Measurements in the in vivo Assessment of the Skin Barrier: Perspective for Dermatology and Cosmetic Sciences.

Skin barrier function, confined to the stratum corneum, is traditionally evaluated using established, noninvasive biophysical methods like transepidermal water loss, capacitance and conductance. However, these methods neither measure skin molecular composition nor its structure, hindering the actual causes of skin barrier change or impairment. At the same time, confocal Raman microspectroscopy (CRS) can directly measure skin molecular composition and structure and has proven itself to be a powerful technique for biomolecular analysis. The aims of this literature review were to evaluate noninvasive biophysical methods in view of CRS and to outline a direction towards more specific and informative skin measurement methods. We address this by investigating, for the first time, the relation between in vivo assessment of the skin barrier using indirect biophysical methods and the actual skin composition and structure as given by CRS, and emphasize the high potential of CRS for dermatology and cosmetic sciences. CRS acceptance in these fields will require close collaboration between dermatologists, skin scientists and spectroscopy experts towards simplifying the technology and creating robust, rapid, easy-to-use and less expensive CRS applications.

Improved depth resolution in near-infrared diffuse reflectance spectroscopy using obliquely oriented fibers.

We demonstrate a significant improvement of depth selectivity when using obliquely oriented fibers for near-infrared (NIR) diffuse reflectance spectroscopy. This is confirmed by diffuse reflectance measurements of a two-layer tissue-mimicking phantom across the spectral range from 1,000 to 1,940 nm. The experimental proof is supported by Monte Carlo simulations. The results reveal up to fourfold reduction in the mean optical penetration depth, twofold reduction in its variation, and a decrease in the number of scattering events when a single fiber is oriented at an angle of 60 deg. The effect of reducing the mean optical penetration depth is enhanced by orienting both fibers inwardly. Using outwardly oriented fibers enables more selective probing of deeper layers, while reducing the contribution from surface layers. We further demonstrate that the effect of an inward oblique arrangement can be approximated to a decrease in fiber-to-fiber separation in the case of a perpendicular fiber arrangement. This approximation is valid in the weak- or absorption-free regime. Our results assert the advantages of using obliquely oriented fibers when attempting to specifically address superficial tissue layers, for example, for skin cancer detection, or in noninvasive glucose monitoring. Such flexibility could be further advantageous in a range of minimally invasive applications, including catheter-based interventions.