Hydradermabrasion through the lens of Line-Field Confocal Optical Coherence Tomography.

BACKGROUND: Hydradermabrasion, also known as "HydraFacial," is an exfoliative cosmetic procedure for skin rejuvenation that has gained popularity. Despite its increasing popularity, clinical studies validating its efficacy with non-invasive assessment of histological changes to the skin, are scarce. In this study, we used Line-Field Confocal Optical Coherence Tomography (LC-OCT), an optical imaging device, to non-invasively visualize microscopic changes to skin anatomy after hydradermabrasion treatment. MATERIALS/METHODS: Eight volunteers (Fitzpatrick skin types II-V) were recruited for this study. Images, using LC-OCT (DeepLive, DAMAE medical) were obtained before and after hydradermabrasion and at 2 weeks post-treatment. A commercially available hydradermabrasion device was utilized to perform the dermabrasion. RESULTS: In the epidermis, initially, a decrease in the average thickness of the stratum corneum, from 9.42 to 6.67 µm was visualized in LC-OCT images after hydradermabrasion. However, at 2 weeks of follow-up, the average stratum corneum thickness was 9.75 µm, resulting in an overall increase in the average thickness after treatment. Improved homogenization of the stratum corneum and decreased number of undulations in the epidermis post-treatment were also visualized. In all the subjects, the superficial dermis appeared stretched, which returned to baseline by the 2-week follow-up. At the 2-week follow-up, there were no visible differences in the quality and quantity of collagen fibers in the dermis. CONCLUSION: In our study, LC-OCT images of the epidermis and dermis demonstrated microscopic features of skin rejuvenation when treated with hydradermabrasion. Thus, not only highlighting the efficacy of hydradermabrasion but also the potential of LC-OCT to serve as a tool for visualizing the microscopic effects of cosmetic procedures on skin anatomy.

Pressure-Insensitive Epidermal Thickness of Fingertip Skin for Optical Image Encryption.

In this study, an internal fingerprint-guided epidermal thickness of fingertip skin is proposed for optical image encryption based on optical coherence tomography (OCT) combined with U-Net architecture of a convolutional neural network (CNN). The epidermal thickness of fingertip skin is calculated by the distance between the upper and lower boundaries of the epidermal layer in cross-sectional optical coherence tomography (OCT) images, which is segmented using CNN, and the internal fingerprint at the epidermis-dermis junction (DEJ) is extracted based on the maximum intensity projection (MIP) algorithm. The experimental results indicate that the internal fingerprint-guided epidermal thickness is insensitive to pressure due to normal correlation coefficients and the encryption process between epidermal thickness maps of fingertip skin under different pressures. In addition, the result of the numerical simulation demonstrates the feasibility and security of the encryption scheme by structural similarity index matrix (SSIM) analysis between the original image and the recovered image with the correct and error keys decryption, respectively. The robustness is analyzed based on the SSIM value in three aspects: different pressures, noise attacks, and data loss. Key randomness is valid by the gray histograms, and the average correlation coefficients of adjacent pixelated values in three directions and the average entropy were calculated. This study suggests that the epidermal thickness of fingertip skin could be seen as important biometric information for information encryption.

Skin dark spot mapping and evaluation of brightening product efficacy using Line-field Confocal Optical Coherence Tomography (LC-OCT).

BACKGROUND: Facial dark spots remain a significant challenge for the cosmetic industry, in terms of providing effective treatment. Using Line-field Confocal Optical Coherence Tomography (LC-OCT), we investigated the internal structural features of photo-aging spot areas and evaluated the efficacy of a skin-brightening cosmetic product. MATERIALS AND METHODS: Twenty-six Asian female volunteers, aged between 29 and 65 years, applied a cosmetic product on their entire face twice a day for 2 months. LC-OCT was used to evaluate the dermal-epidermal junction (DEJ) undulation and the volume density of melanin in the epidermis at D0 and D56. Skin brightening and redness were also assessed by photography (SkinCam). RESULTS: Using LC-OCT technology, various microscopic dark spot morphologies, spanning from minimally deformed DEJ to complex DEJ patterns, were identified. Dark spots characterized by slight deformities in the DEJ were predominantly observed in the youngest age group, while older volunteers displayed a wavier pattern. Furthermore, a total of 44 spots were monitored to evaluate the brightening product efficacy. A statistically significant reduction in melanin volumetric density of 7.3% in the spots and 12.3% in their surrounding area was observed after 56 days of product application. In line with these results, an analysis of color parameters using SkinCam reveals a significant increase in brightening and decrease in redness in both pigmented spots and the surrounding skin following application. CONCLUSIONS: LC-OCT proves to be a valuable tool for in-depth dark spots characterization and assessment of skin brightening products, enabling various applications in the field of dermatological sciences.

The correlation between sub-epidermal moisture measurement and other early indicators of pressure ulcer development-A prospective cohort observational study. Part 1. The correlation between sub-epidermal moisture measurement and ultrasound.

The correlation between sub-epidermal moisture (SEM) and other early indicators of pressure ulcer (PU) development is yet to be determined. This three-part series aims to bridge this knowledge gap, through investigating SEM and its correlation with evidence-based technologies and assessments. This article focuses on the correlation between SEM and ultrasound. A prospective cohort observational study was undertaken between February and November 2021. Patients undergoing three surgery types were consecutively enrolled to the study following informed consent. Assessments were performed prior to and following surgery for 3 days at the sacrum, both heels and a control site, using a SEM scanner and high-frequency ultrasound scanner (5-15 MHz). Spearman's rank (rs ) explored the correlation between SEM and ultrasound. A total of 60 participants were included; 50% were male with a mean age of 58 years (±13.46). A statistically significant low to moderately positive correlation was observed between SEM and ultrasound across all anatomical sites (rs range = 0.39-0.54, p < 0.05). The only exception was a correlation between SEM and ultrasound on day 0 at the right heel (rs = 0.23, p = 0.09). These results indicate that SEM and ultrasound agreed in the presence of injury; however, SEM was able to identify abnormalities before ultrasound.

[Research advances on application of sub-epidermal moisture scanner in monitoring tissue viability of early pressure injuries].

Pressure injury (PI) not only reduces the quality of life of patients but also is expensive to manage, placing a heavy financial burden on patients and their families, and society. Despite the increasing diversity of methods used to identify early PI, there are still few methods that can truly and accurately predict early PI. The sub-epidermal moisture scanner is the first U.S. Food and Drug Administration-authorized PI management device that can predict the occurrence and development of PI by measuring the level of local tissue bio-capacitance and monitoring the tissue viability. As an emerging diagnostic instrument, the sub-epidermal moisture scanner has already shown great advantages in clinical practice, which can promote the informatization, digitization, and intelligent prevention and management of PI. This paper introduces the pathophysiological mechanism of PI, elucidates the working principle and parameter settings of the sub-epidermal moisture scanner, its clinical application in monitoring tissue viability in early PI, and its limitation, and looks forward to its future development.

Quantitative analysis of age-related changes in vascular structure, oxygen saturation, and epidermal melanin structure using photoacoustic methods.

BACKGROUND: Vascular structure, blood oxygen saturation, and melanin status of the epidermis are chromophore factors related to light absorption. Therefore, they are likely to be related to skin appearance. Thus, it is important to measure these internal skin features and understand their characteristics. Thus, we aimed to analyze the individual differences and aging changes in the skin by measuring the internal skin characteristics, such as vascular structure, oxygen saturation, and the 3D distribution of melanin in the epidermis, using a noninvasive photoacoustic (PA) measurement method. MATERIALS AND METHODS: A PA measurement device was used as a noninvasive measurement method. Eighty Japanese women aged between 20 and 60 years were enrolled. The target area was the buccal region of the face. RESULTS: The blood vessel structure showed a decrease in fine vessels with age, with a stronger tendency observed in the dermis layer, and the volume of blood vessels was larger in the dermis layer than in the dermal-subcutaneous fat boundary layer. Oxygen saturation showed a similar decreasing trend with age in all depths examined. Melanin condition as the torus-like pattern structure tended to increase with age. CONCLUSION: PA measurements revealed the characteristics of several chromophores, providing a new skin aging mechanism.

Rapid measurement of epidermal thickness in OCT images of skin.

Epidermal thickness (ET) changes are associated with several skin diseases. To measure ET, segmentation of optical coherence tomography (OCT) images is essential; manual segmentation is very time-consuming and requires training and some understanding of how to interpret OCT images. Fast results are important in order to analyze ET over different regions of skin in rapid succession to complete a clinical examination and enable the physician to discuss results with the patient in real time. The well-known CNN-graph search (CNN-GS) methodology delivers highly accurate results, but at a high computational cost. Our objective was to build a computational core, based on CNN-GS, able to accurately segment OCT skin images in real time. We accomplished this by fine-tuning the hyperparameters, testing a range of speed-up algorithms including pruning and quantization, designing a novel pixel-skipping process, and implementing the final product with efficient use of core and threads on a multicore central processing unit (CPU). We name this product CNN-GS-skin. The method identifies two defined boundaries on OCT skin images in order to measure ET. We applied CNN-GS-skin to OCT skin images, taken from various body sites of 63 healthy individuals. Compared with CNN-GS, our described method reduced computation time by 130 [Formula: see text] with minimal reduction in ET determination accuracy (from 96.38 to 94.67%).

Zinc penetration through the skin barrier in atopic dermatitis and rosacea using reflectance confocal microscopy.

Atopic dermatitis (AD) is a chronic, recurrent eczematous disorder with a complex pathophysiology caused by skin barrier abnormalities. Rosacea is a common chronic immune-mediated inflammatory disorder that results in diminished skin barrier function. Reflectance confocal microscopy (RCM) is a non-invasive method for visualizing the dynamic status of epidermal and upper dermal structures. In this study, we compared skin barrier permeability among normal, AD and rosacea groups. To assess skin barrier permeability, zinc was applied to lesional skin and the RCM reflectance intensity of zinc penetration was measured. Reflectance confocal microscopy revealed that the intensity in patients with rosacea and AD was higher than that in the normal group at depths of 8-24 µm in both the face and forearm, which were considered as the stratum corneum (SC) and tight junction (TJ) level (p < 0.0001). When comparing AD and rosacea, the intensity of rosacea was higher than that of AD at a depth of 8 µm in the face (p < 0.0001). The intensity of AD was higher than that of rosacea at a depth of 24 µm (p = 0.009). This suggests that skin barrier permeability is increased in the upper epidermis of patients with AD and rosacea. On the face, patients with rosacea had more SC weakness than did those with AD, whereas patients with AD had more TJ weakness than those with rosacea.

Line-field confocal optical coherence tomography: New insights for psoriasis treatment monitoring.

BACKGROUND: Line-field confocal optical coherence tomography (LC-OCT) is a new, valid means for a rapid and non-invasive in vivo examination of the epidermis and upper dermis, allowing digital interpretation and measurement of high-resolution images on a cellular level. Given these properties, it may represent a valid tool for monitoring psoriasis during treatment, allowing a new method to set a precise objective severity of the disease. OBJECTIVES: We aimed to investigate the potentialities of LC-OCT in the non-invasive monitoring of microscopical changes associated with moderate-severe plaque psoriasis (PP) during the treatment with the most common biological drugs. MATERIALS AND METHODS: We performed LC-OCT imaging of PP lesions from 17 patients before and after 8 weeks of treatment. The clinical severity of the single lesions was evaluated using a lesion score (LS), designed considering three parameters: erythema, desquamation and infiltration. LC-OCT images were segmented by artificial intelligence and evaluated based on three microscopic criteria: the thickness of the stratum corneum, the thickness of the living epidermis and the undulation of the dermo-epidermal junction. RESULTS: Line-field confocal optical coherence tomography digital analysis allowed recognition and quantification of the three microscopic criteria, showing a reduction of all these during the follow-up. Furthermore, a high correlation between change in LS and the thickness of the stratum corneum and the thickness of the living epidermis was found. CONCLUSION: Line-field confocal optical coherence tomography can non-invasively monitor the response of PP to different treatments. Morphometric changes occurring in the psoriatic lesion during the 8-week treatment period were identified by in vivo LC-OCT and measured by using artificial intelligence. Although future studies are required, based on these preliminary results, LC-OCT may represent a valid potential tool for precise monitoring of therapeutic response.

Absorption and reduced scattering coefficients in epidermis and dermis from a Swedish cohort study.

Significance: Knowledge of optical properties is important to accurately model light propagation in tissue, but in vivo reference data are sparse. Aim: The aim of our study was to present in vivo skin optical properties from a large Swedish cohort including 3809 subjects using a three-layered skin model and spatially resolved diffuse reflectance spectroscopy (Periflux PF6000 EPOS). Approach: Diffuse reflectance spectra (475 to 850 nm) at 0.4 and 1.2 mm source-detector separations were analyzed using an inverse Monte Carlo method. The model had one epidermis layer with variable thicknesses and melanin-related absorptions and two dermis layers with varying hemoglobin concentrations and equal oxygen saturations. The reduced scattering coefficient was equal across all layers. Results: Median absorption coefficients (mm-1) in the upper dermis ranged from 0.094 at 475 nm to 0.0048 at 850 nm and similarly in the lower dermis from 0.059 to 0.0035. The reduced scattering coefficient (mm-1) ranged from 3.22 to 1.20, and the sampling depth (mm) ranged from 0.23 to 0.38 (0.4 mm separation) and from 0.49 to 0.68 (1.2 mm separation). There were differences in optical properties across sex, age groups, and BMI categories. Conclusions: Reference material for skin optical properties is presented.

High-frequency ultrasound in anti-aging skin therapy monitoring.

Over the last few decades, high-frequency ultrasound has found multiple applications in various diagnostic fields. The fast development of this imaging technique opens up new diagnostic paths in dermatology, allergology, cosmetology, and aesthetic medicine. In this paper, being the first in this area, we discuss the usability of HFUS in anti-aging skin therapy assessment. The fully automated algorithm combining high-quality image selection and entry echo layer segmentation steps followed by the dermal parameters estimation enables qualitative and quantitative evaluation of the effectiveness of anti-aging products. Considering the parameters of subcutaneous layers, the proposed framework provides a reliable tool for TCA-peel therapy assessment; however, it can be successfully applied to other skin-condition-related problems. In this randomized controlled clinical trial, forty-six postmenopausal women were randomly assigned to the experimental and control groups. Women were treated four times at one-week intervals and applied skin cream daily between visits. The three month follow-up study enables measurement of the long-term effect of the therapy. According to the results, the TCA-based therapy increased epidermal (entry echo layer) thickness, indicating that the thinning process has slowed down and the skin's condition has improved. An interesting outcome is the obtained growth in the intensity of the upper dermis in the experimental group, which might suggest a reduced photo-aging effect of TCA-peel and increased water content. The same conclusions connected with the anti-aging effect of TCA-peel can be drawn by observing the parameters describing the contribution of low and medium-intensity pixels in the upper dermis. The decreased share of low-intensity pixels and increased share of medium-intensity pixels in the upper dermis suggest a significant increase in local protein synthesis.

Gadolinium contrast agents: dermal deposits and potential effects on epidermal small nerve fibers.

Small fiber neuropathy (SFN) affects unmyelinated and thinly myelinated nerve fibers causing neuropathic pain with distal distribution and autonomic symptoms. In idiopathic SFN (iSFN), 30% of the cases, the underlying aetiology remains unknown. Gadolinium (Gd)-based contrast agents (GBCA) are widely used in magnetic resonance imaging (MRI). However, side-effects including musculoskeletal disorders and burning skin sensations were reported. We investigated if dermal Gd deposits are more prevalent in iSFN patients exposed to GBCAs, and if dermal nerve fiber density and clinical parameters are likewise affected. 28 patients (19 females) with confirmed or no GBCA exposure were recruited in three German neuromuscular centers. ISFN was confirmed by clinical, neurophysiological, laboratory and genetic investigations. Six volunteers (two females) served as controls. Distal leg skin biopsies were obtained according to European recommendations. In these samples Gd was quantified by elemental bioimaging and intraepidermal nerve fibers (IENF) density via immunofluorescence analysis. Pain phenotyping was performed in all patients, quantitative sensory testing (QST) only in a subset (15 patients; 54%). All patients reported neuropathic pain, described as burning (n = 17), jabbing (n = 16) and hot (n = 11) and five QST scores were significantly altered. Compared to an equal distribution significantly more patients reported GBCA exposures (82%), while 18% confirmed no exposures. Compared to unexposed patients/controls significantly increased Gd deposits and lower z-scores of the IENF density were confirmed in exposed patients. QST scores and pain characteristics were not affected. This study suggests that GBCA exposure might alter IENF density in iSFN patients. Our results pave the road for further studies investigating the possible role of GBCA in small fiber damage, but more investigations and larger samples are needed to draw firm conclusions.

In vivo detection of healthy skin: Comparison of multiphoton microscopy and reflectance confocal microscopy.

BACKGROUND: Noninvasive skin examination evolved rapidly in recent years, with multiphoton microscopy (MPM) and reflectance confocal microscopy (RCM) being used to image in-vivo skin at high resolution. The aim of this study is to compare the imaging clarity between the two techniques and measure the thickness of the epidermis in different body sites. We also measured the degree of skin aging with noninvasive tools. METHODS: Fifty-six volunteers were evaluated and measured at three different body sites, including the cheek, volar forearm, and back. We used RCM and MPM to evaluate the clarity of each skin layer, including stratum corneum, stratum granulosum, stratum spinosum, dermo-epidermal junction, and dermis. We measured epidermal thickness (ET) at the three body sites in individuals of different ages and genders. We assessed skin aging by the second harmonic to autofluorescence aging index of dermis (SAAID), and multiple linear regression was used to analyze the factors affecting SAAID. RESULTS: MPM had advantages in observation of stratum granulosum, collagen fiber, and elastic fiber (p < 0.001), but RCM provided better observation in dermo-epidermal junction layer (p < 0.001). The epidermis was thicker in the cheek area than the volar forearm and back in both RCM and MPM detection, and the average ET measured by MPM was lower than RCM. ET varied among the three body sites with significant differences (p < 0.05). ET was significantly lower at almost all sites in individuals above 40y (p < 0.05). SAAID decreased with age, and more rapidly in women. Cheeks have lower SAAID scores than other body sites. CONCLUSION: MPM and RCM provide noninvasive methods for imaging skin and each method has its own advantages. Epidermal thickness and SAAID correlated with age, gender, and different body sites. MPM could also assess the degree of skin aging, which could guide the clinical treatment of patients with diffferent ages and genders in the above body sites.

Age-dependent changes in epidermal architecture explored using an automated image analysis algorithm on in vivo reflectance confocal microscopy images.

BACKGROUND: Reflectance confocal microscopy (RCM) allows for real-time in vivo visualization of the epidermis at the cellular level noninvasively. Parameters relating to tissue architecture can be extracted from RCM images, however, analysis of such images requires manual identification of cells to derive these parameters, which can be time-consuming and subject to human error, highlighting the need for an automated cell identification method. METHODS: First, the region-of-interest (ROI) containing cells needs to be identified, followed by the identification of individual cells within the ROI. To perform this task, we use successive applications of Sato and Gabor filters. The final step is post-processing improvement of cell detection and removal of size outliers. The proposed algorithm is evaluated on manually annotated real data. It is then applied to 5345 images to study the evolution of epidermal architecture in children and adults. The images were acquired on the volar forearm of healthy children (3 months to 10 years) and women (25-80 years), and on the volar forearm and cheek of women (40-80 years). Following the identification of cell locations, parameters such as cell area, cell perimeter, and cell density are calculated, as well as the probability distribution of the number of nearest neighbors per cell. The thicknesses of the Stratum Corneum and supra-papillary epidermis are also calculated using a hybrid deep-learning method. RESULTS: Epidermal keratinocytes are significantly larger (area and perimeter) in the granular layer than in the spinous layer and they get progressively larger with a child's age. Skin continues to mature dynamically during adulthood, as keratinocyte size continues to increase with age on both the cheeks and volar forearm, but the topology and cell aspect ratio remain unchanged across different epidermal layers, body sites, and age. Stratum Corneum and supra-papillary epidermis thicknesses increase with age, at a faster rate in children than in adults. CONCLUSIONS: The proposed methodology can be applied to large datasets to automate image analysis and the calculation of parameters relevant to skin physiology. These data validate the dynamic nature of skin maturation during childhood and skin aging in adulthood.

Revealing the Meissner Corpuscles in Human Glabrous Skin Using In Vivo Non-Invasive Imaging Techniques.

The presence of mechanoreceptors in glabrous skin allows humans to discriminate textures by touch. The amount and distribution of these receptors defines our tactile sensitivity and can be affected by diseases such as diabetes, HIV-related pathologies, and hereditary neuropathies. The quantification of mechanoreceptors as clinical markers by biopsy is an invasive method of diagnosis. We report the localization and quantification of Meissner corpuscles in glabrous skin using in vivo, non-invasive optical microscopy techniques. Our approach is supported by the discovery of epidermal protrusions which are co-localized with Meissner corpuscles. Index fingers, small fingers, and tenar palm regions of ten participants were imaged by optical coherence tomography (OCT) and laser scan microscopy (LSM) to determine the thickness of the stratum corneum and epidermis and to count the Meissner corpuscles. We discovered that regions containing Meissner corpuscles could be easily identified by LSM with an enhanced optical reflectance above the corpuscles, caused by a protrusion of the strongly reflecting epidermis into the stratum corneum with its weak reflectance. We suggest that this local morphology above Meissner corpuscles has a function in tactile perception.

In vivo transepidermal water loss: Validation of a new multi-sensor open chamber water evaporation system Tewameter TM Hex.

BACKGROUND: Instrumentation technology for transepidermal water loss measurements has not been substantially modified since its introduction by Nilsson in 1977. Recent progress in sensor development allowed a new sensor arrangement using a matrix of 30 sensors. Raw measurement values are processed with spatial statistical analysis. We aimed to compare the new, multi-sensor probe (Tewameter TM Hex) with the established Tewameter TM 300 probe and to gain reference data for the new parameters of transepidermal energy loss and water vapor concentration on skin. MATERIAL AND METHODS: Baseline measurements and repeated measurements on the volar forearm and assessment on eight different anatomical locations were performed on 24 healthy volunteers (both gender) with the TM Hex and the TM 300. RESULTS: A significant correlation (p < 0.001; R-coefficient = 0.9) between TM Hex and the TM 300 with a low coefficient of variance (CV) 11% for TM Hex and 19% for TM 300, could be assessed. The CV ranged between 7% (right inner upper arm) and 14% (palms). Average transepidermal heat loss ranged from 12 W/m2 on the lower leg to 38.8 W/m2 on the palm. CONCLUSION: The correlation between TM Hex and TM 300 along with the robustness of the measurements with the TM Hex shows that the new probe for assessment of epidermal barrier function is comparable to the TM 300. In most conditions, TM Hex provides more accurate measurements than TM 300. New parameters open the field to studying skin's water and energy balance.

Automatic granular and spinous epidermal cell identification and analysis on in vivo reflectance confocal microscopy images using cell morphological features.

Significance: Reflectance confocal microscopy (RCM) allows for real-time in vivo visualization of the skin at the cellular level. The study of RCM images provides information on the structural properties of the epidermis. These may change in each layer of the epidermis, depending on the subject's age and the presence of certain dermatological conditions. Studying RCM images requires manual identification of cells to derive these properties, which is time consuming and subject to human error, highlighting the need for an automated cell identification method. Aim: We aim to design an automated pipeline for the analysis of the structure of the epidermis from RCM images of the Stratum granulosum and Stratum spinosum. Approach: We identified the region of interest containing the epidermal cells and the individual cells in the segmented tissue area using tubeness filters to highlight membranes. We used prior biological knowledge on cell size to process the resulting detected cells, removing cells that were too small and reapplying the used filters locally on detected regions that were too big to be considered a single cell. The proposed full image analysis pipeline (FIAP) was compared with machine learning-based approaches (cell cutter, different U-Net configurations, and loss functions). Results: All methods were evaluated both on simulated data (four images) and on manually annotated RCM data (seven images). Accuracy was measured using recall and precision metrics. Both accuracy metrics were higher in the proposed FIAP for both real ( precision = 0.720 ± 0.068 , recall = 0.850 ± 0.11 ) and synthetic images ( precision = 0.835 ± 0.067 , recall = 0.925 ± 0.012 ). The tested machine learning methods failed to identify and segment keratinocytes on RCM images with a satisfactory accuracy. Conclusions: We showed that automatic cell segmentation can be achieved using a pipeline based on membrane detection, with an accuracy that matches expert manual cell identification. To our knowledge, this is the first method based on membrane detection to study healthy skin using RCM images evaluated against manually identified cell positions.

Radiation-induced alterations in multi-layered, in-vitro skin models detected by optical coherence tomography and histological methods.

BACKGROUND: Inflammatory skin reactions and skin alterations are still a potential side effect in radiation therapy (RT), which also need attention for patients' health care. METHOD: In a pre-clinical study we consider alterations in irradiated in-vitro skin models of epidermal and dermal layers. Typical dose regimes in radiation therapy are applied for irradiation. For non-invasive imaging and characterization optical coherence tomography (OCT) is used. Histological staining method is additionally applied for comparison and discussion. RESULTS: Structural features, such as keratinization, modifications in epidermal cell layer thickness and disorder in the layering-as indications for reactions to ionizing radiation and aging-could be observed by means of OCT and confirmed by histology. We were able to recognize known RT induced changes such as hyper-keratosis, acantholysis, and epidermal hyperplasia as well as disruption and/or demarcation of the dermo-epidermal junction. CONCLUSION: The results may pave the way for OCT to be considered as a possible adjunctive tool to detect and monitor early skin inflammation and side effects of radiotherapy, thus supporting patient healthcare in the future.

Tattoo Pigments Are Localized Intracellularly in the Epidermis and Dermis of Fresh and Old Tattoos: In vivo Study Using Two-Photon Excited Fluorescence Lifetime Imaging.

BACKGROUND: The knowledge about the location and kinetics of tattoo pigments in human skin after application and during the recovery is restricted due to the limitation of in vivo methods for visualizing pigments. Here, the localization and distribution of tattoo ink pigments in freshly and old tattooed human skin during the regeneration of the epidermis and dermis were investigated in vivo. METHODS: Two-photon excited fluorescence lifetime imaging (TPE-FLIM) was used to identify tattoo ink pigments in human skin in vivo down to the reticular dermis. One subject with a freshly applied tattoo and 10 subjects with tattoos applied over 3 years ago were investigated in the epidermal and dermal layers in vivo. One histological slide of tattooed skin was used to localize skin-resident tattoo pigment using light microscopy. RESULTS: The carbon black particles deposited around the incision have still been visible 84 days after tattoo application, showing delayed recovery of the epidermis. The TPE-FLIM parameters of carbon black tattoo ink pigments were found to be different to all skin components except for melanin. Distinction from melanin in the skin was based on higher fluorescence intensity and agglomerate size. Using TPE-FLIM in vivo tattoo pigment was found in 75% of tattoos applied up to 9 years ago in the epidermis within keratinocytes, dendritic cells, and basal cells and in the dermis within the macrophages, mast cells, and fibroblasts. Loading of highly fluorescent carbon black particles enables in vivo imaging of dendritic cells in the epidermis and fibroblasts in the dermis, which cannot be visualized in native conditions. The collagen I structures showed a higher directionality similar to scar tissue resulting in a greater firmness and decreased elasticity of the tattooed skin. CONCLUSIONS: Here, we show the kinetics and location of carbon black tattoo ink pigment immediately after application for the first time in vivo in human skin. Carbon black particles are located exclusively intracellularly in the skin of fresh and old tattoos. They are found within macrophages, mast cells, and fibroblasts in the dermis and within keratinocytes, dendritic cells, and basal cells in the continuously renewed epidermis even in 9-year-old tattoos in skin showing no inflammation.