63 results on '"Johannes Schleusener"'
Search Results
2. Semantic modeling of cell damage prediction: a machine learning approach at human-level performance in dermatology
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Patrick Wagner, Maximilian Springenberg, Marius Kröger, Rose K. C. Moritz, Johannes Schleusener, Martina C. Meinke, and Jackie Ma
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Medicine ,Science - Abstract
Abstract Machine learning is transforming the field of histopathology. Especially in classification related tasks, there have been many successful applications of deep learning already. Yet, in tasks that rely on regression and many niche applications, the domain lacks cohesive procedures that are adapted to the learning processes of neural networks. In this work, we investigate cell damage in whole slide images of the epidermis. A common way for pathologists to annotate a score, characterizing the degree of damage for these samples, is the ratio between healthy and unhealthy nuclei. The annotation procedure of these scores, however, is expensive and prone to be noisy among pathologists. We propose a new measure of damage, that is the total area of damage, relative to the total area of the epidermis. In this work, we present results of regression and segmentation models, predicting both scores on a curated and public dataset. We have acquired the dataset in collaborative efforts with medical professionals. Our study resulted in a comprehensive evaluation of the proposed damage metrics in the epidermis, with recommendations, emphasizing practical relevance for real world applications.
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- 2023
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3. Topical Peroxisome Proliferator-Activated Receptor Agonist Induces Molecular Alterations Enhancing Barrier Function and Water-Holding Capacity of the Human Stratum Corneum In Vivo
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Maxim E. Darvin, Andrew Salazar, Johannes Schleusener, Jürgen Lademann, and Jörg von Hagen
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Raman microspectroscopy ,lipidomic analysis ,long-chain lipids ,ceramides ,triacylglycerols ,water mobility ,Chemistry ,QD1-999 - Abstract
The peroxisome proliferator-activated receptor (PPAR) ligands modulate a variety of skin functions but are rarely used in cosmetics. The aim of this double-blind, placebo-controlled, in vivo study was to determine the effect of a topically applied 0.1% PPAR ligand on the composition and physiological parameters of the stratum corneum (SC). By comparing verum and placebo groups post-treatment, we demonstrate (via lipidomic analysis of tape strips) an unstatistically significant trend toward an increase in long-chain triacylglycerols (C50–C56) and medium- and long-chain ceramides (C42–C50) at the superficial SC. By comparing treated and untreated skin using confocal Raman microspectroscopy, we found that the changes in lipid composition in the verum group led to a significant increase in the number of trans conformers and orthorhombic organisation of lipids at the exemplary SC depth. An increase in unfolded states in the secondary and tertiary keratin structures results in an increased ability to bind water. The concentrations of tightly and strongly bound water increase, while weakly bound and unbound water decrease in the entire SC, indicating a transformation of water mobility to a state of increased hydrogen bonding. Thus, the topical PPAR ligands improve the water-holding capacity and the barrier function of the SC.
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- 2024
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4. Application of 233 nm far-UVC LEDs for eradication of MRSA and MSSA and risk assessment on skin models
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Paula Zwicker, Johannes Schleusener, Silke B. Lohan, Loris Busch, Claudia Sicher, Sven Einfeldt, Michael Kneissl, Anja A. Kühl, Cornelia M. Keck, Christian Witzel, Axel Kramer, and Martina C. Meinke
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Medicine ,Science - Abstract
Abstract A newly developed UVC LED source with an emission wavelength of 233 nm was proved on bactericidal efficacy and skin tolerability. The bactericidal efficacy was qualitatively analysed using blood agar test. Subsequently, quantitative analyses were performed on germ carrier tests using the MRSA strain DSM11822, the MSSA strain DSM799, S. epidermidis DSM1798 with various soil loads. Additionally, the compatibility of the germicidal radiation doses on excised human skin and reconstructed human epidermis was proved. Cell viability, DNA damage and production of radicals were assessed in comparison to typical UVC radiation from discharge lamps (222 nm, 254 nm) and UVB (280–380 nm) radiation for clinical assessment. At a dose of 40 mJ/cm2, the 233 nm light source reduced the viable microorganisms by a log10 reduction (LR) of 5 log10 levels if no soil load was present. Mucin and protein containing soil loads diminished the effect to an LR of 1.5–3.3. A salt solution representing artificial sweat (pH 8.4) had only minor effects on the reduction. The viability of the skin models was not reduced and the DNA damage was far below the damage evoked by 0.1 UVB minimal erythema dose, which can be regarded as safe. Furthermore, the induced damage vanished after 24 h. Irradiation on four consecutive days also did not evoke DNA damage. The radical formation was far lower than 20 min outdoor visible light would cause, which is classified as low radical load and can be compensated by the antioxidant defence system.
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- 2022
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5. Atopic Dermatitis: Molecular Alterations between Lesional and Non-Lesional Skin Determined Noninvasively by In Vivo Confocal Raman Microspectroscopy
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Michael Zolotas, Johannes Schleusener, Jürgen Lademann, Martina C. Meinke, Georgios Kokolakis, and Maxim E. Darvin
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atopic eczema ,atopic dermatitis ,skin barrier function ,stratum corneum ,lipid ordering ,keratin structure ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Atopic dermatitis (AD)/atopic eczema is a chronic relapsing inflammatory skin disease affecting nearly 14% of the adult population. An important pathogenetic pillar in AD is the disrupted skin barrier function (SBF). The atopic stratum corneum (SC) has been examined using several methods, including Raman microspectroscopy, yet so far, there is no depth-dependent analysis over the entire SC thickness. Therefore, we recruited 21 AD patients (9 female, 12 male) and compared the lesional (LAS) with non-lesional atopic skin (nLAS) in vivo with confocal Raman microspectroscopy. Our results demonstrated decreased total intercellular lipid and carotenoid concentrations, as well as a shift towards decreased orthorhombic lateral lipid organisation in LAS. Further, we observed a lower concentration of natural moisturising factor (NMF) and a trend towards increased strongly bound and decreased weakly bound water in LAS. Finally, LAS showed an altered secondary and tertiary keratin structure, demonstrating a more folded keratin state than nLAS. The obtained results are discussed in comparison with healthy skin and yield detailed insights into the atopic SC structure. LAS clearly shows molecular alterations at certain SC depths compared with nLAS which imply a reduced SBF. A thorough understanding of these alterations provides useful information on the aetiology of AD and for the development/control of targeted topical therapies.
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- 2023
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6. Author Correction: Skin tolerant inactivation of multiresistant pathogens using far-UVC LEDs
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Johannes Glaab, Neysha Lobo‑Ploch, Hyun Kyong Cho, Thomas Filler, Heiko Gundlach, Martin Guttmann, Sylvia Hagedorn, Silke B. Lohan, Frank Mehnke, Johannes Schleusener, Claudia Sicher, Luca Sulmoni, Tim Wernicke, Lucas Wittenbecher, Ulrike Woggon, Paula Zwicker, Axel Kramer, Martina C. Meinke, Michael Kneissl, Markus Weyers, Ulrike Winterwerber, and Sven Einfeldt
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Medicine ,Science - Published
- 2022
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7. Label-free imaging of M1 and M2 macrophage phenotypes in the human dermis in vivo using two-photon excited FLIM
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Marius Kröger, Jörg Scheffel, Evgeny A Shirshin, Johannes Schleusener, Martina C Meinke, Jürgen Lademann, Marcus Maurer, and Maxim E Darvin
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skin ,dermis ,immune cells ,phagocytosis ,confocal microscopy ,non-invasive ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Macrophages (ΜΦs) are important immune effector cells that promote (M1 ΜΦs) or inhibit (M2 ΜΦs) inflammation and are involved in numerous physiological and pathogenic immune responses. Their precise role and relevance, however, are not fully understood for lack of noninvasive quantification methods. Here, we show that two-photon excited fluorescence lifetime imaging (TPE-FLIM), a label-free noninvasive method, can visualize ΜΦs in the human dermis in vivo. We demonstrate in vitro that human dermal ΜΦs exhibit specific TPE-FLIM properties that distinguish them from the main components of the extracellular matrix and other dermal cells. We visualized ΜΦs, their phenotypes and phagocytosis in the skin of healthy individuals in vivo using TPE-FLIM. Additionally, machine learning identified M1 and M2 MФs with a sensitivity of 0.88±0.04 and 0.82±0.03 and a specificity of 0.89±0.03 and 0.90±0.03, respectively. In clinical research, TPE-FLIM can advance the understanding of the role of MФs in health and disease.
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- 2022
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8. Skin tolerant inactivation of multiresistant pathogens using far-UVC LEDs
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Johannes Glaab, Neysha Lobo-Ploch, Hyun Kyong Cho, Thomas Filler, Heiko Gundlach, Martin Guttmann, Sylvia Hagedorn, Silke B. Lohan, Frank Mehnke, Johannes Schleusener, Claudia Sicher, Luca Sulmoni, Tim Wernicke, Lucas Wittenbecher, Ulrike Woggon, Paula Zwicker, Axel Kramer, Martina C. Meinke, Michael Kneissl, Markus Weyers, Ulrike Winterwerber, and Sven Einfeldt
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Medicine ,Science - Abstract
Abstract Multiresistant pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) cause serious postoperative infections. A skin tolerant far-UVC (
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- 2021
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9. Red- and Near-Infrared-Excited Autofluorescence as a Marker for Acute Oxidative Stress in Skin Exposed to Cigarette Smoke Ex Vivo and In Vivo
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Phuong Thao Tran, Parichat Tawornchat, Burkhard Kleuser, Silke B. Lohan, Johannes Schleusener, Martina C. Meinke, and Maxim E. Darvin
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cigarette smoke ,oxidative stress ,Raman spectroscopy ,NIR autofluorescence ,red autofluorescence ,skin fluorophores ,Therapeutics. Pharmacology ,RM1-950 - Abstract
Air pollution is increasing worldwide and skin is exposed to high levels of pollution daily, causing oxidative stress and other negative consequences. The methods used to determine oxidative stress in the skin are invasive and non-invasive label-free in vivo methods, which are severely limited. Here, a non-invasive and label-free method to determine the effect of cigarette smoke (CS) exposure on skin ex vivo (porcine) and in vivo (human) was established. The method is based on the measurement of significant CS-exposure-induced enhancement in red- and near-infrared (NIR)-excited autofluorescence (AF) intensities in the skin. To understand the origin of red- and NIR-excited skin AF, the skin was exposed to several doses of CS in a smoking chamber. UVA irradiation was used as a positive control of oxidative stress in the skin. The skin was measured with confocal Raman microspectroscopy before CS exposure, immediately after CS exposure, and after skin cleaning. CS exposure significantly increased the intensity of red- and NIR-excited skin AF in a dose-dependent manner in the epidermis, as confirmed by laser scanning microscopy AF imaging and fluorescence spectroscopy measurements. UVA irradiation enhanced the intensity of AF, but to a lower extent than CS exposure. We concluded that the increase in red- and NIR-excited AF intensities of the skin after CS exposure could clearly be related to the induction of oxidative stress in skin, where skin surface lipids are mainly oxidized.
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- 2023
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10. Characterization of Collagen I Fiber Thickness, Density, and Orientation in the Human Skin In Vivo Using Second-Harmonic Generation Imaging
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Marius Kröger, Johannes Schleusener, Sora Jung, and Maxim E. Darvin
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papillary dermis ,reticular dermis ,ageing ,collagen ,blood capillary ,immune cells ,Applied optics. Photonics ,TA1501-1820 - Abstract
The assessment of dermal alterations is necessary to monitor skin aging, cancer, and other skin diseases and alterations. The gold standard of morphologic diagnostics is still histopathology. Here, we proposed parameters to distinguish morphologically different collagen I structures in the extracellular matrix and to characterize varying collagen I structures in the skin with similar SAAID (SHG-to-AF Aging Index of Dermis, SHG—second-harmonic generation; AF—autofluorescence) values. Test datasets for the papillary and reticular extracellular matrix from images in 24 female subjects, 36 to 50 years of age, were generated. Parameters for SAAID, edge detection, and fast Fourier transformation directionality were determined. Additionally, textural analyses based on the grey level co-occurrence matrix (GLCM) were conducted. At first, changes in the GLCM parameters were determined in the native greyscale images and, furthermore, in the Hilbert-transformed images. Our results demonstrate a robust set of parameters for noninvasive in vivo classification for morphologically different collagen I structures in the skin, with similar and different SAAID values. We anticipate our method to enable an automated prevention and monitoring system with an age- and gender-specific algorithm.
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- 2021
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11. Retaining Skin Barrier Function Properties of the Stratum Corneum with Components of the Natural Moisturizing Factor—A Randomized, Placebo-Controlled Double-Blind In Vivo Study
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Johannes Schleusener, Andrew Salazar, Jörg von Hagen, Jürgen Lademann, and Maxim E. Darvin
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confocal Raman microspectroscopy ,skin hydration ,water binding ,lipid organization ,lamellae ,Organic chemistry ,QD241-441 - Abstract
The influence of a topically applied formulation containing components of natural moisturizing factor (NMF) on barrier-related parameters of the stratum corneum (SC) was investigated in vivo using confocal Raman microspectroscopy in a randomized, placebo-controlled double-blind study on 12 volunteers for 14 days. This method allowed for the elucidation of subtle differences between the verum and the placebo even though the components of the verum naturally occur in the SC. This differentiation is not possible non-invasively by conventional methods. In this study, we found that the applied verum and placebo formulations disrupted the equilibrium of water, NMF and lipids in the SC. The adverse effects of the formulation could be mitigated by incorporating it into a simplified supplementation of NMF molecules. As a long-term effect, the amount of strongly bound water increases at 30–40% SC depth (p < 0.05) and the amount of weakly bound water decreases at 30–40% SC depth (p < 0.05) for the verum. This supplement was also unexpectedly able to prevent intercellular lipids (ICL) disorganization in selected depths. In the long term, the verum treatment limited the lateral disorganization of the ICL to the upper 20% SC depth. Further research is required to elucidate the interplay of these factors in the SC, to better understand their contribution to the equilibrium and barrier function of the skin. This understanding of the interaction of these naturally occurring components could help in the future to develop and optimize topical treatments for diseases like psoriasis, atopic dermatitis, ichthyosis where the skin barrier is disrupted.
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- 2021
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12. Oxidative stress coping capacity (OSC) value: Development and validation of an in vitro measurement method for blood plasma using electron paramagnetic resonance spectroscopy (EPR) and vitamin C
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Pauline Nibbe, Johannes Schleusener, Silas Siebert, Richard Borgart, Doreen Brandt, Ronja Westphalen, Nadine Schüler, Bastian Berger, Eva M.J. Peters, Martina C. Meinke, and Silke B. Lohan
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Oxidative Stress ,Plasma ,Physiology (medical) ,Electron Spin Resonance Spectroscopy ,Humans ,Pilot Projects ,Ascorbic Acid ,Vitamins ,Oxidation-Reduction ,Biochemistry ,Antioxidants - Abstract
Oxidative stress as a driver of disease is reinforcing the trend towards supplementation with antioxidants. While antioxidants positively influence the redox status when applied at physiological doses, higher concentrations may have pro-oxidative effects. Precise assessment methods for testing the supply of antioxidants are lacking. Using in-situ-irradiation as stressor and electron paramagnetic resonance (EPR) spectroscopy as readout system for formed radicals, a stress response assessment method was developed, using protein solutions and plasma samples from transfusion medicine. The method was validated in a double-blind placebo-controlled in vivo cross-over pilot study in blood plasma samples of individuals before and after vitamin C supplementation. Reference measurements were performed for the exogenous antioxidants β-carotene and vitamin C, and glutathione as an endogenous representative. Malondialdehyde was studied for oxidative stress indication. Protein solutions without antioxidants showed a linear increase in radical concentration during irradiation. The in-vitro-addition of vitamin C or plasma samples from subjects displayed two slopes (m
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- 2023
13. Treatment of the Candida subspecies Candida albicans and Candida parapsilosis with two far‐UVC sources to minimise mycoses in clinical practice
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Johannes Schleusener, Silke B. Lohan, Loris Busch, Kamran Ghoreschi, Neysha Lobo Ploch, Stefanie May, Simone Vogel, Jürgen Eberle, and Martina C. Meinke
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Candida parapsilosis ,Antifungal Agents ,Infectious Diseases ,Mycoses ,Ultraviolet Rays ,Candida albicans ,Humans ,Dermatology ,General Medicine - Abstract
Fungal infections have increased considerably over the last decades, becoming progressively resistant to common drugs. UVC light has shown microbiological eradication effects, whereby the wavelength of 254 nm is strongly carcino- and mutagenic. Therefore, 222 and 233 nm, which do not significantly harm skin cells, were tested for their antifungal effects. Microbicidal doses were reached at 40 mJ/cm
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- 2022
14. Cosmetic Formulations with Melaleuca alternifolia Essential Oil for the Improvement of Photoaged Skin: A <scp>Double‐Blind</scp> , Randomized, <scp>Placebo‐Controlled</scp> Clinical Study
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Victor Hugo Infante, Patricia Maria Maia Campos, Maxim Darvin, Silke Lohan, Johannes Schleusener, Sabine Schanzer, Juergen Lademann, and Martina Meinke
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General Medicine ,Physical and Theoretical Chemistry ,Biochemistry - Published
- 2022
15. tMCR‐ALS method for the determination of water concentration profiles in the stratum corneum of untreated and treated skin in vivo
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Chun Sik Choe, Jin Song Ri, Se Hyok Choe, Pok Sil Kim, Jürgen Lademann, Johannes Schleusener, and Maxim E. Darvin
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General Materials Science ,Spectroscopy - Published
- 2022
16. Tattoo pigments are localized intracellularly in the epidermis and dermis of fresh and old tattoos – in vivo study using two-photon excited FLIM
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Marius Kröger, Johannes Schleusener, Jürgen Lademann, Martina C. Meinke, Sora Jung, and Maxim E. Darvin
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Dermatology - Abstract
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.
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- 2023
17. Quantitative determination of concentration profiles of skin components and topically applied oils by tailored multivariate curve resolution‐alternating least squares using in vivo confocal Raman micro‐spectroscopy
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ChunSik Choe, Johannes Schleusener, JinSong Ri, SeHyok Choe, PokSil Kim, Jürgen Lademann, and Maxim E. Darvin
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General Engineering ,General Physics and Astronomy ,General Materials Science ,General Chemistry ,General Biochemistry, Genetics and Molecular Biology - Abstract
The main components of the stratum corneum (SC), water, lipids, and proteins, are non-homogeneously distributed throughout the depth. The quantitative determination of their concentration profiles and penetration depth of topically applied substances are urgent topics of dermatological and cosmetic research. Confocal Raman micro-spectroscopy has distinct advantages when determining semi-quantitative concentrations of SC components and topically applied substances non-invasively and in vivo. In this work, we applied a tailored multivariate curve resolution-alternating least squares (tMCR-ALS) method to analyze Raman spectra of the SC in the 2000-4000 cm
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- 2022
18. Skin tolerant inactivation of multiresistant pathogens using far-UVC LEDs
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Michael Kneissl, Tim Wernicke, Johannes Schleusener, Martin Guttmann, Martina C. Meinke, Ulrike Winterwerber, Lucas Wittenbecher, Paula Zwicker, Luca Sulmoni, Markus Weyers, Silke B. Lohan, Thomas Filler, Ulrike Woggon, Neysha Lobo-Ploch, Axel Kramer, Claudia Sicher, Johannes Glaab, Hyun Kyong Cho, Frank Mehnke, Sven Einfeldt, Heiko Gundlach, and Sylvia Hagedorn
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Methicillin-Resistant Staphylococcus aureus ,Disease prevention ,Swine ,Ultraviolet Rays ,DNA damage ,Disinfectant ,Science ,02 engineering and technology ,medicine.disease_cause ,Radiation Tolerance ,01 natural sciences ,Article ,Microbiology ,Agar plate ,Postoperative Complications ,Skin Physiological Phenomena ,0103 physical sciences ,medicine ,Animals ,Porcine skin ,Irradiation ,Skin ,010302 applied physics ,Cross Infection ,Microbial Viability ,Multidisciplinary ,biology ,Chemistry ,500 Naturwissenschaften und Mathematik::530 Physik::530 Physik ,021001 nanoscience & nanotechnology ,biology.organism_classification ,Drug Resistance, Multiple ,Disinfection ,Clinical Practice ,Staphylococcus aureus ,Inorganic LEDs ,Infectious diseases ,Medicine ,0210 nano-technology ,Biomedical engineering ,Bacteria ,DNA Damage - Abstract
Multiresistant pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) cause serious postoperative infections. A skin tolerant far-UVC (2. MRSA bacteria in different concentrations on blood agar plates were inactivated with irradiation doses in the range of 15–40 mJ/cm2. Porcine skin irradiated with a dose of 40 mJ/cm2 at 233 nm showed only 3.7% CPD and 2.3% 6-4PP DNA damage. Corresponding irradiation at 254 nm caused 11–14 times higher damage. Thus, the skin damage caused by the disinfectant doses is so small that it can be expected to be compensated by the skin's natural repair mechanisms. LED-based far-UVC lamps could therefore soon be used in everyday clinical practice to eradicate multiresistant pathogens directly on humans.
- Published
- 2021
19. Characterization of radical types, penetration profile and distribution pattern of the topically applied photosensitizer THPTS in porcine skin ex vivo
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I. Streit, Juergen Lademann, Johannes Schleusener, Marius Kröger, Silke B. Lohan, Martina C. Meinke, and Maxim E. Darvin
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Fluorescence-lifetime imaging microscopy ,Porphyrins ,Time Factors ,Infrared Rays ,Swine ,Radical ,medicine.medical_treatment ,Pharmaceutical Science ,Photodynamic therapy ,02 engineering and technology ,Administration, Cutaneous ,030226 pharmacology & pharmacy ,03 medical and health sciences ,Spatio-Temporal Analysis ,0302 clinical medicine ,Stratum corneum ,medicine ,Animals ,Tissue Distribution ,Photosensitizer ,Irradiation ,Penetration depth ,Skin ,Photosensitizing Agents ,Chemistry ,General Medicine ,Penetration (firestop) ,021001 nanoscience & nanotechnology ,medicine.anatomical_structure ,Photochemotherapy ,Biophysics ,Reactive Oxygen Species ,0210 nano-technology ,Biotechnology - Abstract
The topical photodynamic therapy (PDT) is mainly used in the treatment of dermato-oncological diseases. The distribution and functionality of the photosensitizer Tetrahydroporphyrin-Tetratosylat (THPTS) was investigated using microscopic and spectroscopic methods after topical application to excised porcine skin followed by irradiation. The distribution of THPTS was determined by two-photon tomography combined with fluorescence lifetime imaging (TPT/FLIM) and confocal Raman microspectroscopy (CRM). The radicals were quantified and characterized by electron paramagnetic resonance (EPR) spectroscopy. Results show a penetration depth of THPTS into the skin down to around 12 ± 5 µm. A penetration of THPTS through the stratum corneum was not clearly observable after 1 h penetration time, but cannot be excluded. The irradiation within the phototherapeutic window (spectral range of visible and near infrared light in the range ≈ 650–850 nm) is needed to activate THPTS. An incubation time of 10 min showed the highest radical production. A longer incubation time affected the functionality of THPTS, whereby significant less radicals were detectable. During PDT mainly reactive oxygen species (ROS) and lipid oxygen species (LOS) are produced. Overall, the irradiation dose per se influences the radical types formed in skin. While ROS are always prominent at low doses, LOS increase at high doses, independent of previous skin treatment and the irradiation wavelength used.
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- 2021
20. Blind source separation of molecular components of the human skin in vivo: non-negative matrix factorization of Raman microspectroscopy data
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Johannes Schleusener, Maxim E. Darvin, A. V. Venets, Boris P. Yakimov, Juergen Lademann, Evgeny A. Shirshin, and Victor V. Fadeev
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Materials science ,Confocal ,Human skin ,Spectrum Analysis, Raman ,Biochemistry ,Blind signal separation ,Spectral line ,Analytical Chemistry ,Non-negative matrix factorization ,Matrix decomposition ,030207 dermatology & venereal diseases ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,In vivo ,Electrochemistry ,Humans ,Environmental Chemistry ,Spectroscopy ,Skin ,030304 developmental biology ,0303 health sciences ,symbols ,Epidermis ,Raman spectroscopy ,Biological system ,Algorithms - Abstract
Determination of the molecular composition of the skin is crucial for numerous tasks in medicine, pharmacology, dermatology and cosmetology. Confocal Raman microspectroscopy is a sensitive method for the evaluation of molecular depth profiles in the skin in vivo. Since the Raman spectra of most of the skin constituents significantly superimpose, a spectral decomposition by a set of predefined library components is usually performed to disentangle their contributions. However, the incorrect choice of the number and type of components or differences between the spectra of the basic components measured in vitro and in vivo can lead to incorrect results of the decomposition procedure. Here, we investigate an alternative data-driven approach based on a non-negative matrix factorization (NNMF) algorithm of depth-resolved Raman spectra of skin that does not require a priori information of spectral data for the analysis. Using the model and experimentally measured depth-resolved Raman spectra of the upper epidermis in vivo, we show that NNMF provides depth profiles of endogenous molecular components and exogenous agents penetrating through the upper epidermis for the spectra and concentration. Moreover, we demonstrate that this approach is capable of providing new information on the molecular profiles of the skin.
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- 2021
21. [YIA] The influence of skin barrier disruption and melanin content on the formation of DNA lesions and radicals in ex vivo human skin induced by 233 nm far-UVC irradiation from LEDs
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Loris Busch, Johannes Schleusener, Daniela F. Zamudio Díaz, Marius Kröger, Silke B. Lohan, Paula Zwicker, Einfeldt Sven, Michael Kneissl, Anja A. Kühl, Christian Witzel, Holger Klose, Cornelia M. Keck, Axel Kramer, and Martina C. Meinke
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Physiology (medical) ,Biochemistry - Published
- 2022
22. Lycopene, but not zeaxanthin, serves as a skeleton for the formation of an orthorhombic organization of intercellular lipids within the lamellae in the stratum corneum: Molecular dynamics simulations of the hydrated ceramide NS bilayer model
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Jin-Song Ri, Chun-Sik Choe, Se-Hyok Choe, Kwang-Hyok Jong, Song-Nam Hong, Johannes Schleusener, Juergen Lademann, and Maxim E. Darvin
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Lycopene ,Zeaxanthins ,Lipid Bilayers ,Biophysics ,Cell Biology ,Molecular Dynamics Simulation ,Ceramides ,Biochemistry ,Carotenoids ,Skeleton - Abstract
Carotenoids play an important role in the protection of biomembranes against oxidative damage. Their function depends on the surroundings and the organization of the lipid membrane they are embedded in. Carotenoids are located parallel or perpendicular to the surface of the lipid bilayer. The influence of carotenoids on the organization of the lipid bilayer in the stratum corneum has not been thoroughly considered. Here, the orientation of the exemplary cutaneous carotenoids lycopene and zeaxanthin in a hydrated ceramide NS24 bilayer model and the influence of carotenoids on the lateral organization of the lipid bilayer model were studied by means of molecular dynamics simulations for 32 °C and 37 °C. The results confirm that lycopene is located parallel and zeaxanthin perpendicular to the surface of the lipid bilayer. The lycopene-loaded lipid bilayer appeared to have a strong orthorhombic organization, while zeaxanthin-loaded and pure lipid bilayers were organized in a disordered hexagonal-like and liquid-like state, respectively. The effect is stronger at 32 °C compared to 37 °C based on p-values. Therefore, it was assumed that carotenoids without hydroxyl polar groups in their structure facilitate the formation of the orthorhombic organization of lipids, which provides the skin barrier function. It was shown that the distance between carotenoid atoms matched the distance between atoms in the lipids, indicating that parallel located carotenoids without hydroxyl groups serve as a skeleton for lipid membranes inside the lamellae. The obtained results provide reasonable prediction of the overall qualitative properties of lipid model systems and show the importance of parallel-oriented carotenoids in the development and maintenance of the skin barrier function.
- Published
- 2022
23. Cosmetic Formulations with Melaleuca alternifolia Essential Oil for the Improvement of Photoaged Skin: A Double-Blind, Randomized, Placebo-Controlled Clinical Study
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Victor, Hugo Infante, Patricia, Maria Maia Campos, Maxim, Darvin, Silke, Lohan, Johannes, Schleusener, Sabine, Schanzer, Juergen, Lademann, and Martina, Meinke
- Abstract
This aim of this study was to evaluate the penetration depth, antioxidant capacity and the clinical efficacy of Melaleuca alternifolia pure essential oil and in a nanoemulsion to prevent skin photoaging. For this, 2% of pure essential oil or 2% of this essential oil in a nanoemulsion were vehiculated in a formulation. The skin penetration was evaluated using confocal Raman microspectroscopy. The radical protection factor was evaluated using electron paramagnetic resonance spectroscopy. For a clinical study, 40 male participants, aged 18-28 years, were enrolled, being divided into three groups: vehicle formulation, M. alternifolia pure essential oil and M. alternifolia Nanoemulsion. All the participants also received a sunscreen SPF 50 to use during the day. Before and after 90 days of study, skin hydrolipidics and morphological characteristics were performed by skin imaging and biophysical techniques. The nanoemulsion presented a lower antioxidant capacity and a higher penetration through the stratum corneum, reaching the viable epidermis, improving the stratum granulosum morphology. The groups presented an increase in the papillary depth, improving in the dermis echogenicity and the collagen fibers. Melaleuca alternifolia essential provides the potential to improve photoaged skin, being the application of nanoemulsion able to reach deeper skin layers.
- Published
- 2022
24. The Effectiveness of Glycerol Solutions for Optical Clearing of the Intact Skin as Measured by Confocal Raman Microspectroscopy
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Maxim E. Darvin, I. Yu. Yanina, V. V. Tuchin, Johannes Schleusener, and Juergen Lademann
- Subjects
010302 applied physics ,Chromatography ,integumentary system ,Confocal ,Laser ,01 natural sciences ,eye diseases ,Atomic and Molecular Physics, and Optics ,Electronic, Optical and Magnetic Materials ,law.invention ,010309 optics ,symbols.namesake ,chemistry.chemical_compound ,medicine.anatomical_structure ,Distilled water ,chemistry ,law ,0103 physical sciences ,Microscopy ,symbols ,Stratum corneum ,medicine ,Glycerol ,Raman spectroscopy ,Penetration depth - Abstract
The effect of glycerol solutions of different concentrations and exposure times on the optical clearing efficiency in intact pig skin has been studied in the analysis of Raman spectra recorded at different depths. It has been found that a solution of 80% glycerol and 20% dimethylsulfoxide penetrated through the stratum corneum during 45 min. The increase in optical clearing has been achieved by using mixtures of optical clearing agents with dimethylsulfoxide and distilled water. Thus, the greatest optical clearing efficiency was observed when using a mixture of 60% glycerol and 40% water for 45 min (an increase of 3.4 times at a depth of 80 µm). Thus, it has been shown that it is possible to control the skin optical parameters at a depth of about 80 µm using the method of confocal Raman microscopy.
- Published
- 2020
25. Author response: Label-free imaging of M1 and M2 macrophage phenotypes in the human dermis in vivo using two-photon excited FLIM
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Marius Kröger, Jörg Scheffel, Evgeny A Shirshin, Johannes Schleusener, Martina C Meinke, Jürgen Lademann, Marcus Maurer, and Maxim E Darvin
- Published
- 2021
26. Water migration at skin optical clearing
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Anton Yu. Sdobnov, Johannes Schleusener, Jürgen Lademann, Valery V. Tuchin, and Maxim E. Darvin
- Published
- 2021
27. Safety and efficacy of combined essential oils for the skin barrier properties: In vitro, ex vivo and clinical studies
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Victor H. P. Infante, Patrícia M. B. G. Maia Campos, Lorena Rigo Gaspar, Maxim E. Darvin, Johannes Schleusener, Karen C. Rangel, Martina C. Meinke, and Jürgen Lademann
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Aging ,Colloid and Surface Chemistry ,Chemistry (miscellaneous) ,Swine ,Skin Absorption ,Drug Discovery ,Oils, Volatile ,Pharmaceutical Science ,Animals ,Plant Oils ,Dermatology ,Epidermis ,Skin - Abstract
To evaluate the safety and the synergistic effects of tea tree, lavender, eucalyptus and tangerine essential oils in combination on the skin using in vitro, ex vivo and clinical studies.The phototoxicity was predicted using 3T3 neutral red uptake phototoxicity test (OECD TG 432). Skin penetration was evaluated by confocal Raman microspectroscopy using direct application of essential oils to pig ears. For the clinical studies, 40 participants were enrolled and randomized in three groups: (1) lavender, eucalyptus and tangerine, (2) the same essential oils plus melaleuca and (3) placebo group. The skin was evaluated by noninvasive techniques before and after a 90-day period of topical use.The essential oils were non-phototoxic, but the tangerine oil showed dose-dependent cytotoxicity (IC50: 33.1 µg/ml), presenting 35% of penetration in the viable epidermis. On the contrary, 17.7 µg/ml in combination was applied per day in the clinical study and the penetration rate for the combinations (10%, 1.77 µg/ml achieving the viable epidermis) guaranteed the safety, since in the clinical study, the application of the four essential oils improved skin barrier and morphologic skin characteristics, as well as increased skin hydration and decreased sebum levels, with no unwanted effects reported.All essential oils studied were considered non-cytotoxic or non-phototoxic separately except tangerine, which present a dose-dependent cytotoxicity. Finally, the essential oils in combination in an appropriate amount were safe and effective in the improvement of the hydrolipidic balance and morphological properties of the skin.évaluer la sécurité d’emploi et les effets synergiques des associations d’huiles essentielles d’arbre à thé, de lavande, d’eucalyptus et de mandarine sur la peau à l’aide d’études in vitro, ex vivo et cliniques. MÉTHODES: la phototoxicité a été prédite avec le test de phototoxicité de fixation du rouge neutre 3T3 (OCDE TG 432). La pénétration cutanée a été évaluée par microspectroscopie confocale de Raman grâce à l’application directe d’huiles essentielles sur les oreilles de cochons. Pour les études cliniques, 40 participants ont été inclus et randomisés dans trois groupes : (1) lavande, eucalyptus et mandarine, (2) les mêmes huiles essentielles plus melaleuca et (3) un groupe placebo. La peau a été évaluée par des techniques non invasives avant et après une période d’utilisation topique de 90 jours. RÉSULTATS: les huiles essentielles se sont avérées non phototoxiques, mais l’huile de mandarine a montré une cytotoxicité dose-dépendante (CI 50 : 33,1 µg/ml), représentant 35 % de pénétration dans l’épiderme viable. À l’inverse, dans l’étude clinique, une quantité de 17,7 µg/ml par jour en association a été appliquée, et le taux de pénétration des associations (10 %, soit 1,77 µg/ml atteignant l’épiderme viable) a garanti la sécurité d’emploi, puisque dans l’étude clinique, l’application des quatre huiles essentielles a amélioré la barrière cutanée et les caractéristiques morphologiques de la peau, et a entraîné une augmentation de l’hydratation cutanée et une diminution des taux de sébum, sans signalement d’effets indésirables.chacune des huiles essentielles étudiées a été considérée comme non cytotoxique ou non phototoxique, à l’exception de la mandarine, qui présente une cytotoxicité dose-dépendante. Enfin, l’association d’huiles essentielles en quantité appropriée a démontré sa sécurité d’emploi et son efficacité dans l’amélioration de l’équilibre hydrolipidique et des propriétés morphologiques de la peau.
- Published
- 2021
28. Label-free imaging of macrophage phenotypes and phagocytic activity in the human dermisin vivousing two-photon excited FLIM
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Marcus Maurer, Martina C. Meinke, Jörg Scheffel, Johannes Schleusener, Maxim E. Darvin, Marius Kröger, Evgeny A. Shirshin, and Jürgen Lademann
- Subjects
Fluorescence-lifetime imaging microscopy ,medicine.anatomical_structure ,Two-photon excitation microscopy ,Dermis ,In vivo ,Chemistry ,Phagocytosis ,medicine ,Biophysics ,Macrophage ,Sensitivity (control systems) ,In vitro - Abstract
Macrophages (MΦs) are important immune effector cells that promote (M1 MΦs) or inhibit (M2 MΦs) inflammation and are involved in numerous physiological and pathogenic immune responses. Their precise role and relevance, however, is not fully understood because of the lack of non-invasive quantification methods. Here, we show that two-photon excited fluorescence lifetime imaging (TPE-FLIM), a label-free non-invasive method, can visualize MΦs in human dermisin vivo. We demonstratein vitrothat human dermal MΦs exhibit specific TPE-FLIM properties that distinguish them from the main components of the extracellular matrix and other dermal cells. We visualized MΦs, their phenotypes and phagocytosis in the skin of healthy individualsin vivousing TPE-FLIM. Additionally, machine learning identified M1 and M2 MΦs with a sensitivity of 0.88±0.04 and 0.82±0.03 and a specificity of 0.89±0.03 and 0.90±0.03, respectively. In clinical research, TPE-FLIM can advance the understanding of the role of MΦs in health and disease.
- Published
- 2021
29. Characterization of Collagen I Fiber Thickness, Density, and Orientation in the Human Skin In Vivo Using Second-Harmonic Generation Imaging
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Johannes Schleusener, Sora Jung, Marius Kröger, and Maxim E. Darvin
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collagen ,Materials science ,animal structures ,two-photon tomography ,Human skin ,Matrix (biology) ,blood capillary ,Skin Aging ,Extracellular matrix ,immune cells ,Dermis ,medicine ,Radiology, Nuclear Medicine and imaging ,Applied optics. Photonics ,Instrumentation ,reticular dermis ,integumentary system ,Papillary dermis ,Atomic and Molecular Physics, and Optics ,TA1501-1820 ,medicine.anatomical_structure ,ageing ,Reticular connective tissue ,papillary dermis ,intravital imaging ,Reticular Dermis ,Biomedical engineering - Abstract
The assessment of dermal alterations is necessary to monitor skin aging, cancer, and other skin diseases and alterations. The gold standard of morphologic diagnostics is still histopathology. Here, we proposed parameters to distinguish morphologically different collagen I structures in the extracellular matrix and to characterize varying collagen I structures in the skin with similar SAAID (SHG-to-AF Aging Index of Dermis, SHG—second-harmonic generation, AF—autofluorescence) values. Test datasets for the papillary and reticular extracellular matrix from images in 24 female subjects, 35 to 60 years of age, were generated. Parameters for SAAID, edge detection, and fast Fourier transformation directionality were determined. Additionally, textural analyses based on the grey level co-occurrence matrix (GLCM) were conducted. At first, changes in the GLCM parameters were determined in the native greyscale images and, furthermore, in the Hilbert-transformed images. Our results demonstrate a robust set of parameters for noninvasive in vivo classification for morphologically different collagen I structures in the skin, with similar and different SAAID values. We anticipate our method to enable an automated prevention and monitoring system with an age- and gender-specific algorithm.
- Published
- 2021
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30. In vivo Tracking of DNA for Precise Determination of the Stratum Corneum Thickness and Superficial Microbiome Using Confocal Raman Microscopy
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Maxim E. Darvin, Johannes Schleusener, Jürgen Lademann, Jin Song Ri, Chun Sik Choe, and Se Hyok Choe
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Adult ,Male ,Keratinocytes ,0301 basic medicine ,Materials science ,Physiology ,Confocal ,Nucleoid ,Dermatology ,Young Adult ,030207 dermatology & venereal diseases ,03 medical and health sciences ,symbols.namesake ,chemistry.chemical_compound ,0302 clinical medicine ,In vivo ,Microscopy ,medicine ,Stratum corneum ,Humans ,Corneocytes ,Pharmacology ,Microscopy, Confocal ,Corneocyte ,Bacteria ,integumentary system ,Microbiota ,DNA ,General Medicine ,Middle Aged ,Lipids ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Skin barrier function ,symbols ,Female ,Epidermis ,Cell maturation ,Cell nucleus ,Raman spectroscopy ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Biomedical engineering - Abstract
The skin barrier function is mostly provided by the stratum corneum (SC), the uppermost layer of the epidermis. To noninvasively analyze the physiological properties of the skin barrier functionin vivo, it is important to determine the SC thickness. Confocal Raman microscopy (CRM) is widely used for this task. In the present in vivo study, a new method based on the determination of the DNA concentration profile using CRM is introduced for determining the SC thickness. The obtained SC thickness values are compared with those obtained using other CRM-based methods determining the water and lipid depth profiles. The obtained results show almost no significant differences in SC thickness for the utilized methods. Therefore, the results indicate that it is possible to calculate the SC thickness by using the DNA profile in the fingerprint region, which is comparable with the SC thickness calculated by the water depth profiles (ANOVA test p = 0.77) and the lipid depth profile (ANOVA test p = 0.74). This provides the possibility to measure the SC thickness by using the DNA profile, in case the water or lipid profile analyses are influenced by a topically applied formulation. The increase in DNA concentration in the superficial SC (0–2 µm) is related to the DNA presence in the microbiome of the skin, which was not present in the SC depth below 4 µm.
- Published
- 2019
31. The non‐homogenous distribution and aggregation of carotenoids in the stratum corneum correlates with the organization of intercellular lipids in vivo
- Author
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Maxim E. Darvin, Johannes Schleusener, JaeRyong Ri, Juergen Lademann, and ChunSik Choe
- Subjects
Male ,0301 basic medicine ,Sebum secretion ,Radical ,Stratum granulosum ,macromolecular substances ,Dermatology ,Spectrum Analysis, Raman ,Biochemistry ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,polycyclic compounds ,Stratum corneum ,medicine ,Humans ,Molecular Biology ,Carotenoid ,Aged ,chemistry.chemical_classification ,Microscopy, Confocal ,Chemistry ,organic chemicals ,food and beverages ,Penetration (firestop) ,Middle Aged ,Carotenoids ,Healthy Volunteers ,biological factors ,030104 developmental biology ,medicine.anatomical_structure ,Biophysics ,Female ,Epidermis ,Intracellular - Abstract
The human stratum corneum (SC) contains an abundant amount of carotenoid antioxidants, quenching free radicals and thereby protecting the skin. For the precise measurements of the depth-dependent carotenoid concentration, confocal Raman microscopy is a suitable method. The quantitative concentration can be determined by the carotenoid-related peak intensity of a Gaussian function approached at ≈1524 cm-1 using non-linear regression. Results show that the carotenoid concentration is higher at the superficial layers of the SC then decreases to a minimum at 20% SC depth and increases again towards the bottom of the SC. In the present work, two carotenoid penetration pathways into the SC are postulated. The first pathway is from the stratum granulosum to the bottom of the SC, while in the second pathway, the carotenoids are delivered to the skin surface by sweat and/or sebum secretion and penetrate from outside. The carotenoids are aggregated at the superficial layers, which are shown by high correlation between the aggregation states of carotenoids and the lateral organization of lipids. At the 30%-40% SC depths, the ordered and dense lipid molecules intensify the lipid-carotenoid interactions and weaken the carotenoid-carotenoid interaction and thus exhibit the disaggregation of carotenoids. At 90%-100% SC depths, the carotenoid-lipid interaction is weakened and the carotenoids have a tendency to be aggregated. Thus, the molecular structural correlation of carotenoid and SC lipid might be reserved in the intercellular space of the SC and also serves as the skeleton of the intercellular lipids.
- Published
- 2019
32. Influence of polyester spacer fabric, cotton, chloroprene rubber, and silicone on microclimatic and morphologic physiologic skin parameters in vivo
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Johannes Schleusener, Fanny Knorr, Gisela Thiede, Heike Richter, Jürgen Lademann, Sabine Schanzer, Maxim E. Darvin, Simon Gallinger, Marc Kraft, Sora Jung, and Ulrich Wegener
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Adult ,Neoprene ,Materials science ,Polyesters ,Silicones ,Dermatology ,01 natural sciences ,010309 optics ,030207 dermatology & venereal diseases ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Silicone ,Body Water ,Natural rubber ,In vivo ,Skin Physiological Phenomena ,0103 physical sciences ,Stratum corneum ,medicine ,Humans ,Cotton Fiber ,Skin ,Transepidermal water loss ,Moisture ,Chloroprene ,technology, industry, and agriculture ,Microclimate ,Middle Aged ,Water Loss, Insensible ,Healthy Volunteers ,Polyester ,medicine.anatomical_structure ,Epidermal Cells ,chemistry ,visual_art ,visual_art.visual_art_medium ,Skin Temperature ,Biomedical engineering - Abstract
Background Skin diseases can develop upon disadvantageous microclimate in relation to skin contact with textiles of supporting devices. Increased temperature, moisture, mechanical fracture, pressure, and inflammatory processes often occur mutually and enhance each other in their adverse effects. Therefore, the early prevention of skin irritations by improvement of microclimatic properties of skin in contact with supporting devices is important. Materials and methods In this study, the microclimate under occlusion with polyester, cotton, chloroprene rubber, and silicone textiles, used for supporting devices, was analyzed by determining several characteristic physiologic skin parameters in vivo, including temperature, moisture, and transepidermal water loss (TEWL). This is achieved by comparing a miniaturized in vivo detection device with several established optical and sensory methods in vivo. Results A highly significant TEWL decrease was found after polyester, chloroprene rubber, and silicone application. The application of all materials showed highly significant decrease in skin surface temperature, with chloroprene rubber showing the lowest. Similarly, all materials showed highly significant increase in relative moisture, where the highest increase was found for chloroprene rubber and silicone and the lowest increase for cotton. The cutaneous carotenoid concentration of chloroprene rubber, silicone, and polyester decreased. A manipulation of the surface structure of the stratum corneum was recognized for all materials except for cotton by laser scanning microscopy. Conclusion The skin parameters temperature, relative moisture, antioxidant status, and TEWL can effectively characterize the microclimatic environment during occlusion with medical supporting materials. These parameters could potentially be used to develop standardized testing procedures for material evaluation.
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- 2019
33. Confocal Raman imaging of skin sections containing hair follicles using classical least squares regression and multivariate curve resolution–alternating least squares
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S. Guo, V. Carrer, A. Patzelt, Juergen Lademann, Maxim E. Darvin, T. Bocklitz, Johannes Schleusener, and L. Coderch
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Multivariate curve resolution ,Multivariate analysis ,Materials science ,Confocal ,Raman imaging ,Hyperspectral imaging ,Statistical and Nonlinear Physics ,Atomic and Molecular Physics, and Optics ,Regression ,Classical least squares ,Electronic, Optical and Magnetic Materials ,Alternating least squares ,Electrical and Electronic Engineering ,Biomedical engineering - Published
- 2019
34. Bedeutung des follikulären Penetrationswegs für den Wirkstofftransport mittels Nanocarriern
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Juergen Lademann, P Breuckmann, Alexa Patzelt, Martina C. Meinke, Johannes Schleusener, Heike Richter, Maxim E. Darvin, Sabine Schanzer, and V. Carrer
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Gynecology ,medicine.medical_specialty ,business.industry ,Dermatology ,Penetration (firestop) ,Drug application ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,030220 oncology & carcinogenesis ,medicine ,Triggered release ,Nanocarriers ,business - Abstract
Der Haarfollikel stellt einen bedeutenden Penetrationsweg fur topisch applizierte Substanzen dar. Die perkutane Absorption von bestimmten Substanzen kann durch die Beteiligung der Haarfollikel deutlich gesteigert und beschleunigt werden. Ferner haben Nanopartikel die Eigenschaft, besonders tief und effektiv in die Haarfollikel hinein zu penetrieren. Dies bietet die Moglichkeit, den Wirkstofftransport fur topisch applizierte Substanzen zu optimieren, indem die Nanocarrier allein als Transporter fur die Wirkstoffe in den Haarfollikel fungieren. Innerhalb des Haarfollikels muss nach erfolgter Penetration eine Freisetzung des Wirkstoffs vom Nanocarrier erfolgen. Dies kann durch verschiedene Mechanismen getriggert werden. Die freigesetzten Nanocarrier konnen somit unabhangig vom Nanopartikel in das den Haarfollikel umgebende lebende Gewebe ubergehen. Mithilfe dieser innovativen Strategie kann die Bioverfugbarkeit von topisch applizierten Substanzen deutlich verbessert werden. Ein Transport von Wirkstoffen in die Haarfollikel mithilfe von Partikeln und die dortige Wirkstofffreisetzung stellen eine sehr effektive neue Methode dar, Wirkstoffe durch die Hautbarriere zu transportieren.
- Published
- 2019
35. Retaining Skin Barrier Function Properties of the Stratum Corneum with Components of the Natural Moisturizing Factor—A Randomized, Placebo-Controlled Double-Blind In Vivo Study
- Author
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Johannes, Schleusener, Andrew, Salazar, Jörg, von Hagen, Jürgen, Lademann, and Maxim E, Darvin
- Subjects
Adult ,Male ,lipid organization ,Administration, Topical ,Middle Aged ,Lipid Metabolism ,Spectrum Analysis, Raman ,Lipids ,Article ,Dermatitis, Atopic ,water binding ,lcsh:QD241-441 ,Double-Blind Method ,lcsh:Organic chemistry ,lamellae ,Humans ,Female ,Epidermis ,skin hydration ,confocal Raman microspectroscopy ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Skin - Abstract
The influence of a topically applied formulation containing components of natural moisturizing factor (NMF) on barrier-related parameters of the stratum corneum (SC) was investigated in vivo using confocal Raman microspectroscopy in a randomized, placebo-controlled double-blind study on 12 volunteers for 14 days. This method allowed for the elucidation of subtle differences between the verum and the placebo even though the components of the verum naturally occur in the SC. This differentiation is not possible non-invasively by conventional methods. In this study, we found that the applied verum and placebo formulations disrupted the equilibrium of water, NMF and lipids in the SC. The adverse effects of the formulation could be mitigated by incorporating it into a simplified supplementation of NMF molecules. As a long-term effect, the amount of strongly bound water increases at 30–40% SC depth (p <, 0.05) and the amount of weakly bound water decreases at 30–40% SC depth (p <, 0.05) for the verum. This supplement was also unexpectedly able to prevent intercellular lipids (ICL) disorganization in selected depths. In the long term, the verum treatment limited the lateral disorganization of the ICL to the upper 20% SC depth. Further research is required to elucidate the interplay of these factors in the SC, to better understand their contribution to the equilibrium and barrier function of the skin. This understanding of the interaction of these naturally occurring components could help in the future to develop and optimize topical treatments for diseases like psoriasis, atopic dermatitis, ichthyosis where the skin barrier is disrupted.
- Published
- 2021
36. Electrohydrodynamic spray applicator for homogenous application and reduced overspray of sunscreen
- Author
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Heike Richter, Martina C. Meinke, Valentin Langen, Jürgen Lademann, Johannes Schleusener, Charlotte Wille, and Sabine Schanzer
- Subjects
spectroscopy ,Materials science ,Ultraviolet Rays ,Dermatology ,01 natural sciences ,010309 optics ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,Sun protection factor ,Hand rubbing ,0103 physical sciences ,Homogeneity (physics) ,Confocal laser scanning microscopy ,sunscreens ,Humans ,Skin ,Laser Scanning Microscopy ,confocal laser scanning microscopy ,Spectrum Analysis ,Reflectivity ,Aerosol ,Electrohydrodynamics ,Sun Protection Factor ,Sunscreening Agents ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Biomedical engineering - Abstract
Background The recommended amount of sunscreen by hand application (2 mg/cm2 ) is in reality not achieved, which decreases the homogeneity and thereby the effective sun protection factor (SPF). Materials and methods The homogeneity of sunscreen applied by a newly developed spray applicator using an electrostatically charged aerosol, for which a hand rubbing of the formulation is not necessary, is evaluated. In vivo experiments were performed on the volar forearms of human volunteers using the spray applicator compared to the standardized hand application according to ISO 24444. Results The distribution homogeneity was assessed qualitatively using in vivo laser scanning microscopy and quantitatively by absorption spectroscopy after tape stripping and by the standard deviation of multiple spatially displaced reflectance measurements for non-invasive SPF determination below the minimal erythemal dose, which showed a significantly higher homogeneity by 20.9% after spray application compared to hand application. Conclusion Non-invasive SPF determination of multiple spatially displaced reflectance measurements was proven to be a suitable method for the non-invasive determination of the sunscreen distribution homogeneity. Electrostatically charged spray application increased the sunscreen distribution homogeneity on the skin and can reduce the amount of overspray.
- Published
- 2020
37. In vivo sun protection factor and UVA protection factor determination using (hybrid) diffuse reflectance spectroscopy and a multi‐lambda‐LED light source
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Carina Reble, Carolin Maria Throm, Georg Wiora, Jürgen Lademann, Ludger Kolbe, Johannes Schleusener, Martina C. Meinke, Hans Karrer, Sabine Schanzer, and Georg Khazaka
- Subjects
Materials science ,Correlation coefficient ,Diffuse reflectance infrared fourier transform ,Ultraviolet Rays ,non-invasive ,SPF determination by HDRS ,General Physics and Astronomy ,Lambda ,medicine.disease_cause ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,010309 optics ,Light source ,Sun protection factor ,Optics ,In vivo ,0103 physical sciences ,medicine ,Humans ,General Materials Science ,Spectrograph ,sunscreen ,business.industry ,Spectrum Analysis ,010401 analytical chemistry ,General Engineering ,General Chemistry ,fiber probe ,non-erythemal testing ,0104 chemical sciences ,Sun Protection Factor ,business ,Sunscreening Agents ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Ultraviolet - Abstract
The sun protection factor (SPF) values are currently determined using an invasive procedure, in which the volunteers are irradiated with ultraviolet (UV) light. Non-invasive approaches based on hybrid diffuse reflectance spectroscopy (HDRS) have shown a good correlation with conventional SPF testing. Here, we present a novel compact and adjustable DRS test system. The in vivo measurements were performed using a multi-lambda-LED light source and an 84-channel imaging spectrograph with a fiber optic probe for detection. A transmission spectrum was calculated based on the reflectance measured with sunscreen and the reflectance measured without sunscreen. The preexposure in vitro spectrum was fitted to the in vivo spectrum. Each of the 11 test products was investigated on 10 volunteers. The SPF and UVA-PF values obtained by this new approach were compared with in vivo SPF results determined by certified test institutes. A correlation coefficient R-2 = 0.86 for SPF, and R-2 = 0.92 for UVA-PF were calculated. Having examined various approaches to apply the HDRS principle, the method we present was found to produce valid and reproducible results, suggesting that the multi-lambda-LED device is suitable for in-vivo SPF testing based on the HDRS principle as well as for in-vivo UVA-PF measurements.
- Published
- 2020
38. Stratum corneum occlusion induces water transformation towards lower bonding state: a molecular levelin vivostudy by confocal Raman microspectroscopy
- Author
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Maxim E. Darvin, Johannes Schleusener, Jürgen Lademann, Sehyok Choe, ChunSik Choe, and Jin-Song Ri
- Subjects
Adult ,Male ,Aging ,penetration ,Pharmaceutical Science ,Human skin ,Dermatology ,Spectrum Analysis, Raman ,030226 pharmacology & pharmacy ,030207 dermatology & venereal diseases ,03 medical and health sciences ,Young Adult ,swelling ,0302 clinical medicine ,Colloid and Surface Chemistry ,corneocytes ,Drug Discovery ,Keratin ,medicine ,Stratum corneum ,Bound water ,Humans ,skin barrier ,chemistry.chemical_classification ,Transepidermal water loss ,Corneocyte ,integumentary system ,skin physiology ,Water ,Middle Aged ,medicine.anatomical_structure ,chemistry ,Chemistry (miscellaneous) ,Biophysics ,Female ,Swelling ,medicine.symptom ,Epidermis ,Water binding ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit - Abstract
It is conventionally understood that occlusive effects are the retention of excessive water in the stratum corneum (SC), the increase of SC thickness (swelling) and a decrease of the transepidermal water loss. However, the influence of occlusion on water binding properties in the SC is unknown.The action of plant-derived jojoba and almond oils, as well as mineral-derived paraffin oil and petrolatum topically applied on human skin, is investigated in vivo using confocal Raman microspectroscopy. To understand the oils' influence on the SC on the molecular level, the depth-dependent hydrogen bonding states of water in the SC and their relationship to the conformation of keratin, concentration of natural moisturizing factor (NMF) molecules and lipid organization were investigated.A significant SC swelling was observed only in petrolatum-treated skin. The water concentration was increased in oil-treated skin in the intermediate SC region (40-70% SC depth). Meanwhile, the amount of free, weakly and tightly bound water increased, and strongly bound water decreased in the uppermost SC region (0-30% SC depth). The NMF concentration of oil-treated skin was significantly lower at 50-70% SC depth. The lateral organization of lipids in oil-treated skin was lower at 0-30% SC depth. The secondary structure of keratin was changed towards an increase of β-sheet content in mineral-derived oil-treated skin and changed towards an increase of α-helix content in plant-derived oil-treated skin.The occlusive properties can be summarized as the increase of free water and the transformation of water from a more strongly to a more weakly hydrogen bonding state in the uppermost SC, although some oils cause insignificant changes of the SC thickness. The accompanied changes in the keratin conformation at the intermediate swelling region of the SC also emphasize the role of keratin in the SC's water-transporting system, that is the water in the SC transports intercellularly and intracellularly in the intermediate swelling region and only intercellularly in the uppermost non-swelling region. Bearing this in mind, almond, jojoba and paraffin oils, which are not occlusive from the conventional viewpoint, have an occlusion effect similar to petrolatum on the SC.Il est généralement entendu que les effets occlusifs consistent en la rétention d'un excès d'eau dans la couche cornée (stratum corneum, SC), l'augmentation d'épaisseur de la SC (gonflement) et une diminution de la perte d'eau trans-épidermique. Cependant, l'influence de l'occlusion sur les propriétés de fixation de l'eau dans le SC est inconnue. MÉTHODES: L'action des huiles de jojoba et d'amande d'origine végétale, ainsi que des huiles de paraffine et de pétrolatum d'origine minérale appliquées topiquement sur la peau humaine est étudiée in vivo à l'aide de la microspectroscopie Raman confocale. Pour comprendre l'influence des huiles sur le SC au niveau moléculaire, on a étudié les états de liaison hydrogène de l'eau dans le SC en fonction de la profondeur et leur relation avec la conformation de la kératine, la concentration des molécules du facteur naturel d'hydratation (NMF) et l'organisation des lipides. RÉSULTATS: Un gonflement significatif de le SC n'a été observé que dans la peau traitée au pétrolatum. La concentration en eau a été augmentée dans la peau traitée au pétrolatum dans la région SC intermédiaire (40-70% de profondeur du SC). En meme temps, la quantité d'eau libre, faiblement et fortement liée augmentait, tandis que l'eau fortement liée diminuait dans la région SC supérieure (0-30% de profondeur du SC). La concentration en NMF de la peau traitée à l'huile était plus basse d´une manière significative à 50-70% de profondeur du SC. L'organisation latérale des lipides dans la peau huilée était plus basse à une profondeur du SC de 0 à 30 %. La structure secondaire de la kératine a été modifiée pour augmenter la teneur en feuillet-β dans les peaux huilées d'origine minérale et pour augmenter la teneur en hélice α dans les peaux huilées d'origine végétale.Les propriétés occlusives peuvent être résumées comme l'augmentation de l'eau libre et la transformation de l'eau d'un état de liaison hydrogène plus fort à un état de liaison hydrogène plus faible dans le SC supérieure, bien que certaines huiles provoquent des changements insignifiants de l'épaisseur de la SC. Les modifications de la conformation de la kératine dans la zone de gonflement intermédiaire du SC soulignent également le rôle de la kératine dans le système de transport de l'eau du SC, c'est-à-dire que l'eau est transportée du SC de manière intercellulaire et intracellulaire dans la zone de gonflement intermédiaire et seulement de manière intercellulaire dans la zone non gonflée la plus élevée. En considérant cela, les huiles d'amande, de jojoba et de paraffine, qui ne sont pas occlusives du point de vue conventionnel, ont un effet d'occlusion similaire à celui du pétrolatum sur le SC.
- Published
- 2020
39. Response to comment by Puppels et al. on 'A modification for the calculation of water depth profiles in oil‐treated skin by in vivo Raman microscopy'
- Author
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ChunSik Choe, Jürgen Lademann, Johannes Schleusener, Maxim E. Darvin, and Sehyok Choe
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Materials science ,Analytical chemistry ,General Physics and Astronomy ,Spectrum Analysis, Raman ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,010309 optics ,symbols.namesake ,petrolatum ,Raman microscopy ,In vivo ,Raman band ,0103 physical sciences ,Skin surface ,Microscopy ,Stratum corneum ,medicine ,General Materials Science ,skin hydration ,topical application ,Skin ,integumentary system ,010401 analytical chemistry ,cosmetic oils ,General Engineering ,Water ,General Chemistry ,0104 chemical sciences ,Water depth ,medicine.anatomical_structure ,normalization ,Homogeneous ,symbols ,Epidermis ,Raman spectroscopy ,Oils ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit - Abstract
The presence of penetrated oils in the stratum corneum (SC), oil-induced occlusion of the SC and formation of occluding homogeneous film on the skin surface are discussed in relation to their influence on results of water profile calculations using conventional and newly proposed extended methods. It is shown that the conventional method does not determine the water profiles in treated skin correctly due to the superposition of Raman bands of SC's proteins and penetrated and remnant oils.
- Published
- 2020
40. Age related depth profiles of human Stratum Corneum barrier-related molecular parameters by confocal Raman microscopy in vivo
- Author
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Jürgen Lademann, Maxim E. Darvin, Johannes Schleusener, and ChunSik Choe
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Adult ,Male ,0301 basic medicine ,Aging ,Nonlinear Optical Microscopy ,Hydrogen ,Confocal ,chemistry.chemical_element ,030207 dermatology & venereal diseases ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,Stratum corneum ,medicine ,Humans ,Bound water ,Molecule ,Transepidermal water loss ,Microscopy, Confocal ,Chemistry ,Hydrogen bond ,Middle Aged ,Skin Aging ,030104 developmental biology ,medicine.anatomical_structure ,Biophysics ,symbols ,Female ,Epidermis ,Raman spectroscopy ,Developmental Biology - Abstract
In this study, stratum corneum (SC) depth profiles of hydrogen bound water molecule types, intercellular lipid (ICL) ordering, concentration of natural moisturizing factor (NMF) and keratin folding/unfolding properties are investigated in vivo for older (mean 50 years old) and younger (mean 29 years old) human skin using confocal Raman microscopy. The results show that the SC of the older group is modestly thicker (p
- Published
- 2018
41. Confocal Raman microscopy combined with optical clearing for identification of inks in multicolored tattooed skinin vivo
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Jürgen Lademann, Olaf Seidel, Maxim E. Darvin, Franziska Parenz, Jürgen Popp, Christoph Krafft, and Johannes Schleusener
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Swine ,Confocal ,Color ,Human skin ,Spectrum Analysis, Raman ,Tattoo ink ,01 natural sciences ,Biochemistry ,Analytical Chemistry ,010309 optics ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,Dermis ,0103 physical sciences ,Electrochemistry ,medicine ,Stratum corneum ,Animals ,Humans ,Environmental Chemistry ,Coloring Agents ,Spectroscopy ,Skin ,Microscopy, Confocal ,Tattooing ,integumentary system ,Chemistry ,Papillary dermis ,Tattoo removal ,medicine.anatomical_structure ,Ink ,Reticular Dermis ,Biomedical engineering - Abstract
Raman measurements applied on freshly tattooed porcine skin ex vivo showed a possibility of obtaining the ink pigment related information in the skin. Based on these results, confocal Raman microscopy was used to identify the tattoo ink pigments of different colors in multicolored tattooed human skin in vivo. The Raman signatures of tattoo ink pigments were unique. Therefore, it could be shown that the applied method is successful for the identification of the tattoo ink pigments in human skin in vivo down to depths of approx. 50 μm, which is sufficient to screen the entire epidermis and the top of the papillary dermis area on the forearm and leg skin sites. Additional application of the optical clearing technique in vivo by topical application of glycerol, combined with tape stripping removal of the uppermost stratum corneum layers and defatting allows the extension of depths of investigation in tattooed skin down to approx. 400 μm, i.e. to cover the entire papillary dermis and a large part of the reticular dermis. Thus, the tattoo ink pigments were identified in vivo and depth-dependently in human tattooed skin confirming their presence in the papillary and reticular dermis. The proposed non-invasive in vivo Raman screening combined with optical clearing for identifying the tattoo pigments in the dermis can be an important task preceding a laser-based tattoo removal procedure and for determining the optimal laser parameters.
- Published
- 2018
42. Keratin-water-NMF interaction as a three layer model in the human stratum corneum using in vivo confocal Raman microscopy
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Juergen Lademann, Johannes Schleusener, Maxim E. Darvin, and ChunSik Choe
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Male ,0301 basic medicine ,lcsh:Medicine ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit ,Spectrum Analysis, Raman ,Protein Structure, Secondary ,030207 dermatology & venereal diseases ,0302 clinical medicine ,Protein structure ,Keratin ,Disulfides ,lcsh:Science ,chemistry.chemical_classification ,Microscopy, Confocal ,Multidisciplinary ,integumentary system ,Hydrogen bond ,Middle Aged ,Molecular conformation ,medicine.anatomical_structure ,symbols ,Keratins ,Female ,Water binding ,Adult ,macromolecular substances ,Models, Biological ,Article ,Young Adult ,03 medical and health sciences ,symbols.namesake ,Stratum corneum ,medicine ,Humans ,Molecule ,Cysteine ,lcsh:R ,Water ,Hydrogen Bonding ,Protein tertiary structure ,Protein Structure, Tertiary ,Confocal microscopy ,030104 developmental biology ,chemistry ,Biophysics ,Tyrosine ,lcsh:Q ,Epidermis ,Raman spectroscopy - Abstract
The secondary and tertiary structure of keratin and natural moisturizing factor (NMF) are of great importance regarding the water regulating functions in the stratum corneum (SC). In this in vivo study, the depth-dependent keratin conformation and its relationship to the hydrogen bonding states of water and its content in the SC, are investigated using confocal Raman microscopy. Based on the obtained depth-profiles for the β-sheet/α-helix ratio, the stability of disulphide bonds, the amount of cysteine forming disulphide bonds, the buried/exposed tyrosine and the folding/unfolding states of keratin, a “three layer model” of the SC, regarding the keratin-water-NMF interaction is proposed. At the uppermost layers (30–0% SC depth), the keratin filaments are highly folded, entailing limited water binding sites, and NMF is mostly responsible for binding water. At the intermediate layers (70–30% SC depth), the keratin filaments are unfolded, have the most water binding sites and are prone to swelling. At the bottom layers (100–80% SC depth), the water binding sites are already occupied with water and cannot swell substantially. The hydrogen bonding states of water molecules can only be explained by considering both, the molecular structure of keratin and the contribution of NMF as a holistic system.
- Published
- 2017
43. In vivo detection of changes in cutaneous carotenoids after chemotherapy using shifted excitation resonance Raman difference and fluorescence spectroscopy
- Author
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Maxim E. Darvin, Harald Fuss, Felia Elban, Gisela Thiede, Marcel Braune, Juergen Lademann, Johannes Schleusener, Günther Tränkle, Bernd Sumpf, Martin Maiwald, Dunja Kutzer, and Sora Jung
- Subjects
Adult ,Male ,medicine.medical_specialty ,medicine.medical_treatment ,Urology ,Antineoplastic Agents ,Dermatology ,chemotherapy ,Spectrum Analysis, Raman ,01 natural sciences ,Fluorescence spectroscopy ,010309 optics ,030207 dermatology & venereal diseases ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,In vivo ,Neoplasms ,0103 physical sciences ,Stratum corneum ,medicine ,Humans ,Doxorubicin ,shifted excitation resonance ,ddc:610 ,Aged ,Skin ,Chemotherapy ,business.industry ,Cancer ,Middle Aged ,medicine.disease ,Fluorescence ,Carotenoids ,medicine.anatomical_structure ,Spectrometry, Fluorescence ,symbols ,Female ,fluorescence ,Raman spectroscopy ,business ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit ,Raman difference spectroscopy ,medicine.drug - Abstract
Background: Various cutaneous toxicities under chemotherapy indicate a local effect of chemotherapy by secretion after systemic application. Here, changes in the fluorescence and Raman spectral properties of the stratum corneum subsequent to intravenous chemotherapy were assessed. Methods: Twenty healthy subjects and 20 cancer patients undergoing chemotherapy were included. Measurement time points in cancer patients were before the first cycle of chemotherapy (Tbase) and immediately after intravenous application of the chemotherapy (T1). Healthy subjects were measured once without any further intervention. Measurements were conducted using an individually manufactured system consisting of a handheld probe and a wavelength-tunable diode laser-based 488 nm SHG light source. Hereby, changes in both skin fluorescence and shifted excitation resonance Raman difference spectroscopy (SERRDS) carotenoid signals were assessed. Results: Healthy subjects showed significantly (P < .001) higher mean concentrations of carotenoids compared to cancer subjects at Tbase. An increase in fluorescence intensity was detected in almost all patients after chemotherapy, especially after doxorubicin infusion. Furthermore, a decrease in the carotenoid concentration in the skin after chemotherapy was found. Conclusion: The SERRDS based noninvasive detection can be used as an indirect quantitative assessment of fluorescent chemotherapeutics. The lower carotenoid SERRDS intensities at T(base) might be due to cancerous diseases and co-medication.
- Published
- 2019
44. Modified normalization method in in vivo stratum corneum analysis using confocal Raman microscopy to compensate nonhomogeneous distribution of keratin
- Author
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Maxim E. Darvin, Johannes Schleusener, Jürgen Lademann, Sehyok Choe, and ChunSik Choe
- Subjects
chemistry.chemical_classification ,Normalization (statistics) ,Chemistry ,Confocal ,symbols.namesake ,medicine.anatomical_structure ,In vivo ,Keratin ,Microscopy ,Stratum corneum ,medicine ,symbols ,Biophysics ,Distribution (pharmacology) ,General Materials Science ,Raman spectroscopy ,Spectroscopy - Published
- 2019
45. Hydrogen bound water profiles in the skin influenced by optical clearing molecular agents - Quantitative analysis using confocal Raman microscopy
- Author
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Valery V. Tuchin, Johannes Schleusener, Jürgen Lademann, Anton Sdobnov, and Maxim E. Darvin
- Subjects
Optical Phenomena ,Swine ,Confocal ,General Physics and Astronomy ,конфокальная рамановская микроскопия ,дерма ,Spectrum Analysis, Raman ,кожный барьер ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,глицерин ,010309 optics ,symbols.namesake ,chemistry.chemical_compound ,эпидермис ,0103 physical sciences ,Microscopy ,medicine ,Glycerol ,Animals ,Bound water ,General Materials Science ,Dehydration ,Skin ,Microscopy, Confocal ,Chromatography ,Chemistry ,010401 analytical chemistry ,General Engineering ,Water ,General Chemistry ,medicine.disease ,eye diseases ,0104 chemical sciences ,иогексол ,symbols ,Iohexol ,Raman spectroscopy ,Quantitative analysis (chemistry) ,Hydrogen ,medicine.drug - Abstract
Confocal Raman microscopy has been used to measure depth-dependent profiles of porcine skin ex vivo in the high wavenumber region after application of molecular optical clearing agents (OCAs). Glycerol (70%) and iohexol (100% Omnipaque [300]) water solutions were used as OCAs and topically applied to porcine ear skin for 30 and 60 minutes. Using Gaussian function-based deconvolution, the changes of hydrogen bound water molecule types have been microscopically analyzed down to the depth of 200 μm. Results show that both OCAs induced skin dehydration (reduction of total water), which is 51.3% for glycerol (60 minutes), 33.1% for glycerol (30 minutes), 8.3% for Omnipaque (60 minutes) and 4.4% for Omnipaque (30 minutes), on average for the 40 to 200 μm depths. Among the water types in the skin, the following reduction was observed in concentration of weakly bound (51.1%, 33.2%, 7.5% and 4.6%), strongly bound (50.4%, 33.0%, 7.9% and 3.4%), tightly bound (63.6%, 42.3%, 26.1% and 12.9%) and unbound (55.4%, 28.7%, 10.1% and 5.9%) water types on average for the 40 to 200 μm depths, post application of glycerol (60 minutes), glycerol (30 minutes), Omnipaque (60 minutes) and Omnipaque (30 minutes), respectively. As most concentrated in the skin, weakly and strongly bound water types are preferentially involved in the OCA-induced water flux in the skin, and thus, are responsible for optical clearing efficiency.
- Published
- 2019
46. A modification for the calculation of water depth profiles in oil‐treated skin by in vivo confocal Raman microscopy
- Author
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Maxim E. Darvin, Johannes Schleusener, Sehyok Choe, Jürgen Lademann, and ChunSik Choe
- Subjects
Materials science ,Confocal ,Analytical chemistry ,General Physics and Astronomy ,Spectrum Analysis, Raman ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,010309 optics ,symbols.namesake ,In vivo ,0103 physical sciences ,Keratin ,Microscopy ,Stratum corneum ,medicine ,stratum corneum ,General Materials Science ,skin hydration ,topical application ,Skin ,chemistry.chemical_classification ,Microscopy, Confocal ,integumentary system ,cosmetics ,010401 analytical chemistry ,General Engineering ,Water ,General Chemistry ,Water concentration ,0104 chemical sciences ,Water depth ,medicine.anatomical_structure ,chemistry ,skin barrier function ,symbols ,Epidermis ,Raman spectroscopy ,600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit - Abstract
In this study, an extended calculation method for the determination of the water profiles in oil-treated skin is proposed, which is based on the calculation of the ratio between the Raman band intensities of water (3350-3550 cm���1) and keratin Amide I at 1650 cm���1. The proposed method is compared with the conventional method based on the ratio of the Raman band intensities of water (3350-3550 cm���1) and keratin at 2930 cm���1. The conventional method creates artifacts in the depth profiles of the water concentration in oil-treated skin, showing a lower amount of water in the upper and intermediate layers of the stratum corneum, which is due to the superposition of oil- and keratin-related Raman bands at 2930 cm���1. The proposed extended method shows no artifacts and has the potential to determine the water depth profiles after topical application of formulations on the skin.
- Published
- 2019
47. Application of spectroscopic methods for the prevention of side effects on patients during chemotherapy (Conference Presentation)
- Author
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Maxim E. Darvin, Jürgen Lademann, Gisela Thiede, Sora Jung, Thomas Bocklitz, Johannes Schleusener, and Martina C. Meinke
- Subjects
medicine.medical_specialty ,Chemotherapy ,Presentation ,business.industry ,General surgery ,medicine.medical_treatment ,media_common.quotation_subject ,medicine ,business ,media_common - Published
- 2018
48. Nonscanning large-area Raman imaging for ex vivo/in vivo skin cancer discrimination
- Author
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Gisela Thiede, Benito Moralejo, Elmar Schmälzlin, Ingo Gersonde, Maxim E. Darvin, Johannes Schleusener, and Martin Roth
- Subjects
inorganic chemicals ,Materials science ,Skin Neoplasms ,Swine ,Biomedical Engineering ,Raman imaging ,FOS: Physical sciences ,macromolecular substances ,Spectrum Analysis, Raman ,01 natural sciences ,010309 optics ,Biomaterials ,symbols.namesake ,In vivo ,0103 physical sciences ,Image Interpretation, Computer-Assisted ,medicine ,Field spectroscopy ,Animals ,Humans ,ddc:530 ,Nevus ,Instrumentation and Methods for Astrophysics (astro-ph.IM) ,Aged ,Skin ,Aged, 80 and over ,Single exposure ,Phantoms, Imaging ,010401 analytical chemistry ,technology, industry, and agriculture ,Institut für Physik und Astronomie ,Signal Processing, Computer-Assisted ,Equipment Design ,medicine.disease ,Physics - Medical Physics ,Atomic and Molecular Physics, and Optics ,0104 chemical sciences ,Electronic, Optical and Magnetic Materials ,Forearm ,Case-Control Studies ,symbols ,Medical Physics (physics.med-ph) ,Skin cancer ,Astrophysics - Instrumentation and Methods for Astrophysics ,Raman spectroscopy ,Preclinical imaging ,Ex vivo ,Algorithms ,Biomedical engineering - Abstract
Imaging Raman spectroscopy can be used to identify cancerous tissue. Traditionally, a step-by-step scanning of the sample is applied to generate a Raman image, which, however, is too slow for routine examination of patients. By transferring the technique of integral field spectroscopy (IFS) from astronomy to Raman imaging, it becomes possible to record entire Raman images quickly within a single exposure, without the need for a tedious scanning procedure. An IFS-based Raman imaging setup is presented, which is capable of measuring skin ex vivo or in vivo. It is demonstrated how Raman images of healthy and cancerous skin biopsies were recorded and analyzed. (C) The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License.
- Published
- 2018
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49. Human skin in vivo has a higher skin barrier function than porcine skin ex vivo-comprehensive Raman microscopic study of the stratum corneum
- Author
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ChunSik Choe, Jürgen Lademann, Johannes Schleusener, and Maxim E. Darvin
- Subjects
Male ,Swine ,Confocal ,General Physics and Astronomy ,Human skin ,02 engineering and technology ,Spectrum Analysis, Raman ,General Biochemistry, Genetics and Molecular Biology ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,Keratin ,Stratum corneum ,medicine ,Animals ,Humans ,General Materials Science ,chemistry.chemical_classification ,Corneocyte ,integumentary system ,General Engineering ,Hydrogen Bonding ,General Chemistry ,Penetration (firestop) ,021001 nanoscience & nanotechnology ,Lipid Metabolism ,medicine.anatomical_structure ,chemistry ,Biophysics ,Keratins ,Female ,Epidermis ,0210 nano-technology ,Extracellular Space ,Ex vivo - Abstract
Porcine skin is widely used as a human skin model in dermatology. For both, porcine stratum corneum (SC) ex vivo and human SC in vivo, the hydrogen bonding states of water, the secondary and tertiary structures of keratin, the natural moisturizing factor (NMF) concentrations and the intercellular lipids' (ICL) lateral organization are investigated depth-dependently using confocal Raman microscopy. The SC depth profiles show that porcine SC ex vivo is characterized by lower hydrogen bonding states of water (10%-30% SC depth), lower NMF concentration in the whole SC, more β-sheet form of keratin (10%-90% SC depth), more folded tertiary keratin structures (30%-70% SC depth) and higher hexagonal lateral packing order of ICL (10%-50% SC depth) compared to human SC in vivo. The results clearly show a higher value of skin barrier function of human SC in vivo than of porcine SC ex vivo. Thus, the human SC in vivo is less permeable for lipophilic and hydrophilic substances than porcine SC ex vivo. Considering the porcine SC as an ex vivo model of human SC in vivo, these findings should be taken into consideration.
- Published
- 2017
50. Perturbation Factors in the Clinical Handling of a Fiber-Coupled Raman Probe for Cutaneous in Vivo Diagnostic Raman Spectroscopy
- Author
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Carina Reble, Johannes Schleusener, Hans-Joachim Cappius, Martina C. Meinke, Jürgen Helfmann, Joachim W. Fluhr, Patrycja Gluszczynska, and Ingo Gersonde
- Subjects
Skin Neoplasms ,Microscope ,Optical fiber ,Sus scrofa ,Analytical chemistry ,Perturbation (astronomy) ,Human skin ,Spectrum Analysis, Raman ,law.invention ,symbols.namesake ,law ,In vivo ,Animals ,Humans ,Ear, External ,Melanoma ,Instrumentation ,Optical Fibers ,Spectroscopy ,Skin ,Principal Component Analysis ,Chemistry ,Equipment Design ,Carcinoma, Basal Cell ,Carcinoma, Squamous Cell ,symbols ,Fiber probe ,Artifacts ,Raman spectroscopy ,Ex vivo ,Disinfectants ,Hair ,Biomedical engineering - Abstract
The application of fiber-coupled Raman probes for the discrimination of cancerous and normal skin has the advantage of a noninvasive in vivo application, easy clinical handling, and access to the majority of body sites, which would otherwise be limited by stationary Raman microscopes. Nevertheless, including optical fibers and miniaturizing optical components, as well as measuring in vivo, involves the sensibility to external perturbation factors that could introduce artifacts to the acquired Raman spectra and thereby potentially reduce classification performance. In this study, typical perturbation factors of Raman measurements with a Raman fiber probe, optimized for clinical in vivo discrimination of skin cancer, were investigated experimentally. Measurements were performed under standardized conditions in clinical settings in vivo on human skin, as well as ex vivo on porcine ears. Raman spectra were analyzed in the fingerprint region between 1150 and 1730 cm−1 using principal component analysis. The largest artifacts in the Raman spectra were found in measurements performed under the influence of strong ambient light conditions as well as after miscellaneous pre-treatments to the skin, such as use of a permanent marker or a sunscreen. Minor influences were also found in measurements using H2O immersion and when varying the probe contact force. The effect of reasonable variation of the fiber-bending radius was found to be of negligible impact. The influence of measurements on hairy or sun-exposed body sites, as well as inter-subject variation, was also investigated. The presented results may serve as a guide to avoid negative effects during the process of data acquisition and so avoid misclassifcation in tumor discrimination.
- Published
- 2015
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