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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.

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.

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

The epidermal protective functions are closely associated with skin hydration homeostasis. The understanding of different states of water binding is a rising concept in assessing topically applied formulations and their interaction within the stratum corneum (SC). In addition to global water content, primary bound water, partially bound water, and unbound water and barrier-related lipid lateral packing and protein secondary structure can be measured by Raman spectroscopy. This study aimed to establish an in vitro SC model to evaluate differences in the efficacy of a natural sugar-derived complex in combination with glycerol and a botanical extract in modulating SC water binding and structural proteins and barrier lipids. These compounds were selected due to their water-binding and soothing properties. The SC water profiles were assessed at the surface and in 8 μm SC depth. After a 12-hour hyperhydration and subsequent product incubation the measurements were performed during a 6 hours desiccation phase. The maximal water caption and the time until reaching a steady state are measured as well as water retention and resistance against water loss. Global water content, partially bound, and unbound water, as well as lipid and protein structures were assessed with confocal Raman microspectroscopy. Both the natural sugar-derived mixture and more pronounced, the same mixture with additional glycerol increased all three water-binding parameters at the surface and in 8 μm SC depth at the beginning and during the desiccation phase. Further addition of botanical extract did not result in an additional increase of the water-binding. All three formulations showed an increase in the lipid lateral packing values prevented the protein alteration as measured by β-sheets signal compared to blank. The present model is suited for screening studies comparing the specific effects of different compounds on hydration states. The natural sugar-derived mixture Aquaxyl showed evidence for an improvement of all SC hydration states, lipid and protein structure which was further enhanced by the addition of glycerol 5%. This improvement was evidenced at the surface and within the SC for all hydration-related parameters, and the lipid as well the protein structures. The addition of botanical extract phytoessence blue daisy did not show further improvement.

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.

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.

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.

The antioxidant status of the skin shows constant alterations depending on nutrition and other lifestyle factors. Carotenoids can serve as marker substances for the antioxidant status of the epidermis in vivo. The carotenoid concentration of 2 homemade green smoothies and a commercial green smoothie, all containing fruits and vegetables, was assessed by resonance Raman spectroscopy. Furthermore, a pilot study was conducted to investigate changes of the cutaneous carotenoid concentration of 20 healthy volunteers under the daily intake of either a homemade smoothie or the commercial smoothie in vivo using reflectance spectroscopy. We hypothesized that higher carotenoid concentrations in the homemade smoothies compared to those of the commercial one would lead to a faster increase of the cutaneous carotenoid concentration in healthy volunteers. The measurements of the homemade smoothies showed notably higher initial carotenoid concentrations (7.6 ±0.8)*10-4arbitrary units (a.u.) (smoothie A) and (10.4 ± 0.6)*10-4 a.u. (smoothie B) compared to the commercial smoothie (5.8 ± 0.2)*10-4 a.u. Nevertheless, the commercial smoothie showed a higher stability of carotenoids over 24 hours. 8 weeks after daily consumption of the homemade smoothies, volunteers showed an insignificant increase of cutaneous carotenoids from (4.5 ± 0.1)*10-4 a.u. at baseline to (4.8 ± 0.3)*10-4 a.u. (means ± standard error of the mean). The volunteers consuming the commercial smoothie showed an insignificant increase of cutaneous carotenoids from (4.3 ± 0.2)*10-4 a.u. to (4.7 ± 0.2)*10-4 a.u. after 8 weeks. The observed increase of cutaneous carotenoid concentrations could be classified as a strong tendency. Fresh homemade smoothies can exhibit a higher carotenoid concentration compared to commercial smoothies but need to be consumed immediately after preparation.

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.

Background Solar radiation causes skin damage through the generation of reactive oxygen species (ROS). While UV filters effectively reduce UV‐induced ROS, they cannot prevent VIS‐induced (400‐760 nm) oxidative stress. Therefore, potent antioxidants are needed as additives to sunscreen products. Methods We investigated VIS‐induced ROS formation and the photoprotective effects of the Nrf2 inducer Licochalcone A (LicA). Results Visible spectrum of 400‐500 nm dose‐dependently induced ROS in cultured human fibroblasts at doses equivalent to 1 hour of sunshine on a sunny summer day (150 J/cm2). A pretreatment for 24 hours with 1 µmol/L LicA reduced ROS formation to the level of unirradiated cells while UV filters alone were ineffective, even at SPF50+. In vivo, topical treatment with a LicA‐containing SPF50 + formulation significantly prevented the depletion of intradermal carotenoids by VIS irradiation while SPF50 + control did not protect. Conclusion LicA may be a useful additive antioxidant for sunscreens.

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.

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.

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.

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.

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.

ZusammenfassungIn den letzten Jahren sind die Hautkrebsraten weltweit gestiegen. Die Anwendung von Sonnenschutzmitteln ist eine weit verbreitete Strategie in der Prävention gegen Hautkrebs, Sonnenbrand und Photoalterung. Es sollten kombinierte Breitbandfiltersysteme mit hoher Wirkung gegen UV-B- und UV-A-Strahlung eingesetzt werden. Weitere Studien sind notwendig, um die Entwicklung von Sonnenschutzmitteln gegen die ebenfalls schädlichen IRA-Strahlen voranzutreiben. Obwohl die regelmäßige Anwendung von Sonnenschutzmitteln ein wesentlicher Bestandteil in der Hautkrebsprävention ist, sollten insbesondere bei Kindern kombinierte Schutzmaßnahmen angewendet werden: Sonnenkarenz bei starker Sonneneinstrahlung, textiler Sonnenschutz, Aufenthalt im Schatten und die Anwendung von Sonnenschutzmitteln.

The possibility to use oral intake of fluorescent dyes approved on humans for intravital fluorescence in vivo imaging is attractive due to its simplicity and non-invasive character. Here we investigate the potential of solid lipid curcumin particles (SLCP) for imaging of capillary permeability using fluorescence video capillaroscopy. Curcumin fluorescence corresponding to 0.7 μg/mL concentration was observed from blood plasma after oral consumption of 2 g of SLCP. However, this signal was ~15% higher than that of blood plasma intrinsic fluorescence, which was itself at least 10-fold lower than the background signal from pericapillary tissue. By comparing the optical properties and concentration to the case of intravenous injection of sodium fluorescein, we analyzed which parameters of the experiment should be optimized for using oral consumption of fluorescing compounds in intravital in vivo capillaroscopy measurements.

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.

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.

Multiphoton microscopy (MPM) is a method of molecular imaging and specifically of intravital imaging that is characterized by high spatial resolution in combination with a greater depth of penetration into the tissue. MPM is a multimodal method based on detection of nonlinear optical signals - multiphoton fluorescence and optical harmonics - and also allows imaging with the use of the parameters of fluorescence decay kinetics. This review describes and discusses photophysical processes within major reporter molecules used in MPM with endogenous contrasts and summarizes several modern experiments that illustrate the capabilities of label-free MPM for molecular imaging of biochemical processes in connective tissue and cells.

Background: The chronicity of psoriasis often requires continuous topical treatment. Materials and Methods: Here, the radical protection of a cream containing various herbal oils was evaluated in vivo by electron paramagnetic resonance (EPR) spectroscopy and its skin penetration by Raman microscopy in intact and barrier-disturbed skin. Changes in skin barrier properties were evaluated after 4 weeks of daily topical application using in vivo laser scanning microscopy (LSM) and transepidermal water loss in 26 healthy volunteers. A randomized, controlled, double-blind, three-arm parallel clinical study evaluated the efficacy of the herbal oil cream compared to a 0.05% calcipotriol-containing cream and to a vehicle cream, in 135 patients with mild to moderate plaque psoriasis with the change in Psoriasis Area and Severity Index (PASI) from baseline to week 12 as the primary endpoint. Results: EPR spectroscopy disclosed a significantly higher radical formation in untreated than skin treated with the herbal oil cream (p ≤ 0.05). LSM measurements indicated a protective skin barrier effect in treated compared to untreated skin. In the clinical trial, the topical application of herbal oils showed a significant reduction of the PASI score compared to topical calcipotriol at week 12 (p = 0.016). The mean reduction in PASI was 49% for the herbal oil cream, 38% for calcipotriol, and 55% for the vehicle cream. The percentage of patients, who reached PASI 50 and 75 at any time point, was 55.9% and 29.4% for the herbal oil cream, 47.4% and 15.8% for calcipotriol, and 23 (60.5%) and 13 (34.2%) for the vehicle, respectively (p > 0.05). The vehicle, originally designed as a placebo, contained a main ingredient of the herbal oil cream and therefore showed corresponding results. Conclusion: The herbal oil cream demonstrated effectiveness in the treatment of mild to moderate plaque psoriasis.