Cosmetic Formulations with Melaleuca alternifolia Essential Oil for the Improvement of Photoaged Skin: A Double-Blind, Randomized, Placebo-Controlled Clinical Study.

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.

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.

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.

In vivo Skin Penetration, Radical Protection, and Structural Changes after Topical Application of a Herbal Oil Cream Compared to Topical Calcipotriol in Mild to Moderate Psoriasis.

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.

Eco-friendly sunscreen formulation based on starches and PEG-75 lanolin increases the antioxidant capacity and the light scattering activity in the visible light.

Most modern sunscreens contain physical filters, which scatter the sunlight, increasing the photons' pathway in the upper stratum corneum. This effect can lead to a better efficacy of the UV filters and improve the diffuse reflection. However, the addition of nanosized inorganic UV filters reduces the antioxidant capacity of sunscreen formulations. Two cream formulations (F1, F2) which differ in the ingredient PEG75 Lanolin (F2), have been characterized for their radical protection factor (RPF) and their optical properties in vitro using electron paramagnetic resonance (EPR) spectroscopy and UV/VIS spectrometry. The RPF for PEG-75 Lanolin was also determined. Furthermore, their radical protection properties were analyzed on porcine skin ex vivo after visible light irradiation by EPR. The structure of each formulation in the skin surface was determined by reflectance confocal microscopy in vivo. The addition of lanolin increased the reflectance and reduced the transmittance for visible light, improving the scattering drastically. Besides, the antioxidant capacity was also increased for F2, something unpublished until now. F1 presented a lower scattering provided by starches. The sunscreens showed high scattering properties and antioxidant capacity, especially for F2, which presented the lowest radical formation in the skin model. These results are consistent with the RPF measurements where F2 has a higher RPF value (193 ± 3 × 1014 radicals/mg) than F1 (155 ± 4 × 1014 radicals/mg) and for PEG-75 Lanolin (37 ± 1 × 1014 radicals/mg). The combination of starches and PEG-75 Lanolin is the first solution to provide both, light scattering and antioxidant capacity, in sunscreens.

Kinetics of the carotenoid concentration degradation of smoothies and their influence on the antioxidant status of the human skin in vivo during 8 weeks of daily consumption.

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 Oxidation-Induced Autofluorescence Hypothesis: Red Edge Excitation and Implications for Metabolic Imaging.

Endogenous autofluorescence of biological tissues is an important source of information for biomedical diagnostics. Despite the molecular complexity of biological tissues, the list of commonly known fluorophores is strictly limited. Still, the question of molecular sources of the red and near-infrared excited autofluorescence remains open. In this work we demonstrated that the oxidation products of organic components (lipids, proteins, amino acids, etc.) can serve as the molecular source of such red and near-infrared excited autofluorescence. Using model solutions and cell systems (human keratinocytes) under oxidative stress induced by UV irradiation we demonstrated that oxidation products can contribute significantly to the autofluorescence signal of biological systems in the entire visible range of the spectrum, even at the emission and excitation wavelengths higher than 650 nm. The obtained results suggest the principal possibility to explain the red fluorescence excitation in a large class of biosystems-aggregates of proteins and peptides, cells and tissues-by the impact of oxidation products, since oxidation products are inevitably presented in the tissue. The observed fluorescence signal with broad excitation originated from oxidation products may also lead to the alteration of metabolic imaging results and has to be taken into account.

Laser scanning microscopy for control of skin decontamination efficacy from airborne particulates using highly absorbent textile nanofiber material in combination with PEG-12 dimethicone.

BACKGROUND: The decontamination of the skin is indispensable if airborne particulate contaminants deposit on the skin surface. Skin washing can have adverse effects as by skin rubbing the particles can be transferred deeply into the hair follicles, where they can be entrapped for a period of more than 10 days. Thus, alternative skin decontamination strategies are necessary. MATERIALS AND METHODS: For imaging the contaminants in the skin, sodium fluorescein-labeled soot particles of submicron size (≈600 nm) were visualized using laser scanning microscopy. RESULTS: In the present ex vivo pilot study on porcine ear skin, it was shown that sodium fluorescein-labeled soot particles of submicron size (≈600 nm) could be efficiently removed from the skin with highly absorbent textile nanofiber material, whose efficacy could be further increased by spraying the contaminated skin area with the viscous fluid PEG-12 dimethicone before textile application. CONCLUSION: In case of skin contamination with particulates, the contact washing should be avoided due to rubbing particles deeply into the hair follicles, where they can accumulate for a long time and induce negative consequences. Efficient skin decontamination could include pretreatment of skin surface with the viscous fluid PEG-12 dimethicone and subsequent application of highly absorbent textile nanofiber material.

Wound Healing Process After Thermomechanical Skin Ablation.

BACKGROUND AND OBJECTIVES: Energy-based devices have been widely applied for skin ablation. A novel ablation technique based on thermomechanical principles (Tixel© ) has been recently developed. The aim of this study was to examine the wound-healing process and clinical aspects after thermomechanical skin ablation. STUDY DESIGN/MATERIALS AND METHODS: Six female participants were treated with Tixel© on healthy skin of the dorsal side of the right forearm in a single session with a 600 µm protrusion and 12 milliseconds pulse. The treated area was examined with confocal laser scanning microscopy on day 1, 2, 7, and 14 after treatment. Clinical symptoms were evaluated at the same time-points. RESULTS: All patients developed erythema and mild edema on the treated areas, which completely disappeared within 14 days. No post-inflammatory hyperpigmentation or scarring was observed. Thermomechanical skin ablation resulted in the formation of homogeneous micro-ablation zones. Two weeks after ablation, the honeycomb patterns of the epidermis in all examined layers was thoroughly restored. Thus, wound-healing was completed. CONCLUSIONS: Wound healing after thermomechanical skin ablation is much faster compared with other fractionated ablation methods. Treatment intervals of 2-4 weeks could be recommended. Lasers Surg. Med. © 2020 The Authors. Lasers in Surgery and Medicine published by Wiley Periodicals, Inc.

In vivo detection of changes in cutaneous carotenoids after chemotherapy using shifted excitation resonance Raman difference and fluorescence spectroscopy.

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 Tbase might be due to cancerous diseases and co-medication.

Recent advances in follicular drug delivery of nanoparticles.

Introduction: The improvement of percutaneous absorption represents a clear dermatopharmaceutical aim. Recently, the hair follicle was recognized to be an important penetration pathway. Especially nanoparticles show an enhanced intrafollicular penetration and can be utilized to target specific cell populations within the hair follicle.Areas covered: The present review briefly summarizes the recent advances in follicular drug delivery of nanoparticles. Moreover, the particularities of the hair follicle as a penetration pathway are summarized which include its structure and specific barrier properties. Recently, the mechanism of the follicular penetration process has been clarified.In the meantime, different strategies have been developed to successfully improve follicular drug delivery of nanoparticles. One approach is to equip the nanocarriers with a triggered release system enabling them to release their drug load at the right time and place.Expert opinion: Follicular drug delivery with smart nanocarrier-based drug delivery systems represents a promising approach to increase the percutaneous absorption of topically applied substances. Although technical achievements and efficacy proofs concerning an increased penetration of substances are already available, the practical implementation into clinical application still represents an additional challenge and should be in the focus of interest in future research.

Reflectance confocal microscopy for noninvasive examination of nonmelanocytic tumors and virus-associated skin lesions in organ transplant recipients.

BACKGROUND: Drug-induced immunosuppression is necessary to prevent rejection of the foreign organ in transplanted patients, but neoplastic and virus-associated skin diseases are frequent complications. Reflectance confocal microscopy (RCM) recently emerged as a promising tool for the early diagnosis of skin lesions. MATERIALS AND METHODS: A total of 61 skin lesions, among them 20 basal cell carcinomas, six Bowen's diseases, 23 actinic keratoses, and 12 verrucae, were analyzed. All lesions were clinically evaluated followed by RCM evaluation by two independent dermatologists and histological examination. RESULTS: For the diagnosis of basal cell carcinoma, a sensitivity of 100% by both investigators (INV I + II) and a specificity of 100% by INV I and 80% by INV II were achieved. The sensitivity average rate for RCM features reached by both investigators ranged between 60% and 100%, and the specificity between 55% and 90%. For the diagnosis of actinic keratosis, a concordant sensitivity of 94.4% and a specificity of 80% (INV I) and 60% (INV II) were detected. The sensitivity average rate of specific RCM criteria ranged between 72.3% and 97.2%, whereas specificity ranged between 20% and 90%. Regarding verrucae, RCM confirmed the histological diagnosis with a sensitivity of 85.7% (INV I) and 100% (INV II), while specificity was 100% and 80%, respectively. CONCLUSION: Reflectance confocal microscopy resulted to be a reliable tool for the noninvasive diagnosis of neoplastic and virus-associated skin changes in organ transplant recipients. Nevertheless, given the frequency and diagnostic complexity of the hyperkeratotic lesions occurring post-transplantation, larger cohorts of patients are required to confirm and consolidate these findings.

The non-homogenous distribution and aggregation of carotenoids in the stratum corneum correlates with the organization of intercellular lipids in vivo.

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.

Influence of Sorafenib, Sunitinib and Capecitabine on the Antioxidant Status of the Skin.

BACKGROUND/AIM: The aim of the present study was to examine the effect of orally administered sorafenib (Nexavar®), sunitinib (Sutent®) and capecitabine (Xeloda®), which cause palmoplantar erythrodysesthesia (PPE), on the antioxidant status of the skin and the formation of free radicals. PATIENTS AND METHODS: A total of 42 patients were enrolled, of which 36 (85%) completed the study. Overall, 19 received capecitabine (2,000-4,000 mg per day), 10 sunitinib (25-50 mg per day) and 7 sorafenib (400-800 mg per day). Cutaneous carotenoids as markers of the antioxidant status of the skin were measured 1 day before the first oral administration (Tbase) and at day 18 of treatment (T1). RESULTS: The mean antioxidant concentrations increased significantly in patients treated with sunitinib from 3.99±1.01 to 4.68±1.32, p=0.047 and sorafenib from 4.83±0.74 to 5.3±0.78, p=0.007. Treatment with capecitabine did not significantly increase the mean antioxidant concentration. CONCLUSION: Formation of free radicals may not be the underlying patho-mechanism of tyrosine kinase inhibitors (TKI)- and capecitabine-associated PPE.

In vivo optical imaging of the viable epidermis around the nailfold capillaries for the assessment of heart failure severity in humans.

Heart failure (HF) is among the socially significant diseases, involving over 2% of the adult population in the developed countries. Diagnostics of the HF severity remains complicated due to the absence of specific symptoms and objective criteria. Here, we present an indicator of the HF severity based on the imaging of tissue parameters around the nailfold capillaries. High resolution nailfold video capillaroscopy was performed to determine the perivascular zone (PZ) size around nailfold capillaries, and 2-photon tomography with fluorescence lifetime imaging was used to investigate PZ composition. We found that the size of PZ around the nailfold capillaries strongly correlates with HF severity. Further investigations using 2-photon tomography demonstrated that PZ corresponds to the border of viable epidermis and it was suggested that the PZ size variations were due to the different amounts of interstitial fluid that potentially further translates in clinically significant oedema. The obtained results allow for the development of a quantitative indicator of oedematous syndrome, which can be used in various applications to monitor the dynamics of interstitial fluid retention. We therefore suggest PZ size measured with nailfold video capillaroscopy as a novel quantitative sensitive non-invasive marker of HF severity.

From UV Protection to Protection in the Whole Spectral Range of the Solar Radiation: New Aspects of Sunscreen Development.

Sunscreens have been constantly improving in the past few years. Today, they provide an efficient protection not only in the UVB but also in the UVA spectral region of the solar radiation. Recently it could be demonstrated that 50% of all free radicals induced in the skin due to solar radiation are formed in the visible and infrared spectral region. The good protective efficacy of sunscreens in the UV region prompts people to stay much longer in the sun than if they had left their skin unprotected. However, as no protection in the visible and infrared spectral region is provided, high amounts of free radicals are induced here that could easily exceed the critical radical concentration. This chapter describes how the effect of sunscreens can be extended to cover also the visible and infrared spectral region of the solar radiation by adding pigments and antioxidants with high radical protection factors to the sunscreen formulations.

Two-photon autofluorescence lifetime imaging of human skin papillary dermis in vivo: assessment of blood capillaries and structural proteins localization.

The papillary dermis of human skin is responsible for its biomechanical properties and for supply of epidermis with chemicals. Dermis is mainly composed of structural protein molecules, including collagen and elastin, and contains blood capillaries. Connective tissue diseases, as well as cardiovascular complications have manifestations on the molecular level in the papillary dermis (e.g. alteration of collagen I and III content) and in the capillary structure. In this paper we assessed the molecular structure of internal and external regions of skin capillaries using two-photon fluorescence lifetime imaging (FLIM) of endogenous compounds. It was shown that the capillaries are characterized by a fast fluorescence decay, which is originated from red blood cells and blood plasma. Using the second harmonic generation signal, FLIM segmentation was performed, which provided for spatial localization and fluorescence decay parameters distribution of collagen I and elastin in the dermal papillae. It was demonstrated that the lifetime distribution was different for the inner area of dermal papillae around the capillary loop that was suggested to be due to collagen III. Hence, we propose a generalized approach to two-photon imaging of the papillary dermis components, which extends the capabilities of this technique in skin diagnosis.

A comparative study of ex vivo skin optical clearing using two-photon microscopy.

Multiphoton tomography (MPT) is a prospective tool for imaging the skin structure. Aiming to increase the probing depth, a comparative ex vivo study of optical clearing of porcine ear skin was performed by using two optical clearing agents (OCAs), i.e., glycerol and iohexol (OmnipaqueTM ) at different concentrations, which exhibit different osmotic properties. The results show that a topical application of glycerol or OmnipaqueTM solutions onto the skin for 60 min significantly improved the depth and contrast of the MPT signals. By utilizing 40%, 60% and 100% glycerol, and 60% and 100% OmnipaqueTM it was demonstrated that both agents improve autofluorescence and SHG (second harmonic generation) signals from the skin. At the applied concentrations and agent time exposure, glycerol is more effective than OmnipaqueTM . However, tissue shrinkage and cell morphology changes were found for highly concentrated glycerol solutions. OmnipaqueTM , on the contrary, increases the safety and has no or minimal tissue shrinkage during the optical clearing process. Moreover OmnipaqueTM allows for robust multimodal optical/X-ray imaging with automatically matched optically cleared and X-ray contrasted tissue volumes. These findings make OmnipaqueTM more prospective than glycerol for some particular application.

In vivo/ex vivo targeting of Langerhans cells after topical application of the immune response modifier TMX-202: confocal Raman microscopy and histology analysis.

The increased ability of TMX-202 (derivative of imiquimod) to penetrate the intact stratum corneum (SC) and the follicular orifices of porcine ear skin was shown ex vivo using confocal Raman microscopy and laser scanning microscopy. Moreover, to assess whether TMX-202 is able to reach the immune cells, Langerhans cells extracted from pretreated human skin were investigated ex vivo using confocal Raman microscopy combined with multivariate statistical methods. Tracking the Raman peak of dimethyl sulfoxide centered at 690 cm(−1), the absorption of TMX-202 containing formulation by Langerhans cells was shown. To answer the question whether the TMX-202 active ingredient is able to reach Langerhans cells, the attraction of immune cells to TMX-202 containing formulation treated skin was measured in the in vivo rodent model Mastomys coucha. The results show that TMX-202 active ingredient is able to reach Langerhans cells after penetrating through the intact skin and subsequently attract immune cells. Both the intercellular/transcellular as well as the follicular pathways allow the penetration through the intact barrier of the SC.

Confocal Raman microscopy and multivariate statistical analysis for determination of different penetration abilities of caffeine and propylene glycol applied simultaneously in a mixture on porcine skin ex vivo.

Propylene glycol is one of the known substances added in cosmetic formulations as a penetration enhancer. Recently, nanocrystals have been employed also to increase the skin penetration of active components. Caffeine is a component with many applications and its penetration into the epidermis is controversially discussed in the literature. In the present study, the penetration ability of two components - caffeine nanocrystals and propylene glycol, applied topically on porcine ear skin in the form of a gel, was investigated ex vivo using two confocal Raman microscopes operated at different excitation wavelengths (785nm and 633nm). Several depth profiles were acquired in the fingerprint region and different spectral ranges, i.e., 526-600cm(-1) and 810-880cm(-1) were chosen for independent analysis of caffeine and propylene glycol penetration into the skin, respectively. Multivariate statistical methods such as principal component analysis (PCA) and linear discriminant analysis (LDA) combined with Student's t-test were employed to calculate the maximum penetration depths of each substance (caffeine and propylene glycol). The results show that propylene glycol penetrates significantly deeper than caffeine (20.7-22.0µm versus 12.3-13.0µm) without any penetration enhancement effect on caffeine. The results confirm that different substances, even if applied onto the skin as a mixture, can penetrate differently. The penetration depths of caffeine and propylene glycol obtained using two different confocal Raman microscopes are comparable showing that both types of microscopes are well suited for such investigations and that multivariate statistical PCA-LDA methods combined with Student's t-test are very useful for analyzing the penetration of different substances into the skin.

Influence of Topical, Systemic and Combined Application of Antioxidants on the Barrier Properties of the Human Skin.

BACKGROUND: The formation of free radicals in human skin by solar ultraviolet radiation is considered to be the main reason for extrinsic skin aging. The antioxidants in human tissue represent an efficient protection system against the destructive action of these reactive free radicals. In this study, the parameters of the skin, epidermal thickness, stratum corneum moisture, elasticity and wrinkle volume, were determined before and after the treatment with antioxidant- or placebo-containing tablets and creams. METHODS: The study included 5 groups of 15 volunteers each, who were treated for 2 months with antioxidant-containing or placebo tablets, creams or a combination of antioxidant-containing tablets and cream. The skin parameters were measured at time point 0 and at week 8 utilizing ultrasound for the determination of epidermal thickness, a corneometer for stratum corneum moisture measurements, skin profilometry for quantifying the wrinkle volume and a cutometer for determining the elasticity. RESULTS: The verum cream had a positive influence on epidermal thickness, elasticity and skin moisture, but the verum tablets improved the epidermal thickness only. The combined application of verum tablets and creams led to a significant improvement of all investigated skin parameters, whereas the application of placebo tablets or cream did not influence any parameters. CONCLUSION: The topical and oral supplementation of antioxidants can be an instrument to improve several skin parameters and potentially counteract or decelerate the process of extrinsic skin aging.