[Show abstract][Hide abstract] ABSTRACT: Background: Pegylated liposomal doxorubicin (PLD) is a highly efficient chemotherapeutic; however, it induces dermal side effects such as palmar-plantar erythrodysesthesia (PPE) in up to 80% of cases, probably by being emitted with the sweat onto the skin surface. Aim: The aim of the present study was to examine whether a topically applied ointment containing antioxidants with a high radical protection factor is able to prevent the formation of PPE. Methods: Twenty patients suffering from ovarian carcinoma and treated with PLD were observed. Results: 60% of the patients tolerated the regular application of the cream and developed no PPE. The remaining 40% interrupted the application. Six of them developed PPE and resumed ointment application thereafter. In these cases the PPE disappeared or was strongly reduced. Conclusion: The results of the observation clearly demonstrate that topical application of the ointment is an efficient strategy against the development of PPE during chemotherapy with PLD.
[Show abstract][Hide abstract] ABSTRACT: The combination of CARS (coherent anti-Stokes Raman scattering), SHG (second harmonic generation) and TPEF (two photon excited fluorescence) imaging - referred to as multimodal imaging - provides complementary contrast based on molecular vibrations, the structure of various tissue components and endogenous fluorophores, respectively.
A comprehensive overview of the appearance of human skin in multimodal imaging is presented.
Multimodal imaging of unstained skin cross sections of 32 individuals is performed using a Laser scanning microscope (LSM) and picosecond laser pulse for excitation.
The epidermis, dermis and subcutis are distinguishable in all three applied modalities, but is unveiled best in multimodal images. While the subcutis is dominated by CARS signal, predominately SHG and secondary TPEF signal are observed for the dermis. In contrast, no SHG signal is detected in the epidermis, whereas CARS and TPEF show equal contributions. Additionally, the appearance of the major skin appendages, i.e. the hair follicle, sebaceous and sweat gland as well as blood vessels belonging to the vascular system is described. All four investigated functional units show a characteristic morphochemistry in TPEF and CARS allowing to distinguish further subunits, e.g. the major components of the hair follicle, while the SHG signal delineates the localization of the functional units.
Multimodal imaging is a powerful tool to investigate human skin by providing high contrast based on the molecular constitution. It is therefore suggested that multimodal imaging has a high potential in the application for dermatological research and clinical diagnostics of various skin alterations. This article is protected by copyright. All rights reserved.
British Journal of Dermatology 08/2013; 169(4). DOI:10.1111/bjd.12427 · 4.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the clinical setting, optical coherence tomography (OCT) is applicable for the non-invasive diagnosis of skin cancer and may in particular be used for margin definition prior to excision. In this regard, OCT may improve the success rate of removing tumor lesions more effectively, preventing repetitive excision, which may subsequently result in smaller excisions. In this study, we have aimed to evaluate the applicability of OCT for in vivo presurgical margin assessment of non-melanocytic skin tumors (NMSC) and to describe the feasibility of different scanning techniques. A total number of 18 patients planned for excision of lesions suspicious of NMSC were included in this study. Based on OCT, we defined the specific tumor margins on 19 lesions preoperatively using different scanning modalities. Sixty-one margin points and five complete tumor margins were analysed on 18 patients with a total of 19 lesions including 63% basal cell carcinoma (BCC) (n = 12), 16% (n = 3) squamous cell carcinoma (SCC) and 21% of other types of skin tumors (n = 4) were classified. In 84% of the cases (n = 16), the OCT-defined lateral margins correctly indicated complete removal of the tumor. The surgical margins chosen by the surgeon never fell below the OCT-defined margin. Regarding the techniques of marginal definition, punctual tumor border scan in the perpendicular direction, with an extension of free-run scans for unsure cases can hardly be recommended. This study shows that suspected NMSC can effectively be confirmed, and furthermore, resection margin can be minimized under OCT control without reducing the rate of complete removal.
[Show abstract][Hide abstract] ABSTRACT: The treatment of infected wounds is one possible therapeutic aspect of plasma medicine. Chronic wounds are often associated with microbial biofilms which limit the efficacy of antiseptics. The present study investigates two different surface barrier discharges with air plasma to compare their efficacy against microbial biofilms with chlorhexidine digluconate solution (CHX) as representative of an important antibiofilm antiseptic. Pseudomonas aeruginosa SG81 and Staphylococcus epidermidis RP62A were cultivated on polycarbonate discs. The biofilms were treated for 30, 60, 150, 300 or 600 s with plasma or for 600 s with 0.1% CHX, respectively. After treatment, biofilms were dispensed by ultrasound and the antimicrobial effects were determined as difference in the number of the colony forming units by microbial culture. A high antimicrobial efficacy on biofilms of both plasma sources in comparison to CHX treatment was shown. The efficacy differs between the used strains and plasma sources. For illustration, the biofilms were examined under a scanning electron microscope before and after treatment. Additionally, cytotoxicity was determined by the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay with L929 mouse fibroblast cell line. The cell toxicity of the used plasma limits its applicability on human tissue to maximally 150 s. The emitted UV irradiance was measured to estimate whether UV could limit the application on human tissue at the given parameters. It was found that the UV emission is negligibly low. In conclusion, the results support the assumption that air plasma could be an option for therapy of chronic wounds.
PLoS ONE 07/2013; 8(7):e70462. DOI:10.1371/journal.pone.0070462 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Modern sunscreens are well suited to provide sufficient protection in the UV-range, because the filter substances absorb or scatter UV-radiation. Although up to 50% of radicals were formed in the visible and infrared spectral range during solar radiation protection strategies are not provided in this range. Previous investigations of commercially available products have shown that in addition to physical filters, antioxidants are necessary to provide protective effects in the infrared range by neutralizing already formed radicals. In this study the efficacy of filter substances and antioxidants to reduce radical formation in both spectral ranges was investigated after UV/VIS- or IR-irradiation. Optical properties and radical protection were determined for the investigated creams. It was found that organic UV-filters lower radical formation in the UV/VIS-range to 35% compared to untreated skin, independent of the presence of antioxidants. Further reduction to 14% was reached by addition of 2% physical filters, while physical filters alone were ineffective in the UV/VIS range due to the low concentration. In contrast, this filter type reduced radical formation in the IR-range significantly to 65%; similar effects were aroused after application of antioxidants. Sunscreens which contain organic UV-filters, physical filters and antioxidants ensure protection in the complete solar spectrum. This article is protected by copyright. All rights reserved.
Photochemistry and Photobiology 07/2013; 89(5). DOI:10.1111/php.12137 · 2.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Carotenoids are organic pigments that are found in the chloroplasts and chromoplasts of plants and some other photosynthetic organisms like algae, bacteria, and fungi. There are over 600 known carotenoids; they are divided into two classes, xanthophylls (which contain oxygen) and carotenes (which are purely hydrocarbons, and contain no oxygen).
All carotenoids are tetraterpenoids, meaning that they are produced from 8 isoprene (2-methyl-1,3-butadiene) molecules and contain 40 carbon atoms. In humans, four carotenoids (beta-carotene, alpha-carotene, gamma-carotene, and beta-crypoxanthin) have vitamin A activity (meaning they can be converted to retinal). Bixin ((2E,4E,6E,8E,10E,12E,14E,16Z,18E)-20-methoxy-4,8,13,17-tetramethyl-20-oxoicosa 2,4,6,8,10,12,14, 16,18-nonaenoic acid) is an polyunsaturated, norcarotenoid, red dye from the main fruit of annatto, its seeds reduced to powder are widely used to color food and sunscreen. The annatto seeds contain about 5% pigment, which consist of 70-80% of bixin. The bixin is soluble in fats but insoluble in water. When exposed to alkalis, the methyl ester is hydrolyzed and produces the norbixin dicarboxylic acid, a water soluble derivative. It is a chemically unstable compound when isolated and is converted, via
isomerization, in bixin trans-(β-bixin), the cis-trans isomer of bixin. Carotenoids bixin and norbixin have two stereo configurations, i.e., cis and trans. In extracts, under normal conditions, the cis-bixin or cis-norbixin are more unstable. The cis-bixin or cis-norbixin solution under heating are partially converted into the trans configuration, which is more stable and known as isobixin and isonorbixin. Brazil is one of the major producers of annatto and its planting occurs primarily in the North and Northeast regions, although its
cultivation in the last decades expanded to other regions, in the Southeast, especially Sao Paulo and Rio de Janeiro. The international market growing demand is justified by the prohibition of synthetic dyes due to studies that have proven their toxicity. Like many carotenoids, bixin and norbixin also present significant antioxidant properties. Both carotenoids can reduce levels of malondialdehyde (biomarker of lipid peroxidation) when induced by cyclophosphamide (immunosuppressant), as well as protect the DNA from
oxidative damage in vitro. Bixin inhibited in vitro generation of superoxide and the generation of reactive oxygen species such as hydrogen peroxide and hydroxyl radicals and with protective action against mutagenicity in human lymphocytes. Besides the antioxidant property, these carotenoids also present important metabolic actions in lipids and sugars. Bixin acts as PPAR-γ agonist (receptor activated by peroxisome proliferators range), a nuclear receptor that acts in the metabolism of lipids and carbohydrates and has anti-inflammatory, immunomodulatory and anti-atherosclerotic action. This interaction of BIX and the PPAR gamma receptor may increase the sensitivity to insulin in adipocytes so that there is a greater glucose uptake. Furthermore, the interaction with the receptor regulates lipid metabolism improving metabolic syndrome present in diabetic patients.
Carotenoids: Food Sources, Production and Health Benefits, 1 edited by Masayoshi Yamaguchi, 07/2013: chapter Chapter 13 - Bixin and Norbixin: Chemistry, Production and Health Benefits: pages 261-270; Nova Publishers., ISBN: 978-1-62808-622-5
[Show abstract][Hide abstract] ABSTRACT: For many years, several attempts have been made to enhance skin penetration by chemical, physical or mechanical manipulation to reduce the barrier function of the skin. The present study demonstrates the possibility of penetration enhancement for 400 nm sized nanocapsules loaded with a model drug consisting of a fluorescent dye by the application of tissue-tolerable plasma (TTP). Therefore, the stability of the nanocapsules and their penetration through the skin barrier prior to and in combination with TTP application was evaluated. The results revealed that the penetration of the nanocapsules could be effectively enhanced when applied in combination with TTP, hence delivering the model drug unaffected by plasma into deeper skin layers. The stability testing showed no significant structural changes of the nanocapsules after contact with TTP. Thus, the present study introduces a new strategy for the penetration enhancement of substances by the combined utilization of nanocapsules and TTP.
[Show abstract][Hide abstract] ABSTRACT: Hyperforin is well-known for its anti-inflammatory, anti-tumour, anti-bacterial and antioxidant properties. The application of a hyperforin-rich verum cream could strengthen the skin barrier function by reducing radical formation and stabilizing stratum corneum lipids. Here, it was investigated whether topical treatment with a hyperforin-rich cream increases the radical protection of the skin during VIS/NIR irradiation. Skin lipid profile was investigated applying HPTLC on skin lipid extracts. Furthermore, the absorption- and scattering coefficients, which influence radical formation, were determined. 11 volunteers were included in this study. After a single cream application, VIS/NIR-induced radical formation could be completely inhibited by both verum and placebo showing an immediate protection. After an application period of 4 weeks, radical formation could be significantly reduced by 45% following placebo application and 78% after verum application showing a long-term protection. Furthermore, the skin lipids in both verum and placebo groups increased directly after a single cream application but only significantly for ceramide [AP] and [NP]. After long-term cream application, concentration of cholesterol and the ceramides increased, but no significance was observed. These results indicate that regular application of the hyperforin-rich cream can reduce radical formation and can stabilize skin lipids, which are responsible for the barrier function.
European journal of pharmaceutics and biopharmaceutics: official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V 06/2013; 86(2). DOI:10.1016/j.ejpb.2013.06.016 · 3.38 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The objective of the present study was the investigation of hair follicle morphology in eight mammalian species in order to evaluate the species-specific contribution of hair follicles to skin penetration particularly with regard to the utilization of the different animal species as skin models for human skin.
Cyanoacrylate skin surface biopsy method (CSSB), light microscopy and also digital photography were used for the measurements of hair follicle morphology.
The results revealed species-specific differences regarding the pattern of hair follicle distribution and also differences with regard to hair follicle parameters and characteristics. The results also showed that hair follicles generally possess enormous reservoir capacities, regarding the follicular volume. In all examined species, hair follicles reached at least one-fifth of stratum corneum storage capacity. The results were compared with human data obtained in a previous study.
With regard to hair follicle morphology and skin structure, the porcine skin seems to be the most appropriate skin model for human skin analog to previous investigations, whereas the skin of dog, cat, and rabbit showed the most significant differences.
Skin Research and Technology 06/2013; 20(2). DOI:10.1111/srt.12098 · 1.31 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The monitoring of wound-healing processes is indispensable for the therapeutic effectiveness and improved care of chronic wounds. Histological sections provide the best morphological assessment of wound recovery, but cause further tissue destruction and increase the risk of infection. Therefore, it is reasonable to apply a diagnostic tool that allows a non-invasive and reliable observation of morphological changes in wound healing.
Optical coherence tomography (OCT) is an imaging technique for in vivo evaluation of skin diseases with a resolution close to histopathology. The aim of this study was to investigate whether OCT is suited to display the phases of wound healing. For this purpose, six patients with chronic wounds were objectively characterized by OCT during a period of 2 weeks.
Comparable results between histological findings and OCT were achieved. OCT allowed the detection of partial loss of the epidermis, vasoconstriction, vasodilatation and epithelialization.
Consequently, OCT could be a potential non-invasive diagnostic tool for the characterization and monitoring of cutaneous wound-healing processes over time.
Skin Research and Technology 06/2013; 20(1). DOI:10.1111/srt.12077 · 1.31 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The standard diagnostic procedure for basal cell carcinoma (BCC) is invasive tissue biopsy with time-consuming histological examination. To reduce the number of biopsies, noninvasive optical methods have been developed providing high-resolution skin examination. We present direct comparison of a reflectance confocal microscope (RLSM) and a multiphoton tomograph (MPT) for BCC diagnosis. Both systems are applied to nine patients prior to surgery, and the results are analyzed, including histological results. Both systems prove suitable for detecting typical characteristics of BCC in various stages. The RLSM allows large horizontal overview images to be obtained, enabling the investigator to find the regions of interest quickly, e.g., BCC nests. Elongated cells and palisading structures are easily recognized using both methods. Due to the higher resolution, changes in nucleus diameter or cytoplasm could be visualized with the MPT. Therefore, the nucleus diameter, nucleus/cytoplasm ratio, and cell density are estimated for normal and BCC cells using the MPT. The nucleus of elongated BCC cells is significantly longer than other measured normal skin cells, whereas the cell density and nucleus/cytoplasm ratio of BCC cannot be significantly distinguished from granular cells.
[Show abstract][Hide abstract] ABSTRACT: Herd health programs for the maintenance of welfare and productivity in cattle need efficient tools for monitoring the health of individual animals. Recent reports demonstrate that the oxidative status is related to various stress conditions in dairy cows. Biomarkers, among other carotenoids, could serve as indicators of stress originating from the environment (e.g., heat stress or sun radiation) or from the animal itself (e.g., disease). To date, only invasive in vitro tests are available to assess the oxidative status in cattle. The present study compares the results of optical noninvasive in vivo measurements of dermal carotenoids in cattle udder skin using an LED-based miniaturized spectroscopic system (MSS) with those obtained by photometric analysis of beta carotene in whole blood samples using a portable device. Correlations between the concentrations of dermal and blood carotenoids were calculated under consideration of the nutritional status of the animals. Significant correlation (R = 0.86) was found for cattle with a moderate to obese body condition. Thus, the blood and skin concentrations of the marker substance beta carotene are comparable under stable stress conditions of the cattle. This demonstrates that the MSS is suitable for noninvasive assessment of dermal carotenoid concentrations in cattle.
[Show abstract][Hide abstract] ABSTRACT: The results of the risk assessment of the tissue-tolerable plasma (TTP) jet kINPen med® and first results of pilot clinical studies are presented. Producing an atmospheric pressure plasma, this plasma jet entails no risk for humans in terms of temperature increase, UV radiation or free radical formation by the plasma. The antiseptic efficacy in vitro on porcine skin and in vivo on human skin was compared to that of octenidine. TTP could significantly reduce the bacterial load in comparison to untreated skin. However, the slightly reduced antiseptic properties of TTP are attributed to the current parameter set-up and technical limitations.
Clinical Plasma Medicine 06/2013; 1(1):5–10. DOI:10.1016/j.cpme.2013.01.001
[Show abstract][Hide abstract] ABSTRACT: Carotenoids are important substances for human skin due to their powerful antioxidant properties in reaction of neutralization of free radicals and especially reactive oxygen species, including singlet oxygen. Concentration of carotenoids in the skin could mirror the current redox status of the skin and should be investigated in vivo. Optical methods are ideally suited for determination of carotenoids in mammalian skin in vivo as they are both noninvasive and quick. Four different optical methods could be used for in vivo measurement of carotenoids in the human or animal skin: (1) resonance Raman spectroscopy; (2) Raman microscopy; (3) reflection spectroscopy; (4) skin color measurements. The advantages, shortcomings, and limitations of the above-mentioned optical methods are discussed.
[Show abstract][Hide abstract] ABSTRACT: Vitamin C is a potent radical scavenger and a physiological part of the antioxidant system in human skin. The aim of this study was to measure changes in the radical-scavenging activity of human skin in vivo due to supplementation with different doses of vitamin C and at different time points. Therefore, 33 volunteers were supplemented with vitamin C or placebo for 4 weeks. The skin radical-scavenging activity was measured with electron paramagnetic resonance spectroscopy. After 4 weeks, the intake of 100 mg vitamin C/day resulted in a significant increase in the radical-scavenging activity by 22%. Intake of 180 mg/day even resulted in a significant increase of 37%. No changes were found in the placebo group. A part of the study population was additionally measured after 2 weeks: in this group radical scavenging had already reached maximal activity after 2 weeks. In conclusion, orally administered vitamin C increases the radical-scavenging activity of the skin. The effect occurs fast and is enhanced with higher doses of vitamin C.
[Show abstract][Hide abstract] ABSTRACT: This contribution presents an alternative setup for analytical micro-spectroscopy by means of coherent anti-Stokes Raman scattering (CARS). Instead of using separate detectors for forward- and epi-scattered photons, this setup enhances the comparability of the signals by coupling them into a multimodal optical fiber and employing a fiber optical switch for selection of the signal source at a single detector. To facilitate measurements at different spectral positions, which might be hampered by laser power fluctuations, a feedback loop is introduced into the pump beam to ensure stable and pre-defined laser power levels. The scheme of the microscope and the control software is presented. Its potential is evaluated by recording spectral data of model analytes as well as images of human skin.
[Show abstract][Hide abstract] ABSTRACT: Dendritic cells (DCs) are key activators of cellular immune responses through their capacity to induce naïve T cells and sustained effector T cell responses. This capacity is a function of their superior efficiency of antigen presentation via MHC class I and class II molecules, and the expression of co-stimulatory cell surface molecules and cytokines. Maturation of DCs is induced by microbial factors via pattern recognition receptors such as Toll-like receptors, pro-inflammatory cytokines or cognate interaction with CD4(+) T cells. Here we show that, unexpectedly, the PanDR helper T cell epitope PADRE, a generic T helper cell antigen presented by a large fraction of HLA-DR alleles, when delivered in particle-bound form induced maturation of human DCs. The DCs that received the particle-bound PADRE displayed all features of fully mature DCs, such as high expression of the co-stimulatory molecules CD80, CD86, CD83, the MHC-II molecule HLA-DR, secretion of high levels of the biologically active IL-12 (IL-12p70) and induction of vigorous proliferation of naïve CD4(+) T cells. Furthermore, the maturation of DCs induced by particle-bound PADRE was shown to involve sphingosine kinase, calcium signaling from internal sources and downstream signaling through the MAP kinase and the p72syk pathways, and finally activation of the transcription factor NF-κB. Based on our findings, we propose that particle-bound PADRE may be used as a DC activator in DC-based vaccines.
PLoS ONE 05/2013; 8(5):e63039. DOI:10.1371/journal.pone.0063039 · 3.23 Impact Factor