Dermal Layer Of Skin Research Articles

Gamma oryzanol niosomal gel for skin cancer: formulation and optimization using quality by design (QbD) approach

Skin cancer is fifth most diagnosed disease in human population due to ultraviolet radiation (UV) exposure. Gamma oryzanol (OZ) is a natural antioxidant, and it also has skin anti-aging properties. OZ is naturally found in rice bran oil. The main aim of the present work was to optimize OZ niosomal formulation using quality by design approach including one variable at a time and full factorial design. Niosomes were prepared by solvent injection method and characterized for size, polydispersity index, drug entrapment, and transmission electron microscopy. The optimized batch obtained at X1 [drug to span 60 molar ratio (1:5)], X2 [volume of hydration (75 mL)], and X3 [stirring speed (2500 rpm)] to Y1 [average vesicle size (196.6 nm)] and Y2 [entrapment efficiency (78.31%)] as dependent variables. The optimized OZ noisomes were formulated by niosomal gel to provide improved physicochemical stability upon topical application against UV. The niosomal gel was characterized using pH meter, viscometer, Draize test for skin irritancy, ex vivo permeation studies, and stability studies. Ex vivo permeation studies of OZ niosomal gel not only showed fourfold higher permeation but also exhibited better drug retention in dermal layers of skin as compared to OZ gel. Quality Target Product Profile of OZ niosomal formulation was generated. Risk analysis of optimized OZ gel suggested most critical quality attributes (CQAs) and critical process parameters (CPPs) to be characterized as low risk. Thus, γ-oryzanol niosomal gel for topical use can serve as a promising prophylactic treatment in skin cancer, and the developed prototype formulation can be further extended to future newly discovered drugs with similar characteristics.Graphical abstract

Potentiality of Melittin-Loaded Niosomal Vesicles Against Vancomycin-Intermediate Staphylococcus aureus and Staphylococcal Skin Infection.

BackgroundStaphylococcus aureus is an important human pathogen, especially causing skin and soft tissue infections (SSTIs). Over the decades, the infections caused by antibiotic-resistant strains have often become life-threatening. Consequently, exploration and development of competent approaches to combat these serious circumstances are urgently required.MethodsThe antibacterial activity of melittin (Mel) on S. aureus, methicillin-resistant S. aureus (MRSA) and clinical isolates of vancomycin-intermediate S. aureus (VISA) was investigated by minimum inhibitory concentration (MIC) and time-killing assays. The localization of Mel on the bacterial cell was visualized by confocal laser scanning microscopy and its effect on the membrane was indicated based on propidium iodide uptake. The non-ionic surfactant vesicle (NISV) or niosome nanocarrier was established for Mel loading (Mel-loaded NISV) by the thin-film hydration method. Physicochemical and in vitro biological properties of Mel-loaded NISVs were characterized. The cellular uptake of Mel-loaded NISVs was evaluated by holotomography analysis. In addition, an ex vivo study was conducted on a porcine ear skin model to assess the permeation ability of Mel-loaded NISVs and their potential to inhibit bacterial skin infection.ResultsThe effective inhibitory activity of Mel on skin pathogens was demonstrated. Among the tested strains, VISA was most susceptible to Mel. Regarding to its function, Mel targeted the bacterial cell envelope and disrupted cell membrane integrity. Mel-loaded NISVs were successfully fabricated with a nano-size of 120–200 nm and entrapment efficiency of greater than 90%. Moreover, Mel-loaded NISVs were taken up and accumulated in the intracellular space. Meanwhile, Mel was released and distributed throughout the cytosol and nucleus. Mel-loaded NISVs efficiently inhibited the growth of bacteria, particularly MRSA and VISA. Importantly, they not only penetrated epidermal and dermal skin layers, but also reduced the bacterial growth in infected skin.ConclusionMel-loaded NISVs have a great potential to exhibit antibacterial activity. Therapeutic application of Mel-loaded NISVs could be further developed as an alternative platform for the treatment of skin infection via dermal and transdermal delivery.

Comparison of the Biological Behaviour of Human Dermal Fibroblasts seeded on 3D Printed Polylactic acid, Polycaprolactone and Polyethylene Terephthalate Scaffolds in vitro

Regenerative medicine is a scientific field that improves and repairs diseased and injured tissues. Three-dimensional (3D) printing is an innovative technology that provides a new application field for regenerative medicine. 3D printed scaffolds by programming pore sizes and shapes serve as a temporary basis for cells until the natural extracellular matrix (ECM) is reconstructed. Dermal fibroblasts are mesenchymal cells located in the dermal skin layer that produce and organize ECM components. They play an essential role in skin wound healing and fibrosis. The aim of this study is to analyze the viability, adhesion, distribution, and collagen IV expression of human dermal fibroblasts (HDFs) seeded on 3D printed polylactic acid (PLA), polyethylene terephthalate (PET), and poly-ε-caprolactone (PCL) scaffolds in vitro. HDFs were seeded on scaffolds or tissue culture plastic plates as control and were cultured for 1 and 3 days. 3D PLA, PCL, and PET scaffolds were prepared using a custom made fused deposition modeling printer. The cell viability was measured by WST-1 assay on days 1 and 3. The cell adhesion was evaluated by scanning electron microscopy (SEM). The distribution was analyzed by hematoxylin and eosin (H&E) staining. Collagen IV expression was analyzed by immunohistochemical (IHC) staining. On day 1, the viability of HDFs on the 3D PLA scaffolds was significantly higher than PCL scaffolds. On day 3, the viability of HDFs on 3D PLA and PET scaffolds was significantly higher than PCL scaffolds. SEM images showed that HDFs on 3D PLA scaffolds attached the surfaces, filled the interfiber gaps and maintained their tissue specific morphology on day 3 compared to PCL and PET scaffolds. Histological images stained with H&E demonstrated that the distribution of HDFs on 3D PLA scaffolds was uniform on days 1 and 3. Collagen IV staining was more intense in HDFs on 3D PLA scaffolds on days 1 and 3. This study shows that 3D PLA scaffolds create a appropriate environment for cell viability, adhesion, distribution and may provide a high advantage in skin tissue regeneration.

Optimization and evaluation of ciprofloxacin-loaded collagen/chitosan scaffolds for skin tissue engineering.

A novel ciprofloxacin-loaded collagen-chitosan scaffold was developed for the treatment of wound using freeze drying method. The average pore size and porosity of developed scaffold was found to be around 125µm and 91 ± 0.56%. Moreover, swelling, degradation and mechanical tests profile supported the suitability of scaffold for wound healing process. The scaffold has high degree of hemocompatibility towards the blood and promotes the growth, migration and proliferation of fibroblast. The developed scaffold exhibits antibacterial properties and was found to be efficient against the Gram-negative (E.coli) and Gram-positive (Staphylococcus aureus) bacteria hence can be used for wound healing applications. In vivo study demonstrated that the scaffold not only escalated the tissue regeneration time but also accelerated the wound healing process compared to control. The histological studies revealed better granulation, vascularization, and remodeling of extracellular matrix along with regeneration of epidermal and dermal layer of skin. Overall, the obtained results suggested that the developed skin tissue constructs possess the enormous potential for tissue regeneration and might be a suitable biomaterial for skin tissue engineering applications.

Amphotericin B-loaded deformable lipid vesicles for topical treatment of cutaneous leishmaniasis skin lesions.

Cutaneous leishmaniasis (CL), the most common clinical form of human leishmaniasis, is a non-fatal chronic and disabling disease characterized by erythema and nodular or ulcerative skin lesions that may cause permanent scars and disfigurement. Topical drug delivery represents a simple and efficacious approach to treat CL skin lesions. The association of drugs with nanocarrier systems enhances their permeation properties and increases the drug amount available in the dermis. Here, a deformable lipid vesicle (DLV) was optimized for the topical administration of Amphotericin B (AmB), with the aim of studying and understanding the advantages of this type of delivery system in the transport of a drug through the skin layers. AmB-DVL were characterized in terms of incorporation parameters, stability, and elasticity, and evaluated in vitro for their permeation properties, cytotoxicity, and anti-leishmanial activity. The AmB-DVL exhibited a translucent fluid gel-like aspect and a yellow color, a mean size of 132nm (PdI≤0.1), zeta potential values around zero (mV), and an AmB incorporation efficiency of 95%. Permeation and penetration assays suggest that AmB-DLV are suitable for topical administration since AmB was detected in the epidermal and dermal skin layers. AmB-DVL was able to reduce promastigote viability in a dose-dependent manner, as well as the number of intracellular amastigotes in THP-1 macrophages. Selectivity index (SI) value for AmB-DLV was considerably higher than that observed for free AmB. Results suggest that DLV may represent an attractive vehicle for dermal delivery of AmB and a new low-cost and safe therapeutic option in CL treatment.

Electrospun Asymmetric Membranes as Promising Wound Dressings: A Review

Despite all the efforts that have been done up to now, the currently available wound dressings are still unable to fully re-establish all the structural and functional properties of the native skin. To overcome this situation, researchers from the tissue engineering area have been developing new wound dressings (hydrogels, films, sponges, membranes) aiming to mimic all the features of native skin. Among them, asymmetric membranes emerged as a promising solution since they reproduce both epidermal and dermal skin layers. Wet or dry/wet phase inversion, scCO2-assisted phase inversion, and electrospinning have been the most used techniques to produce such a type of membranes. Among them, the electrospinning technique, due to its versatility, allows the development of multifunctional dressings, using natural and/or synthetic polymers, which resemble the extracellular matrix of native skin as well as address the specific requirements of each skin layer. Moreover, various therapeutic or antimicrobial agents have been loaded within nanofibers to further improve the wound healing performance of these membranes. This review article provides an overview of the application of asymmetric electrospun membranes as wound dressings displaying antibacterial activity and as delivery systems of biomolecules that act as wound healing enhancers.

IN VITRO ASSESSMENT OF THE BIOLOGICAL ACTIVITY OF A NEW REGENERATIVE AGENT PREPARED FROM THE CONCENTRATE OF DEPROTEINIZED DERMAL LAYER OF PORCINE SKIN

Background. Presently, a prospective direction for the development of regenerative medicine in the world is the search for regulatory molecules and the identification of molecular targets to stimulate the body's endogenous regenerative potential. The concentrate of the deproteinized dermal layer of porcine skin (СDDLPS) is a new therapeutic agent with restorative properties, the action of which is directed on the induction of the self resources of cells. Aim. The assessment of the effect of СDDLPS on the proliferative activity of mammalian cells of different histogenesis in vitro. Materials and Methods. To determine the amino acid composition of the СDDLPS liquid chromatography and biochemical methods were used. The biological effects and mechanisms of action of the drug were investigated by cell culture and molecular biological methods. The research was carried out using stable cell lines: human keratinocytes (HaCaT cell line), porcine endothelial cells (PAE cell line), bovine kidney cells (MDBK cell line) and mouse fibroblasts (3T3A31 cell line). Results. The cells of the bovine kidney MDBK cell line were the most sensitive to the effect of the CDDLPS. Also, the obtained results suggest that, depending on the concentration, the drug not only stimulates cell proliferation by 10–30 %, but also significantly enhances biosynthetic processes in cells, in particular, protein synthesis by 20–40 %. Conclusions. CDDLPS is an effective and affordable therapeutic agent with restorative properties, the biological activity of which manifests itself in the activation of cell biosynthetic and proliferative potentials and is comparable to effects of some growth factors, in particular epidermal growth factor

Silver Sulfadiazine's Effect on Keratin-19 Expression as Stem Cell Marker in Burn Wound Healing.

BackgroundBurn wounds are one of the causes of cutaneous injury that involve both epidermal and dermal layers of skin. Silver sulfadiazine (SSD) has been widely used to treat burn wounds, however recent studies have found the treatment to have some drawbacks, such as cellular toxicity effects. Cutaneous wound regeneration is known to start from the basal layer of the epidermal epithelial cells, which are enriched with highly proliferative cells. Keratin-19 (K19) is one of the epidermal stem cell biomarkers found in the skin. This study aims to explore the expression of K19 in burn wound tissue and to investigate the effect of SSD on its expression.MethodsWe created a burn wound model in Sprague Dawley rats and randomly divided them into control and SSD groups. Wound closure was evaluated (visitrak) overtime series followed by histological evaluation of K19 expression in the wound tissue (immunohistochemistry staining).ResultsOur model successfully represents full-thickness damage caused by a burn wound. The SSD group showed a faster reduction of wound surface area (wound closure) compared to the control group with the peak at day 18 post wounding (p < 0.05). K19 expression was found in both groups and was distributed on epidermal layers, hair follicles and dermis of granulation tissue showing similar patterns.ConclusionTopical application of SSD on burn wounds showed superiority in wound closure and is likely to have no harmful effect on epidermal stem cells. However, further study is required to investigate the effect of silver species on cell viability and toxicity effects during long term treatment.

Synthesis and Characterization of Fluorescent Surfactants for Studying the Penetration of Cosmetic Surfactants on the Skin

AbstractSurfactants are one of the important ingredients of cosmetics. However, their penetration and penetration‐promoting behavior in the skin has not been well understood. Aiming to explore the surfactant penetration in skin, a novel fluorescent surfactant F‐C16, based on sodium fluorescein, was synthesized in this paper. F‐C16 has a fluorescence maxima (λem) of around 530 nm, and its fluorescence increased with the polarity of solvent. It possesses good emulsifying properties and low cytotoxicity. Hence, it can be used as a quickly detected indicator for surfactants in cosmetics. As demonstrated by this fluorescent surfactant, it was found that the surfactants can penetrate well into skin and cells. It seems that surfactants in cosmetics can enter the cells and reach the dermal layer of skin.

SkinDot: A modified full-skin transplantation technique

Existing epidermal transplantation procedures applied in burn surgery or wound treatment, such as mesh grafting or the Meek method, do not lead to a restoration of all the skin layers. Dermal skin layers are indispensable in ensuring the quality and function of the transplanted skin as a frictional surface and a carrier of skin appendages such as hair, sweat glands, and sebaceous glands, as well as nerve receptors for detecting pressure, vibration, and temperature. Because of the restricted skin surface area that can be provided by the donor, full-skin transplants cannot be transplanted over a large area. Cultured skin procedures, based on skin cells cultivated in a laboratory, have not yet reached a stage of development where a complex full epidermal transplantation is possible. In particular, the introduction of skin appendages with a functional cell-to-cell communication has not been observed thus far in cultivated skin. Based on the Reverdin transplantation method, in which concave skin islands with epidermal and dermal parts are transplanted, Davis in 1910 described the transplantation of multiple 2−5 mm sized full-skin islands as a new method for the treatment of skin lesions. Further modifying this 100-year-old procedure, we developed a miniaturization and automation of the Davis transplantation method that started in 2011 and called it “SkinDot”. In the following article we describe the effectiveness of the full-skin island transplant procedure in two patients. The transplantation of single 2−3 mm full-skin islands results in a full-skin equivalent without any limits on donor area and with a reduced donor morbidity.

Cerium Oxide Nanoparticles Absorption through Intact and Damaged Human Skin.

Cerium oxide (CeO2) nanoparticles (NPs) are used in polishing products and absorbents, as promoters in wound healing, and as organopesticide decontaminants. While systemic bioaccumulation and organ toxicity has been described after inhalation, data on CeO2 NPs’ transdermal permeation are lacking. Our study was an in vitro investigation of the permeation of 17-nm CeO2 NPs dispersed in synthetic sweat (1 g L−1) using excised human skin on Franz cells. Experiments were performed using intact and needle-abraded skin, separately. The average amount of Ce into intact and damaged skin samples was 3.64 ± 0.15 and 7.07 ± 0.78 µg cm−2, respectively (mean ± SD, p = 0.04). Ce concentration in the receiving solution was 2.0 ± 0.4 and 3.3 ± 0.7 ng cm−2 after 24 h (p = 0.008). The Ce content was higher in dermal layers of damaged skin compared to intact skin (2.93 ± 0.71 µg cm−2 and 0.39 ± 0.16 µg cm−2, respectively; p = 0.004). Our data showed a very low dermal absorption and transdermal permeation of cerium, providing a first indication of Ce skin uptake due to contact with CeO2.

The case of non-small cell lung cancer followed by lip lymphoproliferative disease after immunotherapy

Abstract Case 74 y.o. male Chief Complaint Lower lip nodule Present History On July in XXXX, left hilar tumor was detected by chest computed tomography, and the patient was admitted to our hospital for scrutiny and treatment. It was diagnosed as stage IVa non-small cell lung cancer with undecidable histopathological subtype.Tumor proportion score of PD-L1 had shown 75%. Since Aug. in XXXX, Pembrolizumab was started. The tumor soon became smaller, so the therapy was thought to be effective. However, after 4 courses of those therapies, lower lip nodules appeared on October. Clinical Course Lower lip nodule had grown and formed a tumor in 1cm diameter, with scar in the central area and angiogenesis in the peripheral area. Lip tumor biopsy was performed at the end of October. In dermic layer, large cells with strain nucleus had increased. Abnormal cells showing CD3+, CD4+, CD8+, TIA-1+, CD30+, EBER(-), ISH(-), have indicated the possibility of T cell lymphoma. Since the association between immunotherapy and the lip nodule could not be denied, immunotherapy was ceased. In order to establish an accurate diagnosis of the lip tumor, resection of the nodule was carried out. At that time, the tumor had shrunk spontaneously, and the large abnormal cells had disappeared, which were detected in incisional biopsy. Considered with pathological findings and clinical course, primary skin lymphoproliferative diseases (LPD) had been diagnosed. As lung lesion was also enlarged simultaneously, Re-bronchoscopic exam revealed not LPDs but progression of cancer in the lung. Discussion LPD was confined in dermal layer of skin, occurred and disappeared in natural course. As the size of lip tumor decreased after cessation of immunotherapy, it was supposed that immunodeficiency due to the effect of Pembrolizumab might induce LPD.

Distribution of nickel and chromium containing particles from tattoo needle wear in humans and its possible impact on allergic reactions

BackgroundAllergic reactions to tattoos are amongst the most common side effects occurring with this permanent deposition of pigments into the dermal skin layer. The characterization of such pigments and their distribution has been investigated in recent decades. The health impact of tattoo equipment on the extensive number of people with inked skin has been the focus of neither research nor medical diagnostics. Although tattoo needles contain high amounts of sensitizing elements like nickel (Ni) and chromium (Cr), their influence on metal deposition in skin has never been investigated.ResultsHere, we report the deposition of nano- and micrometer sized tattoo needle wear particles in human skin that translocate to lymph nodes. Usually tattoo needles contain nickel (6–8%) and chromium (15–20%) both of which prompt a high rate of sensitization in the general population. As verified in pig skin, wear significantly increased upon tattooing with the suspected abrasive titanium dioxide white when compared to carbon black pigment. Additionally, scanning electron microscopy of the tattoo needle revealed a high wear after tattooing with ink containing titanium dioxide. The investigation of a skin biopsy obtained from a nickel sensitized patient with type IV allergy toward a tattoo showed both wear particles and iron pigments contaminated with nickel.ConclusionPreviously, the virtually inevitable nickel contamination of iron pigments was suspected to be responsible for nickel-driven tattoo allergies. The evidence from our study clearly points to an additional entry of nickel to both skin and lymph nodes originating from tattoo needle wear with an as yet to be assessed impact on tattoo allergy formation and systemic sensitization.

Fabrication of Bioengineered Skin by Injection Molding: A Feasibility Study on Automation.

The use of bioengineered skin has facilitated fundamental and applied research because it enables the investigation of complex interactions between various cell types as well as the extracellular matrix. The predominantly manual fabrication of these living tissues means, however, that their quality, standardization, and production volume are extremely dependent on the technician's experience. Simple laboratory automation could facilitate the use of living tissues by a greater number of research groups. We developed and present here an injection molding technique for the fabrication of bilayered skin equivalents. The tissue was formed automatically by two separate injections into a customized mold to produce the dermal and epidermal skin layers. We demonstrated the biocompatibility of this fabrication process and confirmed the resulting bilayered morphology of the bioengineered skin using histology and immunohistochemistry. Our findings highlight the possibility of fabricating multilayered living tissue by injection molding, suggesting that further investigation into this automation method could result in the rapid and low-cost fabrication of standardized bioengineered skin.

Itraconazole lipid nanocapsules gel for dermatological applications: In vitro characteristics and treatment of induced cutaneous candidiasis

There is a growing clinical demand for topical itraconazole (ITC) delivery systems because of the expanding potential of the drug for topical fungal and non-fungal applications. Lipid-based nanocarriers offer great promise in this respect. In the present study, a new topical ITC gel based on lipid nanocapsules (LNC) was developed. ITC-LNC were compared to ITC-loaded nanostructured lipid carriers (ITC-NLC) with more established benefits as topical vectors. Both nanocarriers showed high entrapment efficiency (EE > 98%). Compared to ITC-NLC, the ITC-LNC showed a significantly smaller particle size (∼50 vs 155 nm), narrower size distribution (0.09 vs 0.38), faster initial release rate under sink conditions and greater in vitro antifungal activity against Candida albicans (C. albicans) (inhibition zone 29.4 vs 26.4 mm). ITC-LNC and ITC-NLC-based gels significantly enhanced the dermal retention of ITC in excised human skin relative to a conventional ITC gel. Histopathological assessment of a 14-day treatment of induced cutaneous candidiasis in a rat model indicated efficacy of the gel preparations. Fungal elements developed in the superficial epidermal skin layer were cleared by the end of treatment. Equally important, no histopathological changes in the epidermal and dermal layers of rat skin were observed. Findings of this study verified efficacy of topical ITC in the treatment of superficial fungal infections as well as effectiveness of LNC as biomimetic nanocarrier for dermal drug delivery. Combining ITC and LNC would present a bioactive nanocarrier system with good potentials for fungal infections and other skin applications.

In vitro transdermal absorption of Al2O3 nanoparticles

Dermal exposure to Aluminium nanoparticles (AlNPs) can occur in occupationally- and non occupationally exposed- population, due to the use of Al salts-based antiperspirants. No AlNPs transdermal permeation data exists. Our study investigated in vitro the permeation of 30–60 nm Al2O3NPs dispersed in synthetic sweat (20 g/L) using excised human skin on Franz cells. Experiments were performed using intact (experiment 1) and needle-abraded skin (experiment 2). After 24 h traces of Al were detectable in receiving solution of exposed cells (35.0 ± 6.0 ng/cm2 for intact and 88.5 ± 34.2 ng/cm2 for damaged skin, mean and SD) and in blank cells (36.3 ± 7.0 ng/cm2), without statistical significance (p = 0.08, Mann-Whitney test). The average amount of Al into intact and damaged skin samples was 3.96 ± 0.20 μg/cm2 for intact and 4.36 ± 0.47 μg/cm2 for damaged skin (p = 0.08). Al content was similar in epidermal and dermal layers of intact and damaged skin (1.95 ± 0.13 μg/cm2 and 2.31 ± 0.12 μg/cm2 epidermal, 2.01 ± 0.25 μg/cm2 and 2.05 ± 0.35 μg/cm2 dermal). Al is a trace element in human body and the amount found in receiving solutions could be due as background impurity. This data suggest a reassuring transdermal permeation profile for Al2O3NPs.

Age-related biophysical changes of the epidermal and dermal skin in Korean women.

The clinical characteristics of skin were investigated to study the inter-relationship and changes in the biophysical properties of the epidermal and dermal layers associated with aging using noninvasive methods. Our study included 100 healthy women aged between the early 20s and late 60s. Biophysical characteristics of skin such as color (brightness and spots), transparency, wrinkle on crow's feet, elasticity, hydration, sebum content, glossiness, and transepidermal water loss measured under controlled conditions. This study performed in a Korean population demonstrated that aging significantly affects human skin in terms of parameters such as wrinkles, skin color, elasticity, and epidermal hydration. Age-related changes in skin hydration showed varying patterns between the epidermis and dermis. Skin color showed heterogeneous characteristics between the upper and lower epidermal layers associated with aging. Skin elasticity and wrinkles were observed to show and inversely proportional relationship in the early 40s. We confirmed the significant influence of aging on the biophysical properties of skin and determined the distinct age-related biophysical changes in the epidermal and dermal layers of skin using noninvasive method. This study indicates the need for further research to investigate the distinctive age-related changes in characteristics of the epidermal and dermal layers of human skin.

High-frequency (20-MHz) high-intensity focused ultrasound (HIFU) system for dermal intervention: Preclinical evaluation in skin equivalents.

High-intensity focused ultrasound (HIFU) for non-invasive treatment of a range of internal pathologies including cancers of major organs and cerebral pathologies is in exponential growth. Systems, however, operate at relatively low frequencies, in the range of 200-2000kHz as required for deep axial penetration of the body. HIFU utilizing frequencies in excess of 15MHz has so far not been explored, but presents an opportunity to extend the HIFU modality to target specific dermal lesions and small animal research. A new 20-MHz HIFU system (TOOsonix ONE-R) with narrow focus corresponding to the dermis was studied in acoustic skin equivalents, for example, in a tissue-mimicking gel and in bovine liver. HIFU lesion geometry, depth, and diameter were determined. The temperature increase in the focal point was measured as a function of acoustic power and the duration of HIFU exposure. The system produces highly reproducible ultrasound lesions with predictable and configurable depths of 1-2mm, thus corresponding to the depth of the human dermis. The lesion geometry was elongated triangular and sized 0.1-0.5mm, convergent to a focal point skin deep. Focal point temperature ranged between 40 and 90°C depending on the chosen setting. Observations were confirmed ex vivo in bovine liver and porcine muscle. Variation of acoustic power and duration of exposure produced linear effects in the range of the settings studied. Thus, effects could be adjusted within the temperature interval and spatial field relevant for clinical therapy and experimental intervention targeting the dermal layer of human skin. The tested 20-MHz HIFU system for dermal applications fulfilled key prerequisite of narrow-field HIFU dedicated to cutaneous applications regarding reproducibility, geometry, and small size of the applied ultrasound lesions. Controlled adjustment of acoustic lesions within the temperature range 40-90°C qualifies the system for a range of non-ablative and ablative applications in dermatological therapy.

The ethanolic extract of red cabbage (Brassica oleracea l. var,capitata f. rubra) in cream preparation to the dermal-thickness of male Wistar mice (Rattus norvegicus) after Ultraviolet-B exposure

Background: Photoaging effect in dermal layer of skin was commonly caused by Ultraviolet B (UV-B) rays exposure This study aims to assess the histological features of the dermis layer in the male Wistar mice after applied with the ethanolic extract of red cabbage following UV-B exposure.Methods: About 25 male Wistar rats were enrolled in this study by using post-test only control group design during 2 weeks. They were divided into 5 groups as follows: control group (P1), basic cream (P2), 5% ethanolic extract of red cabbage cream (P3), 10% ethanolic extract of red cabbage cream (P4), and 20% ethanolic extract of red cabbage cream (P5). The UV-B dosage used are 325 mJ/cm2 among groups prior to study. The dermis-elastic fiber thickness was assessed by Image Raster software. Statistical analysis was carried out using SPSS version 20 in measuring normality test, mean, and ANOVAResults: There was no statistically significant difference in the thickness of dermal layer among groups (P = 0.3). The mean of dermis thickness was 666.60±143.15µm in P5 group, followed by 651.19±189.18µm in P4 group, and 547.43±188.41µm, 544.76±100.67µm, and 504.10±129.40 µm in P3, P2, and P1 group respectively.Conclusion: There was no a significant difference among different dosage of the ethanolic extract cream of red cabbage to the thickness of dermal layer after UV-B exposure.

Potential protective effects of rhEGF against ultraviolet A irradiation-induced damages on human fibroblasts

BackgroundUltraviolet A (UVA) rays reach the dermal skin layer and generate oxidative stress, DNA damage, and cell inflammation, which in turn lead to photo-aging and photo-carcinogenesis. While there have been many studies about the beneficial effects of topical epidermal growth factor (EGF) treatment in the healing of wounds, the effect of EGF on UVA-induced skin irritation remains unknown. To clarify the effects of EGF on UVA-induced skin damage, it was investigated whether EGF signaling can affect intracellular reactive oxygen species (ROS) and DNA damages in UVA-irradiated human dermal fibroblasts.Materials and methodsFibroblasts cultured with or without rhEGF were UVA-irradiated at 40 mJ/cm2 twice per day for 5 days. After the irradiation, the intracellular ROS levels and expression of catalase and superoxide dismutase-1 (SOD-1) in the fibroblasts were ascertained. Further investigation to determine the effects of EGF on UVA-induced DNA damage, including a single cell gel electrophoresis assay and an enzyme-linked immunosorbent assay (ELISA), was carried out. Moreover, the NF-κB activity was ascertained in order to investigate the effects of EGF on UVA-irradiated fibroblasts.ResultsAs a result, it was revealed that recombinant human EGF (rhEGF) inhibited UVA- increased intracellular ROS in the fibroblasts and increased the expression of catalase and SOD-1. Moreover, in UVA-irradiated fibroblasts, the longest DNA-damaged tails were observed, but this phenomenon was not detected in cells cotreated with both UVA and rhEGF. Also, it was observed that DNA damage induction, including that of cyclobutene pyrimidine dimers, pyrimidine (6-4) pyrimidone photoproducts, and 8-hydroxy-2-deoxyguanosine, was caused by UVA irradiation. Similar to previous results, it was downregulated by rhEGF. Furthermore, rhEGF also inhibited NF-κB gene expression and the NF-κB p65 protein level in the nucleus induced by UVA irradiation.ConclusionThese results suggest that EGF might be a useful material for preventing or improving photo-aging.