Untreated Skin Research Articles

Optimal scaling analysis of polymeric microneedle length and its effect on transdermal insulin delivery

Polymeric microneedle is a highly effective, painless and safe method for transdermal drug delivery with broad market prospect and high application potential. In this paper, a novel method for the preparation of microneedle array with flexible substrate using the method of micro-injection compression molding was proposed. The influence of microneedle length on transdermal insulin delivery was investigated basing on the testing results of drug diffusion experiments. The results showed that the polymeric microneedles prepared by micro-injection compression molding had enough mechanical strength to pierce the skin without breaking. The correlations between dimensionless permeation amount of insulin and the ratio of surface area to microneedle length as well as the ratio of skin thickness to microneedle length were both obtained through optimal scaling analysis. The insulin penetration efficiency presented a 2.29 times improvement after treated with 337 μm microneedle compared with that of untreated skin. The modification of polymeric microneedles could enhance the transdermal penetration efficiency of insulin, and also provide technical basis for the study of transdermal penetration.

Evaluation of chemical treatment combined with vacuum and ultrasonication with a water resonance system for reducing Campylobacter on naturally contaminated chicken carcasses

The present study was conducted to evaluate the efficacy of a new technique for removal of campylobacters from naturally contaminated free-range and broiler chicken carcasses. The carcasses, obtained from commercial processing plants after evisceration, were immersed in 0.1% cetylpyridinium chloride (CPC) or 0.01% sodium hypochlorite (NaOCl) and exposed to a vacuum of −0.02 MPa to remove air from feather. The carcasses were then immersed in a disinfectant solution in a steel container and subjected to ultrasonication with a water resonance system (WRS) at 130 kHz. Tap water was also used instead of the disinfectant, and other carcasses were immersed in each chemical alone. The Campylobacter count on breast and back skin was determined by the most-probable-number (MPN) method. This new treatment using CPC and NaOCl significantly reduced the Campylobacter count on back and breast skin. Treatment with 0.1% CPC combined with vacuum and ultrasonication with the WRS was the most effective, achieving a Campylobacter reduction of 1.36–1.64 log10 MPN/10 g and 0.94–1.16 log10 MPN/10 g on free-range and broiler chicken carcasses, respectively. Scanning electron microscopy (SEM) confirmed that this treatment successfully removed debris from breast skin in comparison to untreated skin. These results suggest that the use of sanitizers employing a combination of vacuum and ultrasonication with a WRS is highly effective for reducing the count of campylobacters on chicken carcasses.

Human Epidermal Zinc Concentrations after Topical Application of ZnO Nanoparticles in Sunscreens

Zinc oxide nanoparticle (ZnO NPs) sunscreens are generally considered safe because the ZnO NP do not penetrate through the outermost stratum corneum (SC) layer of the skin. However, cytotoxicity of zinc ions in the viable epidermis (VE), after release from ZnO NP and penetration into the VE, is ill-defined. We therefore quantified the relative concentrations of endogenous and exogenous Zn using a rare stable zinc-67 isotope (67Zn) ZnO NP sunscreen applied to human skin and the cytotoxicity of human keratinocytes (HaCaT) using multiphoton microscopy, zinc-selective fluorescent sensing and laser-ablation inductively-coupled-plasma mass-spectrometry (LA-ICP-MS) methodology. Multi-photon microscopy with second harmonic generation showed that 67ZnO NPs were retained on the surface or within the SC. Zn fluorescence sensing revealed higher levels of labile and intracellular zinc in both SC and VE relative to untreated skin, confirming that zinc permeated through the SC into the VE as ionic Zn and not as ZnO NPs. Importantly, the LA-ICP-MS estimated exogenous 67Zn concentrations in the VE of 1.0 ± 0.3 mg/mL are much lower than that estimated for endogenous VE zinc of 4.3 ± 0.7 mg/mL. Further, their combined total zinc concentrations in the VE are much less than the exogenous zinc concentration of 21 to 31 mg/mL causing VE cytotoxicity, as defined by the from half maximal inhibitory concentration of exogenous 67Zn found in human keratinocytes (HaCaT). This speaks strongly for the safety of ZnO NP sunscreens applied to intact human skin and the associated recent US FDA guidance.

Phytochemical screening and cytotoxicity evaluation of crude extracts: Toxicity comparison of crude extracts and its ethosomal formulations.

Combined plant extracts of Phyllanthus niruri, Croton tiglium, and Zingiber officinale are reported to have potential pharmacological applications. Ethosomes have a unique ability of encapsulating drugs or plant extracts with varying hydrophobicities in the phospholipid bilayer. To explore cytotoxicity of the combined plant extracts and ethosome loaded combined plant extracts for topical delivery. To study effect of ethosomes loaded combined plant extracts using HaCaT cells model treated with testosterone. Dried powder of plant was extracted with ethanol using Soxhlet and cold macerations. Total phenolic and flavonoid contents were also determined using established methods. The combined extract loaded ethosome formulation was prepared by solvent dispersion method. The plant extracts loaded ethosomes formulation with a vesicle size range 1524.6-167.7nm was prepared. HaCaT cells treated with testosterone negative control showed an IC50 value of 27±1.0. Thw standard marketed topical minoxidil (1% solution) treated cells with testosterone showed an IC50 value 33±1.0 and the combined plant extracts loaded ethosomes with testosterone showed an IC50 value 30±1.0. Morphological alterations of rat skin exposed to the combined plant extract loaded ethosomes solution were assessed and compared with untreated skin and negative control. The preclinical safety was investigated employing an in vitro cytotoxicity and histopathological study. The cell line study results confirmed that the combined plant extracts loaded ethosomes inhibits testosterone and increase cell viability closer to that of standard drug minoxidil. According to our histopathological study, the combined plant extract loaded ethosomal formulations did not cause any damage to the rat skin layer.

Improvement with Fractional Ablative Laser and Topical Poly-L-Lactic Acid in an RDEB Patient

Recessive dystrophic epidermolysis bullosa (RDEB) results from mutations in the COL7A1 gene for type VII collagen. Affected patients demonstrate skin fragility and blistering with scarring. Associated morbidity is great, and there are limited therapeutic options. Fractional carbon dioxide (CO2) ablative laser (FABL) treatments have been shown to promote collagen remodeling and demonstrate synergy with topical poly-l-lactic acid (PLLA) applications. We report the case of a young woman with RDEB who experienced decreased blistering, decreased healing time, and improved quality of life in a local skin area treated with FABL followed by topical PLLA. In comparing biopsy specimens from treated and untreated skin, improved collagen organization and collagen maturity were observed by histopathological examination including with Herovici staining. Moreover, type VII collagen, absent in untreated skin, was detected in treated affected skin by immunostaining. This report illustrates a treatment response to FABL and topical PLLA in a patient with RDEB including enhanced healing time, collagen remodeling, and type VII collagen expression. The findings are promising and, with further study, may have broader implications for treating this disorder.

Dendrobium orchid polysaccharide extract: Preparation, characterization and in vivo skin hydrating efficacy

ObjectiveDendrobium spp., the major globally commercializing tropical orchid, has been continuously used in Chinese medicine. However, preparation and standardization of this therapeutic orchid including its clinical evidence for topical application are sparely to be exploited. MethodsMaceration of the white orchid (Dendrobium cv. Khao Sanan) in water was varied on the extraction time. The extraction yields and total polysaccharide content of the extracts were compared. The best extract condition was selected, standardized, and evaluated on safety and skin hydrating efficacy in human volunteers. ResultsThe extraction for 3 h gave the extract with polysaccharide content that significantly (P < 0.001) greater than the other extraction times with the following specification: yield [(35.60 ± 2.51)%], moisture content [(7.65 ± 0.21)%], total polysaccharide, sugar, reducing sugar and ash content [(72.95 ± 2.37)%, (34.38 ± 0.00)%, (16.28 ± 0.00)%, and (0.12 ± 0.00)%], pH (4.42 ± 0.01), viscosity [(15.07 ± 0.12) cps] swelling and water absorption capacities [(10.00 ± 0.00)%, (1.19 ± 0.24) g/g]. This antioxidative polysaccharide caused no skin irritation with a better skin hydrating efficacy than the untreated skin and the benchmark as examined in 22 Thai volunteers. ConclusionWhite Dendrobium is ready to be supplied as a specialty ingredient with a safe and efficient profile for skin dryness therapy. This innovative application of the identified traditional Chinese medicinal herb flows in the main stream of the consumers’ preferences and demand upon natural derived products.

610 Hibiscus abelmoschus seed extract induces remodelling of the papillary dermal fibrillin-rich microfibrils

Hibiscus abelmoschus seed extract is used in anti-ageing cosmetics and improves both the firmness of skin and reduces the appearance of wrinkles. Whilst it promotes collagen production, less is known about its wider impact upon the remodelling of the dermal extracellular matrix (ECM). In this study we investigated its effects upon the expression and deposition of key ECM biomarkers. A formulation containing the botanical derivative was prepared as an oil in water gel cream and applied to photoaged extensor forearm of healthy individuals (n=9; aged 61-79). The test product and vehicle control (30 μl) were re-applied every 4 days over a 12-day period under occlusion; all-trans retinoic acid (RA; 0.025%) was applied for the final 4 days, as a positive control. Skin biopsies (3 mm diameter) were obtained from all sites and the expression of selected biomarkers compared against baseline biopsies derived from an area of untreated but occluded extensor forearm skin. Epidermal thickness was significantly increased in skin treated with RA (63.6 ± 23 μm) compared to baseline (37.1 ± 5.8 μm; p< 0.0001). Epidermal convolution, and elastic fibre deposition, as determined by Weigert’s Fuchsin staining, were found to be unaffected by any of the applications. Using a semi-quantitative ordinal scale to assess de novo ECM deposition, papillary dermal fibrillin-rich microfibrils were found to be significantly increased in response to treatment with H. abelmoschus (3.42 ± 0.53, p<0.01) or RA (3.4 ± 0.46, p<0.01) compared to the baseline (2.48 ± 0.52). There was no significant difference between the baseline and the vehicle control (vehicle, 2.99 ± 0.85). This study suggests that remodelling and new synthesis of papillary dermal fibrillin-rich microfibrils, induced by the H. abelmoschus seed extract, may contribute towards the marked improvement in the appearance of age-related wrinkles when included in over-the-counter anti-ageing cosmetics.

SToP (See, Treat, Prevent) skin sores and scabies trial: study protocol for a cluster randomised, stepped-wedge trial for skin disease control in remote Western Australia

IntroductionSkin is important in Australian Aboriginal culture informing kinship and identity. In many remote Aboriginal communities, scabies and impetigo are very common. Untreated skin infections are painful, itchy and frequently...

Controlled mechanical vibration and impacts on skin biology.

Different biological models have shown how mechanical stimulation may induce physiological responses from solicited cells, tissues, or organs. In models of cultured skin cells, the frequency of the mechanical stress appears to be a paramount parameter, generating a biological response in some cells, particularly from dermal fibroblasts. Our objective was to explore in ex vivo human skin explants the effects of mechanical stimulation. Mechanical stimulations were provided by a torque test device, with different end effectors, able to generate cyclic strains at different frequencies (from 40 to 120Hz). Skin explant samples were stimulated twice daily by the device for one minute, over 10days. At days 0, 5, and 10, samples were processed by immunohistological procedures, allowing some structural dermal proteins to be quantified (fluorescence). As compared to untreated skin explant samples, the stimulation procedure clearly led some proteins of the dermal-epidermal and some dermal proteins to be overexpressed. This stimulation was found to be frequency-dependent, with the greatest overall increases occurring at 60 and 90Hz. For the first time, ultrafast ultrasound imaging in vitro (phantom mimicking skin mechanical properties) was used to analyze mechanical waves transmitted to the skin layers as a function of end effector shape.

Bilosomes as a novel carrier for the cutaneous delivery for dapsone as a potential treatment of acne: preparation, characterization and in vivo skin deposition assay

In our study, the potential of bilosomes as novel vesicular carrier for the cutaneous delivery of the sulphone compound, Dapsone, for topical treatment of acne was investigated. The effect of different formulation variables (type and concentration of bile salt, and molar ratio of Span 60:cholesterol) on the properties of DPS-loaded bilosomes was investigated using a full factorial design. Design Expert software was used for data analysis and optimization of DPS-loaded bilosomes. The optimized bilosomes, chosen on the basis of their superior properties giving maximum entrapment, in vitro release after different time intervals and RE% with minimum vesicle size. Results showed that the bilosome system prepared using Span® 60: Cholesterol (5:1) and containing 0.25 M sodium deoxycholate as the bile salt was found to obey these criteria, with a desirability value of 0.637. Therefore, this system was chosen for further assessment for its morphological properties, zeta potential, thermal analysis using differential scanning calorimetry and X-ray diffractometry. Results revealed that the chosen bilosomes were spherical in shape with no aggregation, and contained DPS in a molecularly dispersed amorphous form. Finally, the capability of the optimized DPS-loaded bilosomes to deliver DPS through rat skin layers will be investigated and compared with that of DPS alcoholic solution. Results showed that the amounts of DPS retained in the skin treated with DPS-loaded bilosomes, and DPS alcoholic solution after 24 h were found to be 170.57 ± 55.12 and 120.24 ± 10.7 µg/mL, respectively, representing about 1.5-fold higher drug retained in the bilosomes-treated skin. Finally, the safety and the tolerability of the prepared bilosomes were assessed using histopathological examination, and revealed that the control untreated skin sections and skin sections treated with DPS-loaded bilosomes showed normal histological structures characterized by absence of defects or inflammation. Such results can be considered a good addition in the field of pharmaceutical drug delivery for effective topical therapy of acne.

A novel water-based anti-aging suncare formulation provides multifaceted protection and repair against environmental aggressors: evidence from in vitro, ex vivo, and clinical studies.

BackgroundUltraviolet (UV) radiation is an established cause of skin aging, and the role of pollution is increasingly acknowledged. In this study, we evaluated the efficacy of an antipollution and anti-aging suncare product in in vitro, ex vivo, and clinical studies.MethodsWe assessed 1) sunburn cell (SBC) and cyclobutane pyrimidine dimer (CPD) formation and gene expression profile in reconstructed human epidermis following solar irradiation, 2) malondialdehyde (MDA) level, Nrf2 immunostaining, and genetic expression in skin explants exposed to pollution, 3) carbon particle adhesion to healthy forearm skin in a clinical study, and 4) skin firmness, elasticity, and pigmentation spots in healthy women following 56 days of application.Results1) The product fully protected against CPD formation, and provided a high protection against SBC formation, with levels close to non-irradiated samples. Expression of genes encoding pro-inflammatory and oxidative stress response markers was lower in product-treated than untreated skin. 2) Compared with pollution-exposed untreated controls, product-treated skin had 23% lower MDA levels (P<0.01), weaker Nrf2 immunostaining, and attenuated upregulation of CYP1A1. 3) The product significantly decreased adhesion of carbon particles to the skin (15.2% less than control; P<0.01). 4) Clinically, product use led to a decrease in brown spots, with a relative reduction in the count of −1.9% (NS), and in area, −5.0% (P<0.01), and decrease in UV spots, with a relative reduction of −6.9% (P<0.01) and −9.3% (P=0.02) for count and area, respectively. Firmness increased significantly by 14.1% (P<0.01). Participants reported skin was more even in tone (80%), more moisturized (93%), and firmer (74%).ConclusionThis water-based anti-aging SPF50 suncare formulation containing photolyase encapsulated in liposome, active biopeptides, antioxidants, and hyaluronic acid provides multifaceted protection and repair action against pollution and UV-induced skin aging, ideal for everyday use.

ATR-FTIR Characterization of Janus Nanoparticles—Part II: Follow-Up Skin Application

Attenuated total reflection by Fourier transform infrared (ATR-FTIR) was used to implement reliable infrared descriptors over time of Janus nanoparticles (JNP), to follow their behavior before and after cutaneous application. In the last study, ATR-FTIR spectroscopic analysis allowed us to identify the evolution of intensity ratio of ν(C=O) at 1739 cm−1 and δ(H-O-H) at 1639 cm−1 as a spectroscopic descriptor, for JNP before cutaneous application (on the CaF2 window). This descriptor can be used to follow the physical stability (presence) of nanoparticles over time. The purpose of this study was to understand the behavior of JNP on the surface of the human skin. Therefore, a comparative study with the untreated skin and the skin after cutaneous application of lipophilic phase (Labrafil) of JNP was conducted using Franz cells. The suitability of the ATR-FTIR descriptor of JNP was evaluated, and a research of other descriptors was performed to understand the interaction that may exist between nanoparticles and the skin.

Ingenol mebutate-mediated reduction in p53-positive keratinocytes in skin cancerization field directly correlates with clinical response in patients with multiple actinic keratoses.

UV radiation represents the main risk factor for non-melanoma skin cancers. Chronic UV exposure induces 'p53 patches', i.e. clonal outgrowths of keratinocytes with high nuclear expression of mutated p53, which might progress to actinic keratosis (AK) and ultimately squamous cell carcinomas (SCCs). Analysis of ingenol mebutate gel (150 and 500mcg/g) effects in the reduction in 'p53 patches' inside skin cancerization field (CF) in patients with multiple AKs of face/scalp or trunk/extremities, in order to investigate whether the expected reduction in p53+ keratinocytes might have a direct role in the long-term AK reduction in treated areas. We enrolled n=10 patients, treated with ingenol mebutate and evaluated at 2 and 6months after treatment. We observed clinical responses in the majority of patients (n=7), with AK reduction or complete clearance (n=6 and n=1, respectively). Notably, two patients did not respond to the treatment, and in one patient, after initial partial response, new lesion was recorded. In untreated skin CF samples (n=3), we observed numerous p53+ keratinocytes, similar to those observed in invasive SCC samples (53.56±8.79 and 74.34±22.05, respectively; P=0.2). After treatment, we observed a variable p53+ keratinocyte reduction in CF samples at 2months (24.67±31.19; P=0.19). Importantly, the amount of p53+ keratinocytes strongly and directly correlated with AK number (R2 =0.81). Untreated skin CF expresses high level of p53+ keratinocytes as invasive SCC. Ingenol mebutate is able to reduce p53+ keratinocytes with variable efficacy, this reduction degree directly correlating with clinical efficacy.

Transdermal absorption of active substances from cosmetic vehicles.

Cosmetic products mean any substance or mixture intended to be placed in contact with the external parts of the human body (eg, epidermis, lips) and should not pass to the lower parts and penetrate to the skin. As a part of evaluation of cosmetic safety, the transdermal absorption of substances should be investigated. In vitro absorption was investigated with Franz diffusion cells on untreated porcine skin and specimens of the same treated with 15%wt. SLS. The integrity of the skin was discerned by gauging transdermal electrical conductivity (TEC), the concentration of caffeine absorbed by the samples of skin membrane by liquid chromatography, which took place by applying an emulsion and/or a gel containing active hydration agents (urea, sodium hyaluronate, and sericin). The greatest extent of caffeine penetration was seen for pretreatment with just SLS; similar results were in skin treated with the base gel with 10%wt. urea. In the skin treated with the base emulsion only, the amount of caffeine absorbed was twofold less; this increased after adding the active hydration substances. The values measured for TEC corresponded with the amount of caffeine absorbed. The gel proved to be the more potent vehicle for the active ingredient, as it demonstrated greater transdermal caffeine penetration than the emulsions, correlating with the degree of damage to the skin as detected by TEC.

Inhibitory effects of Sanguisorba officinalis root extract on HYBID (KIAA1199)‐mediated hyaluronan degradation and skin wrinkling

Hyaluronan (HA), an important constituent of extracellular matrix in the skin, has many biological activities such as hydration that contributes to firmness and bounciness of the skin. We have reported that reduction in HA in the papillary dermis and over-expression of HYBID (HYaluronan Binding protein Involved in hyaluronan Depolymerization, alias KIAA1199 or CEMIP), a key molecule for HA degradation in skin fibroblasts, are implicated in facial skin wrinkling in Japanese and Caucasian women. However, little or no information is available for substances which inhibit the HYBID-mediated HA degradation. Inhibition of Sanguisorba officinalis root extract and ziyuglycoside I, one of the components of Sanguisorba officinalis root extract, to the HYBID-mediated HA degradation was assessed by size-exclusion chromatography of HA depolymerized by stable transfectants of HYBID in HEK293 cells (HYBID/HEK293 cells) or normal human skin fibroblasts (Detroit 551 cells and NHDF-Ad cells). The HYBID mRNA and protein expression was examined by quantitative real-time PCR and immunoblotting in the skin fibroblasts treated with Sanguisorba officinalis root extract, and size distribution of newly produced HA was evaluated by preparing metabolically radiolabelled HA. A double-blind, randomized and placebo-controlled study was carried out in the 21 healthy Japanese women, who were topically treated with the formulation containing Sanguisorba officinalis root extract or the placebo on each side of the face including crow's foot area. Sanguisorba officinalis root extract, but not ziyuglycoside I, abolished HYBID-mediated HA degradation by HYBID/HEK293 cells. Sanguisorba officinalis root extract also inhibited HYBID-mediated HA degradation in skin fibroblasts by down-regulating HYBID mRNA and protein expression. Although control untreated skin fibroblasts produced polydispersed HA, the cells treated with Sanguisorba officinalis root extract produced only high-molecular-weight HA. Treatment with Sanguisorba officinalis root extract-formulated lotion significantly improved skin elasticity, and reduced skin wrinkling scores at the outer eye corner compared with the placebo formulation. Sanguisorba officinalis root extract showed an anti-HYBID-mediated HA degradation activity and anti-wrinkle activity on human facial skin, which is accompanied by the improvement in elasticity. Our study provides the possibility of a new strategy to inhibit HYBID-mediated HA degradation for anti-wrinkle care.

No effect of Turkish bath (Hammam) on sunburn after exposure to natural ultraviolet radiation: A randomized controlled blinded trial.

Scrubbing of the skin during Hammam, a Turkish spa treatment, removes cells from the ultraviolet radiation absorbing stratum corneum. The aim of this study was to assess if skin is more susceptible to sunburn after a Hammam treatment. In this randomized controlled trial with a paired design, healthy adult participants with skin types 1-3 were randomized to receive Hammam treatment on one side of the back while the other side was used as control. Participants had 30minutes of sun exposure on their backs. Erythema and pain were assessed before and 1, 4, 8, and 24hours after sun exposure. Erythema was assessed by a visual scale and by using chromatography on high-resolution photos. Pain was assessed with the Visual Analogue Scale (VAS). The primary outcome was the difference in chromatography-assessed erythema from baseline to the 24-hour assessment, compared between the Hammam- and the control side. Secondary outcomes were differences in pain and visually assessed erythema. Twelve participants were included with complete follow-up. Eight participants had type-3 skin and four had type-2 skin. There was no difference between the Hammam and the control side for any of the outcomes. Even though erythema assessed by chromatography was slightly higher for the Hammam side, the difference was insignificant. Hammam treatment did not increase sunburn, measured by UV-induced skin erythema and pain, compared with untreated skin following 30 minutes of sun exposure.

Shelf life determination of durian (Durio zibethinus) paste and pulp upon highpressure processing

Globally, there has been an increase in demand for durian. Therefore, it is important to extend the shelf life and at the same time, maintain the quality of durian. This study investigated the effect of high-pressure processing (HPP) on the shelf life of durian paste and pulp. Specifically, HPP treatments of 500 MPa for 5 min on nylon- and skin film-packed durian pulp, and 600 MPa for 5 min on nylon-packed durian paste were applied. It was found that throughout the 56-day storage period, the total soluble solids (TSS), pH and titratable acidity of durian paste and pulp showed no significant changes (p>0.05). The colour (L* values) of nylon-packed untreated durian pulp showed no significant changes (p>0.05), while the untreated durian paste and skin film-packed durian pulp showed significant changes (p 0.05) throughout the storage period. The microbial levels of all the high- pressure (HP)-treated durian samples remained below the detection limit till the end of the storage study. HP-treated durian samples showed lower enzymatic (polygalacturonase and pectin methyl esterase) activities compared to untreated sample throughout the storage period. Sensory evaluation showed no significant difference (p>0.05) between untreated and HP-treated samples. In summary, HPP effectively maintained the overall quality of durian for a minimum of 56 days.

Inhibition of HYBID (KIAA1199)-mediated hyaluronan degradation and anti-wrinkle effect of Geranium thunbergii extract.

Hyaluronan (HA) is an essential constituent of extracellular matrix in the skin. HA reduction in the dermis and overexpression of HYBID (KIAA1199), a key molecule for HA degradation in skin fibroblasts, are implicated in facial skin wrinkling. We aimed to obtain anti-wrinkle agent(s) by screening for inhibition of HYBID-mediated HA degradation. Various plant extracts were screened for inhibition of HA degradation in HYBID-stable transfectants in HEK293 (HYBID/HEK293). Inhibition of HA-degrading activity and HYBID mRNA and protein expression by Geranium thunbergii extract was studied in skin fibroblasts and HYBID/HEK293 cells. Size distribution of newly produced HA was evaluated by preparing metabolically radiolabeled HA in skin fibroblasts. A double-blind, randomized, and placebo-controlled study was performed in healthy Japanese women (n=21) by topically treating each side of the face with a lotion formulated with G.thunbergii extract or placebo for 8weeks. Among the plant extracts tested, only G.thunbergii extract abolished HA depolymerization in skin fibroblasts and HYBID/HEK293 cells by down-regulating HYBID mRNA and protein expression and by inhibiting HYBID-mediated HA-degrading activity. Although untreated skin fibroblasts produced polydispersed HA, G.thunbergii extract-treated cells produced high-molecular-weight HA. Treatment with G.thunbergii extract-formulated lotion significantly improved skin elasticity and reduced skin wrinkling scores at the outer eye corner compared with the placebo formulation. Geranium thunbergii extract inhibited HYBID-mediated HA degradation in vitro and showed anti-wrinkle activity in vivo accompanying the improvement in skin elasticity. Our study provides a possible strategy for anti-wrinkle care through inhibition of HYBID-mediated HA degradation.

Protective effects of a novel facial cream against environmental pollution: in vivo and in vitro assessment.

BackgroundThe effects of pollution on health have received increasing attention in recent years. Extrinsic skin aging occurs via multiple processes, and pollution is now recognized as a major component, causing increased pigmentation and wrinkles via oxidative mechanisms. We tested the antipollution efficacy of a cosmetic facial cream (FC) by assessing its effects on carbon particle adhesion to skin and on oxidative and inflammatory pathways in the skin.MethodsIn an in vivo study, FC was applied once to the forearms of healthy subjects. Carbon E153 powder was applied, and the skin was washed under standardized conditions. Images were taken using a dermoscope to determine the area of particle adherence. Each participant served as their own control, with the contralateral forearm being untreated with the FC but otherwise following the same protocol. In a 5-day ex vivo study, skin explants were treated with the FC daily and exposed to vaporized pollutants on day 2 and day 4 via a closed system. Explants were sampled at baseline and day 5 and culture media on day 5. The parameters evaluated were cellular viability on microscopy, Nrf2 immunostaining, malondialdehyde (MDA) levels in culture, melanin levels, and gene expression profile (TYR, IL6, and CYP1A1).ResultsIn the in vivo adhesion study, after standardized washing, carbon particle deposition on skin treated with the FC was significantly lower than that on untreated skin. In the ex vivo study, samples treated with the FC had reduced Nrf2 staining and MDA levels vs polluted controls. Melanin did not change significantly. The FC modulated pollution-induced increases in CYP1A1, IL-6, and TYR.ConclusionThis FC reduces particle adhesion to skin after a single application and protects against pollution-induced oxidative and inflammatory pathways in the skin.

Abstract P236: Sodium Accumulates in the Skin of Patients and Mice With Psoriasis

Sodium can be buffered in the skin, which mechanism is altered during aging and in certain diseases such as hypertension. High salt environment can promote autoimmunity by expanding pathogenic IL-17 producing T helper (Th17) cells. Psoriasis is a relapsing and remitting inflammatory autoimmune disease affecting the skin and joints and involves proinflammatory Th17 cells. Here we tested the hypothesis if psoriatic skin has a higher sodium content in mice and humans. We used two psoriasis mouse models; the K14-IL-17A ind/+ mice overexpressing IL-17A in K14-positive keratinocytes and the imiquimod (IMQ) mouse model by applying 62.5 mg IMQ cream (5%) on the shaved back and ears of FVB/N mice for 5 days daily. End of the study skins of mice were collected, weighted, dried and ashed to measure water and sodium content. Additionally, skin sodium and water content were measured in psoriasis patients and aged matched healthy controls by non-invasive 23 Na-MRI on non-affected flexor site of the lower leg and by 23 Na-spectroscopy to compare affected and non-affected sites of the leg. K14-IL-17A ind/+ mice had significantly higher sodium content compared to control IL-17A ind/+ mice (0.191±0.021 vs. 0.137±0.023 mg/g dry weight) together with an elevated water content. IMQ-treated back skin had significantly higher sodium content compared to untreated ventral skin of the same mice (0.175±0.023 vs. 0.143±0.014 mg/g dry weight), whereas sham mice had a significantly lower content in both regions (0.116±0.010 vs. 0.107±0.005 mg/g dry weight). IMQ treatment led to significant expansion of IL-17 producing γδT cells in the skin, regional lymph nodes and in the spleen with typical skin lesions. Patients with psoriasis area and severity index (PASI) &gt;5 had significantly higher sodium content in the skin compared to those with lower PASI or with healthy controls (17.73±1.52 vs. 14.32±1.54 vs. 14.30±2.59 AU, respectively); this elevation was water coupled. PASI significantly correlated with skin sodium content (Pearson’s r=0.598, P&lt;0.001). Additionally, patients with PASI&gt;5 has higher sodium content in the affected skin compared to non-affected skin of the same patient. Data from animal models and humans argue for higher sodium accumulation in the inflamed skin.