A novel optical coherence tomography-based in vitro method of anti-aging skin analysis using 3D skin wrinkle mimics.

BACKGROUND: Wrinkles represent a characteristic symptom of skin aging. In recent years, various studies have focused on their prevention and/or cure. However, clinical tests are still the only method available to directly detect and evaluate the anti-wrinkle efficacy of various substances. Moreover, no in vitro strategy for such anti-aging skin analysis has been reported. Therefore, in this study, we aimed to develop a novel technology to overcome these limitations. MATERIALS AND METHODS: Full-thickness (FT) skin wrinkle mimics with various widths and depths were fabricated using a collagen stamping method. These were analyzed and compared using 2D and 3D Swept Source-Optical Coherence Tomography (SS-OCT) imaging technologies. RESULTS: SS-OCT demonstrated superficial and cross-sectional images of the wrinkle mimics, and the size of the wrinkles was validated using image analysis. Retinoic acid treatment significantly decreased both the depth and width of wrinkles formed in the FT skin wrinkle mimics. CONCLUSIONS: Using 3D tissue engineering and SS-OCT imaging technologies, we developed a novel in vitro technique that can directly detect skin wrinkles. This significantly efficient method could lead to an alternative strategy for animal experiments and preclinical anti-aging research on the skin.

Synergistic effects of using novel home-use 660- and 850-nm light-emitting diode mask in combination with hyaluronic acid ampoule on photoaged Asian skin: A prospective, controlled study.

BACKGROUND: Recently, light-emitting diode (LED) devices are among those mostly preferred for esthetic application because they improve the appearance of photoaged skin characterized by wrinkles, sagginess, pigmented lesions, and others. In addition, the use of hyaluronic acid (HA) for skin rejuvenation is already well proven. AIMS: This study aims to evaluate the synergistic effects of using home-use LED mask device with HA ampoule. METHODS: The total number of recruited subjects was 48:24 in Group A treated with both home-use LED mask device and HA ampoule and 24 in Group B treated with HA ampoule only, for 4 weeks. To assess the efficacy of the treatment, the following were used: Antera 3D CS, EOS 800D with Image-Pro Plus, DUB-USB, VisioFace Quick, and Visioscan VC98. RESULTS: After treatment, the volume measurement (mm3 ) for prejowl sulci and nasolabial fold flattening as well as the area measurement (pixel) for lower chin firmness improvement was significantly reduced, and the number of pores (ea) for enlarged pores as well as the desquamation index (%) for the amount of corneocytes significantly decreased in both Group A and Group B. Moreover, the percentage of skin density significantly improved. Furthermore, Group A showed a significantly faster and higher rate of improvement than Group B. CONCLUSION: The use of 660- and 850-nm home-use LED mask device can generate synergistic effects on home-use topical applications like HA on photoaged face, and such device can be safely and efficiently used daily in personal environments.

Permanent Chemotherapy-Induced Alopecia in Patients with Breast Cancer: A 3-Year Prospective Cohort Study.

BACKGROUND: Although chemotherapy-induced alopecia (CIA) is considered temporary, some patients report persistent alopecia several years after chemotherapy. There is, however, a paucity of long-term prospective data on the incidence and impact of permanent CIA (PCIA). The objective of our study was to estimate the long-term incidence of PCIA in a cohort of patients with breast cancer whose hair volume and density were measured prior to chemotherapy and who were followed for 3 years after chemotherapy. MATERIALS AND METHODS: Prospective cohort study of consecutive patients ≥18 years of age with postoperative diagnosis of stage I-III breast cancer expected to receive adjuvant chemotherapy at the outpatient breast cancer clinic at the Samsung Medical Center in Seoul, Korea, from February 2012 to July 2013 (n = 61). Objective hair density and thickness were measured using a noninvasive bioengineering device. RESULTS: The proportion of participants who had PCIA at 6 months and 3 years was 39.5% and 42.3%, respectively. PCIA was characterized in most patients by incomplete hair regrowth. Patients who received a taxane-based regimen were more likely to experience PCIA compared with patients with other types of chemotherapy. At a 3-year follow-up, hair thinning was the most common problem reported by study participants (75.0%), followed by reduced hair volume (53.9%), hair loss (34.6%), and gray hair (34.6%). CONCLUSION: PCIA is a common adverse event of breast cancer adjuvant cytotoxic chemotherapy. Clinicians should be aware of this distressing adverse event and develop supportive care strategies to counsel patients and minimize its impact on quality of life. IMPLICATIONS FOR PRACTICE: Knowledge of permanent chemotherapy-induced alopecia, an under-reported adverse event, should lead to optimized pretherapy counseling, anticipatory coping techniques, and potential therapeutic strategies for this sequela of treatment.

"Open-top" microfluidic device for in vitro three-dimensional capillary beds.

We introduce a novel microfluidic device to co-culture a blood vessel network and cell tissues in an in vivo-like niche. Our "open-top" microfluidic device is composed of microchannels with micropores in the ceiling, which provides direct fluid access from reservoir to microchannel. Fluid connections through micropores afford novel advantages, including: i) the long-term culture of large-scale microvessel network, ii) access of different fluids to inner and exterior sides of the microvessel, and iii) co-culturing of the microvessel network and small cell tissue. In this study, we have successfully assembled microvessels with 5 mm channel widths. We were also able to mimic capillary bed conditions by co-culturing microvessels with cancer spheroids. Intimate contact between the cancer spheroid and microvessel caused vessel recruitment and an increase in vessel formation, and affected vessel morphology. We expect this device to be used as a novel platform for vascularized tissue models.

Coumestrol Down-Regulates Melanin Production in Melan-a Murine Melanocytes through Degradation of Tyrosinase.

Pigmentation reflects skin darkening caused by melanin production, but excessive melanin synthesis may cause problems, such as melasma, solar lentigo, dark spots, and freckles. Considerable effort has been devoted to alleviating these undesired symptoms through the development of safe and effective depigmenting agents. Coumestrol, a plant-derived natural isoflavone with an estrogen-like structure and actions, is known to have anti-aging ability, but its potential depigmenting efficacy has not been evaluated. In the present study, we investigated the effects of coumestrol on melanin synthesis in normal melan-a murine melanocytes. Coumestrol significantly reduced melanin synthesis in a concentration-dependent manner up to a concentration of 25 µM without causing cytotoxicity. It also brightened tissue in an artificial skin model (MelanoDerm) that incorporates both human keratinocytes and melanocytes. Interestingly, although coumestrol did not inhibit tyrosinase activity or transcript level in melan-a cells, it clearly decreased the expression level of tyrosinase protein at a concentration of 25 µM. This coumestrol-induced reduction in tyrosinase protein levels was prevented by pretreatment with the proteasome inhibitor MG-132 or the lysosomal proteolysis inhibitor chloroquine. Collectively, our findings indicate that coumestrol exerts an inhibitory effect on melanin synthesis in melan-a cells, at least in part, through degradation of tyrosinase. These findings suggest that coumestrol is a good candidate for use in depigmentary reagents from a cosmetic and clinical perspective.

Comparison of skin properties in individuals living in cities at two different altitudes: an investigation of the environmental effect on skin.

BACKGROUND: Skin properties vary depending on exogenous factors. Various studies have been used for comparing skin properties between cities for studying environment influence on skin properties. However, for comparison of skin properties between cities, various environmental factors have to be considered. OBJECTIVES: The purpose of this study therefore was to compare skin properties in individuals of the same ethnicity and sex (Indonesian women) between different altitudes and to interpret the environmental effect on skin. METHODS: In this study, we reanalyzed the data obtained from previous study. The data were for healthy Sundanese Indonesian females [(n = 136) at Jakarta (n = 49) and Bandung (n = 87)], and the data consisted of published data (skin hydration, sebum level, pH, elasticity, and transepidermal water loss) and unpublished data [skin color (L*, a*, and b*)]. The skin parameters were measured on Indonesian females aged 20-34 using C+K devices (corneometer, sebumeter, pH meter, and cutometer), Delfin vapometer, and Minolta spectrophotometer, respectively. RESULTS: Sundanese Jakarta (low-altitude) females had higher sebum level and greater redness (a*) value in the forehead than Sundanese Bandung (high-altitude) females. In contrast, Bandung females had higher skin pH, brighter skin color, and greater forehead skin elasticity than Jakarta females. CONCLUSIONS: The skin properties can be influenced by changing altitude because different altitudes have different environments such as air temperature, humidity, UV radiation, and so on, and it is also necessary to investigate the factors which can influence with perceived skin condition such as skin type and skin concerning.

Histone H3K27 Demethylase JMJD3 in Cooperation with NF-κB Regulates Keratinocyte Wound Healing.

Histone H3K27me3 demethylase JMJD3 has been shown to be involved in keratinocyte differentiation and wound healing. However, the exact molecular mechanism underlying JMJD3-mediated keratinocyte wound healing has not been fully elucidated. In this study, we report on the biological function of JMJD3 in keratinocyte wound healing using in vitro cell and in vivo animal models. Our results indicate that JMJD3 up-regulation and NF-κB activation occur in the region of the wound edge during keratinocyte wound healing. We also found that JMJD3 interacts with NF-κB, resulting in increased expression of the inflammatory, matrix metalloproteinase, and growth factor genes via demethylation of H3K27me3 at the gene promoters. Consistently, inactivation of JMJD3 or NF-κB resulted in aberrant keratinocyte wound healing. Our study suggests that regulation of JMJD3 may provide a new therapeutic intervention for treating the chronic skin wound.

Different effects of five depigmentary compounds, rhododendrol, raspberry ketone, monobenzone, rucinol and AP736 on melanogenesis and viability of human epidermal melanocytes.

Numerous medications are used to treat hyperpigmentation. However, several reports have indicated that repeated application of some agents, such as rhododendrol (RD), raspberry ketone (RK) and monobenzone (MB), can be toxic to melanocytes. Although these agents had severe side effects in human trials, no current in vitro methods can predict the safety of such drugs. This study assessed the in vitro effects of five depigmentary compounds including leukoderma-inducing agents. In particular, we determined the effects of different concentrations and exposure times of different depigmentary agents on cell viability and melanogenesis in the presence and absence of ultraviolet B (UVB) radiation. Concentrations of RD, RK and MB that inhibit melanogenesis are similar to concentrations that are cytotoxic; however, concentrations of rucinol (RC) and AP736 that inhibit melanogenesis are much lower than concentrations that are cytotoxic. Furthermore, the concentrations that cause toxic effects depend on exposure duration, and prolonged exposure to RD, RK and MB had more cytotoxic effects than prolonged exposure to RC and AP736. The cytotoxic effects of RD and RK appear to be mediated by apoptosis due to increased expression of caspase-3 and caspase-8; UVB radiation increased the cytotoxicity of these agents and also increased caspase activity. Our results indicate that different leukoderma-inducing compounds have different effects on the viability of normal epidermal melanocytes and suggest that the in vitro assay used here can be used to predict whether an investigational compound that induces leukoderma may lead to adverse effects in human trials.

Quantitative changes in skin composition parameters due to chemotherapy in breast cancer patients: a cohort study.

The objective of this study is to evaluate objective changes in water content, sebum content, transepidermal water loss (TEWL), and melanin due to breast cancer chemotherapy, and their association with subjective symptoms. Prospective cohort study of 61 patients 18 years of age or older with a postoperative diagnosis of stage I-III breast cancer, who received adjuvant chemotherapy between February and September 2012 at an outpatient breast cancer clinic in Korea. Objective skin parameters, measured using a noninvasive bioengineering device, and patient-reported dryness and dullness were assessed before chemotherapy, after two cycles of chemotherapy, and 1, 3, and 6 months after completion of chemotherapy. Water content (-6.5 %), sebum (-75.5 %), and TEWL (-22.4 %) significantly decreased during chemotherapy compared to pre-chemotherapy levels (all p values <0.001). These parameters were lowest at 1 month after completion of chemotherapy and recovered thereafter but did not return to baseline levels after 6 months of follow-up. Melanin increased during chemotherapy with respect to pre-chemotherapy levels (8.4 %; p < 0.001) but decreased from the first month after completion of chemotherapy through the end of follow-up (-17.1 %; p < 0.001). The patterns of skin changes were similar in patients with or without hormone therapy. Most of patients reported dryness (57.9 %) and dullness (49.1 %) after chemotherapy, and patient-reported dryness was significantly associated with decreased sebum content. Chemotherapy-induced substantial changes in objective skin composition parameters. These changes persisted after 6 months from completion of chemotherapy and were associated with patient-reported symptoms. Additional research is needed to translate these findings into interventions for improving the dermatologic quality of life of breast cancer patients undergoing chemotherapy.

Engineering a Blood Vessel Network Module for Body-on-a-Chip Applications.

The blood circulatory system links all organs from one to another to support and maintain each organ's functions consistently. Therefore, blood vessels have been considered as a vital unit. Engineering perfusable functional blood vessels in vitro has been challenging due to difficulties in designing the connection between rigid macroscale tubes and fragile microscale ones. Here, we propose a generalizable method to engineer a "long" perfusable blood vessel network. To form millimeter-scale vessels, fibroblasts were co-cultured with human umbilical vein endothelial cells (HUVECs) in close proximity. In contrast to previous works, in which all cells were permanently placed within the device, we developed a novel method to culture paracrine factor secreting fibroblasts on an O-ring-shaped guide that can be transferred in and out. This approach affords flexibility in co-culture, where the effects of secreted factors can be decoupled. Using this, blood vessels with length up to 2 mm were successfully produced in a reproducible manner (>90%). Because the vessels form a perfusable network within the channel, simple links to inlets and outlets of the device allowed connections to the outside world. The robust and reproducible formation of in vitro engineered vessels can be used as a module to link various organ components as parts of future body-on-a-chip applications.

A quantitative evaluation method using processed optical images and analysis of age-dependent changes on nasolabial lines.

BACKGROUND/PURPOSE: Nasolabial lines (NL) and wrinkles of the face are major features of aging. Wrinkles have been studied widely by morphological methods using 3-dimensional (3D) photographic analysis instrument, but NL were evaluated by visual scoring usually. To evaluate NL quantitatively, another method is needed. This study is purposed to find out quantitative method for evaluation of NL. METHOD: One hundred Korean female subjects aged 20 to 60 were recruited in this study. Facial image was taken using light source adjusted VISIA-CR(®) and 3-dimensional wrinkle depth on the NL area was evaluated by Phase shift rapid in vivo measuring of human skin (PRIMOS(®)). The pixel number of NL area and the angle were obtained using processed images. The severity of NL was assessed by visual score. Skin elasticity was measured by Cutometer(®) MPA580. Statistical significance was determined at P < 0.05. RESULT: The optical images obtained by light source adjusted VISIA-CR(®) were easy to distinguish NL and significantly increased age-dependently. And three parameters of elasticity (R2, R5, and R7) on NL area were gradually decreased with age. The Pearson correlation coefficient was -0.756 (P < 0.01) between R7 parameter and ages. Also the pixel number of NL area, angle, wrinkle depth on the NL area (Ra), and visual score were decreased elasticity-dependently. The pixel number of NL area was highly related to Ra (r = 0.567, P < 0.01) and visual score (r = 0.647, P < 0.01). CONCLUSION: This study has shown that NL severity is related to decrease of dermal elasticity and age using quantitative new method by processing optical images.

Mitochondrial dynamics regulate melanogenesis through proteasomal degradation of MITF via ROS-ERK activation.

Mitochondrial dynamics control mitochondrial functions as well as their morphology. However, the role of mitochondrial dynamics in melanogenesis is largely unknown. Here, we show that mitochondrial dynamics regulate melanogenesis by modulating the ROS-ERK signaling pathway. Genetic and chemical inhibition of Drp1, a mitochondrial fission protein, increased melanin production and mitochondrial elongation in melanocytes and melanoma cells. In contrast, down-regulation of OPA1, a mitochondria fusion regulator, suppressed melanogensis but induced massive mitochondrial fragmentation in hyperpigmented cells. Consistently, treatment with CCCP, a mitochondrial fission chemical inducer, also efficiently repressed melanogenesis. Furthermore, we found that ROS production and ERK phosphorylation were increased in cells with fragmented mitochondria. And inhibition of ROS or ERK suppressed the antimelanogenic effect of mitochondrial fission in α-MSH-treated cells. In addition, the activation of ROS-ERK pathway by mitochondrial fission induced phosphorylation of serine73 on MITF accelerating its proteasomal degradation. In conclusion, mitochondrial dynamics may regulate melanogenesis by modulating ROS-ERK signaling pathway.

Depigmentation effect of kadsuralignan F on melan-a murine melanocytes and human skin equivalents.

The development of melanogenic inhibitors is important for the prevention of hyperpigmentation, and, recently, consideration has been given to natural materials or traditionally used ingredients such as Chinese medicine. The aim of this study is the evaluation of a new anti-melanogenic candidate, kadsuralignan F, from the natural plant Kadsura coccinea, as well as the determination of mechanisms of melanogenesis inhibition at a molecular level. Kadsuralignan F significantly reduced melanin synthesis in a dose-dependent manner in a murine melanocyte cell line and human skin equivalents. There was no direct inhibition on mushroom tyrosinase or cell-extract tyrosinase activity, and mRNA expression of tyrosinase and other melanogenic genes such as tyrosinase-related protein-1 (trp-1) or trp-2 were not affected by kadsuralignan F. Interestingly, the protein level of tyrosinase was dramatically downregulated with kadsuralignan F treatment. We found that a decrease of tyrosinase protein by kadsuralignan F was fully recovered by MG132, a proteasome inhibitor, but not by chloroquine, a lysosome inhibitor. In this study, we found that kadsuralignan F, a lignan from an extract of Kadsura coccinea, has an inhibitory activity on melanin synthesis through tyrosinase degradation. These findings suggest that kadsuralignan F can be used as an active ingredient for hyperpigmentation treatment.

Identification of sake extract as a new anti-melanogenic ingredient by in vitro and clinical trials.

Overproduction of melanin is the cause of skin hyperpigmentation, which is related to several skin diseases and cosmetic concerns. Sake is a Japanese alcoholic beverage produced from rice and water by fermentation, but is little known for its effect on melanogenesis. To identify the effect of sake extract on melanin synthesis, a melanin assay was performed in melan-A murine melanocytes. Sake extract treatment significantly inhibited melanin production in a dose-dependent manner, and tyrosinase, the rate-limiting enzyme of melanogenesis, decreased significantly at the protein level. Further investigations were performed with multiple assay systems; a sake extract reduced melanin production in melan-A/SP-1 murine cell co-culture, and also in MelanoDerm, a skin equivalent model of human keratinocytes-melanocytes. Finally, subjects were treated with a formula containing the sake extract. Topical application of the sake extract product improved skin lightness (L*) significantly within 7 days. We identified sake extract as a new anti-melanogenic ingredient through in vitro and in vivo experiments. These results suggest that a sake extract can be used to improve skin hyperpigmentation.

Activation of transient receptor potential melastatin 8 reduces ultraviolet B-induced prostaglandin E2 production in keratinocytes.

Transient receptor potential melastatin 8 (TRPM8) is a member of the TRP family, and is activated at temperatures below 22°C, or by cooling compounds such as menthol. In this study, it was found that a new role of TRPM8 activation on prostaglandin E2 (PGE2), an inflammatory cytokine and dendritogenesis stimulator of normal human melanocytes. Normal human keratinocytes were pretreated with menthol or incubated at 22°C for TRPM8 activation before ultraviolet (UV)-B irradiation. To examine the specificity between TRPM8 activation and PGE2 release, we inhibited TRPM8 with the antagonist (capsazepine), or introduced TRPM8 siRNA for a gene silencing experiment. UV-B irradiation significantly induced PGE2 release in normal human keratinocytes. Interestingly, activation of TRPM8 at 22°C or with menthol inhibited UV-B-induced PGE2 release. The effect of the TRPM8 agonist was completely blocked by pretreatment with the TRPM8 antagonist, capsazepine. When TRPM8 expression was suppressed by siRNA, UV-B irradiation still upregulated PGE2 in keratinocytes, but pretreatment of menthol or low temperature did not inhibit UV-B-induced PGE2. In conclusion, the activation of TRPM8 inhibits UV-B-induced PGE2 production in keratinocytes, and the activation of TRPM8 may reduce inflammatory responses in skin.

Anaphylactic shock caused by the epidurally-administered hyalurinidase.

Hyaluronidase is an enzyme that has temporary and reversible enzymatic effects on the matrix of connective tissue. When added to local anesthetics in pain treatments, it enhances their infiltration and dispersal into tissues. It is widely used in anesthesia for ocular, dental, and plastic surgery. Reports of drug hypersensitivity to hyaluronidase are rare and are usually confined to peribulbar or retrobulbar anesthesia during ophthalmic surgery. However, few reports exist on adverse drug reaction after epidural injection. We have observed two patients experiencing anaphylactic shock caused by hyaluronidase following epidural injection. Most of the patients with a hypersensitivity to hyaluronidase had one previous uneventful injection containing hyaluronidase, implying that sensitization had taken place. However, hypersensitivity occurring at the first administration is possible. A positive skin test can help establish the diagnosis. Although rare, the possibility of an allergic reaction to hyaluronidase should be considered even in patients with no known previous exposure.

Evaluating the nickel content in metal alloys and the threshold for nickel-induced allergic contact dermatitis.

Many patients are currently suffering from nickel (Ni)-induced allergic contact dermatitis (ACD). There have been few Korean studies dealing with the threshold of Ni-induced ACD and quantifying the total amount of Ni in the metal alloys. The aim of this study is to evaluate the amount of Ni leached from metal alloys and Ni contents in metal alloys, and to estimate the threshold of Ni-induced ACD. All the earrings we examined leached below 0.5 microg/cm(2)/week, the upper limit of European Union (EU) regulation, but the other metal alloys leached a much higher amount of Ni than the limit. Likewise, all the earrings we examined contained less than 0.05% Ni (500 microg/g), the upper limit of EU regulation, but the other metal alloys exceeded this limit. Twenty Ni-sensitive subjects, who were patch-tested with various concentrations of Ni sulphate, showed positive reactions to 5% and 1% Ni sulphate, 10 subjects showed positive reactions to 0.01%, and the most sensitive subject showed reaction even to 0.0001%. The subjects in this study were more sensitive to Ni than those in the previous studies done in Europe. Taken together, strictly regulating the Ni-containing alloys that are made in Korea is needed to lower the occurrence of Ni-induced ACD.

The effects of Musk T on peroxisome proliferator-activated receptor [PPAR]-alpha activation, epidermal skin homeostasis and dermal hyaluronic acid synthesis.

Peroxisome proliferators activated receptors (PPARs) are a family of nuclear hormone receptors that heterodimer with the retinoid X receptor and function as transcriptional regulators of genes. Topically Applied PPAR-alpha agonists possess receptor mediated, pro-differentiating/anti-proliferative effects, lipid metabolism stimulation, and anti-inflammatory activity, which suggest that they could be beneficial for the treatment of a variety of cutaneous diseases. Hyaluronan (HA), a high-molecular-weight linear glycosaminoglycan consisting of alternating D: -glucuronic acid and N-acetyl-D: -glucosamine residues, is one of the major extracellular matrix components in skin. Among the family of HA synthase genes (HAS1, 2, 3) so far identified, one group has demonstrated that the expressions of HAS2 and HAS3 play crucial roles in the regulation of HA synthesis in human skin fibroblasts and keratinocytes, respectively, but the precise regulatory mechanisms are still unknown. We examine Musk T called Ethylene brassylate, Astratone or 1,4-Dioxacycloheptadecane-5,17-dione, which used as just a perfume ingredient, plays a role as PPAR-alpha ligand in vitro and stimulates skin barrier recovery, ceramide synthesis, beta-Glucocerebrosidase, involucrin expression in epidermis in vivo; and examine that Musk T stimulates HAS expression and HA synthesis in human skin fibroblast. Through these experiments, we conclude that Musk T is PPAR-alpha ligand, effects on keratinocyte differentiation, intercellular lipid synthesis in epidermis, HA synthesis stimulation in dermis.