A triple-stimulus responsive melanin-based nanoplatform with an aggregation-induced emission-active photosensitiser for imaging-guided targeted synergistic phototherapy/hypoxia-activated chemotherapy.

Multimodal synergistic therapy has gained increasing attention in cancer treatment to overcome the limitations of monotherapy and achieve high anticancer efficacy. In this study, a synergistic phototherapy and hypoxia-activated chemotherapy nanoplatform based on natural melanin nanoparticles (MPs) loaded with the bioreduction prodrug tirapazamine (TPZ) and decorated with hyaluronic acid (HA) was developed. A self-reporting aggregation-induced emission (AIE)-active photosensitizer (PS) (BATTMN) was linked to the prepared nanoparticles by boronate ester bonds. The MPs and BATTMN-HA played roles as quenchers for PS and cancer targeting/photodynamic moieties, respectively. As a pH sensitive bond, the borate ester bonds between HA and BATTMN are hydrolysed in the acidic cancer environment, thereby separating BATTMN from the nanoparticles and leading to the induction of fluorescence for imaging-guided synergistic phototherapy/hypoxia-activated chemotherapy under dual irradiation. TPZ can be released upon activation by pH, near-infrared (NIR) and hyaluronidase (Hyal). Particularly, the hypoxia-dependent cytotoxicity of TPZ was amplified by oxygen consumption in the tumor intracellular environment induced by the AIE-active PS in photodynamic therapy (PDT). The nanoparticles developed in our research showed favorable photothermal conversion efficiency (η = 37%), desired cytocompatibility, and excellent synergistic therapeutic efficacy. The proposed nanoplatform not only extends the application scope of melanin materials with AIE-active PSs, but also offers useful insights into developing multistimulus as well as multimodal synergistic tumor treatment.

Anti-Melanogenic Effects of Korean Red Ginseng Oil in an Ultraviolet B-Induced Hairless Mouse Model.

A 'remedy for all' natural product widely known in the Korean Peninsula is called Panax Ginseng Meyer. Globalization represents a persistent risk to the ozone layer, leading to bountiful amounts of Ultra-Violet B beams (UVB). The variety in human skin hues is ascribed to the characteristic color called Melanin. However, Melanin overproduction due to UVB beams promotes skin staining and tumorigenesis, a process called photo aging, which damages skin quality. To assess the effects of Korean Red Ginseng Oil (KGO) on photo aging, the murine melanoma cell lines B16/F10 were used in vitro and HRM-2 hairless mice exposed to UVB were studied in vivo. Our results revealed that KGO reduced tyrosinase activity and melanin production in B16/F10 cells along with the suppression of upstream factors involved in the melanin production pathway, both transcriptionally and transitionally. In the in vivo studies, KGO suppressed the expression of Matrix Metalloproteinase (MMP) and Interleukins along with a reduction of depth in wrinkle formation and reduced collagen degradation. Moreover, the feed intake and feed efficiency ratio that decreased as a result of UVB exposure was also improved by KGO treatment. In light of our results, we conclude that KGO can have considerable benefits due to its various properties of natural skin enhancement.

Opto-acoustic synergistic irradiation for vaporization of natural melanin-cored nanodroplets at safe energy levels and efficient sono-chemo-photothermal cancer therapy.

Rationale: Insufficient penetration and accumulation of theranostic payloads in solid tumors greatly challenge the clinical translation of cancer nanomedicines. To address this challenge, we synthesized natural melanin-cored and doxorubicin-loaded perfluoropentane nanodroplets with good biocompatibility and self-assembling ability. Methods: We used an opto-acoustic synergistic irradiation (OASI) method that was effective at lower energy levels than ultrasound- or laser-only irradiation to safely vaporize the nanodroplets and to cavitate the generated microbubbles for mechanically enhancing intratumoral delivery. The delivered melanin and doxorubicin inside the tumors mediated secondary chemo-photothermal therapy under laser irradiation to fully kill cancer cells. Results:In vivo animal experiments demonstrated direct mechanical disruption of tumor structures (H&E staining), enhanced intratumoral penetration of melanin (photoacoustic imaging), and efficient intratumoral accumulation of doxorubicin (fluorescent imaging). Anti-tumor experiments demonstrated that the nanodroplets combined with OASI treatment and subsequent laser irradiation could efficiently eliminate melanoma tumors. Conclusion: Melanin-cored and doxorubicin-loaded perfluoropentane nanodroplets hold great promise for translational sono-chemo-photothermal cancer therapy.

Photothermal therapy-induced immunogenic cell death based on natural melanin nanoparticles against breast cancer.

A photothermal and immune co-therapy strategy based on natural melanin nanoparticles was developed for treating primary and abscopal breast cancers.

Evaluation and visualization of facial massage effects by using ultraviolet stereo-image correlation.

BACKGROUND/PURPOSE: This study proposes a technique for visualizing the effect of facial massage using stereo-image correlation with melanin pigment. METHOD: In this method, the melanin pigment of a subject's face is made visible by using an ultraviolet light and utilized as a random pattern for stereo-image correlation. Stereo-pair images of the face with the melanin pigment before and after facial massage are recorded using a desk-sized measurement equipment. Then, the deformation of the face by the massage can be obtained based on the principle of stereovision. The effectiveness of the proposed method is demonstrated by applying it to the massage effect evaluation of eight subjects (females in their 40s). RESULTS: The results show that the massage effect can be visualized from the displacement and strain distributions across the face obtained by the proposed method. In addition, it is observed that the face is displaced significantly by the massage and individual differences between the subjects can be captured. CONCLUSION: The proposed method is effective for evaluating the effect of a facial massage when the painted pattern disappears due to the applied cream during the massage.

Oral supplementation of L-glutathione prevents ultraviolet B-induced melanogenesis and oxidative stress in BALB/c mice.

Dietary antioxidant supplements such as L-glutathione have gained considerable attention in dermatology and cosmeceutical fields. L-glutathione possesses antiaging, antimelanogenic, antioxidant, and anticancer properties. This study aimed to investigate the inhibitory effects of L-glutathione on melanogenesis activity and oxidative stress in ultraviolet B (UVB)-irradiated BALB/c mice. Eighteen female BALB/c mice were randomly divided into 3 groups: a control group (n=6), a group without UVB irradiation and L-glutathione administration; a UVB irradiated group (n=6), a group irradiated with a UVB dose of 250 mJ/cm2 for 3 min; and a treatment group (n=6), a group irradiated with UVB and treated with 100 mg/kg of L-glutathione by oral gavage. Treatment was given for 14 days, and UVB irradiation was given on days 9, 11, and 13. Oral L-glutathione significantly (P<0.05) reduced lipid peroxidation and elevated superoxide dismutase activity the and glutathione level. L-glutathione also inhibited melanin content and tyrosinase activity significantly (P<0.05) as compared with the UVB-irradiated group. Histopathological examination also showed that L-glutathione reduced the deposition of melanin pigment in the basal layer of the epidermis as compared with that in UVB-irradiated mice. All in all, the present study demonstrated that L-glutathione has the potential to be developed as a photoprotection agent against UVB-induced oxidative stress and melanogenesis.

Infrared thermography as control of handheld IPL device for home-use.

Infrared thermography as contactless method for determining the temperature distribution on the surface is used for analyzing the impact of intense pulsed light hair removal device (IPL) on the skin. Depth of light penetration depending of wavelength is described as well as absorption curves and IPL impulse shapes. Energy balance and IPL impulse influence on the skin is analyzed. Melanin temperature rise by different fluence operation and temperature distribution in the modeled hair is used in order to determine overall skin temperature rise. Estimated energy balance provided by mathematical model has been confirmed with experimental results. Performed measurements, beside determination of the right emissivity, required the identification of most significant parameter in the process which proved to be the skin reference temperature and real temperature rise. Practical IPL application with detailed body temperature analysis is comprehensively described and thermal imaging interpretation problem and determination of the temperature rise is observed.

Anti-Pigmentation Effects of Eight Phellinus linteus-Fermented Traditional Crude Herbal Extracts on Brown Guinea Pigs of Ultraviolet B-Induced Hyperpigmentation.

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

Light-emitting diode 585nm photomodulation inhibiting melanin synthesis and inducing autophagy in human melanocytes.

BACKGROUND: Melasma is a common hyperpigmentation skin disease on face. Light-emitting diode (LED) photomodulation (585nm) is reported to be effective for the treatment of melasma. However, whether and how LED photomodulation would influence melanogenesis of human epidermal melanocytes (HEMs) is unknown. OBJECTIVE: To evaluate the effects of LED photomodulation (585nm) on melanogenesis in HEMs. METHODS: HEMs were irradiated with fluences of 0, 5, 10 and 20J/cm2 585nm LED light. After 5-day treatment, cell viability was analyzed by CCK-8 assay, and apoptosis was assessed by Annexin V APC assay. Melanin content and tyrosinase activity were measured by spectrophotometer. Melanosome stage and autophagosomes were determined under transmission electron microscope (TEM). The formation of autophagic punctate structures was observed under confocal microscope. RT-PCR and western blotting were used to assess the expression of relative mRNA and protein levels. RESULTS: Yellow light LED 585nm had no effects on HEMs cell viability and apoptosis. Treatment with LED 585nm from 5J/cm2 to 20J/cm2 inhibited melanosome maturation, decreased melanin content and tyrosinase activity. Inhibition was accompanied by the decreased expression of tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1) and microphthalmia-associated transcription factor (MITF) on both mRNA and protein levels. Autophagosomes were observed under TEM. Autophagic punctate structures of microtubule-associated protein light chain 3 (LC3) proteins were induced by LED 585nm light. The configuration change of LC3 from LC3-I to LC3-II, and the degradation of p62 protein were observed after LED 585nm. Furthermore, we also revealed that the anti-melanogenic effect of LED 585nm photomodulation was reversed by 3-Methyladenine (3-MA), which inhibits autophagy by blocking autophagosome formation via the inhibition of type III Phosphatidylinositol 3-kinases (PI-3K). CONCLUSIONS: Our finding demonstrated that LED photomodulation with 585nm wavelength suppressed melanin content in HEMs, and the effect was caused by its dose-dependent inhibition on melanogenesis and the induction of HEMs autophagy. This may provide new insights into the efficacy of LED photomodulation in the treatment of hyperpigmentation disorders.

Mitigation of epidermal growth factor receptor inhibitor-induced side effects utilizing melanin and vascular-specific lasers: A case report series.

The advent of targeted chemotherapy has led to the emergence of new dermatologic toxicities. We sought to use lasers and light devices to treat recalcitrant cutaneous adverse effects related to cancer treatment. Three stage III or IV cancer patients with cutaneous complications due to epidermal growth factor receptor (EGFR) inhibitors were treated with melanin and vascular-specific laser and light technologies. Two patients reported reduction in papulopustular eruption following pulse dye laser (PDL) treatment. Two patients noted reduction in hair growth following intense pulsed light (IPL) and/or Alexandrite laser treatments. One patient was treated with both the PDL and IPL and reported improvement of both EGFR-induced hypertrichosis and papulopustular eruption. Laser and light devices targeting melanin and hemoglobin can be utilized to mitigate the cutaneous adverse effects associated with EGFR inhibitors in patients who have failed traditional therapies. This represents a new option for the cancer patient who is suffering from chemotherapy-induced side effects.

Treatment of infraorbital dark circles using 694-nm fractional Q-switched ruby laser.

The objective of this study was to evaluate the efficacy and safety of using a 694-nm fractional Q-switched ruby laser to treat infraorbital dark circles. Thirty women with infraorbital dark circles (predominant color: dark/brown) participated in this open-labeled study. The participants received eight sessions of 694-nm fractional Q-switched ruby laser treatment using a fluence of 3.0-3.5 J/cm2, at an interval of 7 days. The melanin deposition in the lesional skin was observed in vivo using reflectance confocal microscopy (RCM). The morphological changes were evaluated using a global evaluation, an overall self-assessment, and a Mexameter. Twenty-eight of the 30 patients showed global improvements that they rated as excellent or good. Twenty-six patients rated their overall satisfaction as excellent or good. The melanin index indicated a substantial decrease from 240.44 (baseline) to 194.56 (P < 0.05). The RCM results showed a dramatic decrease in melanin deposition in the upper dermis. The adverse effects were minimal. The characteristic finding of dark/brown infraorbital dark circles is caused by increased melanin deposition in the upper dermis. The treatment of these infraorbital dark circles using a 694-nm fractional QSR laser is safe and effective.

In vivo microscopic approaches for facial melanocytic lesions after quality-switched ruby laser therapy: time-sequential imaging of melanin and melanocytes of solar lentigo in Asian skin.

BACKGROUND: The quality-switched ruby laser (QSRL) has been widely used for the treatment of pigmented lesions, but clinical evaluations in most studies have been conducted on macroscopic skin color observation comparing the laser-treated skin with its nontreated surrounding area. A few investigations examined skin changes after laser therapy at a cellular level, but almost none did so noninvasively. OBJECTIVE: To elucidate the dynamic changes after QSRL irradiation of facial solar lentigo using noninvasive optical techniques. MATERIALS AND METHODS: Time-sequential imaging of Japanese female patients with a clinical diagnosis of solar lentigo was performed using ultraviolet photography, high-magnification videomicroscopy, and reflectance-mode confocal microscopy to examine pigmentary change after QSRL irradiation. RESULTS: The present study showed that remaining melanocytes were visible in the solar lentigo of all subjects when crusts peeled off, despite hardly observable skin pigmentation to the naked eye. Moreover, noninvasive confocal imaging revealed that pigmented melanocytes varied in each solar lentigo after QSRL treatment, as indicated by melanin reflection level. CONCLUSIONS: Optical techniques facilitate the evaluation of the in vivo dynamics of epidermal-melanocytic changes in solar lentigo after QSRL therapy and may be useful for monitoring outcomes after laser irradiation.

Comparison of short-pulsed and long-pulsed 532 nm lasers in the removal of freckles.

The purpose of this study was to compare the efficacy and safety of the 532 nm long-pulsed laser (10 ms) with that of the 532 nm short-pulsed laser (10 ns) for freckle removal. Currently, the gold standard for treatment is the short-pulsed laser. Recently, several long-pulsed lasers have been introduced for both hair removal and the treatment of freckles. To our investigative team's knowledge, no controlled experiments have been performed to compare the safety and efficacy of long-pulsed versus short-pulsed lasers for the treatment of freckles. This was a 4-week trial, and all patients had three freckles that were randomly allocated to be treated with short-pulse laser, long-pulse laser, or to receive no treatment (control). All patients had three freckles that were randomly selected to be treated with short-pulse 532 nm Medlite IV laser (10 n, 1 J/cm(2)), or long-pulse 532 nm Aura laser (10 ms, 1 J/cm(2)) or to remain as a control (no treatment). The laser treatment was only performed once, followed by a 1-day and a 1-month follow-up visit. Freckle size was determined by a novel surface area measurement technique that was created by our research staff. The study included 17 sets of freckles (three in each set). All of the lesions which received the short-pulsed laser treatment had immediate whitening of the lesions, which turned into dry scabs the next day. None of the freckles treated in the long-pulsed group or control group developed immediate whitening or scabs. No blisters or ulcers developed. The average pain score in the short-pulsed laser group was 2-3 out of 10, while it was 0 out of 10 in the long-pulsed laser group. All scabs that developed in the short-pulsed laser group fell off between days 6 and 12 (average 8 days). The outcome of this study verified the appropriate treatment of freckles. The study confirmed that when the same energy settings, short-pulsed laser is the more effective laser treatment regimen (when compared with the long-pulsed laser), with high tolerability and minimal side effects for patients with skin types I to IV.

Modulation of antioxidant defense by Alpinia galanga and Curcuma aromatica extracts correlates with their inhibition of UVA-induced melanogenesis.

Ultraviolet A (UVA) irradiation is suggested to contribute to melanogenesis through promoting cellular oxidative stress and impairing antioxidant defenses. An overproduction of melanin can be associated with melanoma skin cancer and hyperpigmentation. Therefore, developing effective antimelanogenic agents is of importance. Alpinia galanga (AG) and Curcuma aromatica (CA) are traditional medicinal plants widely used for skin problems. Hence, this study investigated the antimelanogenic effects of AG and CA extracts (3.8-30 microg/ml) by assessing tyrosinase activity, tyrosinase mRNA levels, and melanin content in human melanoma cells (G361) exposed to UVA. The roles in protecting against melanogenesis were examined by evaluating their inhibitory effects on UVA-induced cellular oxidative stress and modulation of antioxidant defenses including antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GPx), and intracellular glutathione (GSH). In addition, possible active compounds accountable for biological activities of the extracts were identified by thin layer chromatography (TLC)-densitometric analysis. Our study demonstrated that UVA (8 J/cm(2)) induced both tyrosinase activity and mRNA levels and UVA (16 J/cm(2))-mediated melanin production were suppressed by the AG or CA extracts at noncytotoxic concentrations. Both extracts were able to protect against UVA-induced cellular oxidant formation and depletion of CAT and GPx activities and GSH content in a dose-dependent manner. Moreover, TLC-densitometric analysis detected the presence of eugenol and curcuminoids in AG and CA, respectively. This is the first report representing promising findings on AG and CA extract-derived antityrosinase properties correlated with their antioxidant potential. Inhibiting cellular oxidative stress and improving antioxidant defenses might be the mechanisms by which the extracts yield the protective effects on UVA-dependent melanogenesis.

Pulsing influences photoradiation outcomes in cell culture.

BACKGROUND AND OBJECTIVES: Skin pigmentation can adversely affect phototherapy outcomes. Delivering pulsed light has been suggested as a means of enhancing efficacy. Suitable pulse frequencies remain indeterminate, often being selected empirically. This study was undertaken to determine whether pulsed light delivery mitigates the filtering effect of melanin pigment on photomodulation in vitro. STUDY DESIGN AND METHODS: Human HEP-2 cells were cultured in complete DMEM media. Photoradiation was delivered through 0.025% melanin filters at 670 nm (5.0 J/cm(2)/treatment/24 hours) for 72 hours at different pulse rates. Group A received no light treatment. Group B received treatments without pulsing. Groups C, D, E, F, and G received treatments at 6, 18, 36, 100, and 600 Hz. Cell proliferation was assessed by MTT assay and oxidative burst was measured using the 2.7 dichloro-fluorescein-diacetate assay. RESULTS: Cell proliferation was maximally stimulated at 100 Hz at 48 and 72 hours (n = 4, P< or =0.05). Oxidative burst was maximally stimulated at 600 Hz (n = 4, P< or =0.05). All frequencies were stimulatory at 48 and 72 hours (n = 4, P< or =0.05). CONCLUSION: This investigation suggests that light pulsing may improve outcomes by mitigating the filtration effects of cutaneous melanin. Further studies to further define these effects are warranted.

The skin-lightening effects of artocarpin on UVB-induced pigmentation.

This study was conducted to evaluate the effects of the prenylated flavonol artocarpin from the heartwood of Artocarpus incisus on ultraviolet (UV)-induced hyperpigmentation of guinea pig skin. An efficient lightening effect was observed following topical application of artocarpin to UV-stimulated hyperpigmented dorsal skins of brownish guinea pigs.

Tanning.

Cutaneous pigment responses to ultraviolet radiation are outlined in this article. The responses induced by different wavebands are compared, and the protection against further exposure that is conferred by different types of tanning is discussed. The term "tanning" is used here to denote the spectrum of adaptive changes in the skin resulting from ultraviolet exposure and is not applied exclusively to increased melanin pigmentation.