Anti-Melanogenic Effects of Ethanol Extracts of the Leaves and Roots of Patrinia villosa (Thunb.) Juss through Their Inhibition of CREB and Induction of ERK and Autophagy.

Patrinia villosa (Thunb.) Juss is a traditional herb commonly used in East Asia including Korea, Japan, and China. It has been administered to reduce and treat inflammation in Donguibogam, Korea. The mechanism for its anti-inflammatory effects has already been reported. In this study, we confirmed the efficacy of Patrinia villosa (Thunb.) Juss ethanol extract (Pv-EE) for inducing autophagy and investigate its anti-melanogenic properties. Melanin secretion and content were investigated using cells from the melanoma cell line B16F10. Pv-EE inhibited melanin in melanogenesis induced by α-melanocyte-stimulating hormone (α-MSH). The mechanism of inhibition of Pv-EE was confirmed by suppressing the mRNA of microphthalmia-associated transcription factor (MITF), decreasing the phosphorylation level of CREB, and increasing the phosphorylation of ERK. Finally, it was confirmed that Pv-EE induces autophagy through the autophagy markers LC3B and p62, and that the anti-melanogenic effect of Pv-EE is inhibited by the autophagy inhibitor 3-methyl adenine (3-MA). These results suggest that Pv-EE may be used as a skin protectant due to its anti-melanin properties including autophagy.

Characterization of Ginkgo biloba Leaf Flavonoids as Neuroexocytosis Regulators.

Ginkgo biloba leaf (GBL) is known as a potential source of bioactive flavonoids, such as quercetin, arresting the neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-zippering. Here, the GBL flavonoids were isolated in two different manners and then examined for their bioactivity, physicochemical stability, and biocompatibility. The majority of flavonoids in the non-hydrolyzed and acidolyzed isolates, termed non-hydrolyzed isolate (NI) and acidolyzed isolate (AI) hereafter, were rich in flavonol glycosides and aglycones, respectively. Glycosidic/aglyconic quercetin and kaempferol were abundant in both NI and AI, whereas a little of apigenin, luteolin, and isorhamnetin were found in AI. NI was more thermostable in all pH ranges than quercetin, kaempferol, and AI. NI and AI both inhibited neurotransmitter release from differentiated neuronal PC-12 cells. NI and AI showed 1/2-1/3 lower EC50/CC50 values than quercetin and kaempferol. The NI and AI exhibited no toxicity assessed by the tests on chorioallantoic membranes of hen's eggs, removing toxicological concerns of irritation potential. Moreover, GBL isolates, particularly AI, showed antioxidant and anti-inflammatory activities in the use below the CC50 levels. Taken together, these results suggest that GBL isolates that are rich in antioxidant flavonoids are effective anti-neuroexocytotic agents with high stability and low toxicity.

Antiphotoaging and Antimelanogenic Effects of Penthorum chinense Pursh Ethanol Extract due to Antioxidant- and Autophagy-Inducing Properties.

Ethnopharmacological Relevance. Penthorum chinense Pursh (Penthoraceae) is a traditional herbal plant that has been used in China for the treatment of jaundice, cholecystitis, edema, and infectious hepatitis. In addition, the Korea Medicinal Plant Dictionary states that Penthorum chinense Pursh can be used to treat contusions and skin bruises by improving blood flow. Recent studies have shown that Penthorum chinense Pursh ethanol extract (Pc-EE) exhibits strong antioxidant effects. In this study, we examined the effects of Pc-EE on UVB-induced or H2O2-induced oxidative stress, as well as its antimelanogenic properties. Cell viability, matrix metalloproteinase (MMP) expression, cyclooxygenease-2 (COX-2), and interleukin-6 (IL-6) expression and moisturizing factors were investigated in keratinocytes. Collagen synthesis induction was measured in HEK293T cells. For melanogenesis, the effects of Pc-EE on melanin content and tyrosinase activity were measured. Additionally, the antimelanogenic- and autophagy-inducing activities of Pc-EE were examined using immunoblotting and confocal microscopy. Pc-EE protected HaCaT cells against death from UVB irradiation- or H2O2-induced oxidative stress. Pc-EE increased the promoter activity of the type 1 procollagen gene Col1A1 and decreased the expression of MMPs, COX-2, IL-6, and hyaluronidase induced by UVB irradiation- or H2O2-induced oxidative stress. Pc-EE showed a strong antioxidant effect in the DPPH assay. In α-melanocyte-stimulating hormone- (α-MSH-) stimulated B16F10 cells, Pc-EE reduced melanin production, decreased tyrosinase expression and microphthalmia-associated transcription factor (MITF) protein levels, and decreased the phosphorylation levels of p38 and JNK. In HEK293T cells, Pc-EE promoted the expression of GFP-LC3B. In B16F10 cells, the LC3B and melanin contents were reduced by Pc-EE and were restored by the autophagy inhibitor 3-methyladenine (3-MA). These results suggest that Pc-EE can be used as a skin protection agent due to its antiapoptotic, antiaging, anti-inflammatory, and antimelanogenic properties.

The Effect of Alkyl Chain Number in Sucrose Surfactant on the Physical Properties of Quercetin-Loaded Deformable Nanoliposome and Its Effect on In Vitro Human Skin Penetration.

Non-invasive skin penetration of a drug is increased by an edge activator, which enhances the nanoliposome deformability. The objective of this study was to investigate the role of the alkyl chain number of sucrose surfactants as an edge activator in elastic nanoliposomes. In addition, the physicochemical properties of the elastic nanoliposomes were characterized and an in vitro human skin permeation study was performed. Elastic nanoliposomes that were composed of sucrose monostearate (MELQ), sucrose distearate (DELQ), and sucrose tristearte (TELQ) were prepared using a thin-film hydration method. Particle size and entrapment efficiency of elastic nanoliposomes increased proportionally with an increase in the amounts and the numbers of the stearate in sucrose surfactant. Deformability of elastic nanoliposomes was indicated as DELQ > MELQ > TELQ and the same pattern was revealed through the in vitro human skin permeability tests. These results suggest that the number of alkyl chains of sucrose surfactant as edge activator affects the physicochemical property, stability, and skin permeability in elastic nanoliposome. Our findings give a valuable platform for the development of elastic nanoliposomes as skin drug delivery systems.

New approach to the immobilization of glucose oxidase on non-porous silica microspheres functionalized by (3-aminopropyl)trimethoxysilane (APTMS).

The immobilization and encapsulation of glucose oxidase (GOD) onto the mesoporous and the non-porous silica spheres prepared by co-condensation of tetraethylorthosilicate (TEOS) and (3-aminopropyl)trimethoxysilane (APTMS) in the water-in-oil (W/O) emulsion system were studied. The terminal amine group was used as the important functionality for GOD immobilization on the silica substrate. When only TEOS is used as a silica source, the disordered mesoporous silica microspheres are obtained. As the molar ratio of APTMS to TEOS (R(AT)) increases, the surface area and pore volume of the silica particles measured by nitrogen adsorption and desorption method and SEM decrease rapidly. Particularly, the largest change of the surface morphology is observed between R(AT)=0.20 and R(AT)=0.25. The amount and the adsorption time of immobilized enzyme were measured by UV spectroscopy. About 20wt% of GOD was immobilized into the silica substrates above R(AT)=0.60 and was completely adsorbed into the substrate of R(AT)=0.80 with lapse of 4h after addition. In the measurement of the thermal stability, GOD dissolved in buffer solution loses nearly all of its activity after 30 min at 65 degrees C. In contrast, GOD immobilized on the surface-modified silica particles still retains about 90% of its activity after the same treatment. At this temperature, the immobilized glucose oxidase retained half of its initial activity after 4h. It is shown that the suitable usage of functionalizing agent like APTMS as well as the control of surface morphology is very important on the immobilization of enzyme.

The stabilization of L-ascorbic acid in aqueous solution and water-in-oil-in-water double emulsion by controlling pH and electrolyte concentration.

This study presents a new approach that can stabilize effectively L-ascorbic acid in water-in-oil-in-water (w/o/w) double emulsions. Basically, the behavior of L-ascorbic acid in the aqueous phase was observed, considering its molecular deformation. Then, it was found that the stability determined in the aqueous phase by high-performance liquid chromatography (HPLC) showed that the collapse of ionization of L-ascorbic acid played a crucial role in protecting the molecular deformation. Then, the stable aqueous system was incorporated into the internal aqueous phase of the double emulsions. From the HPLC analysis, it was observed that the L-ascorbic acid in an appropriate system showed high molecular stability for a long time. Moreover, in the measurement of in vitro skin permeation, the L-ascorbic acid stabilized in this study showed considerable skin permeation ability, indicating its potential applicability in pharmaceutics and cosmetics.

Nano-structured biphasic polymer film on the hair surface from PEGylated polymer latexes.

In this study, biphasic polymer latexes were synthesized by surfactant-free-emulsion polymerization of butyl methacrylate, poly(ethylene glycol) methyl ether methacrylate, and 2-(methacryloyloxy) ethyl trimethyl ammonium chloride. The latexes synthesized were composed of hydrophobic core phase and hydrophilic shell phase. Nano-structured film morphology could be obtained by annealing the biphasic polymer latexes between the two transition temperatures. It was found that the unique film morphology gave a viscoelastic property to the film. Scanning electron microscope and atomic force microscope images revealed that the biphasic polymer latexes deposited effectively onto the entire hair surface upon conditioning with 1 wt% polymer concentration in water. Consequently, they formed a smooth polymer membrane thereon, showing a high potential for a new hair cosmetic ingredient.