Ethosome Containing Ceramide as a Skin Carrier of Active Ingredients.

BACKGROUND: Numerous formulations have been utilized in the cosmetic and pharmaceutical industries to effectively deliver bioactive ingredients. METHODS: We selected a well-known liposomal formulation of bilayer lipid vesicles composed of ceramide NP. Ethosomes contain hydrophilic vanillic acid or lipophilic α-bisabolol, and their physicochemical properties were evaluated. Vanillic acid is encapsulated in the aqueous core while α-bisabolol is engaged with the lipid phase. The formulation was prepared by the high-pressure homogenization method at 800 bar for 5 min. The particle size, polydispersity index and zeta potential of the ethosome dispersion were analyzed by dynamic light scattering. In order to measure the skin absorption efficiency from artificial skin, an in vitro assay was performed using the Franz diffusion cell method for 24 hours. In addition, ultracentrifuges for encapsulation efficiency, dialysis membranes for active ingredient release, and low-temperature transmission electron microscopy (TEM) to evaluate the morphology of vesicles were utilized. RESULTS: The particle size of the ethosome containing ceramide NP and vanillic acid was in the range of 80 ~ 130 nm, whereas the particle size of the ethosome containing ceramide NP and α-bisabolol was 150 ~ 170 nm. In the vanillic acid-containing ethosome, increasing the amount of ceramide NP decreased the particle size, whereas the size of the α-bisabolol ethosome did not change. The stability of the prepared ethosome did not change significantly for 4 weeks at 25°C, 4°C, and 45°C. The skin absorption efficiency of ceramide NP and vanillic acid-containing ethosome was increased by about 15% compared to the control group, whereas the ethosome containing α-bisabolol and ceramide NP showed slightly higher skin absorption efficiency than the control group. In addition, encapsulation efficiency evaluation, active ingredient release measurement and cryo-TEM were taken. CONCLUSION AND PERSPECTIVE: Based on the results of these studies, we suggest that ethosome formulations containing ceramide NP can be widely used in the cosmetic industry together with other cosmetic formulations.

Myristoleic Acid Promotes Anagen Signaling by Autophagy through Activating Wnt/ß-Catenin and ERK Pathways in Dermal Papilla Cells.

Alopecia is a distressing condition caused by the dysregulation of anagen, catagen, and telogen in the hair cycle. Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Myristoleic acid (MA) increases Wnt reporter activity in DPCs. However, the action mechanisms of MA on the stimulation of anagen signaling in DPCs is not known. In this study, we evaluated the effects of MA on anagen-activating signaling pathways in DPCs. MA significantly increased DPC proliferation and stimulated the G2/M phase, accompanied by increasing cyclin A, Cdc2, and cyclin B1. To elucidate the mechanism by which MA promotes DPC proliferation, we evaluated the effect of MA on autophagy and intracellular pathways. MA induced autophagosome formation by decreasing the levels of the phospho-mammalian target of rapamycin (phospho-mTOR) and increasing autophagy-related 7 (Atg7) and microtubule-associated protein 1A/1B-light chain 3II (LC3II). MA also increased the phosphorylation levels of Wnt/ß-catenin proteins, such as GSK3ß (Ser9) and ß-catenin (Ser552 and Ser675). Treatment with XAV939, an inhibitor of the Wnt/ß-catenin pathway, attenuated the MA-induced increase in ß-catenin nuclear translocation. Moreover, XAV939 reduced MA-induced effects on cell cycle progression, autophagy, and DPC proliferation. On the other hand, MA increased the levels of phospho (Thr202/Tyr204)-extracellular signal regulated kinases (ERK). MA-induced ERK phosphorylation led to changes in the expression levels of Cdc2, Atg7 and LC3II, as well as DPC proliferation. Our results suggest that MA promotes anagen signaling via autophagy and cell cycle progression by activating the Wnt/ß-catenin and ERK pathways in DPCs.

Preparation and Evaluation of Pluronic Lecithin Organogels in Cosmetics.

This study was performed to investigate the application of pluronic lecithin organogel (PLO gel) in cosmetics as a topical drug delivery system. PLO gel was known as transdermal drug delivery systems. It has a very interesting system, owing to their biocompatibility; their amphiphilic nature, facilitating dissolution of various drug classes; and their permeation enhancement properties. We realized that PLO gel has a critical shortcoming of flowability at low temperatures to be used as a cosmetic ingredient. To improve this drawback, this study aimed to find an appropriate quantity of three main compositions of PLO gel, including aqueous phase (poloxamer 407 and water), polyol phase (PEG-400), and oil phase (lecithin and oil), and applied an experimental design using the response surface methodology (RSM). We assessed the elapsed time change by temperature in each PLO gel formulation, observed the morphology of PLO gel using field emission scanning electron microscope (FE-SEM), and determined the gelation point by using differential scanning calorimetry (DSC). Rheology measurements to assess viscoelastic properties were determined by using a rheometer, and skin permeation efficiency was assessed by diffusion system. It was confirmed that three main factors (hydrogenated lecithin, PEG-400, and poloxamer 407) of PLO gel should be balanced without flowability even in cold temperature. Through the RSM, it was assumed that the most effective ingredient was PEG-400 at PLO gel formulation and physical properties. The PLO gel formulation (hydrogenated lecithin 5.0%, PEG-400 20.0%, and poloxamer 407 15.0%) was evaluated as the most suitable formulation for use in cosmetics due to its viscosity and elasticity results. The shape was observed through FE-SEM, and it was confirmed that the PLO gel forms a polymeric bicontinuous microemulsion structure. Regarding the applicability of PLO gel in cosmetics, we verified that PLO gel can be used in a delivery system for active substances. The study findings suggest that PLO gel can be used as one of the ingredients in cosmetic formulations.

Determination of Required HLB Values for Citrus unshiu Fruit Oil, Citrus unshiu Peel Oil, Horse Fat and Camellia japonica Seed Oil.

In the present study, the required hydrophilic lipophilic balance (HLB) values of Citrus unshiu fruit oil (CUFO), Citrus unshiu peel oil (CUPO), horse fat (HF), and Camellia japonica seed oil were determined empirically by preparing oil-in-water (o/w) emulsions. Lipophilic and hydrophilic surfactants were prepared in various ratios in o/w emulsion. The droplet size of the emulsion was measured using a particle size analyzer, and the turbidity was measured using a turbidity meter and a ultraviolet (UV)-vis spectrophotometer. According to the Orafidiya-Oladimeji method, the HLB value of the emulsion having the minimum dispersion ratio, the minimum droplet size, and the maximum turbidity degree was determined as the required HLB value for each essential oil. Based on these methods, the required HLB values of CUFO, CUPO, HF, and Camellia japonica seed oil were determined as 14.75-14.90, 15.35-15.40, 6.30-7.06, and 5.94-6.30, respectively.

Protective effect of Cornus walteri Wangerin leaf against UVB irradiation induced photoaging in human reconstituted skin.

ETHNOPHARMACOLOGICAL RELEVANCE: Cornus walteri Wangerin has been used in oriental traditional medicine for the treatment of antidiarrheal and inflammation. AIM OF THE STUDY: The efficacy of Cornus walteri Wangerin on skin anti-photoaging was investigated. MATERIALS AND METHODS: Hydrolyzed Cornus walteri Wangerin leaf was tested for the anti-photoaging effects against ultraviolet B (UVB)-induced matrix metalloproteinase (MMP)-1, pro-inflammatory cytokines using human reconstituted skin (KeraSkin™-FT) and also tested for elastase activity in vitro. The MMP-1 and pro-inflammatory cytokine levels of the extract were evaluated by enzyme-linked immunosorbent assay (ELISA). RESULTS: The extract of hydrolyzed Cornus walteri Wangerin leaf (CWE) had the elastase inhibitory activity (IC50: 0.457mg/mL). CWE inhibited MMP-1 expression up to 61% in comparison with the control group which was not treated using CWE, but exposed to UVB. CWE also showed an inhibitory effect on releasing pro-inflammatory cytokines (IL-6 and IL-8) in KeraSkin™-FT (30% and 57% inhibition at dose of 50µg/mL, respectively). CONCLUSION: CWE is a promising anti-photoaging agent for the treatment of UVB-induced skin.

Visible-to-near IR quantum dot-based hypermulticolor high-content screening of herbal medicines for the efficacy monitoring of hair growth promotion and hair loss inhibition.

There is a growing interest in alopecia prevention strategies, as the number of alopecia patients is increasing. We examine the efficacy of herbal medicine for hair growth promotion/hair loss inhibition in two cell lines via Western blot and high-content screening (HCS). Nine herbal extracts were obtained from three different herbal medicine mixtures using 3 different extraction methods. Five target proteins-IGF-1 (insulin-like growth factor-1), TGF-ß2 (transforming growth factor-ß2), VEGF (vascular endothelial growth factor), DKK-1 (Dickkopf-1), and Wnt5α-were observed for the assessment of hair growth promotion/hair loss inhibition efficacy. The efficacies of nine extracts were compared with minoxidil as control. Efficacy was defined as a rise in the expression levels of IGF-1, VEGF, and Wnt5α but a decrease in DKK-1 and TGF-ß2. Intracellular concurrent imaging of these proteins was successfully achieved using HCS, employing visible-to-near infrared probing based on quantum-antibody conjugates and hypermulticolor imaging.

Photoprotective effects of methoxycinnamidopropyl polysilsesquioxane.

A new sunscreen ingredient, methoxycinnamidopropyl polysilsesquioxane (MCP-PSQ), which contains an UV-absorbing p-methoxycinnamoyl group, has been developed synthetically and evaluated using in vitro and in vivo approaches. Previous studies revealed that MCP-PSQ has a raising or boosting effect on the sun protection factor (SPF) of other sunscreen agents. In this study, we demonstrated that MCP-PSQ, an organic/inorganic hybrid compound, has photoprotective effects for human fibroblasts, and for hairless mouse and human skin. MCP-PSQ increases cell viability and suppresses the expression of p53 protein in fibroblasts after UV exposure. In addition, the numbers of sunburn cells and mast cells are reduced by topical application of MCP-PSQ on hairless mouse skin after UV irradiation. A 10% MCP-PSQ cream has higher and similar effects on SPF values for human skin compared to 5% titanium dioxide (TiO(2)) and 5% ethylhexyl methoxycinnamate (EHMC), respectively. The SPF value obtained using the MCP-PSQ cream did not drop after UV irradiation of the cream itself. However, higher dose of UV irradiation is required to guarantee the stability or photostability of the formulation. Further, there were no side effects such as erythema, edema, itch or tingling, suggesting that MCP-PSQ is a good sunscreen agent.

MKK6 increases the melanocyte dendricity through the regulation of Rho family GTPases.

BACKGROUND: Melanocyte dendrites serve as the principal conduit for melanosome transfer. The dendrite formation requires actin polymerization mediated by Rho family GTPases including RhoA, Rac1 and Cdc42. OBJECTIVE: The aim of this study is to investigate and explore the involvement of p38 MAPK in melanocyte dendrite formation. METHODS: We transduced melanoma cells with adenovirus harboring the expression cassette for constitutive active form of MKK6, an upstream MAPKK for p38 MAPK. RESULTS: We investigated the effect of melanogenic inducers on melanocyte dendricity, using SK-mel-24 melanoma cells because that this cell line is refractory to several melanogenic inducers in terms of melanogenesis. TPA-induced the phosphorylation of p38 MAPK and the elongation of dendrite length, suggesting that MKK6 may be involved in this process. Overexpression of the constitutive active form of MKK6 resulted in significant elongation of dendrites in the melanoma cell line SK-mel-24. Moreover, overexpression of MKK6 ultimately led to the upregulation of Cdc42 and Rac1, suggesting that MKK6 acts as a crucial upstream signaling molecule for Rho family GTPases. When overexpressed in normal human epidermal melanocytes, MKK6 led also the increase of dendrite length. CONCLUSION: These results suggest that MKK6 is an authentic regulator for melanocytes dendricity, through the modulation of Rho family GTPases.

Anti-inflammatory effects of YeongyoSeungma-tang.

ETHNOPHARMACOLOGICAL RELEVANCE: YeongyoSeungma-tang which includes Fructus Forsythia, has been used in oriental traditional medicine for treatment of early smallpox and atopic dermatitis. AIM OF THE STUDY: YeongyoSeungma-tang was carried out to investigate for anti-inflammatory effects. MATERIALS AND METHODS: YeongyoSeungma-tang was tested for anti-inflammatory effects against lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E(2) (PGE(2)), and tumor necrosis factor alpha (TNF-alpha) releases as well as nuclear factor kappa B (NF-kappaB) expression using RAW264.7 macrophage cells. RESULTS: YeongyoSeungma-tang significantly inhibited generation of NO (42% and 59% inhibition at doses of 5 microg/mL and 10 microg/mL, respectively), PGE(2) (46% and 80% inhibition at doses of 5 microg/mL and 10 microg/mL, respectively) and TNF-alpha (6% and 23% inhibition at doses of 5 microg/mL and 10 microg/mL, respectively) on LPS-stimulated RAW 264.7 cells in a concentration-dependent manner. Consistently in these observations, the expression of inducible NO synthase (iNOS) enzyme was also inhibited by YeongyoSeungma-tang. However, YeongyoSeungma-tang did not show any influence on the expression of cyclooxygenase-2. The cream containing 0.075% YeongyoSeungma-tang showed good skin moisturizing effect without any irritation. CONCLUSION: The present study may support the fact that YeongyoSeungma-tang can have the good possibility as an anti-inflammatory agent for troubled skins.

Antioxidant and antimelanogenic activities of polyamine conjugates from corn bran and related hydroxycinnamic acids.

The antioxidant activity of three major polyamine conjugates, N,N'-dicoumaroyl-putrescine (DCP), N-p-coumaroyl-N'-feruloylputrescine (CFP), and N,N'-diferuloyl-putrescine (DFP) isolated from corn bran, and their related hydroxycinnamic acids, p-coumaric acid and ferulic acid, were evaluated by three antioxidant in vitro assay systems, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide and hydroxyl radicals generated by enzymatic and nonenzymatic reactions. Additionally, five phenolic compounds were evaluated for melanogenesis inhibitory activity using mushroom tyrosinase and B16 melanoma cells. Most of the phenolic compounds significantly scavenged DPPH, superoxide, and hydroxyl radicals in a dose-dependent manner. Particularly, DFP showed potent DPPH (IC50 = 38.46 microM) and superoxide (IC50 = 291.62 microM) radical scavenging activities, while DCP exhibited the strongest hydroxyl radical scavenging activity (IC50 = 120.55 microM). CFP also exerted moderate DPPH, superoxide, and hydroxyl radical scavenging activities. Meanwhile, DCP (IC50 = 181.73 microM) showed potent tyrosinase inhibitory activity toward l-tyrosine as the substrate, whereas DFP (IC50 = 733.64 microM) significantly inhibited melanin synthesis in B16 melanoma cells. These current results indicate that these three polyamine conjugates from corn bran may be useful potential sources of natural antioxidants and skin-whitening agents.