Alstonia scholaris R. Br. Significantly Inhibits Retinoid-Induced Skin Irritation In Vitro and In Vivo.

Topical retinoids inhibit matrix metalloproteinases and accelerate collagen synthesis, thereby triggering antiaging effects in the skin. However, topical retinoids can cause severe skin reactions, including scaling, erythema, papules, and inflammation. The present study demonstrates that the ethanolic bark extract of Alstonia scholaris R. Br. can significantly inhibit all-trans retinoic acid-induced inflammation in human HaCat keratinocyte cells. Furthermore, two representative retinoid-induced proinflammatory cytokines, monocyte chemoattractant protein-1 and interleukin-8, were significantly suppressed by A. scholaris extract (by 82.1% and 26.3% at 100 ppm, and dose-dependently across the tested concentrations) in vitro. In a cumulative irritation patch test, A. scholaris extract decreased retinol-induced skin irritation, while strengthening the ability of retinoids to inhibit matrix metalloproteinase-1 expression, which is strongly associated with aging effects. These results suggest that A. scholaris is a promising compound that may increase the antiaging function of retinoids while reducing their ability to cause skin irritation.

Effects of ortho-dihydroxyisoflavone derivatives from Korean fermented soybean paste on melanogenesis in B16 melanoma cells and human skin equivalents.

In this study we investigated the inhibitory effects and possible mechanisms of action of 8'-hydroxydaidzein and 3'-hydroxydaidzein, two ortho-dihydroxyisoflavone derivatives from Korean fermented soybean paste, on melanogenesis in B16 murine melanoma cells. The two hydroxydaidzeins reduced melanin synthesis comparably to treatment with kojic acid, a proven whitening agent, in B16 melanoma cells. Furthermore, when in vitro human skin equivalents were treated with the hydroxydaidzeins, the levels of melanogenesis were significantly reduced relative to a kojic acid control. The RT-PCR results demonstrated that depigmentation was due to transcriptional repression of several melanogenesis genes, including microphthalmia-associated transcription factor (MITF), by the hydroxydaidzeins. The immunoblotting results confirmed that diminution of MITF expression subsequently decreased expression of tyrosinase, and tyrosinase-related proteins 1 and 2. Cumulatively, these results suggest that hydroxydaidzeins would be potent attenuators of melanin synthesis as well as effective inhibitors of hyperpigmentation in human skin.

Amentoflavone inhibits UVB-induced matrix metalloproteinase-1 expression through the modulation of AP-1 components in normal human fibroblasts.

Amentoflavone is a well-known biflavonoid that has diverse biological effects. Previously, we reported that amentoflavone suppressed UVB-induced matrix metalloproteinase-1 (MMP-1) expression in normal human fibroblasts (NHF). We investigated the effects of amentoflavone on UVB-induced MMP-1 expression in order to elucidate its mode of action. NHF were treated with amentoflavone for indicated times and doses with UVB irradiation. The expressions of MMP-1 gene and protein were determined by RT-PCR and ELISA, respectively. MAP kinase phosphorylation and the expression of c-Fos protein were determined by Western blot. The treatment of amentoflavone completely blocked the upregulation of MMP-1 which is induced by UVB irradiation in HaCaT-NHF co-culture in a dose-dependent manner as well as in NHF monoculture. Also, amentoflavone inhibited UVB-induced activation of extracellular signal-regulated kinase (ERK) without changing total ERK protein level, and did not affect p38 or JNK activation. Finally, AP-1 transcription factor components, phospho-c-Jun and c-Fos protein expressions were decreased by amentoflavone treatment. The major finding of this study shows that amentoflavone inhibits intracellular cell signaling ERK pathway leading to the prevention of MMP-1 expression in human skin fibroblasts. Therefore, these results strongly suggest that amentoflavone should be investigated as a potential agent for the prevention and the treatment of skin photoaging.

Inhibition effect of Gynura procumbens extract on UV-B-induced matrix-metalloproteinase expression in human dermal fibroblasts.

ETHNOPHARMACOLOGICAL RELEVANCE: Gynura procumbens Merr. (Asteraceae) has been used as a traditional remedy for various skin diseases in certain areas of Southeast Asia. AIM OF THE STUDY: In order to evaluate the protective activity of Gynura procumbens extract on skin photoaging and elucidate its mode of action. MATERIALS AND METHODS: Matrix-metalloproteinase (MMP)-1 and -9 expressions were induced by UV-B irradiation in human primary dermal fibroblasts. MMP-1 expression level was measured by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Zymography was employed for evaluating the enzymatic activity of MMP-9. Anti-inflammatory activity and anti-oxidative capacity of the extract were evaluated by ELISA and dichlorodihydrofluorescein diacetate (DCF-DA) assay. RESULTS: The ethanolic extract of Gynura procumbens inhibited MMP-1 expression up to 70% compare to negative control group. The enzymatic activity of MMP-9 was inhibited around 73% by the treatment of 20µg/mL of the extract. The extract markedly reduced the production of reactive oxygen species (ROS). Gynura procumbens extract showed an inhibitory effect on releasing pro-inflammatory cytokines (IL-6 and IL-8) in human HaCat keratinocyte. CONCLUSION: The ethanolic extract of Gynura procumbens inhibited MMP-1 and MMP-9 expressions induced by UV-B irradiation via inhibition of pro-inflammatory cytokine mediator release and ROS production.

An improved method for measurement of change in skin roughness caused by cleansing products under mild application conditions.

BACKGROUND: The aim of this study was to develop a method for the evaluation of subtle change in skin roughness caused by cleansing products under mild application conditions using a non-invasive three-dimensional (3D) analysis system. METHODS: A double-blind comparative study of the modified soap chamber test was performed using two soap bars and a syndet bar. Skin changes were evaluated by visual scoring [mean cumulative irritation index (MCII)] and by bioengineering measurements [transepidermal water loss (TEWL), skin capacitance, and skin surface roughness]. RESULTS: MCII of the syndet bar was statistically higher than that of one soap bar, and TEWL increase after application of the syndet bar was statistically higher than that of both soap bars. Skin capacitance decreased significantly only after application of the syndet bar. The change in the average roughness of the skin surface was significantly greater after the application of the syndet bar than with classic soap bars. CONCLUSION: A simple, fast, and objective evaluation of skin surface topography was performed using a modified soap chamber test and a non-invasive 3D analysis system. The results suggest that measurement of skin roughness using a non-invasive 3D analysis system might be a good method for the evaluation of a subtle change caused by cleansing products under mild application conditions.

Inhibitory effect of panduratin A isolated from Kaempferia panduarata Roxb. on melanin biosynthesis.

Hyperpigmentation disorders are associated with abnormal accumulation of melanin pigments, thus melanin synthesis inhibitors have been of great interest as target molecules for cosmetic and medicinal purposes. The aim of this study was to investigate the in vitro inhibitory effect of panduratin A, isolated from Kaempferia pandurata Roxb., on melanogenesis and its related enzymes such as tyrosinase, tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2) in melan-a murine melanocytes. The IC(50) values of panduratin A for melanogenesis and tyrosinase were 9.6 µm and 8.2 µm, respectively, while those of arbutin as a positive control were 990 µm and 660 µm, respectively. In western blot analysis, panduratin A also significantly decreased tyrosinase, TRP-1 and TRP-2 protein levels. These results indicate that panduratin A effectively inhibits melanin biosynthesis, thus creating the possibility of developing a new skin-whitening agent.

Nanosized emulsions stabilized by semisolid polymer interphase.

We introduce a new approach for stabilizing oil-in-water nanoemulsions using a semisolid interphase formed by the phase separation of amphiphilic block copolymers from the organic phase. This system is illustrated using an amphiphilic diblock copolymer, poly(ethylene oxide)-block-poly(epsilon-caprolactone) (PEO-b-PCL), with commonly used oils. PEO-b-PCL can be miscible with oil at elevated temperatures (70-80 degrees C); however, polymer/oil demixing occurs as the temperature drops below the melting temperature of PEO-b-PCL (approximately 55 degrees C). A homogeneous polymer/oil mixture was dispersed in water at 80 degrees C to generate embryonic emulsions, and then the emulsion size was reduced to a nanometer range through microfluidic homogenization. The structure of the generated nanoemulsions is irreversibly frozen as they are cooled down to ambient temperature. The nanoemulsions stabilized by PEO-b-PCL show the excellent colloidal stability against thermal and chemical stresses, exhibiting no significant changes in the size distribution during incubation for 4 months at ambient temperature or 10 days at 60 degrees C. This study demonstrates that PEO-b-PCL is an attractive emulsifying material for practical nanoemulsion formulations requiring structural stability under a broad range of conditions.

Silicone oil emulsions stabilized by semi-solid nanostructures entrapped at the interface.

Oil-in-water (O/W) emulsions are typically stabilized using water-soluble surfactants, which anchor to the surface of oil droplets dispersed in an aqueous solution. The structure of the anchored surfactants is often susceptible to physical and chemical stresses because of their highly mobile properties. Here we introduce a new approach to prepare stable silicone oil emulsions under various external stresses using a water-insoluble amphiphilic block copolymer, poly(ethylene oxide)-b-poly(epsilon-caprolactone) (PEO-b-PCL). Above the melting temperature (around 60 degrees C) of the hydrophobic segment (PCL), PEO-b-PCL can be dissolved in silicone oil. When the polymer/oil mixture is dispersed in water, PEO-b-PCL is irreversibly reorganized into solid nanostructures at the interface of the aqueous/organic phases. The resulting interfacial structures provide a robust physical barrier to the emulsion coarsening processes. Accordingly, the prepared emulsions exhibit excellent structural tolerance against external stresses, including variations in pH, ionic strength, and temperature.

Morphological effect of lipid carriers on permeation of lidocaine hydrochloride through lipid membranes.

We have studied how the transdermal delivery of lidocaine hydrochloride (LHC) is affected by the morphology of lipid carriers, liposomes and micelles, having the same lipid composition of 1-stearoyl-sn-glycero-3-phosphocholine (LPC) and cholesteryl hemisuccinate (CHEMS). In vitro drug permeation study, carried out on guinea pig skin, has revealed that the liposomes made of LPC and CHEMS significantly enhance the permeation rate of entrapped LHC; by contrast, the mixed micelles with the same composition decrease the degree of delivering co-existing LHC. Basically, we have also investigated the release kinetics of LHC through the cellulose membrane and found that both liposomes and micelles have a similar releasing profile. To experimentally demonstrate this unique behavior, we have observed the fluidity of stratum corneum liposomal membranes in the presence of either our liposomes or micelles. From this study, we have found that LPC/CHEMS liposomes fluidize the lipid membrane of stratum corneum lipids; however, lipid micelles rather make the membrane rigid. These findings highlight that controlling the morphology of drug carriers provides us with a means to modulate the permeability of encapsulated drug molecules.

MAP17 is associated with the T-helper cell cytokine-induced down-regulation of filaggrin transcription in human keratinocytes.

In the meta-analysis of public microarray databases for different skin diseases, we revealed seven commonly up-regulated genes, DSG3, KRT6, MAP17, PLSCR1, RPM2, SOD2 and SPRR2B. We postulated that the genes selected from the meta-analysis may be potentially associated with the abnormal keratinocyte differentiation. To demonstrate this postulation, we alternatively evaluated whether the genes of interest in the meta-analysis can be regulated by T-helper (Th) cell cytokines in normal human epidermal keratinocytes (NHEK). We found that MAP17 was significantly up-regulated in response to interferon-gamma, interleukin 4 (IL-4), IL-6, IL-17A or IL-22 in NHEK. Interestingly, MAP17 was originally reported to interact with PDZK1; in turn, the PDZK1 gene is localized within the atopic dermatitis-linked region on human chromosome 1q21. In an attempt to evaluate whether MAP17 regulates the expression of cornified envelope-associated genes at the 1q21 locus, such as filaggrin, loricrin and involucrin, we found that the over-expression of MAP17 in HaCaT keratinocytes significantly decreased the expression of filaggrin. Taken together, the Th cell cytokine-induced up-regulation of MAP17 expression may be linked to the down-regulation of filaggrin in NHEK, which may be associated with the abnormal epidermal differentiation observed in the dermatological diseases.

Comparative study of the ocular irritation potential of various alkyl polyglucoside surfactants.

In the present work, we assessed the relationship between alkyl carbon chain length and ocular irritation potentials using the hen's egg test-chorioallantoic membrane (HET-CAM) and bovine corneal opacity and permeability (BCOP) assays using 5 commercial alkyl polyglucoside surfactants with different compositions of alkyl chain lengths (C(6)-C(16)). With HET-CAM, there was a good correlation between the proportion of C(10) alkyl polyglucoside and the eye irritation potential Q score (r(2) = 0.912, p = .011). There were no significant differences between the proportion of C(10) alkyl polyglucoside and corneal opacity in BCOP assays; however, there was a relatively high positive correlation between the proportion of C(10) alkyl carbon chain lengths and corneal permeability (r(2) = 0.736, p = .063).

Natural ortho-dihydroxyisoflavone derivatives from aged Korean fermented soybean paste as potent tyrosinase and melanin formation inhibitors.

Natural o-dihydroxyisoflavone (ODI) derivatives with variable hydroxyl substituent at the aromatic ring of isoflavone and three known isoflavones were isolated from five-year-old Korean fermented soybean paste (Doenjang) and evaluated as potent inhibitors on tyrosinase activity and melanin formation in melan-a cells comparing with other known isoflavones, 7,8,4'-trihydroxyisoflavone (1) and 7,3',4'-trihydroxyisoflavone (2) inhibited tyrosinase by 50% at a concentration of 11.21+/-0.8 microM and 5.23+/-0.6 microM (IC(50)), respectively, whereas, 6,7,4'-trihydroxyisoflavone (3), daidzein (4), glycitein (5) and genistein (6) showed very low inhibition activity. Furthermore, those compounds significantly suppressed the cellular melanin formation by 50% at a concentration of 12.23+/-0.7 microM (1), 7.83+/-0.7 microM (2), and 57.83+/-0.5(6) and show more activity than arbutin. But, compounds 3, 4, and 5 showed lower inhibition activity. This study shows that the position of hydroxyl substituent at the aromatic ring of isoflavone plays an important role in the intracellular regulation of melanin formation in cell-based assay system.

Development of a novel method for quantitative evaluation of sensory skin irritation inhibitors.

BACKGROUND/AIMS: Sensory skin irritation is regarded as one of the most serious side effects of cosmetic use. Thus, it is desirable to develop good inhibitors of sensory skin irritation. However, it is difficult to quantify the effect of sensory skin irritation inhibitors. METHODS: We investigated the possibility of using an electrical current perception threshold (CPT) measurement for the quantitative evaluation of these inhibitors. We divided study populations into stinger and non-stinger groups based on their response to 5% lactic acid and assessed CPT values at 2000, 250, and 5 Hz on the cheek. Stingers showed significantly lower CPT values than non-stingers did at 250 and 5 Hz. We also measured CPT values before and after the application of nine materials with inhibitory effects on sensory skin irritation. To investigate the relationship between the change in CPT values and the effect of each material in the clinical stinging test, we conducted the stinging test with the test materials in a 5% lactic acid solution and with a 5% lactic acid solution (positive control). RESULTS: There was a positive correlation between the change in CPT values and the inhibitory effect that each material had on the stinging test. CONCLUSION: The change in CPT values can be used for the quantitative evaluation of sensory skin irritation inhibitors .

Expression of surface markers on the human monocytic leukaemia cell line, THP-1, as indicators for the sensitizing potential of chemicals.

BACKGROUND: Evaluation of skin sensitization potential is an important part of the safety assessment of cosmetic ingredients and topical drugs. Recently, evaluation of changes in surface marker expression induced in dendritic cells (DC) or DC surrogate cell lines following exposure to chemicals represents one approach for in vitro test methods. OBJECTIVE: The study aimed to test the change of expression patterns of surface markers on THP-1 cells by chemicals as a predictive in vitro method for contact sensitization. METHODS: We investigated the expression of CD54, CD86, CD83, CD80, and CD40 after a 1-day exposure to sensitizers (1-chloro-2,4-dinitrobenzene; 2,4-dinitrofluorobenzene; benzocaine; 5-chloro-2-methyl-4-isothiazolin-3-one; hexyl cinnamic aldehyde; eugenol; nickel sulfate hexahydrate; potassium dichromate; cobalt sulfate; 2-mercaptobenzothiazole; and ammonium tetrachloroplatinate) and non-sensitizers (sodium lauryl sulfate, benzalkonium chloride, lactic acid, salicylic acid, isopropanol, and dimethyl sulphoxide). The test concentrations were 0.1x, 0.5x, and 1x of the 50% inhibitory concentration, and the relative fluorescence intensity was used as an expression indicator. RESULT AND CONCLUSION: By evaluating the expression patterns of CD54, CD86, and CD40, we could classify the chemicals as sensitizers or non-sensitizers, but CD80 and CD83 showed non-specific patterns of expression. These data suggest that the THP-1 cells are good model for screening contact sensitizers and CD40 could be a useful marker complementary to CD54 and CD86.

(-)-Catechin promotes adipocyte differentiation in human bone marrow mesenchymal stem cells through PPAR gamma transactivation.

Green tea intake has been shown to confer various health benefits to patients suffering from metabolic disorders. Here, we studied the effect of several major green tea polyphenols on adipocyte differentiation in human bone marrow mesenchymal stem cells (hBM-MSCs) and compared it to the effect of representative antidiabetic drugs. (-)-Catechin was the most potent of the eight green tea polyphenols evaluated in promoting adipocyte differentiation in hBM-MSCs, and this effect was dose-dependent. (-)-Catechin increased the mRNA levels of various adipogenic markers, such as adiponectin, peroxisome proliferator-activated receptor gamma (PPARgamma), FABP4, and LPL, as measured during adipocyte differentiation in hBM-MSCs. In addition, (-)-catechin upregulated the secretion of adiponectin in hBM-MSC culture. Using a reporter gene assay and a competitive ligand binding study, (-)-catechin also significantly activated PPARgamma in a dose-dependent fashion; however, (+)-catechin, the enantiomer of (-)-catechin, was not effective as a PPARgamma agonist, which seems to imply that the effect of (-)-catechin on PPARgamma is stereospecific. In conclusion, our data suggest that (-)-catechin promotes adipocyte differentiation and increased sensitivity to insulin in part by direct activation of PPARgamma, which could be at the basis of the observed pharmacological benefits of green tea intake in reducing the risk of type 2 diabetes.

ortho-dihydroxyisoflavone derivatives from aged Doenjang (Korean fermented soypaste) and its radical scavenging activity.

One new ortho-dihydroxyisoflavone, 7,3',4'-trihydroxyisoflavone (2), and two known ortho-dihydroxyisoflavone derivatives were isolated from 5-year-old Doenjang (Korean fermented soypaste), and evaluated as potent antioxidant by comparing with other known isoflavones. 7,8,4'-Trihydroxyisoflavone (1), 7,3',4'-trihydroxyisoflavone (2), and 6,7,4'-trihydroxyisoflavone (3) inhibited DPPH (Diphenyl-1-picryl hydrazyl) formation by 50% at a concentration of 21.5+/-0.2, 28.7+/-0.4 and 32.6+/-0.6 (IC(50)), respectively, whereas three isoflavones showed weak DPPH radical scavenging activity. In xanthine oxidase (XO) system, in which both inhibition of xanthine oxidase and superoxide scavenging effect were measured in one assay. Compound 1 (IC(50)= 6.6+/-0.4 microM) and 2 (IC(50)=16.8+/-1.2 microM) show significant inhibitory activity and greater effect than allopurinol. But, compound 3 and other isoflavones showed lower inhibition activity. This study shows that the position of hydroxyl substituent at the aromatic ring of isoflavone plays an important role in radical scavenging effect.