Investigation of symptoms of hand skin changes with aging in Korean women and development of a new standard grading scale for hand aging.

BACKGROUND: Skin aging, particularly facial skin, has been actively studied. However, hand skin research is limited. METHODS: Aging symptoms of 100 hands of Korean women aged from 20s to 60s were measured by noninvasive and bioengineering methods. Standard grade images were produced. RESULTS: As people got older, skin wrinkles were getting worse and skin tone was uneven with the occurrence of structural flexion. For each symptom, a suitable standard photograph of the skin of the hand was chosen and a new grading scale was made. CONCLUSIONS: The new grading scale developed in the present study could be employed in studies to explore aging of hand skin as one of objective indicators.

Analysis of the temporal change in biophysical parameters after fractional laser treatments using reflectance confocal microscopy.

BACKGROUND: Fractional photothermolysis is a popular treatment option for photorejuvenation. Previous literature studies have demonstrated the clinical effectiveness of fractional photothermolysis on cutaneous photoaging; however, the associated changes in biophysical properties of the skin following fractional photothermolysis have not been fully elucidated. This study was conducted to investigate the temporal changes in biophysical parameters after fractional laser treatment on Asian skin. MATERIALS AND METHODS: Eleven female subjects underwent a single treatment with an erbium glass fractional laser. Skin roughness, elasticity, transepidermal water loss (TEWL), dermal thickness were evaluated before and immediately after treatment and 3 days, 1 week, 2 weeks, and 4 weeks after treatment. The changes in the dermal papilla were analyzed using a reflectance confocal microscopy (RCM). RESULTS: Skin roughness showed the greatest improvement at the first week and net elasticity was most improved at the second week. TEWL and the percentage of melanized and active dermal papillae (DP) were mostly increased for 3 days. At 4 weeks after treatment, the number of total dermal papillae showed a significant increase compared with pretreatment. CONCLUSION: This is the first study of the characterization and quantification of dermal papilla reflecting the dermal repair process after fractional photothermolysis through an RCM.

Use of non-melanocytic HEK293 cells stably expressing human tyrosinase for the screening of anti-melanogenic agents.

Tyrosinase (TYR) from mushrooms has been inappropriately used in the screening assay for hypopigmenting agents even though its biochemical properties are different from those of human TYR. Cell-free extracts of human epidermal melanocyes (HEMs) could be another choice for the assay, but HEMs grow too slowly to get a sufficient amount of cell-free extracts. In the present study, human embryonic kidney (HEK) 293 cells were transfected with a human TYR construct to establish a cell line that grows rapidly and expresses human TYR constitutively. Cell-free extracts of the established cell line, HEK293-TYR, were tentatively used in the screening assays for 11 phenylpropanoids that have chemical structures similar to that of L-tyrosine, the substrate of TYR. Of the 11 compounds, the strongest inhibition of TYR activity was shown by p-coumaric acid (IC50, 3 µM), followed by 3-(4-hydroxyphenyl)propionic acid (50 µM) and 3-(4-hydroxyphenyl)lactic acid (70 µM). The results indicate that p-coumaric acid has an optimal chemical structure for the inhibition of TYR. The effects of these phenylpropanoids on melanin synthesis in HEMs correlated well with their effects on TYR activity in vitro. This study demonstrated that HEK293-TYR cells can be a good source of the human TYR enzymes needed in the screening assay of anti-melanogenic agents.

Acanthopanax koreanum fruit waste inhibits lipopolysaccharide-induced production of nitric oxide and prostaglandin E2 in RAW 264.7 macrophages.

The Acanthopanax koreanum fruit is a popular fruit in Jeju Island, but the byproducts of the alcoholic beverage prepared using this fruit are major agricultural wastes. The fermentability of this waste causes many economic and environmental problems. Therefore, we investigated the suitability of using A. koreanum fruit waste (AFW) as a source of antiinflammatory agents. AFWs were extracted with 80% EtOH. The ethanolic extract was then successively partitioned with hexane, CH(2)Cl(2), EtOAc, BuOH, and water. The results indicate that the CH(2)Cl(2) fraction (100 microg/mL) of AFW inhibited the LPS-induced nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in RAW 264.7 cells by 79.6% and 39.7%, respectively. These inhibitory effects of the CH(2)Cl(2) fraction of AFWs were accompanied by decreases in the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins and iNOS and COX-2 mRNA in a dose-dependent pattern. The CH(2)Cl(2) fraction of AFWs also prevented degradation of IkappaB-alpha in a dose-dependent manner. Ursolic acid was identified as major compound present in AFW, and CH(2)Cl(2) extracts by high performance liquid chromatography (HPLC). Furthermore using pure ursolic acid as standard and by HPLC, AFW and CH(2)Cl(2) extracts was found to contain 1.58 mg/g and 1.75 mg/g, respectively. Moreover, we tested the potential application of AFW extracts as a cosmetic material by performing human skin primary irritation tests. In these tests, AFW extracts did not induce any adverse reactions. Based on these results, we suggest that AFW extracts be considered possible anti-inflammatory candidates for topical application.

p-coumaric acid not only inhibits human tyrosinase activity in vitro but also melanogenesis in cells exposed to UVB.

Tyrosinase (TYR) catalyzes rate-limiting steps of melanogenesis and thus its inhibitors are potentially useful as hypopigmenting agents. Recently, p-coumaric acid (p-CA) has been suggested to interfere with the pro-melanogenic actions of tyrosine due to its structural similarity with tyrosine (An SM et al., Br J Dermatol 2008. 159: 292). In this study, we compared the inhibitory effects of p-CA and two other well known TYR inhibitors used in cosmetics--arbutin and kojic acid--on the catalytic activities of mushroom, murine and human TYRs in vitro, using tyrosine and 3,4-dihydroxyphenylalanine (DOPA) as substrates. The results showed that p-CA is a weaker inhibitor of mushroom TYR but much stronger inhibitor of human or murine TYR in comparison with kojic acid and arbutin. In addition, p-CA inhibited human TYR at much lower concentrations than those required for the inhibition of murine or mushroom TYRs. Enzyme kinetics analysis indicated that p-CA is a mixed type (for tyrosine) or competitive inhibitor (for DOPA) of human TYR. Potent antimelanogenic effects of p-CA were observed in human epidermal melanocytes exposed to UVB. The present study demonstrated that p-CA is a potent and selective inhibitor of human TYR and is potentially useful as a hypopigmenting agent.

Effect of Camellia japonica oil on human type I procollagen production and skin barrier function.

Type I collagen is the primary component of the skin dermis. Both the quantity and quality of extracellular collagen are primarily related to skin ageing. In this study, we investigated the possibility that Camellia japonica oil (CJ oil) may be introduced as an anit-wrinkle agent. As a first step to this end, human COL1A2 promoter luciferase assay was performed in human dermal fibroblast cells. CJ oil was determined to activate human COL1A2 promoter in a concentration-dependent manner. In consistency with this result, while matrix metalloproteinase (MMP)-1 activity was inhibited by CJ oil, human type I procollagen synthesis was also induced by CJ oil. These results suggest the possibility that CJ oil may be involved in the skin ageing. For the evaluation of CJ oil's safety and efficiency on human skin, human skin primary irritation test and trans-epidermal water loss (TEWL) were performed. Transepidermal water loss (TEWL) was measured before treatment then, 1h and 2h after treatment; the forearm site was selected to measure TEWL. Also, a human skin primary irritation test was performed on the normal skin (upper back) in 30 volunteers to see if a certain material included in CJ oil has irritation or sensitization potential. In these assays, CJ oil reduced trans-epidermal water loss (TEWL) and did not induce any adverse reactions. Therefore, based on these results, we suggest the possibility that CJ oil may be considered as possible wrinkle-reducing candidates for topical application.

Effects of resveratrol, oxyresveratrol, and their acetylated derivatives on cellular melanogenesis.

Resveratrol and oxyresveratrol are naturally occurring phenolic compounds with various bioactivities, but their uses in cosmetics have been partly limited by their chemical instabilities. This study was performed to examine the anti-melanogenic effects of the acetylated derivatives from resveratrol and oxyresveratrol. Resveratrol and oxyresveratrol were chemically modified to triacetyl resveratrol and tetraacetyl oxyresveratrol, respectively. The acetylated compounds were less susceptible than the parent compounds to oxidative discoloration. The acetylated compounds inhibited the activities of tyrosinases less than parent compounds in vitro, but they were as effective at cellular melanogenesis inhibition, indicating bioconversion to parent compounds inside cells. Supporting this notion, the parent compounds were regenerated when the acetylated compounds were digested with cell lysates. Although resveratrol and triacetyl resveratrol inhibited tyrosinase activity less effectively than oxyresveratrol and tetraacetyl oxyresveratrol in vitro, they inhibited cellular melanogenesis more effectively. This discrepancy was explained by strong inhibition of tyrosinase expression by resveratrol and triacetyl resveratrol. Experiments using a reconstituted skin model indicated that resveratrol derivatives can affect melanin synthesis and cell viability to different extents. Collectively, this study suggests that acetylated derivatives of resveratrol have great potential as anti-melanogenic agents for cosmetic use in terms of efficacy, safety, and stability.

Comparison of the antimelanogenic effects of p-coumaric acid and its methyl ester and their skin permeabilities.

BACKGROUND: p-Coumaric acid (PCA) inhibits human tyrosinase (TYR) activity and melanin synthesis in human epidermal melanocytes. OBJECTIVE: The purpose of the current study was to examine the potential of PCA and its hydrophobic derivative, methyl p-coumarate (MPC), as hypopigmenting agents for topical use. METHODS: PCA and MPC were comparatively tested against in vitro human TYR enzyme activity and cellular melanin synthesis in human epidermal melanocytes. Permeation studies were undertaken using an artificial lipophilic membrane and an excised porcine skin. In vivo hypopigmenting efficacy was assessed on the skin of melanin-possessing hairless mice exposed to UVB. RESULTS: Although PCA was a stronger inhibitor than MPC against TYR activity in vitro, the former inhibited cellular melanin synthesis less effectively than the latter. A non-cell based permeability assay indicated that PCA was practically impermeable through the lipophilic barrier while MPC was highly permeable. In contrast, an ex vivo skin permeation study demonstrated that topically applied PCA in the form of a cream can diffuse into the aqueous medium underneath the skin. No MPC was released from a MPC cream but PCA was released instead as a bio-converted product. Topical application of PCA cream attenuated the UVB-induced erythema formation and pigmentation in mice models, more effectively compared with MPC cream. CONCLUSION: PCA may be useful as an active ingredient for topical applications for a hypopigmenting effect. MPC has potential as a hypopigmenting agent but requires rather invasive methods for its delivery to the target cells.

Inhibition of melanogenesis by tyrosinase siRNA in human melanocytes.

Tyrosinase (TYR) plays a critical role in cellular melanogenesis and, thus, has been the major target of pharmacological approaches for the control of skin pigmentation. This study examined an alternative molecular approach using TYR-small interfering RNA (siRNA) to control melanogenesis in the human melanocytes. Both the mRNA and protein levels of TYR were significantly lowered by TYR-siRNA treatment, whereas TYR-related protein 1 and TYR-related protein 2 displayed no such changes. TYR-siRNA treatment inhibited the cellular melanin synthesis from the externally supplied TYR substrate L-tyrosine. TYR-siRNA also suppressed melanin synthesis and decreased the viability of cells exposed to ultraviolet radiation, supporting a critical role of melanin in protection against ultraviolet radiation. These results suggest that molecular approaches using siRNA targeted to the enzymes of melanogenic pathway may provide a novel strategy for the control of cell pigmentation.

Oenothera laciniata inhibits lipopolysaccharide induced production of nitric oxide, prostaglandin E2, and proinflammatory cytokines in RAW264.7 macrophages.

We elucidated the pharmacological and biological effects of Oenothera laciniata extracts on the production of inflammatory mediators in macrophages. The CH(2)Cl(2) fraction of O. laciniata extract effectively inhibited LPS-induced NO, PGE(2), and proinflammatory cytokine production in RAW264.7 cells. These inhibitory effects of the CH(2)Cl(2) fraction of O. laciniata were accompanied by decreases in the expression of iNOS and COX-2 proteins and iNOS, COX-2, TNF-alpha, IL-1beta, and IL-6 mRNA. Asiatic acid and quercetin were present in the HPLC fingerprint of the O. laciniata extract. We tested the potential application of O. laciniata extract as a cosmetic material by performing primary skin irritation tests. In New Zealand white rabbits, primary irritation tests revealed that application of O. laciniata extracts (1%) did not induce erythema or edema formation. Human skin primary irritation tests were performed on the normal skin (upper back) of 30 volunteers to determine if any material in O. laciniata extracts had irritation or sensitization potential. In these assays, O. laciniata extracts did not induce any adverse reactions. Based on these results, we suggest that O. laciniata extracts be considered possible anti-inflammatory candidates for topical application.