• Title/Summary/Keyword: Human keratinocyte HaCaT cell line

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In Vitro and in Vivo Wound Healing Properties of Plasma and Serum from Crocodylus siamensis Blood

  • Jangpromma, Nisachon;Preecharram, Sutthidech;Srilert, Thanawan;Maijaroen, Surachai;Mahakunakorn, Pramote;Nualkaew, Natsajee;Daduang, Sakda;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1140-1147
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    • 2016
  • The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

The Anti-bacterial Effects and Epidermal Permeability Barrier Function of Red Onion Juice Produced in Jeon-Nam province in Korea (전남 함평산 자색양파 추출물의 항균효과와 피부장벽 보호효과)

  • Youn, Dae-Hwan;Shin, Heon-Tae
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.1
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    • pp.43-56
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    • 2012
  • Objective : In this study, we assessed the anti-bacterial effects and epidermal permeability barrier function of red onion juice comparing to yellow onion juice and $Houttuynia$ $cordata$ extract $in$ $vitro$. Methods : 3types of red and yellow onion juice were prepared as antibacterial agent candidates with Houttuynia cordata hot water extract using 4 different bacterial strains ($Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$, $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$) by colony counting method. The expression of filaggrin, a marker of keratinocyte differentiation, and serine palmitoyl transferase (SPT), a marker of the formation of the stratum corneum lipid barrier, in human HaCat keratinocytes were analyzed using HaCaT cell line. The expression of COX-2 and AP-1 which is a factor of COX-2 transcription were also analyzed by western blotting method. Results : There was detectable anti-bacterial effects on $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$ among 1%, 5%, 10% extracts of yellow and red onion.(81%-100%) The bacteriocidal effects were not shown on $Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$ among $Houttuynia$ $cordata$, yellow onion and red onion extracts. The in vitro results showed the concentration-dependent effects on the expression of both filaggrin and SPT in HaCat cells among 0.01%, 0.05%, 0.1%, 0.5% extracts in Houttuynia cordata and red onion, reflecting the notion that $Houttuynia$ $cordata$ and red onion can induce epidermal keratinocyte differentiation and improve the recovery of skin barrier functions. The concentration-dependent effects also have been shown on the expression of both COX-2 and AP-1 among 0.01%, 0.05%, 0.1%, 0.5% extracts in $Houttuynia$ $cordata$ and red onion, while slight effect in yellow onion. Conclusion : Red onion juice could be a potential candidate enhanser for the skin care and cosmetology.

Development of Natural Antioxidants and Whitening Agents for Cosmeceuticals

  • Kim, Jong-Pyung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.11a
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    • pp.79-92
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    • 2007
  • Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

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