A Melanocyte–Keratinocyte Coculture Model to Assess Regulators of Pigmentation in Vitro

Lei, Tiechi; Virador, Victoria; Vieira, Wilfred D.; Hearing, Vincent J.

doi:10.1006/abio.2002.5665

Cited by 87 publications

(75 citation statements)

References 43 publications

(27 reference statements)

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“…5, it is known that MSH-induced melanogenesis in mouse melanoma cells occurs by an increase of intracellular cyclic AMP content, an accumulation of tyrosinase mRNA, and a stimulation of trosinase activity. 35) Diarylheptanoids from Alnus hirsute Turcz showed melanogenesis inhibitory activities in B16 cells, 36) and cucurbitacins from Trichosanthes kirilowii showed inhibitory effects for tyrosinase activity and melanin synthesis in B16 cells. 37) These inhibitory effects reported by others in B16 system were similar to what found the current study.…”

Section: Discussionmentioning

confidence: 99%

4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

Kim¹,

Lee

et al. 2005

Biological & Pharmaceutical Bulletin

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Tyrosinase (monophenol, dihydroxy-L-phenylalanin: oxygen oxidoreductase, EC 1.14.18.1) is a copper-dependent protein widely distributed in nature.2) The enzyme catalyzes two different reactions: the hydroxylation of monophenols to o-diphenols (monophenolase activity) and the oxidation of odiphenols to o-quinones (diphenolase activity), both utilizing molecular oxygen.3)The enzymatic browning of fruits and vegetables is mostly related to the oxidation of endogenous phenolic containing compounds catalyzed by polyphenol oxidase. 4) Tyrosinase also is responsible for melanization in animals, and is the key enzyme for the regulation of melanogenesis in mammals. 5)Melanin is synthesized in epidermal melanocytes, and then is transferred into epidermal keratinocytes via the melanocytes' dendrites. 6) Melanogenesis is the process by which melanin is produced (and subsequently distributed) by melanocytes within the skin and hair follicles.7) This process results in the synthesis of melanin pigments, which plays a protective role against skin photocarsinogenesis.8) The main physiological stimulus of melanogenesis is the ultraviolet radiation of solar light, which can act directly on melanocytes or indirectly through the release of keratinocyte-derived factors such as MSH (a-melanocyte stimulating hormone). 9)One of the biggest causative agents of hyperpigmentaion is probably UV light. 10) However, skin darkening can be suppressed, at least partially, by deactivating of tyrosinase. 11)Therefore, tyrosinase inhibitors have become increasingly important in the cosmetic and medicinal products used in the prevention of hyperpigmentaion.12) Many compounds, such as hydroquinone, 13) kojic acid, 14) and benzaldehyde-O-alkyloximes 15) have been reported as tyrosinase inhibitors. The established murine B16F10 melanoma cell (B16 cells) line offers a model system with readily quantifiable markers that are characteristic of differentiating, including melanogenesis. 16) In B16 cells, a-tocopheryl ferulate was shown to have depigmenting effect. 17) Although 4,4Ј-dihydroxybiphenyl (44Ј-BP), a bisphenol derivative was known as a radical scavenger for methacrylate polymerization, 18) but its efficacy on tyrosinase inhibition and melanin biosynthesis has not been reported.The aim of this study was to characterize inhibition mode of 44Ј-BP on mushroom tyrosinase. We also examined the inhibitory effect of 44Ј-BP on melanin biosynthesis, and cell viability in B16 cells as a biomarker for the potential cytotoxicity. Cell Culture B16 cells (from Korean Cell Line Bank) were cultured in DMEM with 10% fetal bovine serum (FBS; Gibco) and penicillin/streptomycin (100 IU/50 mg/ml) in a humidified atmosphere containing 5% CO 2 in air at 37°C. B16 cells were cultured in 24-well plates for the melanin quantification and enzyme activity assays. MATERIALS AND METHODS MaterialsDetermination of 44-BP for Cell Viability The cell proliferation assay was carried out by the method of Tada et al. using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (...

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Section: Discussionmentioning

confidence: 99%

4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

Kim¹,

Lee

et al. 2005

Biological & Pharmaceutical Bulletin

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“…Determination of Melanin Content in Vitro-Melanin content was determined as described previously (40,41) with some modifications. Cells were homogenized with 1 N NaOH.…”

Section: Methodsmentioning

confidence: 99%

Suppression of Melanin Production by Expression of HSP70

Hoshino

Matsuda

Yamashita

et al. 2010

Journal of Biological Chemistry

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Skin hyperpigmentation disorders due to abnormal melanin production induced by ultraviolet (UV) irradiation are both a clinical and cosmetic problem. UV irradiation stimulates melanin production in melanocytes by increasing intracellular cAMP. Expression of heat shock proteins (HSPs), especially HSP70, is induced by various stressors, including UV irradiation, to provide cellular resistance to such stressors. In this study we examined the effect of expression of HSP70 on melanin production both in vitro and in vivo. 3-Isobutyl-1-methylxanthine (IBMX), a cAMP-elevating agent, stimulated melanin production in cultured mouse melanoma cells, and this stimulation was suppressed in cells overexpressing HSP70. IBMX-dependent transcriptional activation of the tyrosinase gene was also suppressed in HSP70-overexpressing cells. Expression of microphthalmia-associated transcription factor (MITF), which positively regulates transcription of the tyrosinase gene, was up-regulated by IBMX; however, this up-regulation was not suppressed in HSP70-overexpressing cells. On the other hand, immunoprecipitation and immunostaining analyses revealed a physical interaction between and co-localization of MITF and HSP70, respectively. Furthermore, the transcription of tyrosinase gene in nuclear extract was inhibited by HSP70. In vivo, UV irradiation of wild-type mice increased the amount of melanin in the basal layer of the epidermis, and this increase was suppressed in transgenic mice expressing HSP70. This study provides the first evidence of an inhibitory effect of HSP70 on melanin production both in vitro and in vivo. This effect seems to be mediated by modulation of MITF activity through a direct interaction between HSP70 and MITF.

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“…Earlier studies have shown that the melanin production in melanocytes is influenced by keratinocytes and fibroblasts. [23,24] Primary human melanocytes and keratinocytes have some drawbacks such as the limited proliferate capacity and the donor variability. Thus the use of immortalized cell lines would be more suitable for investigating the effects of pigmentation regulators.…”

Section: Discussionmentioning

confidence: 99%

Effects of hydroxybenzyl alcohols on melanogenesis in melanocyte-keratinocyte co-culture and monolayer culture of melanocytes

Liu

Chu

Pan

2008

Journal of Enzyme Inhibition and Medicinal Chemistry

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In mammalian skin, melanocyte proliferation and melanogenesis can be stimulated by keratinocytes, fibroblasts and other regulatory factors. To determine whether hydroxybenzyl alcohols (HBAs) show more inhibitory in melanocytes cultured alone or in melanocytes co-cultured with keratinocytes, we developed a murine melanocyte -keratinocyte co-culture model to investigate the pigmentation regulators in company with other melanogenic inhibitors and stimulators. It was found that the effects of HBAs and melanogenic factors were more evident in melanocytes co-cultured with keratinocytes. Keratinocytes may play a synergistic role in melanocyte melanogenesis and influence the pigment production. The tests in the co-culture model also imply that the inhibitory effects of HBAs on melanogenesis are due to the direct inhibition of melanosomal tyrosinase activity. HBAs showed a low cytotoxicity. The eventual results proved that HBAs are promising and safe agents for skin whitening in melanocyte alone and in co-culture systems. The co-culture model provides a more physiologically realistic condition to study the interaction between melanocytes and keratinocytes, which enables a reliable screening system for depigmenting compounds.

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A Melanocyte–Keratinocyte Coculture Model to Assess Regulators of Pigmentation in Vitro

Cited by 87 publications

References 43 publications

4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

Suppression of Melanin Production by Expression of HSP70

Effects of hydroxybenzyl alcohols on melanogenesis in melanocyte-keratinocyte co-culture and monolayer culture of melanocytes

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A Melanocyte–Keratinocyte Coculture Model to Assess Regulators of Pigmentation in Vitro

Cited by 87 publications

References 43 publications

4,4&prime;-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

4,4&prime;-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

Suppression of Melanin Production by Expression of HSP70

Effects of hydroxybenzyl alcohols on melanogenesis in melanocyte-keratinocyte co-culture and monolayer culture of melanocytes

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Product

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4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor

4,4′-Dihydroxybiphenyl as a New Potent Tyrosinase Inhibitor