• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.718-722
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• 2015

Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO₂) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO₂ under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO₂. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO₂ pretreated with a divalent, positively charged ion, Ca²⁺, and the melanin treatment activity of immobilized lysosomal enzymes on TiO₂ pretreated with Ca²⁺ was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.

• Journal of Pharmacopuncture
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• v.9 no.1
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• pp.75-82
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• 2006

Objective : This research was carried out for the development of medicine for vitiligo treatment and focused on the effect of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells. Methods : Acitivity of tyrosinase playing a vital role in synthesis of Melanin and the quantity of Melanin, which is the final product in cultured B16 melanoma cells, effects of Hominis Placenta extract were measured. Results: The results indicated that Hominis Placenta extract increased both the amount of Melanin and the activity of tyrosinase according to the concentration, and they also supported by western blot analysis. Conclusion : The results suggests that Hominis Placenta extract has an advantageous effect on the promotion of Melanin synthesis and will contribute to the development of vitiligo treatment through further related studies.

• YAKHAK HOEJI
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• v.45 no.6
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• pp.724-729
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• 2001

Melanogenesis is a physiological process resulted in the synthesis of melanin pigments, which has a role in protecting skin front the damaging effect of ultra-violet (UV) radiation. The main aim of the present study was to examine the effect of Ulmus davidiana var. japonica(UL) on Melanogenesis. Cells were cultured in the presence of various concentrations of Ulmus davidiana var. japonica for 48 h, and there were estimated total melanin contents as a final product and activity of tyrosinase, a key enzyme, in Melanogenesis. Among the four solvent extracts tested, EtOAc extract mostly increased tyrosinase activity, And EtOAc extract increased the melanin contents and tyrosinase activity in a dose-dependent manner. Especially It was observed that 100$\mu\textrm{g}$/ml EtOAc extract promotes melanin secretion in B16/F10 melanoma cells by 140% at 48 h treatment and activity of tyrosinase increased by 180% in the presence of same concentration. In conclusion, as for EtOAc extract treatment, there was no effect on the viability of B16/F10 cell, only to stimulate Melanogenesis.

• YAKHAK HOEJI
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• v.57 no.1
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• pp.24-31
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• 2013

Although statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, have been shown to increase melanin synthesis, the exact mechanism of this action is not fully understood. In this study we investigated the possible involvement of intracellular $Ca^{2+}$ signal in the mechanism of stimulation of melanin synthesis induced by lovastatin in B16 cells. Lovastatin stimulated the production of melanin in a dose-dependent manner in the cells. Treatment with mevalonate, FPP and GGPP, precursors of cholesterol, did not significantly suppress the lovastatin-induced melanin production, suggesting that inhibition of cholesterol synthesis may not be involved in the mechanism of the action of lovastatin. In addition, lovastatin did not significantly alter the cAMP concentration and the stimulated production of melanin by lovastatin was not significantly changed by treatment with H89, a potent inhibitor of protein kinase A, which demonstrates that cAMP pathway may not be involved. However, lovastatin increased intracellular $Ca^{2+}$ concentration in a dose-related fashion. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the lovastatin-induced intracellular $Ca^{2+}$ increase and melanin synthesis, whereas intracellular $Ca^{2+}$ reduction with BAPTA/AM and intracellular $Ca^{2+}$ release blockers (dantrolene and TMB-8) completely blunted these actions of lovastatin. Taken together, these results suggest that the intracellular $Ca^{2+}$ release may play an important role in the lovastatin-induced stimulation of melanin synthesis in B16 cells. These results further suggest that lovastatin may be useful for the treatment of hypopigmentation disorders, such as vitiligo.

• The Journal of Korean Medicine
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• v.26 no.3 s.63
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• pp.55-65
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• 2005

Objectives : This study was carried out for the development of medicine for vitiligo treatment and focused on the effect of Psoraleae fructus extract on melanin synthesis of B16 melanoma cells. Methods : Activity of tyrosinase playing a vital role in the synthesis and quantity of melanin, which is the final product in cultured B16 melanoma cells, the effects of Psoraleae fructus extract were measured. Results : The results indicated that Psoraleae fructus extract increased beth the amount of melanin and the activity of tyrosinase according to concentration, also supported by western blot analysis. Conclusions : The results suggest that Psoraleae fructus extract has an advantageous effect on the promotion of melanin synthesis and will contribute to the development of vitiligo treatment through further related studies.

• Journal of Life Science
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• v.27 no.12
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• pp.1452-1461
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• 2017

Melanin production by melanocytes in human hair follicles decreases with time and leads to the graying process, which is a phenotype of human aging and an index of aging. The reduction in melanin production is the result of decreased tyrosinase activity in hair follicles and an accumulation of active oxygen species, such as hydrogen peroxide. This study investigated antioxidant effects and melanin-promoting effects in B16F1 cells treated with black sesame ethanolic nonpolar-soluble extract (SBEEO) and black sesame ethanolic polar-soluble extract (SBEEP). In antioxidation experiments, both SBEEP and SBEEO did not eliminate 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical, but SBEEO at $64{\mu}g/ml$ showed low reducing power. SBEEP exerted cytotoxic effects at concentrations greater than $8{\mu}g/ml$, whereas SBEEO showed cytotoxic effects at concentrations greater than $4{\mu}g/ml$. SBEEP and SBEEO induced melanin synthesis, tyrosinase activity, and DOPA oxidase activity in vitro. In live cells, melanin synthesis was greater in the SBEEP treatment group as compared with that in the SBEEO treatment group. SBEEP stimulated melanin synthesis by modulating the expression of tyrosinase-related protein-2 (TRP-2), which is an important enzyme in melanin synthesis. These results imply that SBEEP obtained from black sesame ethanolic extract may have the potential to improve melanin synthesis.

• Journal of Microbiology and Biotechnology
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• v.28 no.3
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• pp.375-380
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• 2018

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

• KSBB Journal
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• v.32 no.3
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• pp.187-192
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• 2017

In this study, we investigated the effect of Rumex axetosella, Sonchus oleraceus and Euphoibia jolkini extracts on tyrosinase activity and melanin production as natural products of whitening functional cosmetics. To measure the melanin production, 50, 100, $200{\mu}g/mL$ of Rumex axetosella, Sonchus oleraceus and Euphoibia jolkini extracts were treated on ${\alpha}-MSH$ treated B16F10 melanoma cells, respectively. Melanin contents in ${\alpha}-MSH$ treated B16F10 melanoma cells were decreased by 41.5, 51.11, and 61% in $200{\mu}g/mL$ treatment compared to none treatment, respectively. In addition, the intracellular tyrosinase activity was decreased after treatments with all extracts. Furthermore, $100{\mu}g/mL$ of Euphoibia jolkini extract was decreased 81.5% of melanin production in B16F10 melanoma cells. When the three extracts were compared, Euphoibia jolkini extract was considered to be the most functional material for whitening effect.

• Journal of Periodontal and Implant Science
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• v.40 no.4
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• pp.201-205
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• 2010

Purpose: The purpose of this study was to evaluate the clinical effectiveness of and patient's satisfaction with treatment of gingival melanin hyperpigmentation with a Nd:YAG laser and a high speed rotary instrument. Methods: Three patients with melanin hyperpigmentation in the anterior parts of the gingiva were chosen for this case study. Clinical photographs were taken at the preoperative state and three patients were treated under local anesthesia. In the maxilla, the gingival deepithelization was conducted with a high speed diamond bur, whereas, in the mandible with a Nd:YAG laser. Clinical photographs were taken immediately after the procedures and at the 1st, 2nd, and 4th week to evaluate clinical color changes. A week after the procedure, the patients filled out a questionnaire about any pain or discomfort. At the 4th week after the procedure, the patients filled out questionnaires about esthetic aspects of the results of treatment. Results: In all cases, both anterior gingival areas were depigmented with satisfaction and the patients did not complain of severe pain or discomfort. At the 1st week of healing, the gingiva showed moderate to fast epithelization. Two weeks after the procedure, clinically, the gingiva showed almost complete healing. Four weeks after the procedure, there was significant improvement in gingival melanin hyperpigmentation. Conclusions: The Nd:YAG laser and the high speed rotary instruments seem to be effective for the esthetic treatment of gingival melanin hyperpigmentation.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.69-74
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• 2013

Objectives : Since hypopigmentation is known to increase the risk of skin cancer, melanogenesis in the skin needs to be regulated. Here, we evaluated the melanogenesis stimulatory effects of a modified Goagium herbal remedy (HR) and HR+ox bile (Bos taurus domesticus) extract (OBE) to address hypopigmentation disorders. Methods : B16F10 melanoma cells were treated with different dosages of HR and HR+OBE for 24 to 48 h after 1 h of 10 nM ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH). After the treatment, cell viability, tyrosinase activity, melanin synthesis and the expression of genes related to melanin synthesis were measured and the regulation of the ${\alpha}$-MSH signalling through cAMP responding element binding protein (CREB) was determined. Results : HR and HR+OBE with the ranges of $15{\sim}100{\mu}g/mL$ did not affect cell viability in melanoma cells. The 1 h treatment of HR+OBE (50 and $100{\mu}g/mL$) potentiated the phosphorylation of CREB by enhancing ${\alpha}$-MSH signaling and its 24 h treatment increased CREB expression. Consistent with CREB potentiation, their treatment for 24 h, the expression of microphthalmia-associated transcription factor (MIFT), tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 were increased in realtime PCR. Ultimately, the 48 h treatment of HR+OBE (50 and $100{\mu}g/mL$) increased tyrosniase activity and melanin contents in the melanoma cells in comparison to the control. Conclusions : HR+OBE (50 and $100{\mu}g/mL$) increases melanin synthesis in B16F10 melanoma cells via the stimulation of tyrosinase activity and expression of MIFT, tyrosinase, TRP-1 and TRP-2. HR+OBE can be used as the a possible treatment for hypopigmentation of the skin.