Skin-Lightening Agent with Different Pathways of Action ... - Dermage

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COSMETICS SKIN-LIGHTENING To sum up, ET-1 as well as α-MSH control dendricity and have an influence on phagocytosis - the transfer of melanosomes from melanocytes to keratinocytes. In keratinocytes the melanosomes form the secondary lysosomes around the keratinocyte nucleus. Melanosomes in dark skin differ from those in skin types I and II, they are larger and packaged as single units, whereas the melanosomes in fair skin are smaller and packaged in groups. During differentiation of keratinocytes in the Stratum corneum the degradation of the melanosome unit is much stronger in skin types I and II than in dark skin, which generates a kind of »melanin dust effect« and make these skin types appear less pigmented (20). To study the influence of Belides on melanosome transfer we used a Fluosphere beads model. For simulation of melanosome uptake HaCaT cells were irradiated for phagocytosis stimulation with UV light. It could be shown that 1.0 or 1.5% Belides (1 DIV) significantly reduced the Fluosphere uptake (25.9 and 40.9% related to cell count), compared to control cultures (set to 100% uptake), when the phagocytic activity was monitored (Fig. 9). Furthermore, microscopic images taken from control cultures and cultures preincubated with Belides showed that a lower number of FluoSphere beads were accumulated around the cell nuclei when the keratinocytes were treated with Belides (Fig. 10). The in vitro results demonstrate that Belides acts as a functional ingredient at different stages of melanogenesis. By influencing pathways of action before, during and after melanin synthesis Belides effectively reduces skin pigmentation. Finally, to demonstrate that Belides produces a noticeable lightening effect on human skin an initial in vivo study was carried out on 5 volunteers from the Philippines (age: 19-39; 4 females, 1 male) over a period of 4 weeks. The results show a significant lightening effect in comparison to the untreated and placebo areas when oil-in-water formulations (O/W) containing 2 or 5% Belides were applied twice daily (Fig. 11). Already after 14 days of application a tremendous effect could be observed. Skin lightening was increased to 20.3 and 30.8% while after 28 days to 19.3 and 31.5% (p < 0.05 versus untreated). These results show that the whitening process is complete already after 14 days and continued application of the active does not further lighten the skin. Moreover, a complete knock-out of melanogenesis could be risky for melanocyte survival. Therefore, Belides acts as a gentle modulator of skin pigmentation. Fig. 10 Microscopic images showing (a) the Fluosphere uptake (red FluoSphere beads, 1.0 µm size) in control and (b) in Belides (1%) treated HaCaT cultures (1 DIV). For visualization of the phagocytosis cells were counterstained with DAPI (blue nuclei). Picture images were taken using a Leica DMIL fluorescence microscope (Leica, Germany; PHAGO 3 oil lense 100 x, excitation filter: 515 - 560 nm) Fig. 11 Efficacy test on human skin (pilot study) demonstrate the skin-lightening activity of Belides (chromameter readings). Formulations were applied by 5 volunteers twice daily on the inner forearm. Values are related to initial conditions (p
COSMETICS SKIN-LIGHTENING � Conclusion Belides has proven itself as a very efficient active that successfully interferes with every stage of the melanin synthesis. The extraordinary composition of this active helps to remove blemishes in a very gentle and reliable manner. Belides, a water-soluble active produced without any organic solvent, meets the ever-growing demands of natural beauty care. References (1) Briganti S., Camera E. and Picaro, M., Chemical and Instrumental Approaches to Treat Hyperpigmentation. Pigment Cell. Res. 16 (2003) 101-110 (2) Imokawa G., Miyagishi M., Yada Y., Endothelin-1 as new melanogen: coordinated expression of its gene and the tyrosinase gene in UVB-exposed human epidermis. J. Invest. Dermatol. 105 (1995) 32-37 (3) Hara M., Yaar M., Gilchrest B.A., Endothelin- 1 of keratinocyte origin is a mediator of melanocyte dendricity. J. Invest. Dermatol. 105 (1995) 744-748 (4) Bohm M, Luger T.A., Alpha-melanocyte-stimulating hormone. Its current significance for dermatology. Hautarzt. 55 (2004) 436-445 (5) Seiberg M., Paine C., Sharlow E., Costanzo M., Andrade-Gordon P., Eisinger M., Shapiro S.S., Inhibition of melanosome transfer results in skin lightening. J. Invest. Dermatol. 115 (2000) 162-167 (6) Seiberg M., Keratinocyte-melanocyte interaction during melanosome transfer. Pigment Cell Res. 14 (2001) 236-242 (7) Blaschek W., Ebel S., Hackenthal E., Holzgrabe U., Keller K., Reichling J., HagerROM – Hagers Handbuch der Drogen und Arzneistoffe, Springer Verlag Berlin, Heidelberg (2003) (8) Glensk M., Wray V., Nimtz M., Schöpke T., Triterpenoid saponins of Bellis perennis. Sci. Pharm. 69 (2001) 69-73 (9) Analytical methods collection (SOP-Nr. 1999. 14), University of Hamburg (Germany), Institute for Biochemistry and Food Chemistry (10) Lorenz P., Zeh M., Martens-Lobenhoffer J., Schmidt H., Wolf G., and Horn T.F.W., Natural and newly synthesized hydroxy-1-aryl-isochromans: A class of potential antioxidants and radical scavengers. Free Rad. Res. 39 (2005) 535-545 (11) Jeong C.H., Shim K.H., Tyrosinase Inhibitor Isolated from the Leaves of Zanthoxylum piperitum. Biosci. Biotechnol. Biochem. 68 (2004) 1984-1987 (12) Fullerton A., Fischer T., Lahti A., Wilhelm K.P., Takiwaki H., Serup J., Guidelines for measurement of skin colour and erythema. A report from the Standardization Group of the European Society of Contact Dermatitis. Contact Dermatitis 35 (1996) 1-10 (13) Hori, I., Nihei, K., Kubo, I., Structural criteria for depigmentation mechanism of arbutin. Phytother. Res. 18 (2004) 475-479 (14) Yada Y., Higuchi K., Imokawa G. Effect of Endothelin on Signal Transduction and Proliferation in Human Melanocytes. J. Biol. Chem. 266 (1991) 18352-18357 (15) Kadono S., Manaka I., Kawashima M., Kobayashi T., Imokawa G., The role of the epidermal endothelin cascade in the hyperpigmentation mechanism of lentigo senilis. J. Invest. Dermatol. 116 (2001) 571-577 (16) Imokawa G., Yada Y., and Kimura M., Signalling mechanisms of endothelin- induced mitogenesis and melanogenesis in human melanocytes. Biochem. J. 314 (1996) 305-312 (17) Jamal S. and Schneider R.J., UV-induction of keratinocyte endothelin-1 downregulates Ecadherin in melanocytes and melanoma cells. J. Clin. Invest. 110 (2002) 443-452 (18) Tsatmali M., Ancans J., Thody A. J., Melanocyte Function and its Control by Melanocortin Peptides. J. Histochem. Cytochem. 50 (2002) 125- 134 (19) Lu D., Willard D., Patel I.D., Kadwell S., Overton L., Kost T., Luther M., Chen W., Woychik R.P., Wilkinson W.O., and Cone R.D., Agouti protein is an antagonist of the melanocytestimulating hormone receptor. Nature 371 (1994) 799-802 (20) Jimbow K., Sugijama S., Melanosomal translocation and transfer. In The Pigmentary System: Physiology. a. Pathophysiology. (1998), Oxford Uni. Press. Chemisches Laboratorium Dr. Kurt Richter GmbH (CLR) Bennigsenstr. 25 12159 Berlin Germany Email: info@clr-berlin.com SÖFW-Journal | 131 | 7-2005 49 �
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Page 7: Fig. 8 Inhibition of α-MSH binding

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