Whitening Effect of Eucheuma cottonii Extract in

Phorbol-12-myristate-13-acetate induced Melanocytes

Monica Drestia
2011-080-074

Faculty of Biotechnology
Atma Jaya Catholic University of Indonesia
Jakarta
2014
 Introduction

img2.timeinc.net
(Costin & Hearing 2007).
→ The Factors...

(Costin & Hearing 2007).

 (Selvan et al. 2014).

B16F10 Murine
Melanocytes
 Methods
Sample Extraction

Cell Cultivation

Melanogenesis Induction & Sample

Treatment
Measurement of Tyrosinase Activity

Cytotoxicity Assay

RNA Isolation & qRT-PCR

Sample Extraction

+ → →

←
(Hindritiani et al. 2013).

Cell Cultivation

→
Melanogenesis Induction

Sample Treatment

(Lee et al. 2010).

Cytotoxic Assay
37oC
3 hr

+ → 590 nm

MTT reagent
Treated cells (20 µL)
(Hindritiani et al. 2013).

Measurement of Tyrosinase Activity

37oC
1 hr

→ →
+ L-DOPA 475 nm

Treated cells,
lysed
RNA Extraction & qRT-PCR

→
+ Trizol

treated cells
Kappa SYBR FAST One-Step qRT-PCR

 Primer :
- GAPDH
- Mus musculus microphthalmia-associated
transcription factor (MITF)
 Results
Figure 1 Cell viability percentage of B16F10 cells after PMA
induction and 24h incubation of samples treatment.

• From this result, it is known that all the used concentrations of E. cottonii extract, likewise
ascorbic acid, didn’t display significant cytotoxic activity that may affect cell proliferation.
The best concentration of E. cottonii extract which had highest cell viability percentage
(78.6%) was 100 ug mL-1.
• E. cottonii extracts increased B16F10 viability percentage due to the presence of many
compounds, since it has been reported that marine macroalgae components (such as α-
tocopherol, fatty acids, phenolic compound and its derivatives, including simple phenols,
flavonoids, phenyl proponoids, tannins, and lignins) have antioxidant properties that protect
cells from any damage and improve cell viability (Kumar et al. 2007).
 Results
Figure 2 The effect of E. cottonii extract on the mRNA expression of
melanogenic transcription factor MITF of treated B16F10 cells.

• E. cottonii extract decreased the level of MITF expression in a dose-independent manner.

The optimum concentration of E. cottonii extract with the lowest MITF expression (about
0.11%) was 100 ug mL-1.
• It was reported that marine macroalgae polysaccharides (such as carrageenan and
fucoidan) significantly decreased MITF expression of melanocytes. MITF is an important
regulating factor of melanogenesis. Inhibition of MITF expression can reduce tyrosinase
activity and inhibit melanin formation of the B16F10 cells (Song et al. 2015).
 Results
Figure 3 The effect of E. cottonii extract on the tyrosinase activity
of treated B16F10 cells.

• The tyrosinase activity assay indicated that E. cottonii extract significantly reduced
tyrosinase level of the cells.
• As mentioned in previous slide, it was reported that E. cottonii contains polysaccharide
(carrageenan) which can reduce the expression of MITF gene. Since the tyrosinase level
is regulated by MITF gene, downregulation of MITF gene can lead to lower tyrosinase
level in melanocytes, which also inhibits melanogenesis mechanism (Song et al. 2015).
 Conclusion

• E. cottonii extract significantly reduced tyrosinase activity

and MITF expression level of the cells without any
significant cytotoxic activity on cell proliferation
• E. cottonii extract can be considered as natural and safe
skin whitening agent
 References
Costin GE, Hearing VJ. 2007. Human skin pigmentation: melanocytes modulate skin color
in response to stress. FASEB.21:976-994. 
Hindritiani R, Dhianawaty D, Sujatno M, Sutedja E, Setiawan. 2013. Penurunan aktivitas
tirosinase dan jumlah melanin oleh fraksi etil asetat buah malaka (Phyllantus emblica) pada
mouse melanoma B16 cell-line. MKB.45(2):118-124.
Kumar KS, Ganesan K, Rao PVS. 2007. Antioxidant potential of solvent extracts of
Kappaphycus alvarezii (doty) doty- an edible seaweed. J Food Chem. 107:289-295.
Lee CW, Kim HS, Kim JW, Yoon JH, Cho Y, Hwang JK. 2010. Inhibitory effect of panduratin
a isolated from Kaempferia panduarata Roxb. on melanin biosynthesis. Phytother
Res. 24:1600–1604.
Selvan BK, Piriya PS, Chandrasekhar M, Vennison SJ. 2014. Macro algae (Eucheuma cottonii
and Sargassum sp) are reservoirs of biodiesel and bioactive compounds. J Chem
Pharm. 2:62-70.
Song YS, Balcos MC, Yun HY, Baek KJ, Kwon NS, Kim MK, Kim DS. 2015. ERK activation
by fucoidan leads to inhibition of melanogenesis in Mel-Ab cells. Korean J Physiol
Pharmacol. 19:29-34.
 Many thanks to:
• Lord Jesus Christ and Mother Mary
• Ms.Yanti, PhD as my advisor
• Mr. Adi Yulandi S.Si, M.T as today’s moderator
• Yonsei University
• Faculty of Biotechnology Atma Jaya Catholic University 0f Indonesia
• Mom, Dad, and my family
• The Lovely Skin Team (Patrice, Keren, Margareth, Halim, Alwi)
• PSSP Team (Bu Diah, Bu Silmi)
• Yudith and Willchan as today’s awesome partner
• Kak Rika, mami Teteph, Kimberly, Oline Anggasta, Edu
• Penghuni lab kuljar (Grace, Disa, Kent, Boni, Dira, Laudus, Yunita, Edbert,
Tjong, Sur, Natan), Odi, Merci
• Penghuni lab mikro (*Vege, Mike, Pau), penghuni lab biokim (*Kak Fani, Stella,
Olga, Ocil, Hux, Debi, geng bioplastic)
• Technisians
• FTb 2011 family, FTb 2012, FTb 2013, FTb 2014
• All of today’s seminar attendants 