BREADFRUIT EXTRACT AS SKIN LIGHTENER 53 (A) (B) Figure 9. Morphology of melanocyte B16Fl melanoma cells treated with (A) 0.1% DMSO (control) and (B) 10 µg/ml of hydroquinone for three days. Magnification: 400x.

54 JOURNAL OF COSMETIC SCIENCE morphology of melanocyte B 16F 1 melanoma cells was shorter in length compared to the control cells. These obtained results indicate the cytotoxicity of hydroquinone at the concentration of 10 µg/ml used in this study, which was similar to the effect observed previously (17,18). At a concentration of 10 µg/ml, kojic acid did not show a significant inhibitory effect on melanin production compared to the control as shown in Figure 6. Although kojic acid showed stronger tyrosinase inhibition than A. incisus extract based on the IC 50 values from the DOPAchrom assay (IC 50 values of kojic acid and the extract were 7 .89 and 10.26 µg/ml, respectively), kojic acid showed lower inhibitory activity on melano­ genesis than the ether extract in the cell culture model. These results correspond with those from the previous study that indicated a low- or non-inhibitory effect of kojic acid on the production of melanin in Melan-a, and MelAb cells (18-20). The lipophilic system of cell membrane may decrease the penetration of kojic acid through the cell, resulting in lower inhibitory action on the intracellular tyrosinase enzyme. In addition, as shown in Figure 10, we found changes in the morphology of the kojic acid-treated cells, in losing dendritics, according to the microscopic observation. The effects of 10 µg/ml of A. incisus ether extract and 4.5 µg/ml of artocarpin on the proliferation and morphology of melanocyte cells are shown in Figures 8 and 11. We found that artocarpin significantly decreased the proliferation of cells, whereas the extract did not affect cell proliferation. The obtained micrographs of cells treated with artocarpin indicated more evidence of the effect of artocarpin on melanocyte morphology by losing dendrites but not destroying the cells. Furthermore, a higher concentration of artocarpin (�4.5 µg/ml) caused cell death. Surprisingly, A. incisus crude extract con­ taining a similar artocarpin amount did not show cytoxicity in the melanocyte cells. Additionally, at higher concentration, the morphology of cells treated with the extract was not changed. This indicates that the extract can suppress melanin production without affecting cell morphology or cell growth. It is possible that the extract contains some components that are contributing to cell structure stabilization and/or protecting the cell from damage in response to external stimuli. CONCLUSIONS Nowadays the botanical extract plays an increasingly important role in cosmetics. In the research for new natural lightening agents, plant extracts with depigmenting effects, including tyrosinase-inhibitory activity, melanogenesis-inhibitory activity, and antioxi­ dant activities have been investigated. As applied to cosmetics, isolation and purification of the active ingredient from the crude extract are sometimes not needed because such isolation and purification may lead to a loss of the biological activity and may lead to toxicity. Therefore, in this study, the crude extract of A. incisus was clarified in its melanogenesis-inhibitory and antioxidant activities. In the past, some studies reported the constituents and effects of A. incisus's heartwood methanol extract on melanogenesis inhibition. However, as far as we know, there has never been a report on the effects of the ether extract of A. incisus's heartwood on tyrosinase enzyme, melanogenesis, or oxidation activities.