ARECA CATECHU L. EXTRACT 293 Figure 5 Skin section incubated with elastase (1 lag/ml) in the presence of CC-516 (100 lag/ml) Table III Effect of CC-516 on Cell Proliferation at Various Concentrations Concentration (%) Materials 1 x 10 -2 1 x 10 -3 1 x 10 -4 1 x 10 -5 1 x 10 -6 Ascorbic acid 105 130 150 140 105 CC-516 110 143 185* 120 95 Control 100 100 100 100 100 Data in the table are the cell viabilities (%) compared to control (100%). *p 0.05 vs control and ascorbic acid. Table IV Effect of CC-516 on Collagen Synthesis (monolayer and three-dimensional culture) Concentration Monolayer 3-D Culture (%) Materials l x 10 -3 1 x 10 • 1 x 10 5 I x 10 • 1 x 10 -2 I x 10 3 Ascorbicacid 102 104 118 95 98 92 CC-516 108 140' 120 120 100 104 Control 100 100 100 100 100 100 Data shown in the table are the synthesis ratio (%) compared to control (100%). *p 0.05 vs control and ascorbic acid. thesis by a factor of 1.2, at 0.01% and 0.001% concentrations. This stimulation is almost the same as that of the control (100%). The elasticity of the skin depends on its water content and on the swelling capacity of the fibers of its connective tissue. Evalu- ation of stratum corneum hydration by the capacitance method showed that skin mois- turizing increased about 16.5% against untreated cream. In the skin hydration experi- ment, all measurements were done in triplicate and the average values were used for

294 JOURNAL OF COSMETIC SCIENCE statistical evaluation. Both in the cheek region and the eye region, there was no sig- nificant difference between the control (base cream) and treated cream (base cream + 3% CC-516) on all the measuring points. From the results, which are represented in Table V as means of five test persons, it can be concluded that after six weeks of treatment with a test sample, a smaller depth of penetration of the skin into the tube, and thus elasticity, increased by 35%. In the case of the control, no modification was observed. Table V shows the anti-wrinkle effect by three-dimensional image analyzing. The average values were used for statistical evalu- ation. We applied 2 mg/cm 2 of a control cream or a cream containing 3% of CC-516 twice a day for six weeks to twenty volunteers. The results obtained from the treated side show significant reductions of roughness (-16.7%) and of the average depth of the cutaneous relief (23%), but the untreated skin (control) shows no significant difference. The study of wrinkle depth using SEM reveals apparent reduction in depth of the wrinkles against untreated skin (Figures 6,7), but the control cream showed no signifi- cant difference between before and after treatments (not shown). CONCLUSIONS The enzyme elastase has received significant attention, primarily because of its reactivity and unspecificity. It is able to attack all major connective tissue matrix proteins, e.g., elastin, collagen, and keratin (43). In contrast to elastase, collagenase is a specific proteinase with a limited number of substrates (44). One major target of cosmetologic research is to find effective elastase inhibitors. Such inhibitors could find their applica- tion in anti-wrinkle, anti-aging creams and more generally in numerous compositions intended for skin care. The object of this natural extract is not only to provide an active ingredient that inhibits elastase enzymatic activities (protective role) but also provides the necessary supplies for cells to maintain an appropriate energetic level and to produce their constituents (repairing role). The cells synthesize, secrete, and deposit the proteins of the extracellular matrix made up by collagens, elastin, fibronectin, etc. These proteins ensure the cell inking at the basal plate in the dermal structure. Protecting the skin, its elasticity is the major theme of cosmetic research. The action of a product can be Table V Anti-Wrinkle Effect on Roughness and on Mean Depth of Wrinkles and Skin Elasticity of Cream Treated With CC-516 Anti-wrinkle effect Elasticity Roughness Mean depth (arbitrary units) (peak depth, pm) ds (mm) Before 32.7 40.5 0.0475 Untreated cream After 28.3 40.0 0.0480 Before 33.4 52.0 0.0575 Treated cream After 28.0 40.0 0.0375 The control is untreated cream before and after six weeks of treatment. Data for skin to which the cream treated with CC-516 has been applied is shown before and after six weeks of treatment. The results obtained on the treated side show significant reduction (-16.7% of roughness and 23% of average depth of wrinkles) compared to before treatment. Skin elasticity increased about 35% compared to before treatment.