A. INCISUS EXTRACT AND WRINKLE REDUCTION 321 the concentration used here (50 μg/ml) was found to inhibit elaboration of this key enzyme, MMP-1. Concomitantly, the same extract concentration markedly stimulated type I procollagen synthesis by wrinkled-skin fi broblasts. Such a stimulation of procollagen synthesis might be caused, at least partially, by an increasing cell proliferation, since enhancement of wrinkle-skin fi broblast proliferation by the extract was found. From these combined effects, the extract could improve collagen metabolism by dermal fi broblasts. EFFECT OF THE EXTRACT ON CONTRACTION OF THE FIBROBLAST-EMBEDDED COLLAGEN LATTICE In addition to improved collagen metabolism, the reorganization and reorientation of collagen fi bers are necessary to minimize the development of skin wrinkles (12,32). Such remodeling of collagen fi bers is governed by fi broblasts and, in turn, is shown to affect the locomotion and attachment of fi broblasts (11,33). Figure 7A shows the extent of contrac- tion (initial diameter decreased by 56%) of lattices populated with nonwrinkled-skin fi - broblasts vigorously for the fi rst three days after that, the contraction progressed more slowly. In comparison, the lattices containing the wrinkled-skin fi broblasts produced a slight contraction (the initial diameter decreased by only 12% after three days), but most importantly, the extract restored the capacity of the lattice with the wrinkled-skin fi bro- blasts to contract in a manner similar to that of the lattices containing nonwrinkled-skin Figure 6. Effects of A. incisus extract on (A) type I procollagen or (B) MMP-1 production by nonwrinkled- skin and wrinkled-skin fi broblasts. Fibroblasts were treated with 0.1% DMSO (control) or 50 μg/ml of ex- tract for 72 h. Each bar represents mean ± S.D. of triplicate study. *p 0.05 and **p 0.01, when compared with control values (Student’s t-test).

JOURNAL OF COSMETIC SCIENCE 322 fi broblasts (by 53% at three days). Furthermore, the contraction was uniform as shown by the disk shape of the lattices (Figure 7B). These observations show that our extract im- proved the contractile ability and reorganization of the collagen fi bers shown by the wrinkled-skin fi broblasts. Taken together with the above-mentioned results, these obser- vations suggest that the extract affects not only enhancement of dermal collagen produc- tion, but also the restoration of dermal collagen produced by fi broblasts. However, it is important to note that a limitation of this study is the source of the fi broblasts used. As the study was performed with fi broblasts obtained from only one individual, further investigation in fi broblasts from various sources is necessary to ensure the effi cacy of the extract on the restoration of fi broblast functions. CONCLUSIONS Nowadays, the botanical extract is playing an increasingly important role in cosmetics. The isolation and purifi cation of active ingredients is sometimes unnecessary for cosmet- ics, and purifi cation may lead to a loss of biological activity and may cause toxicity. Our Figure 7. Effect of A. incisus extract on the contractile capacity of nonwrinkled-skin and wrinkled-skin fi - broblasts. Fibroblasts embedded in the collagen lattice were treated with 0.1% DMSO (control) or 50 μg/ml extract. (A) The diameter of each lattice was measured over seven days of culture. Each point represents mean ± S.D. of three samples in duplicate. (B) Photographs of collagen lattices fl oating singly in the culture dishes at day 3. In dishes marked ‘XX’, the boundary of the lattice is diffi cult to discern because it fi lls of the dish.