211 J. Cosmet. Sci., 61, 211–223 (May/June 2010) Anti-wrinkle activity of Platycarya strobilacea extract and its application as a cosmeceutical ingredient YOUNG HEUI KIM, KI HO KIM, CHANG SUNG HAN, HONG CHUL YANG, SUN HEE PARK, HYE-IN JANG, JIN-WAN KIM, YUN-SUN CHOI, and NAM HO LEE, R&D Center, Bioland Ltd., Songjeong, Byongchon, Cheonan, Chungnam 330-863 (Y.H.K., K.H.K., C.S.H., H.C.Y., S.H.P.), Instiute of Life Science, Univera, Inc, Econet Center 302-4, Sungsu-dong 2, Sungdong, Seoul 133-120 (H.-I. J., J.-W.K., Y.-S.C.), and Department of Chemistry, College of Natural Science, Cheju National University, Ara 1-Dong, Jeju 690-756 (N.H.L.), Republic of Korea. Accepted for publication February 23, 2010. Presented at the 2008 IFSCC Congress, Barcelona, Spain, October 6–9, 2008. Synopsis In order to investigate the potential of Platycarya strobilacea fruit extract as an active ingredient for cos- metics, we measured their free-radical scavenging activity, elastase inhibitory activity, the expression of MMP-1 (matrix metalloproteinase-1), and type I collagen synthesis in normal human fi broblast cells. To isolate the main component compounds from P. strobilacea fruit extract, we purifi ed the extract through solvent fractionation, column chromatography, and recrystallization. The component compounds were identifi ed as ellagic acid and 4-O-xyloside of ellagic acid (ellagic acid 4-O-xylopyranoside). P. strobilacea fruit extract and ellagic acid increased the expression of type I collagen mRNA in a dose-dependent man- ner (up to 37% and 41% at 20 μg/ml and 1.0 μg/ml, respectively), comparable to that of ascorbic acid (up to 39% at 500 μM). A clinical study of measurements using visual evaluation and image analysis showed a statistically signifi cant difference (p 0.05) between the effects of the test and placebo prod- ucts. This result suggests that P. strobilacea fruit extract could be used as an active ingredient for anti- aging cosmetics. INTRODUCTION Skin aging is characterized by a progressive deterioration of the skin's functional proper- ties, linked to alterations of dermal connective tissue due to the changes at the cell, gene, and protein levels. Skin aging can be divided into two basic processes: intrinsic aging and photoaging. Intrinsic skin aging describes the irreversible physiological process that starts in age groups from around 17 to 25 years, as soon as physical maturation is accom- plished. Photoaging is the term given to the superposition of chronic sun damage on the intrinsic aging process (1–3).

JOURNAL OF COSMETIC SCIENCE 212 The dermis is the thicker, deeper layer of the skin underlying the epidermis, and is mainly composed of such connective tissues as collagen and elastic fi bers. Among these, collagen fi ber is the main component of the extracellular matrix (ECM), as the repre- sentative connective tissue that comprises about 90% of the dermis collagen has a di- rect infl uence on skin tension. Maintenance of the collagen structure is related to the intrinsic aging and photo-aging processes of the skin (4,5). Therefore, a variety of in- vestigations have been focused on protection against skin aging through the inhibition of collagenase activity, which disintegrates the ECM proteins (6). Matrix metallopro- teinases (MMPs) are an enzyme family to digest basal membranes and ECM. MMPs can be divided into four categories according to the preferred substrate: collagenases (MMP- 1,8,13,18), gelatinases (MMP-2,9), stromelysins (MMP-3,10), and membrane-type MMPs (MMP-14,15,16,17) (7). It is therefore suggested that an evaluation of the in- hibitory effi cacy of materials for MMP-1 gene expression could be used as a screening method to uncover promising candidates that would inhibit the degradation of colla- gens (8–10). Platycarya strobilacea Sieb. et Zucc. (family: Juglandaceae) is widely distributed through- out Korea, Japan, Taiwan, and China (11). It is commonly called Kool-pee tree in Korea. The root bark of this species have been used in traditional oriental medicine for the treat- ment of infl ammation and diarrhea. Its leaves have also been used for the treatment of acute suppuration or abscess on the skin and head region (12). The constituents of the bark and leaves have been studied. It is known that euglon and tannin exist in the bark and fruit, respectively (13). We have previously screened various plants for free-radical scavenging activity and the inhibition of elastase activity (14), and have also reported the anti-wrinkle activity of zi- yuglycoside I isolated from a Sanguisorba offi cinalis root extract, which showed a signifi - cant inhibitory effect against aging (15). In this study, we investigated free-radical scavenging activity, elastase inhibition activity, reduction of MMP-1 mRNA expression in vitro, and type I collagen synthesis in normal human fi broblasts for the development of potential anti-wrinkle ingredients as raw materials for use in cosmetic products. We elucidated the main components from the P. strobilacea fruit extract. MATERIALS AND METHODS REAGENTS AND EQUIPMENT All reagents were purchased from Sigma-Aldrich (St. Louis, MO). Other commer- cially available reagents and solvents were used as received. Human fi broblasts were acquired from ATCC (American Type Culture Collection, CRL-2076). The com- pound 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was used as received. Medium, fetal bovine serum, and antibiotics were purchased from Life Technologies (Grand Island, NY). The elastase inhibitory activity assay was per- formed with a UV-spectrophotometer (Hewlett-Packard, HP-8453). An ELISA reader (Tecan, A-5082, Austria) and PCR (Bio-Rad, MycyclerTM thermal cycler) were used in the cytotoxicity assay and in the assay on the inhibition of MMP-1 expression.