COSMECEUTICAL PROPERTIES OF LEVAN 80 �----------------------, - 70 8 60 QJ r/) ca � 50 tS I �l [ I J (a) r--1 I r - I I 1 (b) (c) (d) Concentration (w/v%) 405 Figure 8. Anti-inflammation effect of levan in 3-D artificial skin: (a) control (b) 0.05% SDS (c) 0.05% SDS + 0.01 mg/ml of levan (d) 0.05% SDS + 0.05 mg/ml of levan. The data are expressed as mean values (± standard deviations) of four experiments. of hyaluronic acid, and also exhibited effects on the proliferation of human fibroblast and keratinocytes reminiscent of hyaluronic acid. Moreover, in our cell proliferation test on 3-D bio-artificial skin, after the induction of primary skin inflammation using 0.05% SLS, cell proliferation on the 3-D artificial skin in the presence of levan was determined to be much more pronounced than in the absence of levan (SLS treatment only). In our anti-inflammation test, the quantity of IL-la. secreted in the 3-D artificial skin treated with levan was shown to be less than that of the 3-D artificial skin treated only with SLS. As a result of these studies, levan was found to exert an anti-inflammatory effect against inflammatory reactions to skin irritants. Levan was also shown to exert a cell prolifera­ tion effect in bio-artificial skin. We also evaluated levan with regard to its safety and lack of toxicity in a series of safety tests that involved fibroblasts. Our results indicate that levan might, indeed, prove to be a useful and safe cosmeceutical agent. REFERENCES (1) R. Dedonder, Levansucrase from Bacillus subtiliJ, Methods Enzymol., 8, 500-505 (1966). (2) G. D. Bonnett, I. M. Sims, R. J. Simpson, and A. J. Cairns, Structural diversity of fructan in relation to the taxonomy of the Poaceae, New Phytologist, 136, 11-17 (1997). (3) N. C. Carpita, J. Kanabus, and T. L. Housley, Linkage structure of fructan and fructan oligomers from Triticum aestivum and Festuca arundinacea leaves,]. Plant Physiol., 134, 162-168 (1989). (4) G. A. F. Hendry and R. K. Wallace, "The Origin, Distribution, and Evolutionary Significance of Fructan," in Science and Technology of Fructans, M. Suzuki and N. J. Chatterton, Eds. (CRC Press, Boca Raton, FL, 1993), pp. 119-139. (5) E. A.H. Pilon-Smits, M. J.M. Ebskamp, M. J. Paul, M. J. W. Jeucen, P. J. Weisbeek, and S. C. M. Smeekens, Improved performance of transgenic fructan-accumulating tobacco under drought stress, Plant Physiol., 107, 125-130 (1995). (6) M. J. Pabst, Levan and levansucrase of Actinomyces viscosus, Infect. Immun,, 15, 518-526 (1977). (7) T. N. Warner and C.H.Miller, Cell-associated levan of Actinomyces viscosus, Infect. Immun., 19, 711-719 (1978). (8) K. Tanaka, T. Karigane, S. Fujii, T. Chinzaka, and S. Niagamura, Intermolecular fruccosyl and

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