214 JOURNAL OF COSMETIC SCIENCE Alpha-bisabolol is a sesquiterpene alcohol which was demonstrated to be able to block two of the three possible pathways of the formation of inflammatory-active leukotrienes and prostaglandins 6l . Pseudopterosins were found to inhibit prostaglandin E-2 and leukotriene C production showing that they mediate their anti-inflammatory effect via inhibition of eicosanoid release from inflammatory cells (7) . Pseudopterosins were also found to be involved in receptor mediated inflammatory events, suggesting involvement in the cellular arm of inflammation 8l Summary The Skin inflammation process is a protection mechanism where the body utilizes the immune system to fight off invaders. Although perceived symptoms may look the same, there are different types of inflammation that involve various mediators. While acute allergic inflammation is mediated by mast cells and immunoglobulin E, for example, cytotoxic inflammation is mediated by cytotoxic antibody and complement. Therefore, an anti­ .inflammatory compound should possess activity in major crossroads in the cascade to be able to prevent them from occurring. When designing a formula, thought should be put into its composition in terms of penetrability of its compounds, both in order to substantiate activity claims and prevent irritation. When irritation is expected, or when a product is designed for sensitive skin/skin areas, the addition of anti-inflammatory compounds can make the difference between a safe and unsafe f�rmula. References I) Gordon K.B. and Chan L.S., Inflammation and Immunity in: The Biology of the Skin, !'' Edition, 233-254, (2000). 2) Rougier A., In Vivo Percutaneous Absorption in: Percutaneous Absorption, 3rd Edition, 193- 211, (/999) 3) Fujisawa Y., Sakamoto M., Matsushita M., Fujita T., Nishioka K., Glycyrrhizin Inhibits The lytic Pathway q[Complement- Possible Mechanism of its Anti-!n.flammato,y Effect on liver Cells in Viral Hepatits, Microbial lmmunol. 44, 799-804, (2000). 4) Oh C., Kim Y., Eun J., Yokoyama T., Kato M., Nakashima I., Induction qf T lymphocyte Apoptosis by Treatment with Glycyrrhizin. Am J Chin Med 27, 217-26, (/999). 5) Van Uum S.H., Walken B.R., Hermus A.R., Sweep C.G., Smits P., de Leeuw P.W., Lenders J.W., E/fect of Glycyrrhetinic Acid on / I-Beta-Hydroxy Steroid Dehydrogenase Activity in Normatensive and Hypetensive Subjects. Clin. Sci 102, 203-11, (2002). 6) Stanzl K., Vollhardt J., Pickenhagen W., Nissen H.P., Protective and Curative in Vivo Efficacy qf Bisabo/ol against Sodium Hydroxide on SLS-induced Irritation on Human Skin - Indication for a Non-linear Dose Response Cure. I FSCC Congress 1998 Cannes France paper P004 I 06. 7) Mayer A.M.S., Jacobson P.B., Fenical W., Jacobs R.S., Glaser K.B., Pharmcological Characterization of Pseudopterosins. Novel Anti-Inflammatory Natural Products Isolated from the Caribbean Soft Coral, Pseudopterogorgia Elisabethae, Life Science 62(26) PL40 I­ PL407, (1998). 8) Dayan N., Ortega L., Riemer J., Moya C., Jacobs R. S., Pseudopterosins - Solubility Characteristics and Anti-Inflammatory Activity. 28th Inter. Symp. Control Rel. Bioact. Mater. Control Release Society, Inc. Proceed. USA, #5098, 2001.

2003 ANNUAL SCIENTIFIC MEETING 215 THE FIBRONECTIN NETWORK DURING AGING: A MISSING CELL CONNECTIVITY Eric Perrier, F. Pivard, S. Grenier and V. Andre Coletica, F-69007, Lyon, France The connective tissue of the skin has been extensively studied but there is still main information missing. The main scaffolds, namely the collagen and the elastic networks, are now more or less elucidated as far as their three-dimensional organization and functions are concerned. For instance, collagens molecules are able to be assembled into collagen fibrils and then fibers that are responsible for mechanical properties of the skin. On the other hand, the complex elastic fibers network is also tremendously important for the plasticity of the skin, and plays a pivotal role in the intrinsic and UV-related aging phenomena. A third network has been poorly studied until now but is also essential to the connective tissue organization as well as the cell-cell and cell-matrix interactions: the fibronectin network. Fibronectin designates a family of glycoproteins (about twenty members) synthesized by alternative splicing. Produced in insoluble form by the connective tissue's fibroblasts, it is found within cells at the cell surface and on extra-cellular levels. Fibronectin is capable of contracting lesions, not only with most of the other connective tissue molecules, but also with numerous cellular types through the integrins present on their surface. It also combines with the components of the cytoskeleton such as actin, through surface proteoglycans, to promote cellular adhesion (Ruoslahti, 1989). Poorly studied until now, we have decided to investigate the expression of this protein using monolayer and 3D fibroblast-based cell cultures such as reconstructed dermis and reconstructed full skin, using cells extracted from human biopsies coming with "young" and "mature" donors. How fibronectin is expressed while aging? Are there some age-related relationships concerning the level of fibronectin, that are relevant and comparable between in vitro and ex vivo experiments? If we modulate the fibronectin content of the extracellular matrix, what are the main biological properties observed, and which type of cosmetic activity could we demonstrate in vivo, in final cosmetic formulations? Working with an hospital research team, we have first analyzed on a standardize way, the evolution of the fibronectin expression in skin human biopsies from donors with different ages. Usir.g immunohistological techniques, we have observed that fibronectin was slowly reduced and disappeared progressively in human tissue with age. This work was in good accordance with the work of Karttunen et al. (1986) conducted on human biopsies of renal cortex, which demonstrated the same decrease. This evaluation shows that the fibronectin loss with age is following the strong reduction of total proteins content observed during the same time. We have then evaluated the fibronectin synthesis in three different cell culture models: fibroblasts in monolayers, Equivalent Dermis (Mimederm®) and Reconstructed Skin models (Mimeskin®). Each model was composed of cells from young or mature subjects (young fibroblasts, mature fibroblasts, young Mimederm®, mature Mimederm®, young Mimeskin®, mature Mimeskin®). In all of the cases, the quantities of fibronectin produced were quantified in incubatory environments using a specific sensitive ELISA dosage. The differential expression of the fibronectin gene in the young and mature models was also analyzed by Northern Blot. It has been discovered that fibronectin should be studied while using reconstructed skin models only, the other models being irrelevant compared to the results obtained ex vivo. Using such models able to more closely mimic the skin, we have observed that a reduction of 20% of fibronectin was observed using a sensitive and specific ELISA method. Fibronectin protein but also RNAm coding for this protein was strongly reduced using the same model (-43% after Northern Blot analysis). Using such observation, it has been then possible to build a miniaturized test able to screen best ingredients, plant extract or pure chemicals, able to stimulate fibronectin concentration in order to counteract effects of aging. We have demonstrated the ability of active compounds selected, to stimulate fibroblast migration (up to +65% vs the control), such ability being even stronger than what is able to be observed with TGFbeta (10ng/ml) and FGF (10µg/ml). Resulting selected products have then been used in a 3D, reconstructed cellular model, where artificial wounds have been created to mimic the loss of matrix components observed in a wrinkle