210 JOURNAL OF COSMETIC SCIENCE Contour Alterations Contour alterations, more commonly known as wrinkles, can be due to cutaneous dehydration, facial expression, or redundant skin. Cutaneous dehydration wrinkles are the most common and can be rapidly corrected with moisturizing formulations containing occlusives (petrolatum, silicone) and humectants (glycerin). Wrinkles of facial expression are present when the face is in motion. Botox Cosmetic, which denervates the facial muscles, can relax wrinkles that are present with facial motion. Wrinkles of the face that disappear with pulling on the skin can be minimized through the use of filler substances, such as autologous fat, collagen, and hyaluronic acid. These substances expand the skin filling the void left by lost endogenous collagen. Commonly, facial wrinkles are skin creases that cannot be removed with filler substances. These are facial folds commonly around the mouth and lower face present both at rest and with facial movement where cosmetics migrate only increasing the appearance of the folds. Enhanced film forming abilities and skin substantivity accompanied by enhanced light reflection to minimize the depth of the fold are required to improve the facial appearance. Future Directions There are needs that have not been met in the cosmetics, skin care, and treatment market. Colored cosmetics should provide better film-forming characteristics with increased adherence to adnexal structures, such as the pores and follicular ostia, to prevent migration into redundant skin folds of the face. Skin care products that absorb sebum and reflect light while providing increased coverage and sun protection would create the appearance of improved skin texture. Treatment products with anti-inflammatory effects would not only reduce facial redness, but inhibit the cumulative inflammatory effects physically observed as extrinsic UV-induced aging. Summary To keep abreast of changes in the appearance-related market, new cosmetics and skin care products must compliment the wide array of consumer driven antiaging procedures. This presentation has discussed cosmetic dermatology research and presented issues regarding opportunities for new products as they relate to medical procedure induced alterations in the physiology of skin performance and appearance.

2003 ANNUAL SCIENTIFIC MEETING BIOCHEMICAL AND BIOENGINEERING ANALYSIS OF THE SKIN'S NATURAL MOISTURIZING FACTORS Marisa H. Robinson and R. Randall Wickett, Ph.D. University of Cincinnati College of Pharmacy, Cincinnati, Ohio Introduction 211 The natural moisturizing factor (NMF) of the skin is important for maintaining proper moisture levels in the stratum corneum. NMF is largely a degradation product of filaggrin, a histidine-rich protein found in the upper epidermis, which aggregates keratin filaments in the cells of the stratum granulosum. In the stratum corneum, filaggrin is degraded into a number of low molecular weight, hygroscopic molecules such as urea, pyrrolidone carboxylic acid, and individual amino acids. These molecules comprise the NMF. In these studies, we are using free amino acids as markers for NMF levels, as they are the major components. NMF levels can be expected to be altered by common activities such as bathing, soaking and surfactant damage. There is surprisingly little information on the effects these activities have on the skin. Our group has done several studies on soaking and extraction of the skin to uncover the effects of these treatments on NMF. We have developed a sensitive method to quantify the components of NMF using High­ Performance Liquid Chromatography (HPLC). Materials and Methods Analytical Method: A D-Squame® adhesive disc is removed from the skin and placed in a microcentrifuge tube. 300 µL of 0.006 M perchloric acid and 10 µL of 2 mmol/mL AABA are added to each tube. The samples sit for 3 hours at room temperature, after which the liquid extract is removed by pipetter to a fresh microcentrifuge tube, and the tube containing the D-Squame® is retained for protein analysis by the Pierce BCA protein assay. 20 µL of the extract is placed in a HPLC microvial, along with 30 µL of borate buffer and 10 µL of Waters AccQ-Fluor derivatizing reagent (6-a-quinolyl-n­ hydroxysuccinimidyl carbamate) and immediately vortexed for 5 seconds. Amino acid analyses are performed on a 15 cm C-18 reversed-phase column at 40° C under gradient conditions. The run time is 40 minutes. Peaks are eluted in order of increasing hydrophobicity with the most hydrophilic amino acids eluting earliest. Fluorescence detection is used, with excitation at 250 nm and emission at 395 nm. We have selected four amino acids to represent all of the free amino acids for quantification: serine, histidine, citrulline, and phenylalanine. Experimental Method: Volunteers with normal skin on the forearms were selected. After a one­ week no-product washout on the test areas, two sites were marked on each volar forearm, with treatments matched between contralateral sites as controls. Baseline biophysical measurements (MAT, TEWL, pH, sorption/desorption) were taken after a twenty-minute acclimation period. One site perarm was extracted with 1: 1 acetone/ether for 5 minutes, and the extracted sites re-measured. One arm was soaked in a 40 +/- 2.5 degree Celsius bath for 10 minutes, and then blotted dry. Measurements were collected 0.25, 0.50 and 4 hours after bathing. 15 or 20 D-Squames® were taken sequentially on each site for measurement of NMF levels 0.50 and 4 hours after soaking. Three studies of this variety were performed, the second and third of which omitted the solvent extraction. The second and third studies included D-Squame® collection on the soaked sites at both 0.50 hours and 4 hours after soaking, to measure NMF recovery over this time period. In the third study, samples were also collected on the outer calf after a one-week washout with no lotion in order to generate dry skin. The sites were equilibrated for 20 minutes, then biophysical measurements and 15 D-Squames® were collected.