ABSTRACTS 45 Integration of the above information into product design and treatment regimens should substantially improve our ability to care for healthy and diseased skin of the elderly. SCIENTIFIC SESSION III FRONTIERS OF ALTERNATIVE TESTING Surfactant-induced primary irritancy in an in vitro model Vincent A. DeLeo, M.D., Columbia University, 530 West 168th St., New York, NY 10032 Primary irritancy induced in human and animal skin by surfactants is characterized by an inflamma- tory reaction, mediated in part by membrane-de- rived arachidonate metabolites. We have developed an in vitro model for investigating the membrane- related mechanism of this reaction. Human epi- dermal keratinocytes and mouse fibroblasts (C3H 10T «) were pre-labelled with [3H] arachidonic acid (AA) or [3H] choline (Ch) the cells were treated with selected surfactants the media were re- moved and assayed for release of labelled metabo- lites. Release of label correlated positively with sur- factant concentration. In addition, when groups of different surfactants were assayed, the rank order of response (release of label) was similar to the rank order irritancy of these agents in animal skin. Systems such as the one described may prove useful as an in vitro alternative to whole animal testing for cutaneous primary irritancy. In vitro toxicology--What is the next step? Alan M. Goldberg, Ph.D., The Johns Hopkins University, 615 N. Wolfe St., Baltimore, MD 21205 The presentations that are included in this sympo- sium will put into perspective the major advances that have been made in developing in vitro method- ology for skin and eye irritation. We have a unique opportunity. Clearly, these methods are in the de- velopment stage. It is our responsibility to examine these methods as possible approaches to providing interpretable data to assess toxicity. We have the additional opportunity to begin standardization and validation of these methods so that they will be ac- cepted by both the scientific and regulatory com- munities. By providing 9alidatable methodology in this area we will not only provide alternatives to currently established methods but we will be able to establish in vitro methods as usable in product safety evaluation. Acute toxicity testing John M. Frazier, Ph.D., The Johns Hopkins Uni- versity, 615 N. Wolfe St., Baltimore, MD 21205 The objective of acute toxicity testing is to obtain reliable data to access the toxicological potential of chemical agents. Historically this has been accom- plished by whole animal testing using an LD50 protocol or its equivalent. Such an approach has been relatively successful but has required a high investment of resources, including time, space, money, and live animals. Recently, substantial ef- forts have been exerted in developing alternative methods for performing these tests which poten- tially have the advantages of being more reliable, more economical, and utilize non-whole animal test systems. The current status of in vitro alternatives to acute toxicity testing will be reviewed as well as the outlook for the future. Several examples will be presented to illustrate the nature of in vitro test systems and to provide a basis to discuss their strengths and weaknesses. In vitro detection of skin photosensitizers Irene E. Kochevar, Ph.D., Department of Derma- tology, Massachusetts General Hospital, Harvard • Medical School, Boston, MA 02114 The overall purpose of this research was to develop an in vitro method for evaluating the phototoxic po- tential of chemicals. The assay developed is based on the inhibition of stimulated 3H-thymidine in- corporation into the DNA of human lymphocytes by exposure of the cells to the test chemical and the appropriate wavelengths of ultraviolet or visible ra- diation. This assay has been validated by testing more than 30 photosensitizing chemicals. The one false negative appears to be due to in vivo metabo- lism of the drug which produces the phototoxic agent. The animal rights movement and alternatives Andrew N. Rowan, Ph.D., Tufts University School of Veterinary Medicine, 207 Harrison Ave., Boston, MA 02111 The animal rights movement has grown in power and sophistication over the past fifteen years. At the same time, pressures to develop and use alternatives to animals have increased. In 1970 the concept of alternatives had little or no credibility or visibility among the scientific community. Animal welfare groups continued to push for government interven- tion of some sort on alternatives (usually defined as those techniques which could replace animals, re- duce the number used, or reduce animal distress and suffering). However little happened until 1980 when a) the Research Modernization Act was intro- duced into Congress and b) the Coalition against the Draize Test was formed. The Research Moderni- zation Act galvanized public pressure in support of alternatives and Congress began to pay some atten- tion to the issue. The Coalition against the Draize

46 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Test put substantial pressure on the cosmetic in- dustry, and several million dollars have now been set aside for research on alternatives. The combined pressure on Congress, the regulatory agencies, and the industry has led to much greater attention being paid to alternatives. A tissu• • culture model of ocular irritancy Marcia M. Jumblatt, Sandra J. Simmons, and Ar- thur H. Neufeld, Ph.D., Eye Research Institute of Retina Foundation, 20 Staniford St., Boston, MA 02114 We have developed an in vitro model of ocular irri- tancy based on the ability of cultured corneal epi- thelial cells to migrate and re-cover a standard wound. Rabbit corneal epithelial cells derived from primary cultures are grown to confluency in a 24- well plate. A preliminary cytotoxicity screen is used to select an appropriate range of toxicant concentra- tions. Cell number is determined and standard 7-mm diameter wounds are produced. Quadrupli- cate wells are then exposed to various concentrations of the test agent or control medium. After 24 hrs the cultures are fixed and stained and the remaining wound areas are quantitated. A variety of ocular ir- ritants and industrial formulations have been evalu- ated using this method. When the dose necessary to inhibit closure is determined and used to rank com- pounds, there is a positive correlation with ocular irritancy scores. This test may provide a useful al- ternative to the Draize test. SCIENTIFIC SESSION IV NEW RAW MATERIALS/TECHNOLOGY PPG-20 methyl glucose ether distearate: A nat- urally derived skin moisturizer and emollient Joseph P. Pavlichko, Amerchol Corporation, 136 Talmadge Rd., P.O.B. 4051, Edison, NJ 08818- 4051 PPG-20 methyl glucose ether distearate (Glucam © P-20 Distearate), patent pending, is the stearic acid ester of the 20 mole propoxylate of methyl gluco- side. It is one of a series of esterified alkoxylated glucose derivatives which are multifunctional and exhibit excellent safety and mildness. The moistur- izing properties of Glucam P-20 Distearate have been demonstrated by means of transepidermal water loss studies (TEWLS). Glucam P-20 Dis- tearate imparts a highly efficient, semi-occlusive barrier which provides optimum rather than exces- sive water retention. The TEWLS data indicate that the use of 5% Glucam P-20 Distearate in a simple nonionic, o/w lotion significantly reduces water loss from the stratum corneum versus a con- trol lotion. Glucam P-20 Distearate possesses out- standing lubricant and smoothing properties, both key attributes of an emollient. These properties provide a highly desirable afterfeel to the skin sur- face in skin care emulsions. Its lubricating quality also enables it to be utilized as a highly effective binding agent, aiding compression in pressed powders and as a pigment dispersant in makeup and lipstick. Development of novel amino acid derived cat- ionic surfactant with mild characteristics Kazatami Sakamoto, Ajinomoto USA, Inc., 9 West 57th St., New York, NY 10019 Although cationic surfactants are indispensable in- gredients for many cosmetic formulations, their po- tential irritating properties on skin and eyes have restricted their applications at times for practical use. Many derivatives of amino acids have been in- vestigated and characteristic properties of salt of No•-fatty acid acyl-L-arginine esters (AAE) were found. Surface activities (foaming, wetting, emulsi- fying, etc.), antistatic properties, and antimicrobial properties for bacteria such as Pseudomonas aeru- ginosa, E. coli, Bacillus subtilis, Staphylococcus epidermidis and Pityrosporum ovale and anti-viral properties of AAE are measured in comparison with benzethonium chloride and other alkyl am- moniums. AAE is characterized as a mild substance such as water in animals and humans for primary skin and eye irritation tests and no phototoxicity and sensitization were found for guinea pigs. As a result, AAE showed potential properties as a cat- ionic surfactant or preservative for cosmetic and toi- letry formulations with mild properties as well as surface active and antimicrobial properties. PLURONIC © ethylene oxide-propylene oxide block copolymer surfactants as eye irritation mitigants and foam enhancers Jerry Burke and Robert Roelofs, BASF Wyandotte Corporation, Performance Chemicals Division, Central Research, 1419 Biddle Ave., Wyandotte, MI 48192 Various types of cosmetic ingredients have been used in the past to mitigate the eye irritation in- duced by many anionic surfactants used in hair shampoo compositions. Unfortunately, many of these ingredients adversely affect other performance criteria such as foaming. The major objectives of this study were to quantitatively determine the ef- fect of PLURONIC © ethylene oxide-propylene oxide block copolymer surfactants on the eye irrita- tion and foam performance of specific anionic sur- factant systems. A lather generation test procedure, with sebum, was used to assess the foaming properties of alkyl