Numerous biochemical reactions take place during the transition from basal layer to the stratum corneum (SC), including the synthesis of keratins and cornified envelope-associated proteins. Epidermal differentiation complex (EDC) is constituted through cross-linking of involucrin, loricrin, profilaggrin, among others, on intracellular surface of plasma membrane in upper spinous and granular layers of epidermis. Studies have shown that variation in EDC results in the pathogenesis of at least three common skin disorders, ichthyosis vulgaris, atopic dermatitis and psoriasis. In this study, we evaluated the effects of a moisturizing cosmetic (MC) on the production of some envelope proteins and keratins, using an in vitro model of human keratinocytes cultures. Incubation of KC with MC promoted significant increases in the synthesis of envelope proteins filaggrin, involucrin and loricrin. We also evaluate the ability of MC to stimulate the production of K10 and K14. A significant rise of 35.1 and 23.7 % was elicited in the production of keratin 10 and 14, respectively, in relation to control group. Both, water content of the SC and skin surface lipids are important factors in the appearance and function of skin barrier. Optimized skin care cosmetics should be developed to supply a proper moisturizing effect and the same time being capable to balance the synthesis of skin barrier proteins. In this study, we demonstrated the ability of a moisturizing cosmetic to increase the levels of all proteins evaluated. The results indicate a biological activity of this skin care product in the promotion of skin barrier integrity.