Background: The stratum corneum (SC) consists of corneocytes, intercellular lipids, and corneodesmosomes. Most of the previous studies have focused on a ceramide with a single chain fatty acid, not with diverse chain lengths. Objective: We evaluated whether a lipid mixture enriched by ceramide NP with fatty acids of diverse chain lengths can restore the skin barrier impaired by topical corticosteroid. Methods: 27 healthy adult males were recruited. Topical corticosteroid was applied on both the forearms and, either the test cream that contains a lipid mixture or a vehicle cream was applied on the left or right forearm, respectively. The functional parameters of the skin barrier, epidermal differentiation markers, hyaluronic acid synthase 3 (HAS3), cytokines, and the lipid profiles in the SC were analyzed. Results: The barrier recovery rate, SC integrity, and SC hydration were significantly improved in the test cream-applied site. Filaggrin and HAS3 levels were significantly higher, Interleukin-1α (IL-1α) levels were increased, and IL-2, IL-6, IL-10, and IL-13 levels decreased in the test cream-applied site. Lipid analysis showed that C18, C20, and total ceramide NP levels increased in the test cream-applied site. C16, C18, C20, C24, and total ceramide NP levels were elevated in the test cream-applied site after acute barrier disruption. Conclusion: A lipid mixture enriched by ceramide NP with fatty acids of diverse chain lengths could recover the barrier impaired by topical corticosteroid.