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Background: Atopic dermatitis (AD) is a chronic, inflammatory skin disease characterized by pruritus, xerosis, and skin barrier dysfunction. Skin barrier alteration is associated with an increase in trans-epidermal water loss (TEWL) and reduction in skin hydration. Dupilumab is a monoclonal antibody targeting interleukin-13 modulating pro-inflammatory signal transduction, which has been approved for moderate to severe AD. The aim of this study is to evaluate the effects of Dupilumab on skin barrier functions, using non-invasive instruments and clinical evaluation., Materials and Methods: Thirty patients affected by moderate-severe AD, who had been administered dupilumab, were evaluated by clinical examination and through the instrumental measurements of TEWL and corneometry at the baseline (T0) and 8 weeks (T1) on lesional skin. The clinical evaluation was performed using the Eczema Area and Severity Index (EASI) score. Moreover, a Dermatology Life Quality Index (DLQI) and 7-day numeric rating scale (NRS) questionnaires were administered to each patient., Results: The instrumental parameters of skin barrier recovery confirmed the clinical improvement outcomes with a statistically significant reduction of TEWL at T1., Conclusion: Our data confirm the clinical outcomes already reported in the literature and show that there was an inverse proportional correlation between TEWL levels and clinical severity after 8 weeks of treatment with dupilumab., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)