Most patients with immediate-type-hypersensitivity to natural rubber latex (NRL) give a history of hand eczema. Susceptibility to allergens is increased by irritant hand eczema due to damage to the skin barrier. Therefore, especially for employees in medical or paramedical professions, reduction in skin irritation is of importance. The present study reports the effect of glove powder on skin roughness, one feature of skin irritancy. Skin replicas, performed before and after wearing different types of gloves, were evaluated by laser profilometry. Significant alteration of skin roughness was evident and determined by different factors. Use of unpowdered Biogel gloves on prepowdered hands results in an increase in skin roughness, in contrast to the use of Biogel gloves on hands without any pre-application. On the other hand, powdered Manex neoderm gloves show no influence on the skin structure, whereas powdered Peha taft gloves also increase the roughness of the skin. Other potential candidates causing skin morphology alteration could be the glove pH, rubber chemicals with irritant potential and other characteristics of gloves, which are already under investigation.