Controversy exists on whether or not the epidermal keratinizing squamous epithelium of skin grafts and flaps applied to line a neovagina changes histologically to a nonkeratinizing mucosal type squamous epithelium after vaginoplasty in male-to-female transsexuals. To end this discussion, the aim of this study was to objectify the short-term and long-term histologic aspect of this neovaginal epithelial lining. Biopsies were taken from the epithelium lining of the neovagina of 9 male-to-female transsexuals, from the moment of vaginoplasty up to 14 years after. These were stained with hematoxylin-eosin and periodic acid-Schiff stain for histologic comparison to normal vaginal biopsies. Because no changes that might have been induced by local influences or hormonal therapy were found, we concluded that short-term and long-term changes in the histologic aspect of inverted skin flaps do not occur after penile and scrotal skin vaginoplasty in male-to-female transsexuals.