Antimicrobial peptides (AMPs) form a part of the skin’s innate immune system. Their primary activity is to provide antimicrobial benefits and hence protect from infections. During the course of an infection or an injury, these AMPs become activated and provide protection against pathogens. Major AMPs that are present constitutively on human skin include psoriasin, RNAse7, and lysozyme while other AMPs like LL-37, HBD2, and HBD3 are induced after infection, injury or inflammation. Niacinamide is a well-known cosmetic ingredient that has been used traditionally for skin lightening, antiaging and skin barrier building benefits. Recent data indicate that niacinamide treatment can boost AMPs in gut epithelial cells and in neutrophils. Treatment with niacinamide also provided protection from skin infections through the boost of AMPs. The objective of the current study was to investigate if niacinamide formulations provide enhanced skin hygiene benefits, via protection against microbes and also increased AMPs secretion from skin cells. Healthy subjects provided informed consent to participate in in vivo studies. Formulations were applied to the subjects’ forearms and then E.coli survival was monitored following standard protocols. Psoriasin secretion was measured by ELISA after treating primary skin keratinocytes with niacinamide. Data from primary skin keratinocytes showed that niacinamide boosts the level of AMPs, such as psoriasin. Further, human studies demonstrated that niacinamide-containing formulations can provide protection from transient pathogens like E coli.