Nasal colonization of Staphylococcus aureus (S.aureus) is known as a significant risk factor for nosocomial infections, and clearance of its nasal colonization greatly reduces the risk. In the present study the preparation and characterizations of the chitosan-o/w cream incorporated with lysostaphin (CCL) were described. It showed that the concentration of incorporated lysostaphin had a direct relationship with its release behavior from the cream. It was rapid at 2 and 3.5 mg lysostaphin/g cream and of a more sustained pattern at 5 mg lysostaphin/g cream. Efficacy of lysostaphin released from the CCL cream to inhibit S.aureus growth was higher than that of lysostaphin delivery routinely treated, as demonstrated by the reduction of the mucociliary transport rate (MTR) in contrast to the control graphite particles (p < 0.05). Therefore, it is concluded that drug delivery by the CCL holds its potential as a local nasal anti-S.aureus infection.