The aim of this study was to prepare chlorhexidine acetate (CA)/montmorillonite intercalation composites and its antibacterial potential was evaluated with pathogenic bacteria, Staphylococcus aureus and Pseudomonas aeruginosa. The CA/MMT was characterized by X-ray diffraction (XRD), Fourier transformed infrared (FT-IR), and thermogravimetric analysis (TGA). CA was successfully intercalated into the interlayer of MMT and in vitro release properties of the intercalated CA have been investigated in phosphate buffered saline media (pH 7.4) at 37+/-0.5 degrees C. At drug release study, CA showed initial burst effect for 24 h and then continuously released for 72 h. Their antibacterial activity was assayed by the inhibitory zone method. The CA/MMT was tested for antimicrobial activity against S. aureus and P. aeruginosa. The CA/MMT strongly inhibited the growth of a wide variety of microorganisms, including gram-positive bacteria, gram-negative bacteria.