In this study, organic montmorillonite (OMMT) is a modified form of montmorillonite (MMT) in which chitosan (CS) intercalated MMT by ion exchange of sodium ions from Na/MMT with –NH3 + of CS. The structural analysis confirmed intercalation of CS into MMT layers, indicating that CS molecular chains incorporated into the MMT layers. The interlayer distance of the MMT layered was 1.128nm and in the OMMT layers enlarged to 2.365 nm. Antibacterial activity analysis showed that unmodified MMT could not inhibit the growth of bacteria. Nevertheless, after addition of the CS molecules, an increase in the interlayer distance of MMT was observed. No difference was observed between the viability of the human dental pulp stem cells (hDPSCs) contacted to different concentrations (ranging from 0.5 to 2mg/ml) of MMT and OMMT in all time intervals, when compared with the control samples. Furthermore, neither MMT nor OMMT showed apoptosis and cytotoxicity effect on the cells. The strong antibacterial activity of the synthesized OMMT nanocomposite was also confirmed against E. coli, S. aureus, K. pneumonia and P. aeruginosa, suggesting its high potential for the prevention of post-surgical infections.