Chronic wounds are commonly colonized with bacteria in a way that prevents full healing process and capacity for repair. Nano-chitosan, a biodegradable and nontoxic biopolymer, has shown bacteriostatic activity against a wide spectrum of bacteria. Effectively, pulsed electromagnetic fields are shown to have both wound healing enhancement and antibacterial activity. This work aimed to combine the use of nano-chitosan and exposure to a pulsed electric field to overcome two common types of infectious bacteria, namely and . Here, bacteria growing rate, growth kinetics and cell cytotoxicity (levels of lactate dehydrogenase, protein leakage and nucleic acid leakage) were investigated. Our findings confirmed the maximum antibacterial synergistic combination of nano-chitosan and exposure against than using each one alone. It is presumed that the exposure has influenced bacteria membrane charge distribution in a manner that allowed more chitosan to anchor the surface and enter inside the cell. Significantly, cell cytotoxicity substantiates high enzymatic levels as a result of cell membrane disintegration. In conclusion, exposure to pulsed electromagnetic fields has a synergistic antibacterial effect against and with maximum inhibitory effect for the last one. Extensive work should be done to evaluate the combination against different bacteria types to get general conclusive results. The ability of using pulsed electromagnetic fields as a wound healing accelerator and antibacterial cofactor has been proved, but in vivo experimental work in the future to verify the use of such a new combination against infectious wounds and to determine optimum treatment conditions is a must.