Chitosan is a natural polymer with antibacterial property, that is biodegradable, extremely abundant and non-toxic. This study aimed to develop and characterize chitosan hydrogels in combination with nerolidol, in order to optimize the antimicrobial and healing properties. The hydrogels were prepared using a reaction of the chitosan with acetic acid solution, followed by the addition of 2 or 4% of the nerolidol. Using thermogravimetry, differential scanning calorimetry and infrared spectroscopy, the incorporation of nerolidol in the hydrogel was confirmed. Direct contact tests using hydrogels and Staphylococcus aureus showed a synergistic effect in the materials, enabling total inhibition of bacterial growth. The hydrogel containing 2% nerolidol showed excellent healing effects. The beginning of re-epithelialization and reorganization of collagen was already observed on the 7th day of treatment. The material created proofed to be promising as a healing and antibacterial agent.