Artemisia judaica (ArJ) is a Mediterranean aromatic plant used traditionally to treat gastrointestinal ailments, skin diseases, atherosclerosis, and as an immuno-stimulant. This study describes ArJ essential oil constituents and investigates their wound healing activity. The in vitro antioxidant and antibiofilm activities of ArJ essential oil were investigated. The in vivo pro/anti-inflammatory and oxidative/antioxidant markers were compared with standard silver sulfadiazine (SS) in a second-degree skin burn experimental rat model. The gas chromatography-equipped flame ionization detector (GC-FID) analysis of ArJ essential oil revealed the major classes of compounds as oxygenated monoterpenes (>57%) and cinnamic acid derivatives (18.03%). The antimicrobial tests of ArJ essential oil revealed that Bacillus cereus, Candida albicans, and Aspergillus niger were the most susceptible test organisms. Two second-degree burns (each 1 inch square in diameter) were created on the dorsum of rats using an aluminum cylinder heated to 120 °C for 10 s. The wounds were treated either with ArJ or SS ointments for 21 days, while the negative control remained untreated, and biopsies were obtained for histological and biochemical analysis. The ArJ group demonstrated a significant increase in antioxidant superoxide dismutase (SOD) and catalase (CAT) enzymatic activities, while lipid peroxide (LP) levels remained insignificant compared to the negative control group. Additionally, ArJ and SS groups demonstrated a significant decrease in inflammatory levels of tumor necrosis factor α (TNF-α) compared to the negative group, while interleukin 1 beta (IL-1b) and IL-6 were comparable to the negative group. At the same time, anti-inflammatory IL-10 and transforming growth factor beta 1 (TGF-b1) markers increased significantly in the ArJ group compared to the negative control. The ArJ results demonstrated potent wound healing effects, comparable to SS, attributable to antioxidant and anti-inflammatory effects as well as a high proportion of oxygenated monoterpenes and cinnamate derivatives.