Background and ObjectivesHypertrophic scar (HS), a common complication in wound healing, is characterized by the disarrangement of collagen, fibers, and extracellular matrix. Artesunate (ART) can inhibit the abnormal formation of fibroblasts and collagens. Fractional CO2 laser (FCO2L) can facilitate tissue remodeling and the absorption of drugs into ablative microthermal columns in HS. So far, no research has investigated the efficacy of ART combined with an FCO2L in treating HS. To investigate the theoretical basis and clinical significance of this combination, we established a rabbit model of HS to observe the change in the expression of transforming growth factor β1 (TGF‐β1) and proliferating cell nuclear antigen (PCNA).Study Design/Materials and MethodsForty New Zealand white rabbits were randomly divided into four groups: control group, ART group, FCO2L group, and ART + FCO2L (combination) group. Four wounds were surgically established in the ear of each rabbit and allowed to develop into HS. ART (20 μL/cm2) was injected in ART and combination groups, and FCO2L (combo mode, deep energy:10m J, super energy: 50 mJ) in FCO2L and combination groups on the 28th day after HS occurred. Three rounds of treatment were applied (once every 14 days). HS samples were measured by hematoxylin and eosin staining, Van Gieson staining, immunohistochemistry, and Western blot analysis on the 70th day.ResultsThe morphological and histopathological changes in HS were significant. HSs were smoother and smaller and the collagen fibers were thinner and less disordered in the combination group than those in ART and FCO2L groups. Meanwhile, the hypertrophic index (HI), fiber density (NA), and collagen fiber content (AA) were lower in the combination group (1.54 ± 0.15, 3.30 ± 0.22, 30.37 ± 1.41%) than in the ART group (2.51 ± 0.22, 4.69 ± 0.16, 44.68 ± 2.30%) and FCO2L group (1.99 ± 0.14, 4.13 ± 0.12, 37.74 ± 1.38%) (P < 0.01). Additionally, the expressions of TGF‐β1 and PCNA protein were suppressed in the ART group (0.30 ± 0.03, 0.25 ± 0.03) and FCO2L group (0.35 ± 0.03, 0.32 ± 0.05), and the suppression was more significant in the combination group(0.07 ± 0.02, 0.07 ± 0.02) (P < 0.01).ConclusionsThe combination of ART and FCO2L can effectively reduce HS in the rabbit model. This is the first report about this combination in the treatment of HS. A novel treatment is expected to be based on our findings. Lasers Surg. Med. © 2021 Wiley Periodicals LLC