The healing of skin wounds remains an important concern in medicine, especially in chronic wounds caused by various diseases such as diabetes. Using herbs or herbal products to heal skin wounds is a therapeutic challenge for traditional medicine. In this context, the main aim of our work was to highlight the in vitro healing potential of Stellaria media (L.) Vill. (SM) extract using the scratch assay on normal human dermal fibroblasts (NHDF). The ability to stimulate cell migration and proliferation under the influence of different concentrations of SM extract (range between 12.5 and 200 µg/mL) was determined compared to the control (untreated in vitro-simulated wound) and positive control (allantoin 50 µg/mL). Our results showed that the concentration of 100 µg/mL SM extract applied on the simulated wound recorded the strongest and fastest (24 h) migration (with wound closure) and proliferation of NHDF compared with the control. In addition, the SM extract was characterized in terms of bioactive compounds (total phenols and flavonoids content), antioxidant capacity (FRAP (The Ferric-Reducing Antioxidant Power) assay and electrochemical method), and antimicrobial activity. The results show that the SM extract contains a considerable amount of polyphenols (17.19 ± 1.32 mg GAE/g dw and 7.28 ± 1.18 mg QE/g dw for total phenol and flavonoid content, respectively) with antioxidant capacity. Antimicrobial activity against Gram-positive bacteria (S. aureus) is higher than E. coli at a dose of 15 µg/mL. This study showed that Stellaria media is a source of polyphenols compounds with antioxidant capacity, and for the first time, its wound healing potential was emphasized.