Oxidative stress is a destructive phenomenon that affects various cell structures including membranes, proteins, lipoproteins, lipids, and DNA. Oxidative stress and inflammation owing to lifestyle changes may lead to serious diseases such as Cancers, Gout, and Arthritis etc. These disorders can be prevented using different therapeutic strategies including nanomedicine. Biosynthesized gold nanoparticles (GNPs) because of their anti‐inflammatory and antioxidant bioactivities can be key player in reversal of these ailments. This study was carried out to evaluate the anti‐inflammatory and antioxidant potential of bio fabricated GNPs with Sarcococca saligna (S. saligna) extract on injured human adipose‐derived Mesenchymal stem cells (hADMSCs). GNPs were characterized by ultraviolet–visible (UV–Vis) spectroscopy, Scanning Electron Microscopy (SEM), x‐ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and energy dispersive x‐ray (EDS). Phytochemical screening of biosynthesized GNPs exhibited a significant release of polyphenols, that is, total phenolic content (TPC) and total flavonoid content (TFC). GNPs priming amended the in vitro injury caused by Monosodium Iodoacetate (MIA) as exhibited by improved cell viability, wound closure response and superoxide dismutase activity (SOD). The anti‐inflammatory conduct assessed through NF‐κB pathway and other associated inflammatory markers reported down‐regulation of TNF‐α (0.644 ± 0.045), IL‐1β (0.694 ± 0.147) and IL‐6 (0.622 ± 0.112), apoptosis causing genes like Caspase‐3 (0.734 ± 0.13) and BAX (0.830 ± 0.12), NF‐κB pathway, p65 (0.672 ± 0.084) and p105 (0.539 ± 0.083) associated genes. High SOD activity (95 ± 5.25%) revealed by treated hADMSCs with GNPs also supported the antioxidant role of GNPs in vitro model. This study concludes that S. saligna bio fabricated GNPs priming may improve the therapeutic potential of hADMSCs against chronic inflammatory problems by regulating NF‐κB pathway.