Callus elicitation is advantageous for metabolite production due to its ability to increase yield, provide controllable conditions, and allow for genetic manipulation, offering a sustainable and scalable alternative to traditional plant-based extraction methods for the production of bioactive substances. In this research, in vitro callus cultures (CCs) of the wild medicinal plant Aerva sanguinolenta were used to evaluate the efficacy of various elicitation regimes of silver nanoparticles (AgNPs) and salicylic acid (SA) to evoke an increased production of secondary metabolites, such as aervine and antioxidant metabolites. Three concentrations of SA (i.e., 20, 50, and 100 µM) and three concentrations of AgNPs (i.e., 30, 60, and 90 µg/L) were used on shoot explant cultures using MS (Murashige and Skoog) media. All the SA and AgNP elicitation treatments significantly increased the production of antioxidant metabolites, total phenolic contents (TPCs), and total flavonoid contents (TFCs) compared to the control treatment experiments. The contents of aervine were increased significantly upon elicitation compared to the control trial. Furthermore, the antioxidant potential of the test extract was enhanced compared to the control treatment. Comparatively, the AgNPs were more beneficial as elicitors than the SA treatments. The elicitation treatments with about 90 µg/L AgNPs and 100 µM SA were the best among all elicitation regimes. Callus elicitation with SA and AgNPs can stimulate increased metabolite production and be used as a sustainable practice in the welfare and service industries for drug development and drug discovery.