Eco-Friendly Synthesis and Characterization of Calotropis gigantea-Derived Silver Nanoparticles for Combating Antibiotic-Resistant Helicobacter pylori and Gastric Cancer Cells

<b>Background:</b> The eco-friendly synthesis of silver nanoparticles (AgNPs) utilizing medicinal flora presents a viable strategy for the development of multifunctional agents exhibiting antimicrobial, antioxidant, anti-inflammatory, and anticancer properties. This investigation aims to elucidate the phytochemical composition of <i>Calotropis gigantea</i> and its contribution to the synthesis of CG-AgNPs that demonstrate efficacy against <i>Helicobacter pylori</i> and gastric cancer cell lines. <b>Methods:</b> The aqueous plant leaf extract of <i>C. gigantea</i> underwent comprehensive analysis via gas chromatography-mass spectrometry (GC-MS), identifying a total of 25 bioactive constituents, including oleic and oxalic acid derivatives. The fabrication and analysis of silver nanoparticles (AgNPs) were performed utilizing methodologies including ultraviolet-visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), high-resolution transmission electron microscopy (HR-TEM), dynamic light scattering (DLS), and assessments of zeta potential. Antibacterial efficacy was evaluated through methods including agar well diffusion, time-kill kinetics, and biofilm assays. The cytotoxic impact on AGS gastric cancer cells was investigated using MTT assays, DAPI staining, and acridine orange/ethidium bromide (AO/EtBr) staining techniques. The assessment of antioxidant potential was performed utilizing DPPH and ABTS assays. The anti-inflammatory properties were analyzed through protein denaturation and membrane stabilization tests. <b>Results</b>: CG-AgNPs exhibited a spherical morphology (11-17 nm) with commendable stability, denoted by using zeta potential analysis measurement of -30.2 mV. The antibacterial activity showed a significant inhibition zone of 16.00 ± 0.17 mm at a concentration of 50 µg/mL against <i>H. pylori</i>, in addition to notable biofilm disruption. The viability of AGS cells was reduced by 61% at a concentration of 100 micrograms per milliliter, with apoptosis being confirmed through relevant assays. The antioxidant potential varied from 18% to 83% (DPPH) and reached 74% (ABTS) at a concentration of 100 µg/mL. The anti-inflammatory assays indicated a BSA denaturation inhibition ranging from 45% to 80% and a membrane stabilization effect between 54% and 85%. <b>Conclusions:</b> CG-AgNPs exhibit substantial antibacterial, antioxidant, anti-inflammatory, and anticancer activities, underscoring their pharmaceutical potential, particularly for combating antibiotic-resistant pathogens and gastric malignancies.